bluuebunny/crossref_metadata_2025 huggingface dataset:
Middle School Females Have More Efficient Multitasking Abilities than Males
Authors: Rascon Jozelyn , Pena Francisco J. , Ramirez Juliana A. , Pena Marcelo , Trujillo Elizabeth , Melendez Gladys , Lazcano Cameron , Morales Acuna Francisco J. , Gurovich Benjamin , Valencia Carolina , Gurovich Alvaro N.
Published: June 2021

This project was created to educate seventh grade students about their capacity of multitasking. This was a part of the American Physiological Society's PhUn week activities and we used reaction time (RT) as a way to illustrate the physiology behind multitasking. Sixteen students (10 boys and 6 girls) from a seventh‐grade science class at Dr. Hornedo Middle School, El Paso, TX were able to interact with the Biopac MP40 system. The Biopac MP40 is an interactive system that collects electrical impulse data, physiological response, via electromyograph electrodes and calculate RT based on the time of the response from an auditive stimulus. In order to carry out the reaction time activity, the researchers demonstrated how to properly execute the procedure using the Biopac MP40. An acoustic signal was produced from the Biopac MP40 system and was heard by the students through the use of headphones. Once the students heard the acoustic signal they lifted their finger producing an electrical signal released through the nerves and picked up via electrodes placed on the inner forearm of the student's dominant arm.

This procedure was repeated twice. During the first trial the students had their eyes closed and were able to focus on the signal. However, during the second trial, the students had their eyes open and were simultaneously asked questions while they reacted to the acoustic signal. The data gathered demonstrated that, overall, the slowest and the mean RT were higher with eyes open and distraction (0.5±0.0 vs. 0.4±0.1 ms, 0.4±0.0 vs. 0.3±0.1 ms, respectively, both p<0.001). Similar results were observed in boys but not in girls. In fact, girls did not have any differences when having their eyes open and distracted. Overall, these results showed that distraction did affect RT, which could indicate a lower multitasking capacity. However, girls were able to focus better than boys, showing better multitasking abilities than boys.

Support or Funding Information

This work was partially supported by the National Institute of General Medicine Sciences of the National Institute of Health under linked Awards Numbers RL5GM118969, TL4GM118971, and UL1GM118970.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Human Cadaver Observational Anatomy Outreach Activity for TWU Undergraduates
Authors: Sola Maryvi Gonzalez , Rosario Martin G.
Published: June 2021

The outreach initiative was to target undergraduate students with interest in learning advance human anatomy in Texas Woman's University (TWU) Denton Campus, in collaboration with Dallas Campus. The primary purpose was to enhance student learning of anatomy through observation and identification of human cadavers, as well as encourage students to pursue Graduate or Health Allied Careers. We had the participation of 42 female students in total. Our methods included 1) having students take a 10 question pre‐test (10 points total) before going to the anatomy laboratory, and a brief orientation. Then 2) the students were further allocated into smaller groups to observe human cadavers (guided by Dr. González and Dr. Rosario), and identified the structures related to the following anatomical systems: muscular, skeletal, nervous system (among others). After the anatomy laboratory experience, 3) students answered a post‐test [same questions in pre‐test (10 points total)] with the purpose of quantifying learning parameters. Finally, to measure the attainment of the activity, 4) students were asked to respond to a ten question survey (no points assigned) self‐reporting their experience of the activity. Results of the comparisons between pre‐test (average of 5.88 out of 10 points) and post‐test (average of 7.78 out of 10 points) indicate that there was a significant increase in learning (P Value p<.05) after the activity took place. Survey results indicated the following; 38 out of 42 students assigned an A grade (excellence) to the activity and 35 out of 42 students indicated an interest in applying to graduate school, with 25 students who reported that definitely will apply. Thus, we conclude that the outreach activity assisted students to understand the different anatomical systems in a human body in a real setting, measured by obtaining a better grade at post‐test and satisfactory survey responses. Our future goals are to establish the outreach as the TWU Anatomy Week one time during the school year and to encourage and inspire students, especially minority scholars, to continue further graduate studies or health allied careers.

Support or Funding Information

Physical Therapy Department, Texas Woman's University, Dallas Campus

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Meeting the Medical Students: Transformation and Closure for Donor Families
Authors: Moon Mary Brennan , O'Donoghue Daniel L. , Halliday Nancy
Published: June 2021

Beginning in 2000, our institution inaugurated the Anatomical Donor Luncheon. This event occurs during orientation week for all incoming first‐year medical students. The families of our anatomical donors are invited to lunch where they meet the medical students who have been assigned to their deceased family member to learn human anatomy through dissection. Both students and donor families are informed that the venue provides an opportunity to promote the humanistic aspects of medicine. The families are asked to provide the medical students with the social history of their deceased family member, but are not given specific instructions on how to do so. The effect of this encounter on student attitudes has been previously reported. In the current study, we evaluated the reactions and attitudes of the donor families after meeting the medical students. Focus group questions were developed to explore the experience of donor families who met with first year medical students during the luncheon. The focus group comments were digitally recorded and transcribed, keeping the identities of responders anonymous. The audio data was subsequently destroyed. The constant comparative method was used for coding the textual data. One meta‐theme and three sub‐themes were identified. The meta‐theme was Donor Family Participants Experience Transformation and Closure. The subthemes included Motivating Factors for Donor Luncheon Participation, Impressions of the Venue, and Positive or Negative Experiences. Our data suggests that the Donor Luncheon was a mechanism for families to find closure on the death of their loved one and transformed their hesitation regarding body donation into excitement and appreciation. The institutional review board of the University of Oklahoma Health Sciences Center approved this study.

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 rapid histology station in the gross laboratory can assist students in understanding the value of microanatomy
Authors: Rae Guenevere
Published: June 2021

The integration of gross anatomy and histopathology is a recent trend in anatomical sciences education. For this study, a rapid histology station was set up in the gross anatomy laboratory that allowed medical students to create and stain microscope slides directly from cadaveric tissues. The students (n=200) were instructed on the purpose behind tissue preparation and routine hematoxylin and eosin staining methods. Students were also given access to a microscope to examine the crude architecture of the tissue immediately after slide preparation. The slides were utilized during the next semester histology course for clinical application and self‐directed activities. A survey was given to participants to determine the impact that slide preparation activities have on medical students' understanding of microanatomy and also to assess whether using a rapid histology station can assist in horizontal integration of the anatomical sciences. The surveys included both restricted response items (n=20) and open‐ended items (n=5) that necessitated a quantitative and qualitative analysis. Thematic analysis of the qualitative data resulted in several key themes being identified and suggest that student involvement in histology slide preparation can benefit students in unique educational ways beyond the traditional identification of anatomic structures.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Incorporation of Student Peer‐Teaching in a 1st Semester Medical Gross Anatomy Course
Authors: Rarey Kyle E. , Nonabur Venkatesh , Maxey Amanda , Wright Ashleigh , Hagen Melanie
Published: June 2021

The first semester of anatomy at the University of Florida College of Medicine is based upon teaching relevant anatomy as it relates to the physical exam and the utilization of prosections in the anatomy laboratory to see and learn the assigned anatomy. With a class size of ~140 students and with 4 hours allotted to an anatomy laboratory experience per week, students are divided into 10 teams; each team of students are further divided into an orange group and a blue group. Students of 2 groups of the 20 groups decide which of 14 lab sessions they will serve as peer teachers. To prepare for their peer‐teaching experience, they meet a week prior to the selected laboratory session with the teaching faculty to learn the assigned prosected anatomy. Students are expected to practice teaching the assigned anatomy and present one or more 15 minute teaching sessions to the faculty. On the day of an anatomy lab, there were 4 stations of student teachers. Members of the first‐year class rotate to different stations every 15 minutes. The students' perspective of the value of the peer‐teaching has been very positive. An online survey taken this past academic year by students in the first 3 years of training identified that the student peer‐teaching: 1) was beneficial to their learning of assigned laboratory anatomy (84% strongly agreed or agreed) and 2) facilitated their learning of the assigned anatomy by rotating to different stations with different peer teachers (84% strongly agreed or agreed). Quantitative data of the survey and course debriefings further support the view that student peer‐teaching positively augments student learning of anatomy. While some students stated they would be more comfortable with a member of the faculty to teach them, a greater number of students thoroughly enjoyed being taught by their peers; they believed it was the best part of anatomy. In fact, student peer‐teaching is used as a recruitment feature by first and second years when they interact with perspective candidates.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Andragogical Integration of Wet & Dry Simulation in the Modern Anatomy Lab
Authors: Chen Wilson , Benninger Brion Leeroy
Published: June 2021

INTRODUCTION

Dissection has been traditionally heralded as the epitome of anatomical learning, that it cannot be replaced by any other approach. However, this is only partially true in the contemporary timeline in teaching; dissection cannot be replaced, but as a stand‐alone modality it is no longer the best method of cultivating future physicians. The objective of this study is to integrate wet, dry, and mixed simulations in a systematic, andragogical approach into the modern day anatomy curriculum.

METHODS

Literature search was conducted regarding multimodal stimulation, cognitive load, and hands‐on learning. 7 modalities were added to dissection learning for the multimodal stimulation of today's medical students, including clay & 3D models for palpation, X‐ray for superimposed 3D structure, CT/MRI for cross‐sectional view of 3D structure, SECTRA 3D‐CT for 3D reconstruction, ultrasound for dynamic 2D breakdown of 3D structures, clinical examination to highlight important pathology, and exam/assessment to validate simulation learning. Students were asked to provide feedback on the perceived quality, effectiveness, and enjoyment of the dynamic and integrated learning environment.

DISCUSSION

Studies have shown that students prefer multimodal learning over single‐mode learning, and prefer kinesthetic (hands‐on) learning above the other visual, auditory, and verbal inputs. The majority of the students reported high satisfaction with the quality, effectiveness, and enjoyment of learning during integrated anatomy lab. The simulation‐integrated learning environment steps away from traditional pedagogical methods and mirrors the technology‐saturated behavior of the millennial student.

CONCLUSION

With the amount of technological resources available to the modern day medical school, a paradigm shift is needed to evolve from premium but outdated methods of anatomical teaching to one in which dissection is no longer the single centerpiece for the millennial medical student.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using Digital Multimedia to Enhance the Learning of Gross Anatomy and Integrating with Clinical Science
Authors: Doroudi Majid , Johnson Krisitne , Shuckett Rhonda , Fejtek Monika
Published: June 2021

Background

With the introduction of reformed curricula in medicine, most schools have reduced the total hours allocated for anatomy teaching and laboratory practical hours. These changes have been a continuous debate and triggered the emergence of innovative teaching and learning strategies in order to maximize students' learning of anatomy in the new context. Interactive modules allow for students to learn at their own pace and engage with their learning through quizzes and media to best supplement learning. The purpose of this project was to develop a module that can help support the learning of second year students in the new curriculum of UBC's Faculty of Medicine. This module was based on the content of a week‐long topic on osteoarthritis (OA) and hand anatomy and focused on the integration of clinical knowledge with the concurrent instruction of pertinent anatomy.

Methods

An interactive module on hand anatomy and osteoarthritis was developed to reflect curricular content. The module was made available to students on the curriculum website and completion of a voluntary survey of ten subjective questions was encouraged at the end of the module.

Results

94% of the participants felt their clinical knowledge of OA was strong or very strong after completion of the module, compared to 44% before completion of the module. Anatomical knowledge of the hand and wrist improved from 5 feeling strong or very strong to 83% of the participants. 88% of the participants felt that the integration of a clinical case of OA with the anatomy was either effective or very effective in assisting learning of the curricular content. 94% of the participants would like to see more modules like this for curricular content.

Conclusions

The development of an interactive module that explored clinical and anatomical content curricular objectives improved students' subjective knowledge of clinical and anatomical knowledge. Students found it useful for their learning to integrate cases into the core concepts of anatomy and would like to see more modules like this in the future.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anatomy e‐learning and virtual dissection as aids to achieving learning outcomes in a self‐directed‐learning focused graduate entry medical school in Ireland
Authors: McDermott kieran W. , Allardyce Joanna M. , Goggin Martina M.
Published: June 2021

Traditionally, anatomy has been taught didactically and predominantly with the use of cadavers. However, with the introduction of graduate entry medical programmes of shorter duration and the ever increasing need for space, didactic teaching and dissecting facilities are often sacrificed and novel teaching modalities and technologies have been introduced in their place. The relatively recent development of interactive 3D learning technologies to complement anatomy learning has been rapid, and current research in this area is active. However, the literature has yet to establish whether or not there is definitive evidence to support the benefits and limitations of virtual dissection.

The Graduate Entry Medical School (GEMS) at the University of Limerick (UL) is a relatively new medical school which does not offer cadaveric dissection as part of its four year medical degree programme. Instead it employs a wide range of teaching and learning modalities in the delivery of the anatomy curriculum, with a strong emphasis on small group and self‐directed learning. These modalities include problem based learning, provision of anatomy learning outcomes and related questions prior to flipped classroom style tutorials, free access to anatomical models, Aclands dissection videos, anatomy apps, web‐based anatomy e‐learning platforms and Anatomage tables. Weekly tutor‐facilitated Anatomage sessions are also provided. Anatomage promises to provide access to 3D human anatomy to rival that of a cadaver and is the most recent addition to our anatomy learning toolkit.

Our aims are firstly, to establish how best way to incorporate Anatomage into the graduate entry medicine curriculum in UL and secondly, in the longer term, to investigate the benefits and limitations of Anatomage, compared with cadaveric dissection and other innovative anatomy learning technologies in medical education. Here we present some preliminary data, on the opinions of first and second year graduate entry medical students about the efficacy of the anatomy resources and how Anatomage may enhance the anatomy learning experience. Students complete short surveys and the data collected from each cohort, first, second and third year students with differing exposure to the resources, is compared, to look for differences between students who had just commenced the programme to those nearing its completion. These studies will potentially inform international best practice in innovative approaches to anatomy learning and medical education in general.

Support or Funding Information

Graduate Entry Medical School, University of Limerick

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Analysis of Student Growth in Both Spatial Ability and Study Skills in Gross Anatomy.
Authors: Peterson Diana Coomes , Hamel Renee , Carriker Colin
Published: June 2021

Higher spatial ability has been previously shown to positively correlate with anatomy knowledge assessment scores. It has been hypothesized that students utilizing study materials that necessitate spatial aptitude will strengthen both spatial capabilities as well as student performance on curriculums that necessitate spatial awareness. We hypothesized that students would perform better on spatial ability tests at the end of a gross anatomy course than at the beginning. To test this hypothesis, all students took a spatial ability quiz at the beginning and end of the semester. Results indicate that students performed significantly better on the spatial tests after a semester of gross anatomy (p<0.01). This increased performance was much more evident with students who originally performed low on the test (p=3.37e‐6). Performance for both beginning and end spatial tests also correlated strongly with the overall score that each student received for the course (P>0.01). We conclude that gross anatomy curriculum helps to develop spatial aptitude in first year clinical students.

We also hypothesized that students would improve their study skills utilizing a variety of pedagogies and thus perform better on quizzes after short durations of study time. To test this hypothesis, students were divided into three groups. Each group was assigned to study deep cervical anatomy utilizing either notes, a prosected cadaver, or a virtual dissector program (VH Dissector). Students were directed to study for 10 minutes and then take a quiz over the materials. This procedure was performed on both the first and last days of the semester.

Examination scores at the beginning of the semester showed that the group studying with notes performed significantly better than those that used either cadaveric or virtual dissector computerized study methods (p<0.01). No distinction was observed between examination performance for groups who utilized either the virtual anatomy or cadaveric study materials. At the end of the semester, groups that used computer dissector program or the cadaveric study methods performed at equal proficiency as those that studied with notes.

The end of the semester examination performance correlated strongly with improvements in spatial ability for the students utilizing virtual anatomy software as a study method. However, no correlation was observed with spatial ability changes for the groups using either notes or cadaveric study materials. Because the overall correlations between spatial ability and student performance were low, we conclude that student practice and familiarity with various pedagogies are likely to be the primary factor for the observed improvement in performance. Therefore, all forms of pedagogy may be beneficial if students are given appropriate training with the learning techniques.

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 Relationship between the Marginal Tubercle and the Sphenozygomatic Suture: Implications for Neurosurgery
Authors: Pancake Jacob P. , Russell Michelle L. , Zdilla Matthew J.
Published: June 2021

The marginal tubercle, otherwise known as the processus marginalis, is an inconsistent structure that exists along the posterior aspect of the frontal process of the zygomatic bone. The marginal tubercle can be drilled in standard mini‐pterional craniotomy to enhance exposure. Surgical difficulty has been reported to occur when encountering tubercles greater than 13 mm wide (anterior‐to‐posterior dimension), in which case drilling of the tubercle is required for adequate surgical exposure. Despite the surgical importance of the marginal tubercle, few studies have assessed its anatomy. This study analyzed 744 frontal processes, classifying them by the presence of a marginal tubercle and the relationship with the sphenozygomatic suture into four categories based (A, B, C, and D, respectively). Type A had an average width of 11.18±2.27 mm (mean±SD), type B averaged 12.85±2.10 mm, type C averaged 12.09±2.19 mm, and type D averaged 13.51±2.40 mm. There were statistically significant differences in width between all types. In the context of mini‐pterional craniotomies, type D marginal tubercles are implicated as the most likely bony obstacle in the region. Moreover, type D marginal tubercles were found to exist in 31.2% of zygomatic bones. The information from this study will help with the preoperative planning of mini‐pterional craniotomies.

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.
Obesity and Diabetes Affects Growth Plate Cartilage Histomorphometry
Authors: Hung Jun , Castro Monica , Al‐Nakkash Layla , Broderick Tom L. , Plochocki Jeffrey H.
Published: June 2021

Childhood obesity and diabetes are strongly associated with decreased stature and poor bone health in adulthood. Similarly, ob/ob mice that have a leptin gene mutation causing it to overeat and develop the type 2 diabetes mellitus phenotype exhibit significantly reduced limb bone length and fracture resistance. In previous studies, we found decreased growth plate cartilage thickness in conjunction with increased bone fracture risk in obese diabetic mice in comparison with lean counterparts. Here, we use confocal laser scanning microscopy with post‐capture thresholding to assess 3D histological differences in z‐stacks of proximal tibia growth plate cartilage 80–100 μm in depth in juvenile lean and ob/ob mice. We find lean mice have significantly greater numbers of chondrocyte columns in growth plate cartilage with greater chondrocyte cell volumes in comparison with obese diabetic mice (P < 0.05). These data explicate the role of growth plate cartilage in bone health of obese diabetic juveniles. Our findings suggest obesity and diabetes may impair growth plate cartilage development by adversely affecting chondrocyte proliferation and hypertrophy.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Creating A 3D Histological Atlas of Subcortical Nuclei using Ultra‐High Field MRI Registration: A Model for DBS Surgical Planning
Authors: Demarco John P. , Lau Jonathan C. , Khan Ali R.
Published: June 2021

Introduction/Objectives

Deep Brain Stimulation (DBS) is an effective neurosurgical intervention that is used for the treatment of Parkinson's Disease (PD) and other movement disorders. Through the development of higher resolution templates and atlases, DBS targeting can be performed with higher accuracy. The aim of this study is to create a histological atlas of the subcortex and evaluate its ability to inform DBS surgical planning.

Methods

The histological template was creating through in‐situ 7 Tesla (T) Magnetic Resonance Imaging (MRI) of a cadaveric brain, followed by ex‐vivo 7T MR imaging and histological processing of the subcortex. Histology‐to‐MRI registration allowed for the mapping and reconstruction of histological sections back into the MRI space. Ten subcortical structures were directly visualized and segmented based on MRI and histology data.

Results

Through direct visualization of subcortical nuclei we demonstrated that these anatomical structures can be segmented with higher accuracy using histological data as opposed to 7T MRI data.

Conclusion

We hope that further development of this model can be applied to clinically significant anatomical structures, such as the Subthalamic Nucleus and Globus Pallidus Interna, when planning DBS surgery for PD patients.

Support or Funding Information

Canadian Institutes of Health Research, Natural Sciences and Engineering Research Council, BrainsCAN, Brain Canada Foundation

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
3D Histological Evaluation of Growth Plate Cartilage using Confocal Laser Scanning Microscopy
Authors: Castro Monica , Hung Jun , Al‐Nakkash Layla , Plochocki Jeffrey H.
Published: June 2021

Traditional histological processing techniques can subject tissue to distortion and artifacts and provide a limited view of cell dynamics. An evolving histological technique is the use of optical microscopy to acquire high resolution image stacks to visualize tissue in‐depth. Using the confocal laser scanning microscope (CLSM), it is possible to collect an image stack to process into a 3‐D perspective. In bone, autofluorescence is largely due to the collagen component. We aimed to use the autofluorescent properties of the collagen in the growth plate of murine femurs to characterize and evaluate the resting, proliferating and hypertrophic zones of articular cartilage. Murine distal femurs were sectioned along their longitudinal axis. The bones were allowed to incubate in refractive index mating solution (RIMS) media for two days prior to imaging. Z stacks of approximately 80–100 μm in depth were captured using ACS APO 40x/1.15 oil on Leica SPE confocal microscope. Acquisition of the z‐stacks were obtained in the blue‐green and green‐yellow emission spectrum. Incubation of tissue in RIMS solution reduced the opacity of the tissue and facilitated in‐depth optical imaging. Imaging was focused on the growth plate (Fig 1). Sequential image stacks were acquired with confocal microscope and Leica LAS software algorithms were used to compose 3‐D images. Image stacks were further manipulated in ImageJ and ICY to isolate and count the chondrocytes, respectively. 3D histology can be accomplished with minimal processing cost and can provide an alternate perspective of cell dynamics. Intrinsic biomolecules in tissue possess endogenous fluorophores that can at times be advantageous when evaluating intact tissue architecture. Confocal microscopy in conjunction with a refractive index matching solution rendered the murine femurs less opaque and allowed for greater photon penetration and thus resolution of spatial‐temporal structures in the bone matrix. Examination of the matrix and morphology of the cells within, provided an indication of the development of the cells and progression of ossification.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Understanding the Development, Variations, and Defects of the Muscular System in Normal Human Embryos, Fetuses, and Newborns
Authors: Alghamdi Malak A. , Ziermann Janine M. , Diogo Rui
Published: June 2021

There has been, until now, a limited knowledge on the development of the human muscular system, as most textbooks and atlases on human development described in detail the development of the skeletal system and other structures such as the internal organs without mentioning ‐ or doing it in a very brief way ‐ the muscular system. Moreover, within the few published works that did focus on the ontogeny of human muscles, most of them were restricted to a single muscle, a muscle group, a region, or a specific period of time (e.g. before 7 weeks of gestation). This makes the study of muscle anomalies, as well as of variations, very difficult due to the lack of good comparative models about normal development. We recently dissected 17 phenotypically ‘normal’ human newborns, fetuses and embryos between 7 and 39 weeks of development, and compared our results with the scarce literature available on the normal development of the muscular system in humans. Importantly, in the individuals dissected by us there was an indirect relationship between the ages of the fetuses and the number of seemingly fused (i.e. undifferentiated) muscles. That is, muscle anlagen in the early embryonic development often give rise to several muscles even after 7 weeks of gestation. This would contradict what is often described in the literature that by end of week 7 of gestation, all muscles resemble those of the adults (i.e. are differentiated). In addition, the number of the variations (i.e. phenotypes that deviate from the norm, within the ‘normal’ human population) per upper limbs dissected is significantly higher than the number of variations per lower limb studied, a pattern that is strikingly similar to that often reported in the literature about birth defects. Therefore, these and other results support Alberch's (1989) ill‐named “logic of monsters” theory that predicts a parallel between the variant phenotypes of the ‘normal’ populations and the defective phenotypes of genetically ‘abnormal’ individual due to the strong internal constraints that limit the number of possible outcomes. Furthermore, we present, for the first time, detailed data about the development of each head and limb human muscle from late embryonic stages to newborn configuration, and discuss the broader implications of these new data within a broad comparative and developmental framework.

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 the Role of DNA Polymerase IV in the Resolution of R‐loops
Authors: Nguyen Brian , Tashjian Tommy , Godoy Veronica
Published: June 2021

As the replication fork progresses through the DNA template, it may encounter a number of obstacles, one of which is the RNA transcription machinery. This encounter occurs because both processes utilize the same template and because the rate of replication (~1000 nt/s) is much faster than the rate of transcription (40–80 nt/s). The encounter of the two machineries, in either a co‐directional or a head‐on orientation, results in replication‐transcription collisions (RTCs). Failure to resolve these collisions leads to replication fork stalling and eventually cell death. One of the events required to resolve RTCs is the removal of RNA: DNA hybrids, known as R‐loops. In E. coli, RNase H1, coded by rnhA, is a member of a family of highly conserved endonucleases mediating resolution of R‐loops by cleaving the RNA strand on RNA: DNA hybrids. If not resolved, R‐loops cause genomic instability in the form of double‐stranded breaks through the collapse of the replication fork and accumulation of DNA lesions.

In E. coli, both replication stalling and accumulation of DNA lesions leads to induction of the SOS response. Among the first genes to be upregulated is dinB, encoding DNA polymerase IV (DinB), becoming the most abundant DNA polymerase in the cell (~2500 nM). DinB main role is to synthesize DNA over template lesions. There is evidence to support the notion that DinB is involved in recombination, a mechanism that is used to repair double‐stranded breaks. In stressful conditions, DinB, due to its abundance, outcompetes other polymerases for the recombination intermediate and synthesizes DNA from it. Because of its abundance during SOS induction and its role in recombination, DinB may also be involved in the resolution of R‐loops. Here, we investigate the possible role of DinB in R‐loop resolution in a rnhA‐deletion strain. Loss of rnhA results in slow growth that is rescued by expression of DinB from a low copy plasmid. This data suggest DinB is multicopy suppressor of the rnhA deletion and that DinB plays a role in the resolution of R‐loops.

Support or Funding Information

National Institute of General Medical Sciences (RO1GM088230 to VG)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ataxia Telangiectasia and Rad3‐Related Kinase (ATR) May Prevent Replication Stress In Planarian During Regeneration
Authors: Tirgar Reyhaneh , Shamoon Ushna , Akpati Lois , Sawyer Nathaniel , Nguyen Lillian , Nam Edward
Published: June 2021

S. mediterranea is a planarian with extensive regenerative ability which requires substantial cell division. However, DNA replication stress occurs every cell division and must be resolved. From yeast to man, an evolutionary conserved DNA replication stress response (RSR) is essential for life. The RSR is a signal transduction pathway regulated by Ataxia telangiectasia and Rad3‐related kinase (ATR) and Checkpoint 1 kinase (CHK1) to respond to replication stress. However, the RSR has not been established in planarian nor do we know if it functions in regeneration. We hypothesize that ATR may be essential for regeneration. Preliminary results show reduced regenerative ability in the presence of hydroxyurea (HU), a replication stress inducing agent. However, low concentrations (2 mM) did not attenuate regeneration, indicating that ATR and the RSR may be present and able to overcome replication stress. To test this, we treated amputated tails with HU, ATR inhibitor (ATRi), and a combination of both. In 2mM HU, tails regenerated similarly to control. We observed only a one day delay in regeneration with ATRi alone. However, ATRi and 2mM HU in combination prevented planarian regeneration completely. This synergistic effect suggests that ATR is significant in the response to replication stress during regeneration. To obtain genetic evidence, we are in the process of knocking down ATR as well as other RSR genes like CHK1, WRN, etc. Further determining the importance of RSR genes in planarian will elucidate the conserved components and allow us to understand how ATR and the RSR contributes to the incredible regenerative abilities in planarian.

Support or Funding Information

U.S. Department of Education HSI STEM Articulation Grant P031C160118

University of St. Thomas Committee of Student 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.
High‐resolution Maps of Genome‐wide Human Damage and Repair
Authors: Adebali Ogun , Hu Jinchuan , Sancar Aziz
Published: June 2021

Our DNA is continually damaged by cellular and environmental factors. Some of the most common mutagens, including ultraviolet light and platinum‐based drugs, cause bulky adduct formation; these are almost exclusively corrected through nucleotide excision repair. Although the basic mechanisms of excision repair are reasonably well understood, it was previously not known which genomic loci are preferentially damaged and repaired. Using the novel technologies, Damage‐seq and XR‐seq, we were able to decipher the genome‐wide kinetics of damage formation and excision repair following ultraviolet irradiation and cisplatin treatments. The single‐nucleotide resolution maps revealed cellular components affecting the DNA damage formation and repair. Despite some local exceptions, the overall damage formation profiles at initial time points exhibited a uniformity. However, the heterogenous repair resulted in an imbalanced distribution of the remaining damages after time. The repair bias is mostly caused by the transcription‐coupled repair as well as the chromatin structure of the DNA.

Support or Funding Information

National Institutes of Health Grants GM118102 and ES027255.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Insights into the molecular mechanism of alkylation‐induced mutagenesis
Authors: Lee Seongmin
Published: June 2021

The genomic DNA is persistently attacked by endogenous and exogenous alkylating agents (e.g., S‐adenosyl methionine, cisplatin), which gives rise to a wide variety of alkylated DNA lesions such as N7‐MeG, N3‐MeA, O6‐MeG, and cisplatin‐GG intrastrand cross‐links. These lesions, if not removed by DNA repair pathways, can be bypassed by error‐prone translesion synthesis DNA polymerases, which could cause mutations and cancers. The structural basis for promutagenic replication past these lesions remains elusive. To elucidate the alkylation‐induced mutagenesis mechanism, we conducted kinetic and structural studies of the bypass of these lesions by various human DNA polymerases. N7‐MeG formed a Watson‐Crick‐like base pair, rather than a wobble base pair, with thymine, suggesting promutagenicity of N7‐MeG. N3‐MeA formed a Watson‐Crick base pair with thymine but strongly deterred nucleotide incorporation opposite the lesion. O6‐MeG formed a Watson‐Crick‐like base pair with thymine, which was consistent with the reported high mutagenicity and carcinogenicity of O6‐MeG. Cisplatin‐GG intrastrand cross‐links engage in favorable interactions with adenine in non‐instructional fashion, suggesting that the predominant cisplatin‐induced G to T mutations may follow an “A‐rule”. Taken together, these studies revealed the features of promutagenic base pairings of N7‐MeG, O6‐MeG and cisplatin‐GG cross‐links, thereby providing new insights into the molecular mechanisms of alkylated‐induced mutagenesis and carcinogenesis.

Support or Funding Information

Welch Foundation F‐1741

National Institutes of Health ES26676

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 tumor suppressor phosphatase PHLPP1 suppresses inflammatory signaling by regulating the phosphorylation state and activity of STAT1
Authors: Katsenelson Ksenya Cohen , Stender Joshua D. , Uchiyama Satoshi , Nizet Victor , Glass Chris K. , Newton Alexandra C.
Published: June 2021

Post‐translational modifications, such as phosphorylation, play a key role in regulating gene expression. Although much is known about the kinases that control epigenetic mechanisms, much less is known about the phosphatases. Here we report that the PH domain Leucine Rich Repeat Protein Phosphatase 1 (PHLPP1) regulates the transcription of a large number of genes by a mechanism that depends, in part, on the dephosphorylation of the transcription factor STAT1. Using non‐biased high‐throughput RNA‐sequencing, we analyzed genome‐wide mRNA levels of genes in lipopolysaccharide (LPS)‐treated bone marrow‐derived macrophages from wild‐type and Phlpp1−/− mice and found that approximately 1600 transcripts were up or down‐regulated 2‐fold or more in the Phlpp1−/− macrophages. Gene ontology analysis revealed that many of the genes that were upregulated upon loss of PHLPP1 are involved in proinflammatory signaling. De novo motif analysis of the promoters for those genes revealed that 46% of them have recognition motifs for STAT1, a key regulatory transcription factor of inflammatory signaling. Biochemical analysis revealed that PHLPP1 regulates the phosphorylation of STAT1 on Ser727, a site that is important for full transcriptional activity of STAT1, but not Tyr701, a site that recruits STAT1 to the nucleus. Importantly, PHLPP1 deletion resulted in enhanced downstream inflammatory signaling as assessed by a luciferase reporter assay of the Interferon‐Gamma Activated Sequence (GAS) promoter. In addition to the new finding that PHLPP1 is controlling inflammatory signaling, Phlpp1−/− mice are protected from sepsis‐induced death. Our data support a model in which PHLPP1 plays a previously undescribed role in regulating inflammation which has potential therapeutic implications for sepsis‐induced death.

Support or Funding Information

NIH/NCI T32 CA009523 (K.C.K.)

NIH Grant R01 GM067946 (A.C.N.)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Zika Viral Polymerase: a promising therapeutic target
Authors: Wells Grace S. , Bernatchez Jean , Coste Michael , Luna Lucas , Neto Jair Siqueira , Purse Byron , Sohl Christal State
Published: June 2021

Zika virus, a member of the Flaviviridae family, has garnered global attention due to the link of recent outbreaks with increased risk of fetal microcephaly and Guillan‐Barré syndrome in adults. As of yet, there are no approved therapies for treatment or prevention of Zika virus. In this research, we analyze inhibition of Zika virus replication by nucleoside analogs – a therapeutic strategy that has been successful levied against other members of the Flaviviridae family. We have recombinantly expressed and purified the RNA‐dependent RNA polymerase (RdRP) domain of the ZIKV NS5 protein. The purified protein is used in an in vitro primer extension assay to characterize incorporation of native nucleotides and inhibitory analogs opposite a Zika specific template. Of a pool of nucleoside analogs, we have identified a compound that incorporates at approximately 1/5 the rate of native nucleoside. We plan to combine biochemical, cellular, and animal model studies to determine efficacy of these compounds in inhibiting Zika viral replication and infection.

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 Biophysical Analysis of Malate Dehydrogenase and Citrate Synthase Protein‐Protein Interaction
Authors: Ghebreigziabher Daniel , Provost Joseph
Published: June 2021

The metabolon theory describes the protein interactions of enzymes regulating a metabolic pathway. Through these interactions proteins can share substrates through direct channeling. One set of enzymes that are involved in metabolon complexes are citrate synthase and malate dehydrogenase. This study attempts to understand the interactions regulating the metabolon of malate dehydrogenase (MDH) and citrate synthase (CS), which are integral intermediate metabolic enzymes. Using size exclusion chromatography and sedimentation, previous studies identified the interaction between CS and MDH to be isoform specific, but no further work has been conducted to obtain affinity values or the biophysical nature of the interaction. Codon optimized His tagged recombinant proteins were expressed and purified using affinity chromatography. These enzymes include: glyoxysomal malate dehydrogenase (gMDH), human cytosolic malate dehydrogenase (MDH1), human mitochondrial malate dehydrogenase (MDH2), and human mitochondrial citrate synthase (CS). We analyzed the protein interactions using fast thermal melts which further explain the thermal stability of the the dimerization of the metabolons. We first show that protein concentration did not induce oligomerization of CS or any MDH isoform, indicating that any changes are not from MDH or CS homodimerization. Next, we investigated specific CS‐MDH interactions using fast thermal melts. The results show that from the initial melting temperature of CS, the addition of mitochondrial MDH had a greater shift in its melting temperature than the addition of cytosolic MDH, by 3–4°C. The melting point of CS was not altered in the presence of the plant MDH isoform. This indicates a specific and dramatic set of contacts for mitochondrial MDH and CS. However unlike the information found in the literature there was still a measurable shift in melting point when cytosolic MDH was incubated with CS. Overall this shows for the first time a quantifiable interaction between proteins in a well‐established pathway that will allow the further dissection of these interactions.

Support or Funding Information

Support for this project was from NSF REU 146045

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
E‐cadherin interact with Ajuba‐LIM during Drosophila larval brain development
Authors: Tadros Tiffany , Viera Astrid , Underwood Taylor , Fung Siaumin
Published: June 2021

Brain development is an orderly process, but the fundamental steps at its earliest stages remain unclear. Drosophila is an ideal model to characterize these critical maturational mechanisms because they possess neuroblasts that generate neurons and glia throughout the developing brain. These neuroblasts are capable making new cells through a process known as the asymmetrical cell division. In each division, neuroblasts creating one cell that continues to be a neuroblast (maintaining/renewing themselves) and the other cells, the ganglion mother cells will differentiation into neurons and glia (differentiation).

E‐cahderin expression can be visualized by Western blot throughout all stages of pupal development. The pattern of E‐cadherin expression switches from dividing cells to the glial cells at 72 hrs APF. A screen was done to identify proteins interact with E‐cadherin. This screen was done with co‐IP of GFP fusion proteins with E‐cadherin from Drosophila larval brain. Preliminary results showed that Ajuba‐LIM interact with E‐cadherin in the larval brain of Drosophila

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