bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reproducibility of Phosphoinositide Lipid Extraction & Presentation in Bioanalytical Assays
Authors: Branch Angie M. , Neilsen Paul O. 
Published: June 2021

Phosphatidylinositol 4‐phosphate (PI4P) produced by Phosphatidylinositol 4‐Phosphate kinases is an abundant Golgi phosphoinositide functioning in lipid exchange and vesicle transport within mammalian cells. Researchers wanting to quantify PI4P use the Bligh and Dyer method for extracting and separating phospholipids in cells and biological tissues. This common lipid extraction procedure is known to have inherent variation. Using a human promyelocytic leukemia cell line and an ELISA that is sensitive and specific for PI4P, we examine sources of variation in bioassays including sample (HL‐60 cells), the extraction procedure, and lipid detection. We determined a PI4P Mass ELISA has low coefficient of variation (<10%) if the extracted sample volume is held constant and samples are prepped and lipids presented with standardized procedures. Thus, the organic solvent extraction process is the major source of variation in typical Bligh‐Dyer preparations.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 Use of Pharmacogenetics in Targeting SGLT2 and its Variants
Authors: Fassler Amy 
Published: June 2021

When the contents of the bloodstream are filtered by the kidney, glucose is reabsorbed. Familial glycosuria, a benign disorder often attributed to genetic variants in the gene SLC5A2 which encodes the protein Sodium Glucose Transporter 2 (SGLT2), prevents reabsorption of glucose leading to an excess amount in urine. SGLT2 is used for glucose reabsorption, and when functioning properly, about 90% of glucose from the urine is reabsorbed into the bloodstream. Upon glucose and sodium binding, SGLT2 undergoes a conformational change, where the structure shifts from an outward to an inward facing orientation, allowing glucose to be released. Recently, two novel variants in SLC5A2 have been identified in the Marshfield Clinic patient population; further research is being conducted to determine whether these variants impact glucose transportation and how they affect physiological output. To research this, homology models of WT SGLT2 were constructed and analyzed based on a previously solved structure of a bacterial SGLT protein (vSGLT). It can be concluded that SGLT2's structure is composed of 14 transmembrane helices which surround both galactose/glucose and sodium binding sites. Recently, SGLT2 has become a popular target for Type II diabetes therapeutics. These drugs inhibit SGLT2, as they prevent glucose to be reabsorbed; thus, glucose is excreted out of the body. Further understanding how genetic variants effect SGLT2 structure and function may allow practitioners to properly prescribe medications and prevent adverse drug reactions when treating patients with Type II diabetes.Support or Funding InformationMarshfield Clinic Research FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Constructing a 3‐D Molecular Model to Highlight the Conversion of the Normal Protein PrPC into the Mutated PrPSC in a Prion Disease
Authors: Schmitt Lavin Emily F. , Barraza Allan , Bui Hoang (Gwen) , Speth Robert 
Published: June 2021

Gerstmann‐Straussler‐Scheinker (GSS) is a rare, genetically inherited prion disease caused by a mutation in the prion protein gene (PRNP). Patients who are carriers begin to experience GSS symptoms (similar to Parkinson's and Alzheimer's) during their 30s and 40s. This prion protein was modeled to reflect the changes in protein structure that leads to this disorder. While molecular details concerning the progression of this condition remain largely unknown, the alteration in shape of the prion protein could be directly involved in the aggregation of plaques contributing to the progression of GSS. The Milwaukee School of Engineering (MSOE) Center for Biomolecular Modeling ‐ Students Modeling A Research Topic (CBM‐SMART) team at Nova Southeastern University used 3‐D modeling and printing technology to examine structure and function relationships between this normal and mutated prion protein. Details of the normal GSS prion structure (PrPC) from the Protein Data Bank (PDB) File, 3HAK, were imported into Jmol, a protein visualization software. Next, the mutated form of the GSS prion protein, PrPSC (PDB file 3HER) was imported into Jmol. The differences between the two proteins were studied. Jmol commands were used to highlight these differences and a 3‐D molecular model was printed to explain the transition from normal to mutated protein. The transitions highlighted included a mutation in which phenylalanine becomes serine at positon 198, a loop from residues 193–197 which becomes deleted, and the alpha helix structure at residues 164 – 170 which becomes malformed due to the formation of a salt bridge between arginine 164 and aspartic acid 167. The N‐ and C‐ termini of the prion protein were also highlighted. This model can be used by genetic counselors, physicians, and researchers to educate the public on the molecular changes associated with prion diseases like GSS.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hemolymph Glucose Levels as a Measure of Crayfish Stress: A Methodology Using a Human Glucometer
Authors: Caldari‐Torres Cristina , Banta‐Long Wendy , Bruss Andrew , Choi Edric , Fiegel Haley , Jollis Margot Sherman , Ly Stephanie , Viswanathan Sruthi 
Published: June 2021

The objective of this study was to develop a methodology for measuring hemolymph glucose levels in crayfish of the genus Orconectes using a human glucometer. A secondary objective was to confirm through the use of this methodology that subjecting crayfish to acute stress in the form of a short‐term exposure to a basic solution or to chronic stress in the form of prolonged exposure to a nitrate‐containing solution, would increase hemolymph glucose levels. To measure the effects of acute exposure to a basic solution on crayfish hemolymph glucose levels, crayfish were placed in a beaker containing a basic solution (Ringer's buffer, pH=8) or a control solution (Ringer's buffer, pH=7) for a period of 10 seconds, after which time they were removed from the beakers and hemolymph was collected with a syringe. A ReliOn Prime glucometer (Wal‐Mart Stores, Inc.) was used to measured glucose levels (mg/dl) directly from the hemolymph. In order for the glucose levels to be within the sensitivity of the glucometer, hemolymph was mixed with high glucose Dulbecco's Modified Eagle Medium (DMEM; Gibco) in a 3:1 ratio. Glucose levels were determined by subtracting the levels of glucose measured in the DMEM solution from the glucose levels measured in the DMEM+hemolymph solution and correcting for the dilution factor. Crayfish acutely stressed through immersion in a basic solution had significantly higher hemolymph glucose levels compared to control crayfish (45.8 ± 5.9 vs 12.4 ± 3.8 mg/dl, two‐tailed t‐test; p<0.0001). To measure the effects of chronic exposure to a solution containing nitrate on crayfish hemolymph glucose levels, crayfish were housed in control tanks (no nitrate) or tanks containing 14 mg/L nitrate for one week, after which time hemolymph was drawn and glucose levels measured using the methodology described above. Crayfish chronically stressed through exposure to a nitrate‐containing solution had significantly higher hemolymph glucose levels compared to control crayfish (45 ± 14.3 vs 28.4 ± 8.8 mg/dl, two‐tailed t‐test; p=0.045). These results show that implementing this methodology human glucometers can be used to determine crayfish hemolymph glucose levels, making them an easy, inexpensive, and reliable alternative for this type of measurement. We were also able to show that hemolymph glucose levels are increased in both chronically and acutely stressed crayfish compared to control crayfish. These experiments can be used in physiology classrooms to demonstrate and discuss the role of the autonomic nervous system in an animal's response to stressful environmental conditions.Support or Funding InformationThis research was funded by the Denison University Department of Biology.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Engaging Students in Authentic Research in Lab‐based Courses Increases Student Competency in Applying the Scientific Method and Increases Collaboration Between Teaching and Research Faculty
Authors: Parks Lisa , Meitzen John 
Published: June 2021

In response to our TH!NK program, designed to engage students in critical and creative thinking across the campus, and the need to provide more students with authentic research experiences, we have designed and integrated several course‐based research labs into our curriculum. These courses have allowed undergraduates to engage in meaningful research beyond the classroom without taxing the space, time, and resources of the current research faculty. Our newest cell biology lab, using cell culture, Western blots, and immunofluorescent chemistry allows students to learn advanced lab techniques and data analysis typically reserved for faculty research labs. Course‐based research allows student to receive credit toward their degree and allows them the opportunity to design and implement original experiments within a framework for potential publication. Since the development of these courses, students who have taken one or more of these labs have scored higher on post assessments related to applying scientific methods and scientific communication – a key departmental learning outcome ‐ than students who did not take these labs. (92% vs 74% competency on post‐assessments).An added, and somewhat unforeseen benefit, has been the strengthening of the faculty learning community, particularly between teaching‐focused and research‐focused faculty. At many R1 institutions, the teaching mission has taken a backseat to the research‐dominated culture. As these labs have been developed, teaching and research faculty have engaged with each other ‐ becoming a more involved community that has led to increased team teaching, research projects, and publications. Teaching faculty has had another mechanism for staying current and engaged in research and literature, making them better instructors and giving them another outlet for potential scholarly work. Research faculty has had another mechanism for exploring side projects that they may not have had the time or funds to pursue in their labs. In some cases, research faculty have provided a one page proposal for a research project to pursue in class along with necessary protocols. This is modified for a student lab of up to 24 students working in groups of 4. We are hopeful that these labs will eventually increase in number to accommodate all students who wish to enroll in a research‐based lab course and that numbers of publications from undergraduates and collaborations between teaching and research faculty will increase.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Instruction Across an Educational Spectrum as a Tool for Building Teaching Skills in the Biomedical Sciences
Authors: Gwathmey TanYa M. , Howlett Allyn C. 
Published: June 2021

The ability to convey knowledge to individuals at diverse levels of education is an important skill that is requisite for effective teaching, but often lacking in science educators. Post‐doctoral scholars supported by an NIGMS‐funded Institutional Research and Academic Career Development Award were first trained in current pedagogical techniques and educational philosophy through a semester‐long course, coupled with workshops on innovative and transformative instruction. These activities were essential in helping the scholars to develop skills necessary to teach in undergraduate, graduate, medical and allied health professional schools. However, in these settings, the audiences often possess a basic understanding of the foundational principles being presented, and may be capable of filling in “gaps” of information not presented by the instructor, in order to complete the learning process. In an effort to develop well‐trained teachers who are highly effective communicators of biomedical and health sciences, we developed an arrangement with several K‐12 institutions within the local area, where post‐doctoral teaching scholars provided instruction to students through the development of teaching modules that were modified for age‐appropriate instruction at high‐school, middle‐school and elementary‐school levels. These activities required scholars to communicate complex technological concepts to individuals who lacked the basic understanding of the relevant foundational principles being presented, and challenged the teaching scholars in their communication of scientific principles to lay‐person populations. Improvements in communication skills, augmentations in lecture style and testing strategies, as well as the ability to create an active learning environment were reassessed by the scholars' teaching mentors at the completion of these activities.Support or Funding InformationK12‐GM102773This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Skeletal muscle non‐shivering thermogenesis in hibernating Arctic Ground Squirrels
Authors: Oliver Scott Ryan , Krishnan Jishnu , Rogers Jace , Hunstiger Moriah 
Published: June 2021

All mammals depend on heat producing mechanisms to maintain homeostasis. Skeletal muscle shivering and brown adipose tissue (BAT) uncoupling through the expression of uncoupling protein 1 (UCP‐1) are the identified major sources of heat production, though recently, skeletal muscle has also been highlighted as a thermogenic source. Skeletal muscle nonshivering heat production occurs via sarcolipin (SLN) uncoupling of sarcoendoplasmic reticulum (SR) calcium ATPase (SERCA)‐mediated ATP hydrolysis. SLN may play a significant role in hibernating animals due to the wide temperature range that requires regulation. A thermoregulatory mechanism needs to be in place to suppress heat production during hibernation and also initiate rapid thermogenesis during arousal. We hypothesized that skeletal muscle uncoupling driven by SLN activity is a significant source of thermogenesis and inhibiting this pathway can alter the metabolic rate during hibernation in Arctic ground squirrels (AGS). Specifically, this work aimed to elucidate the role of skeletal muscle SLN uncoupling and its connection with BAT heat production in thermoregulation and metabolic control during temperature fluctuations. AGS were treated with inhibitors of BAT‐uncoupling or skeletal muscle‐uncoupling during hibernation to assess skeletal muscle and BAT ability to aid in heat production. Rate of temperature increase and changes in VO2 were compared to internal controls during inter‐bout arousals to quantify the effect of suppressing BAT or skeletal muscle uncoupling throughout hibernation. Results show that suppressing skeletal muscle uncoupling by SLN causes a significant reduction in metabolic rate during AGS rewarming. Interestingly, inhibition of BAT thermogenesis did not reduce metabolic rate. Inhibition of thermogenic sources highlighted the importance of SLN uncoupling and metabolism in thermogenesis and systemic thermoregulation. Thermogenesis control is directly linked to metabolic rate and energy loss regulation, and thus influencing thermoregulation processes has the potential for translation to innovative treatments for obesity and other diseases and conditions relevant to metabolism.Support or Funding InformationResearch reported in this publication was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103395. The content is solely the responsibility of the authors and does not necessarily reflect the official views of the NIH.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 Research about Single‐Point Ligated Ischemic Heart Failure Rabbits in Physiology and Biomechanics
Authors: Zhao Dongliang , Huo Yunlong , Tan Wenchang 
Published: June 2021

ObjectiveAnimal models are important to study basic mechanism and biomechanics of ischemic heart failure (HF). Among a four heart chambers, the left ventricular (LV) plays the most significant mechanical part of pumping blood to the entire body. Biomechanical and morphological analyses can reflect the function of LV. The single‐point ligated HF model is a typical rabbit heart failure model, which can reveal the mechanism of heart failure. Left ventricular myofiber stress and strain analyzing is an effective biomechanical method in HF study. We aim at studying the physiological and biomechanical behave of the ischemic heart failure model.MethodBy single‐point ligation method, we establish the ischemic heart failure model. We ligate the left circumflex coronary arteries of 6 New Zealand white rabbits. The rabbits are scanned by Computed Tomography (CT), with short‐axis slice separated by 1.25 mm, after 2‐weeks and 4‐weeks. To determine regional stress and strain distributions in rabbit ventricular myocardium, an anatomically detailed nonlinear finite element model was used, which can solve the equations of stress equilibrium during passive filling of the left ventricle. At same time, the paraffin sections have be surveyed in pathology.ResultThe volume can be calculated by 3‐D reconstructed model, which range from 1528.33 mm3 (SD 286.23) at 2‐weeks to 1990.72 mm3 (SD 178.71) at 4‐weeks. The strain and stress get bigger from 2‐weeks to 4‐weeks. The huge change can be found at the pathology sections.ConclusionsThis research find the changing regulation of the strain and stress. Based on biomechanics and physiology, a new method for evaluating the function of the left ventricular is presented.Support or Funding InformationThis research is supported in part by the National Natural Science Foundation of China Grant 11372010(Y Huo); Shenzhen Science and Technology Innovation Institution (China) Grant JCYJ20160427170536358 (Y Huo).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exercise in a warm condition enhances plasma volume expansion compared to a cool condition with an increased skin but not core temperature
Authors: Okazaki Kazunobu , Takeda Ryosuke , Imai Daiki , Kawai Eriko , Ota Akemi , Hanno Genta , Morita Emiko , Yokoyama Hisayo 
Published: June 2021

Plasma volume (PV) expansion is a fundamental physiological adaptation to exercise‐heat acclimation. We investigated the effects of an increased body core temperature and skin temperatures on PV expansion after intense exercise.METHODSEight healthy young men underwent two identical trials which differs only ambient temperature (Ta) during exercise. On day 1, baseline blood sample was taken in a thermoneutral condition, then they performed a 72‐min intense‐intermittent exercise (8 sets of 4 min at 80% VO2max – 5 min at 20% VO2max) under cool (Ta of 20°C) and warm (Ta of 30°C) conditions. On day 2, after 23 hours after exercise, blood sample was taken again in the thermoneutral condition. We measured hematocrit, hemoglobin concentration, and plasma albumin and protein concentrations, and from these data, percent change in PV (Δ%PV) and in plasma albumin and total protein content (Δ%Albcont and Δ%TPcont, respectively) from baseline were determined. Esophageal temperature (Tes) and mean skin temperature (Tsk) were measured during exercise.RESULTSDuring exercise, Tsk were higher (p < 0.05) in warm than cool condition while Tes was comparable between trials. Δ%PV was higher in warm condition than cool condition (6.8 ± 1.3 % and 3.3 ± 1.3 %, means ± SE, p < 0.05), Δ%Albcont and Δ%TPcont were also higher in warm condition than cool condition (both, p < 0.05).CONCLUSIONSExercise in a warm condition enhances plasma volume expansion with plasma protein content after intense exercise compared with a cool condition, which is associated with an increased skin temperatures but not core temperature. Thus, increase in skin temperatures would be a possible target to enhance physiological adaptation to exercise‐heat acclimation.Support or Funding InformationThis work was supported by a Grant‐in‐Aid for Scientific Research (B) and a Grant‐in‐Aid for Challenging Exploratory Research (K. Okazaki) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
An Asymmetry Fractal Tree Model of the Spatial Heterogeneity of Myocardial Flow
Authors: Li Jiahang , Huo Yunlong 
Published: June 2021

ObjectiveThe blood flow in the myocardium exists obvious spatial heterogeneity, and the spatial heterogeneity has significant fractal features. The objective of this work is to develop an asymmetry fractal tree model based on the branch features calculated from anatomical data to explain the fracture of myocardium flow heterogeneity.MethodIn this work, we use a developed asymmetry fractal tree model in a previous work to analyze the fractal features of the myocardial flow heterogeneity based on the anatomical data of mice of different ages. In addition, the model predictions are compared with experimental results in mice hearts grouped by ages using nonradioactive fluorescent microsphere measurements.ResultsThe results from the experimental measurements show good agreement with the prediction from our model. The fractal dimensions (D) of the myocardium heterogeneity in our model and the experimental measurement are 1.22 and 1.25, respectively.ConclusionsThis validated asymmetry fractal tree model will implement a persuasive elucidation of the fractal nature of myocardium heterogeneity, and give a theoretical view for the understanding of the interaction between branching features and coronary hemodynamics.Support or Funding InformationThis research is supported in part by the National Natural Science Foundation of China Grant 11372010, 11672006 (Y Huo); Shenzhen Science and Technology Innovation Institution (China) Grant JCYJ20160427170536358 (Y Huo).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.