bluuebunny/crossref_metadata_2025 huggingface dataset:
LTP Nanoparticles Seeded with Plasmid for Vascular Endothelial Growth Factor Receptor 2 Improve Fetal Morphometrics in a Rat Model of Preeclampsia
Authors: Balser Brtittany A , Novak Jacqueline , Yun Yang H , Kisha Annika F , Ramirez Rolando JJ
Published: June 2021

Recent studies by our lab have demonstrated that the delivery of L‐Tyrosine Phosphate nanoparticles seeded with plasmid for Vascular Endothelial Growth Factor Receptor 2 (LTP VEGFR2) normalizes mean arterial pressure, uterine vessel myogenic reactivity, and fetal growth restriction in the reduced uterine perfusion pressure (RUPP) animal model. The goal of this study was to investigate fetal morphometrics in RUPP+LTP VEGFR2 dams. For the RUPP procedure, silver clips are placed on the abdominal aorta (0.2mm i.d.) below the kidneys and the utero‐ovarian arteries (0.1mm i.d.) on day 14 of pregnancy in Sprague‐Dawley rats. SHAM pregnant rats undergo the same surgery without the clip placement. On the same day as surgery, LTP nanoparticles encapsulated with plasmid DNA encoding for VEGF Receptor 2 (VEGFR2), or nonsense were injected into the uterine wall. On day 21 of gestation, the uterus was removed and fetal weights, placental weights, and head circumference were recorded on three fetuses from each dam. The fetuses were selected based on position in the uterus in relation to the site of injection of the LTP nanoparticles. Fetal position was designated as the following: Site 1 was selected as the closest to the injection site, Site 6 was distal in the same uterine horn, and Site 14 was contralateral to Site 6. Fetal weight for pups at Site 1 was significantly increased in RUPP rats treated with VEGFR2 nanoparticles compared to RUPP treated with nonsense nanoparticles (4.70g ± 0.44 vs. 3.89g ± 0.31; p<0.05). Placental weight at Site 1 was significantly increased in RUPP rats treated with VEGFR2 nanoparticles compared to RUPP treated with nonsense nanoparticles (0.635g ± 0.06 vs. 0.491g ± 0.04; p<0.05). Head circumference was also significantly increased in pups at site 1 in RUPP rats treated with VEGFR2 nanoparticles as compared RUPP rats treated with nonsense nanoparticles (2.37cm ± 0.32 vs. 1.45cm ± 0.07; p<0.05). There were no significant differences in fetal weights, placental weights and head circumference in the SHAM group at any of the fetal positions. Furthermore, there were no significant differences when comparing between the SHAM and RUPP groups. The fetal measurements (fetal weight, placental weight and fetal head circumference) from RUPP treated with nonsense nanoparticles tended to be lower compared to the other groups (SHAM treated with VEGFR2 nanoparticles, SHAM treated with nonsense nanoparticles or RUPP treated with VEGFR2 nanoparticles; however, it failed to reach significance in this preliminary study. In conclusion, uterine injection of LTP nanoparticles with plasmid DNA encoding for VEGFR2 improved fetal morphometrics in RUPP animals.

Support or Funding Information

American Heart Association‐18AIREA3396032

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activating the MasR/Ang1‐7 Pathway Reduces Muscle Atrophy and Function Loss Following Denervation
Authors: Albadrani Hind , Renaud Jean‐Marc
Published: June 2021

Muscle atrophy is defined as a decrease in muscle mass that eventually leads to partial or complete muscle wasting. Muscle atrophy occurs in a variety of conditions including chronic diseases such as heart failure, limb immobilization and following a complete loss of muscle activity after spinal cord injury (SCI) and peripheral nerve injury. A consequence of muscle atrophy is large force decrease associated with muscle mass loss as well as contractile components. As a result of the force loss, atrophy decreases quality of life as the patient becomes unable to perform certain tasks as well as worsens the risk of accident while performing those tasks. Preventing the atrophy following peripheral nerve injury and thus muscle denervation will improve muscle physiology, especially, when a long time is necessary for nerve regeneration to be completed. Recent studies have provided evidence that activation of the MasR/Ang1‐7 signaling pathway reverses muscle atrophy induced by Ang II in chronic disease and immobilization. The objective of this study was to determine whether Ang 1‐7 also reduces the atrophy process and force loss following muscle denervation. Mouse extensor digitorum longus (EDL) and soleus were denervated by removing 3–4 mm of the sciatic nerve at the thigh level. Denervated mice were then divided into 4 groups: i) denervated; ii) denervated and treated with Ang 1‐7 (osmotic pump 100 ng/kg body weight/min); iii) denervated and treated with diminazene aceturate (DIZE, oral gavage 15 mg/day/kg body weight), a ACE2 activator, to locally increase Ang 1‐7 levels, and iv) denervated and receiving sucrose (oral gavage 1 mg/day/kg body weight). A 5th group of mice was used as control group, i.e., normal innervated muscle. The results show that Ang 1‐7 partially prevented muscle mass loss while fully preventing a loss of normalized force to CSA up to 4 week denervation period. The maintenance of the normalized maximum force by Ang 1‐7 suggests that muscle integrity, i.e., t‐tubules, sarcoplasmic reticulum (SR), and sarcomere, and functionality, i.e., membrane excitability, Ca2+ release and force, are better maintained. This study is thus providing evidence that Ang 1, 7 has therapeutic potential to prevent function loss following a denervation.

Support or Funding Information

This study was supported by the National Science and Engineering Research Council of Canada (NSERC)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Hypothermia on Skeletal Muscle Contractile Function: Comparison between Hibernators and Non‐hibernators
Authors: Krishnan Jishnu K S , Hunstiger Moriah , Rogers Jace , Oliver Scott Ryan
Published: June 2021

The primary function of skeletal muscle (SKM) is to implement locomotion, but muscles also serve other important functions such as generating heat through shivering (and non‐shivering), which involves continuous contraction and relaxation. Arctic ground squirrels (AGS), Spermophilus parryii, undergo extreme temperature fluctuations during the hibernating season (body temperature as low as −3 °C), where they mostly remain physically inactive. Despite prolonged periods of immobilization and starvation during hibernation, AGS and other hibernating mammals are protected from muscle atrophy, weakness, and loss of protein content. In contrast, non‐hibernators (like Sprague Dawley (SD)) are susceptible to muscle atrophy, rhabdomyolysis and myotonia etc. due to muscle inactivity. To our knowledge no study has shown the effects of temperature on muscle contractile function compared between hibernators and non‐hibernators. We hypothesized that hibernators maintain their muscle contractile function and performance, over a wider temperature range compared to non‐hibernators. We characterized and compared the alterations in SKM contractile function during hypothermic temperature stress. An Ex vivo functional assay was implemented, utilizing a tissue organ bath system (TOBS) and involved extracting diaphragm muscle from both AGS and SD, create strips made from SKM while keeping it in oxygenated Ringers solution. Following transfer of strips to respective organ bath chambers, muscles were exposed to 37°C for 30 mins (control/initial phase), 30 mins of experimental temperature (4 °C; 15 °C; 25 °C and 37 °C), and then finally back to 37 °C for 30min (recovery phase). Real time data (contractile function via force transducer) was recorded from the TOBS using LabChart Pro, and a force frequency curve was plotted. Preliminary data suggest that SKM of both hibernators and non‐hibernators during the experimental hypothermia phase showed suppressed (and slow) contractile function in both species, with the lowest force values at 4 °C. However, during the recovery phase AGS displayed higher functional recovery from hypothermic conditions compared to SD. This evidence supports the fact that SKM of hibernating animals is expected to have higher adaptability and increased contractile functional recovery from thermal stress compared to SKM of non‐hibernating animals.

Support or Funding Information

This work was supported by the INBRE IDeA Faculty Pilot Grant (University of Alaska Fairbanks ) from NIH NIGMS grant number P20GM103395.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Helicobacter pylori chemotaxis on gastric epithelial repair
Authors: Hanyu Hikaru , Engevik Kristen A. , Matthis Andrea L. , Ottemann Karen M. , Montrose Marshall H. , Aihara Eitaro
Published: June 2021

Background

Helicobacter pylori (H. pylori) is a pathogen that contributes to the development of gastric inflammation. H. pylori chronically colonizes the stomach of approximately half of the world population. We previously demonstrated in vivo that H. pylori preferentially colonizes injury sites of the stomach. In this study, we used a reductionist model to determine if H. pylori chemotaxis was in response to molecular attractants intrinsic to the epithelium, and to investigate the necessary components of the H. pylori chemotaxis mechanism. We used gastric organoids (gastroids), a 3D primary culture of mouse gastric epithelial cells that mimics in vivo gastric epithelium.

Methods

Gastroids were generated from isolated corpus tissue of human GFP‐actin (HuGE) transgenic mice, and cultured 6–7 days prior to experiments. GFP fluorescence allowed tracking of epithelial repair, and Hoechst33342 (10 μg/mL) was added to visualize cell nuclei. Sydney strain 1 of wild‐type (WT) H. pylori or mutants defective in motility (ΔmotB), chemotaxis (ΔcheY), or individual chemoreceptors (ΔtlpA, ΔtlpB, ΔtlpC or ΔtlpD), were labeled with CellTrace™ Far Red fluorescent dye (1 μM). Brucella broth containing 106 H. pylori was microinjected (50 nL) into the lumen of gastroids. Brucella broth containing fluorescent beads (105) was microinjected into control gastroids.

Results

High intensity 730 nm light (two‐photon photodamage: PD) caused single cell damage in the gastroid, and repair of damage completed within ~10 min. WT H. pylori began to accumulate at the damage site 3–4 min after gastric damage induction, resulting in delayed repair. The H. pylori mutants ΔtlpA, ΔtlpC, and ΔtlpD also accumulated at the damage site and delayed repair. Fluorescent beads or H. pylori mutants ΔmotB, ΔcheY and ΔtlpB did not accumulate at the damage site, with only ΔtlpB and fluorescent beads not causing delayed repair.

Conclusion

H. pylori sense the damage site via chemotaxis, while motility is essential to complete this preferential accumulation. Data also suggests that H. pylori chemoreceptor tlpB plays an important role in sensing the damage site. As tlpB is reported to have a high affinity for urea, the damaged cell may be a source of high urea concentration.

Support or Funding Information

Funded by NIH RO1 DK102551 (Montrose;Aihara) and Ryuji Ueno Award. Supported by “Tobitate! Study abroad scholarship”

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
HuR Regulates Functions of Paneth Cells in the Intestinal Epithelium by Altering Apical TLR2 Distribution through Control of CNPY3 Expression
Authors: Xiao Lan , Li Xiaoxue , Cai Jiazhong , Chung Hee K. , Jaladanki Rao N. , Wang Jian‐Ying
Published: June 2021

The mammalian intestinal epithelium is a vigorously self‐renewing tissue in the body and its homeostasis and functions depend on a dynamic balance among multiple cell activities. Paneth cells in the gut epithelium are localized at the base of the crypt area and play an important role in the mucosal renewal and defense. Toll‐like receptors (TLRs) regulates Paneth cell functions, but the underlying mechanism remains largely unknown. RNA‐binding protein HuR is highly expressed in the intestinal epithelium and acts as a key biological regulator of mucosal homeostasis by controlling stability and translation of the target mRNAs. Conditional deletion of HuR in intestinal epithelial cells (IECs) inhibits epithelial renewal and delays mucosal healing after acute injury. The current study was to test the hypothesis that HuR regulates Paneth cell functions in the intestinal epithelium via TLRs.

Methods

Studies were conducted in IEC tissue‐specific HuR knockout (IE‐HuR−/−) mice, primary intestinal organoids, and IECs (lines of Caco‐2 and IEC‐6). HuR levels in vitro and ex vivo were decreased by transfection with specific siRNA (siHuR) or increased by its gene overexpression. TLR levels were determined by Western blotting analysis and their cellular distribution was examined by immunohistochemistry.

Results

The HuR‐deficient intestinal epithelium exhibited obvious abnormalities in Paneth cell functions in vivo and ex vivo. The numbers of Paneth cells (lysozyme‐positive cells) in the small intestinal mucosa decreased in mice with ablated HuR compared with those in control littermates. Similarly, primarily cultured HuR‐deficient organoids isolated from IE‐HuR−/− mice also displayed a significant reduction in Paneth cells. HuR deletion also specifically disrupted cellular distribution of TLR2 in the small intestinal epithelium, as shown by a decrease in the levels of apical TLR2 but not TLR4, although it failed to alter total TLR levels. HuR silencing by transfection with siHuR also reduced the levels of membrane TLR2, along with an increase in cytosolic TLR2 in cultured IECs. Mechanistically, HuR directly interacted with the mRNA encoding CNPY3 that was necessary for TLR trafficking to cell surface. HuR silencing inhibited CNPY3 translation without effect on its total mRNA levels.

Conclusions

These results indicate that 1) HuR is crucial for Paneth cell functions; 2) HuR maintains the apical surface distribution of TLR2; and 2) HuR regulates subcellular trafficking of TLR2 by altering CNPY3 expression posttranscriptionally.

Support or Funding Information

NIH‐NIDDK (JYW) and Department of Veterans Affairs (JYW&RNJ)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Influence of Genistein and Exercise on Lipid Metabolism in High Fat‐High Sugar Fed Mice
Authors: Fisher Amy , St Aubin Chaheyla , Broderick Thomas , Al‐Nakkash Layla
Published: June 2021

Intake of diets high in saturated fat with sugar have been associated with chronic diseases including type 2 diabetes and Alzheimer's disease (AD). This study varied diet and exercise and assessed the changes in adipose metabolism by measuring serum markers, hepatosteatosis, and skeletal muscle lipid metabolism. Male and female 5–6 week old C57BL/6J mice were randomly assigned one of the following groups for 12 weeks (n=8–10/group): lean control (Ln), high fat diet, HFD (60% of calories from fat), and high sucrose/fructose (42 g combined/L drinking water) (HFHS), HFHS with genistein (Gen), HFHS with exercise (Ex), or HFHS with genistein and exercise (Gen‐Ex). Exercise consisted of 150 min/week forced treadmill running. Genistein dose was 600 mg genistein/kg HFD. We hypothesize that Ex and Gen will improve deficits in lipid metabolism associated with diabetes and obesity. Males and females had significant weight increase in HFHS groups compared to Ln groups (40.8% and 27.5% respectively, P<0.05). In males, body weight was significantly lower in Ex (8.9%), Gen (19.2%) and Gen‐Ex mice (32.6%, P<0.05) compared to HFHS controls. In females, body weight was significantly lower in the Gen‐Ex (20.7%, P<0.05) compared to HFHS controls. Weight loss was not attributed to reduced food intake, nor correlated to comparable changes in serum triglyceride. Male liver weight/body weight was increased in HFHS (31.8%, P<0.05) versus Ln, and subsequently reduced by Ex (23.5%), Gen (39%) and Gen‐Ex (52%, P<0.05) compared to HFHS controls. Female liver/body weight was comparable between groups. Glucose tolerance tests (GTT) in subgroups of mice (n=3–5/group) showed rescue of serum glucose in Gen‐Ex males (not females), which reflected concomitant changes in serum insulin. Protein expression of the skeletal muscle lipid metabolism marker, carnitine palmitoyltransferase 1b (CPT1b) was greater (80.5%, n=5/group, P<0.05) in males fed HFHS compared to Ln controls, with no differences in treatment interventions. We aim to provide mechanistic evidence for sex‐dependent effects and to demonstrate an association between genistein‐ and exercise‐mediated improvements in diet induced diabetic‐obesity. Current studies continue to evaluate other metabolic markers. Data currently suggests significant weight changes are not correlated with concomitant changes in skeletal muscle lipid metabolism markers.

Support or Funding Information

Layla Al‐Nakkash, and Tom Broderick were supported by Midwestern‐Arizona Alzheimer's Consortium and Midwestern Intramural funds. Amy Fisher and Chaheyla St Aubin were supported by the Department of Biomedical Sciences.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The role of Vitamin K and spinach rich in Vitamin K on functional recovery of injured rabbit sciatic nerve
Authors: Meraiyebu Ajibola Bart , Iho Herbert
Published: June 2021

The aim of the present study was to evaluate functional recovery in injured rabbit sciatic nerve using vitamin k and spinach. A total of 4 matured rabbits were used for the study. The rabbits were first trained to walk in the walking tract apparatus after which the normal Sciatic Function Index was obtained and calculated. A Cut‐induced peripheral nerve injury was performed on the sciatic nerves of rabbits in the four study groups after which the Sciatic Function Index of the rabbits were obtained using a walking tract apparatus a day after injury and subsequently one to three weeks after treatment with vitamin k and spinach. Rabbit 1 was not injured and was not treated, rabbit 2 was injured without treatment, rabbit 3 was injured and treated with 3ml/kg of spinach juice and rabbit 4 was injured and treated with 2g/kg vitamin k supplement. The Sciatic Function Index results show a percentage recovery in rabbit 3 an 4 after injury and treatment with vitamin k and spinach, while they were no significant change in the percentage recovery of the rabbit 2 (injured without treatment). The results obtained from rabbit 2 (injured without treatment) show recovery index of 0.18 and this was used to calculate the percentage recovery obtained to be 0.3%. The rabbit 3 (injured and treated with spinach) showed recovery index of 15.05 and this was used to calculate the percentage recovery obtained to be 21%, the result however justify some conclusion that spinach has functional recovery on injured sciatic nerve. The rabbit 4 (injured and treated with vitamin k) showed recovery index of 31.11 and this was used to calculate the percentage recovery obtained to be 39%, the result however support the role of vitamin K in sphingolipid metabolism and its modulation status and also the activation of vitamin k dependent proteins. The percentage recovery continued to increase in the injured rabbit treated with vitamin k and spinach and these results suggest and support the use of vitamin k for acceleration of peripheral nerve regeneration.

Support or Funding Information

Nil

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Use of Surface Anatomy in Emergency Interventional Procedures: A Mixed Methods Analysis Comparing the Procedural Approaches of Clinicians
Authors: Fillmore Erin
Published: June 2021

A good knowledge of surface anatomy forms an essential part of safe and effective clinical practice for many healthcare specialties. Understanding and using surface anatomy is key to interpreting normal anatomy on medical images and being able to describe the location, appearance and relationships of palpable anatomical structures. It is the definition of usable anatomy. However, evidence shows that even though clinicians rank surface anatomy as an essential aspect of anatomy needed to succeed in training, the way in which qualified clinicians use surface anatomical landmarks for common procedures varies greatly. This two‐phase sequential explanatory mixed methods study explored how practising emergency medicine and surgical clinicians, at various levels of qualification, use surface anatomy in order to perform a cricothyroidotomy, thoracostomy and/or thoracotomy. It examined what surface anatomy is used, how it is used, why it was used, and explored clinicians' perceptions of the importance of surface anatomy in performing these procedures. Using this case study example, the findings illustrate that inconsistencies exist in the use and application of surface anatomy in practice. It highlights the need to more fully understand how surface anatomy is being used across specialities, across procedures and across levels of training. Finally, this study has shown there is a need for improved applied training in landmarking for core procedures that require a comprehensive knowledge of surface anatomy. Only in understanding the most common misconceptions and inconsistencies in surface anatomy's use and application, can current guidelines help shape future teaching of the subject.

Support or Funding Information

None

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Structure and Function of Mammalian SWI/SNF Chromatin Remodeling Complexes in Human Disease
Authors: Kadoch Cigall
Published: June 2021

Recent exome‐ and genome‐wide sequencing studies in human cancer have unmasked a striking frequency of mutations in the genes encoding subunits of the mammalian SWI/SNF (mSWI/SNF) family of ATP‐dependent chromatin remodeling complexes. Our laboratory uses biochemistry, structural biology, systems biology, and functional genomics‐based approaches to define the mechanisms of chromatin and gene regulation carried out by the mSWI/SNF family of regulators. Using a collection of approaches, we recently defined the architecture and assembly pathway across three mSWI/SNF complex classes: canonical BAF, PBAF, and a newly‐defined complex, ncBAF, and defined the requirements of each subunit for complex formation and stability. In addition, we have studied rare, genetically well‐defined pediatric cancers including synovial sarcoma, Ewing sarcoma, malignant rhabdoid tumor and others, all of which involve BAF complex perturbations as critical drivers of their oncogenic programs. These studies have informed the diverse mechanisms underlying mSWI/SNF complex targeting and function and have provided new foundations for therapeutic development.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Histology of the Internal Reproductive Organs of the Female Oryx (Oryx leucoryx)
Authors: Rodriguez‐Sosa Jose Rafael , Ruiz Saul R , Townsend Kathryn Elizabeth
Published: June 2021

The Arabian oryx has been listed as a vulnerable species. Its conservation and propagation are aided with the deep knowledge of the anatomy of the reproductive system. Nowadays, there is a description of the gross anatomy of this system, but not on its microanatomy. In order to describe the histological features of the internal reproductive organs of the Oryx leucoryx, the ovaries, uterine tubes, uterus, and vagina were collected at necropsy of an adult (6‐year‐old) female oryx and immersed in 4% buffer formalin. Representative samples from each organ were taken and processed for routine histology. Tissue sections of 5 μm were mounted, stained with H&E, and evaluated under light microscope. The ovaries (16 mm in length) are covered by a simple cuboidal epithelium, the germinal epithelium. Beneath this epithelium is a layer of dense connective tissue, the tunica albuginea. The tunica covers the cortex, which in turn encloses the medulla. The cortical stroma merges indistinctly with the medulla and embeds few follicles at several stages of development and a corpus luteum. A large number of corpus albicans are located in the cortex. A rich bed of blood vessels of several sizes are present in the medulla, especially at the hilum zone. The uterine tubes (80 mm in length) are muscular ducts, which are comprised of a serosa, muscularis, lamina propria, and epithelium. The muscularis is mainly constituted of circular smooth muscle covered by a thin layer of longitudinal fibers. The mucosa has longitudinal folds and is made of a pseudostratified epithelium with ciliated and non‐ciliated cells. The bicornuate uterus lacks a body and each horn opens in a cervix. The uterine horns (140 mm in length) have several layers, from outer to inner: a serosa, muscularis, lamina propria, and mucosa. The muscularis contains two layers of smooth muscle, an inner circular layer and an outer longitudinal one. A rich vascular stratum separates these layers. The mucosa is lined by a pseudostratified epithelium. Simple coiled glands are present in the lamina propria. Nonglandular regions with highly cellular elevations are present in the mucosa, the caruncles. The mucosa of each cervix contains four dense rings lined by longitudinal folds made of a stratified squamous epithelium. Numerous glands are located in the lamina propria and contain simple columnar epithelium. The vaginal wall was made of a serosa (cranial region) or adventitia (caudal region), muscularis, lamina propria and mucosa. The inner layer of the muscularis consisted of circular smooth muscle fibers enclosed by longitudinal fibers. The mucosa is made of stratified squamous epithelium. In summary, the histology of the internal reproductive organs of the oryx is similar to that of other mammalian species, especially ruminants.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Medical Students First Experience in Human Cadaver Dissection Across Culture
Authors: Gemechu Jickssa M. , Hussein Inaya Hajj , Wasserman Jason A. , Meguid Eiman Abdel , Barremkala Mallikarjuna , Jurjus Abdo
Published: June 2021

While the teaching of anatomy has evolved over the centuries, the cadaver remains its major resource at most medical schools. Approaching the embalmed human body for the first time induces a range of feelings which develop and change throughout an anatomy course. However, the background, cultural views, and experiences students bring to the dissection experience likely influence their reactions and the trajectory of their acclimation to dissection. This survey assessed the self‐rated importance of cultural views and emotional reactions among medical students from different cultural backgrounds. First year medical students from Oakland University William Beaumont School of Medicine (US), American University of Beirut (Lebanon), and Queens University Belfast (Northern Ireland) completed a survey one week before exposure to cadaver dissection (Time 1) and one week after cadaver dissection (Time 2). In terms of self‐reported effect of cultural views on experiences during dissection, the three groups manifested significant differences from each other, with students in the U.S. having the highest levels at Time 1 (Mean=2.30), followed by students in Lebanon (Mean=1.81), and then students in Northern Ireland (Mean=1.46; F=13.07; p<0.001). In terms of the emotions at Time 1, students in Northern Ireland reported the lowest level (Mean=2.28), followed by students in the U.S. (Mean=3.00), and those in Lebanon (Mean=3.16). F= 11.96; p<001). However, the three groups did not show any significant differences on this measure at Time 2. Finally, with respect to dissection as an anxiety‐provoking experience, students in Northern Ireland showed the lowest scores at Time 1 (Mean=1.70), compared to US (Mean=2.45) and Lebanon (Mean=2.36). While both students in the U.S. (Mean=2.11) and Northern Ireland (Mean=1.56) showed some decrease in reported anxiety at Time 2, students in Lebanon showed a much more dramatic decrease (Mean=1.60). Aggregate differences in reported anxiety at Time 2 were significant (F=6.78;p=0.001), driven by the pairwise difference between U.S. students and the other groups. In conclusion, this study suggests that students self‐perceive their cultural views as relatively unimportant in the dissection experience, yet significant group differences between students from different cultural context were observed. There appear to be differences between groups nested these different cultural contexts, not only prior to the dissection experience, but also in terms of how they acclimate to the emotional experiences during dissection. Additionally, while often the U.S. is considered acultural compared to other regions, the U.S. group reports significantly higher levels of importance on this measure than those from Northern Ireland or Lebanon.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Drawing and Peer Teaching Through Illustrations in a Cadaveric‐based Undergraduate Human Anatomy Program
Authors: Robson Naomi Ying , Albabish William , Jadeski Lorraine
Published: June 2021

The University of Guelph offers dissection‐based human anatomy courses to a large cohort of over 1000 undergraduate students per academic year. The third‐year class is the largest, comprised of students enrolled in the Human Kinetics and Bio‐Medical Sciences degree programs. Students attend weekly lectures that introduce the anatomy in a regional‐based approach. Lectures are taught using PowerPoint slides supplemented with interactive activities, including progressive schematic drawings demonstrating anatomical structures and relationships. Weekly cadaver‐based laboratories follow.

Literature indicates extensive benefits to drawing in human anatomy education. Its integration has been positively associated with visual literacy and hand‐eye coordination. Drawing improves student awareness of anatomical details, by facilitating reasoning and deeper understanding of anatomical concepts. Additionally, drawing could promote peer teaching, for example, by encouraging students to create anatomical illustrations for their peers to use as study resources, therefore mutually benefiting both the illustrator‐instructor and learner.

Registered course students with interest in drawing were invited to attend weekly sessions (n=48). The first half an hour involved a surface anatomy review lesson in tandem with the weekly laboratory goals. Participants were then allotted two hours to explore and draw from various cadaveric prosections related to the discussed region. Moreover, to promote peer teaching, students worked one‐on‐one with a graduate student to design an illustration‐based learning resource for their fellow peers of a region the student perceived as challenging.

Preliminary results suggest a positive correlation between drawing and the understanding of anatomical structures, depth, relationships, and overall concepts. Additionally, the production and use of peer‐developed learning tools presents similar benefits.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐Inflammatory effect and bone protection of kefir peptides in a rat model of adjuvant‐induced rheumatoid arthritis
Authors: Lai Yun‐Wen , Ko Chi‐Hua , Chen Hsiao‐Ling , Chen Chuan‐Mu
Published: June 2021

Rheumatoid arthritis (RA) is a chronic autoimmune disease affecting 1% of people in the world. The pathological characteristics include synovial inflammation, bone erosion, and joint structure damage. Furthermore, the inflammatory responses of rheumatoid arthritis attract immune cells to aggregate in joints and release cytokines to cause bone erosion. Kefir is a kind of dairy product that contains a variety of Lactobacillus and yeast. Our previous studies have shown that kefir peptides derived from kefir fermented product can reduce lung inflammation by inhibiting the NF‐κB pathway, but the anti‐inflammatory effect of kefir peptides on RA is still unknown. To investigate the anti‐inflammatory effect of kefir peptides on RA, kefir peptides were utilized to treat the rat model of adjuvant‐induced arthritis. The animals were divided into normal, arthritis‐induced, kefir‐treating and drug control groups. Each group was treated with water, kefir peptides, and Tofacitinib, respectively, for 14 days and the severity of arthritis was tracked by ultrasound scanning. The kefir‐treating group has lower arthritis score than the arthritis‐induced group, and the ultrasonography showed decreased severity of the bone erosion of the ankle joint after kefir peptides treatment. The histological severity score of arthritis in hind paws also showed the kefir‐treating group has lower than the arthritis‐induced group. These results indicate that kefir peptides may have an anti‐inflammatory effect and reduce the bone erosion of ankle joint. This study confirmed that kefir peptides is a potentially promising substance for the treatment of rheumatoid arthritis. In the future, the serum levels of TNF‐α, IL‐6, RANKL, and NF‐κB will be determined to verify the mechanisms of the anti‐inflammatory effect of kefir peptides.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Epigenetic Changes in Pancreatic Beta Cells in Diabetes
Authors: Alhazzaa Rasha , Antonei Csoka
Published: June 2021

Introduction

Diabetes mellitus is a condition in which either the pancreas does not produce enough insulin, or the cells of the body do not respond properly to the insulin produced, resulting in high levels of sugar in the bloodstream. We hypothesized that epigenetic changes in pancreatic beta cells in the pancreas, the main insulin‐producing cells in the body, may contribute to the etiology of diabetes.

Methods

To test this hypothesis, we treated human pancreatic beta cells derived from induced pluripotent stem cells (iPSCs) with either high (20 mM) or low (2 mM) glucose for 14 days.

Results

We found epigenetic changes in several hundred genes, including those involved in many signaling pathways, especially glucose metabolism and insulin secretion. Other pathways affected were those involved in oxytocin metabolism, gastric acid secretion, calcium signaling, and adrenergic signaling.

Conclusion

Our study suggests that diabetes may, at least in part, be caused by epigenetic changes in pancreatic beta cells.

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This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anatomy of the Proboscideal Nerve in the Proboscis of the African Elephant (Loxodonta africana)
Authors: Nabavizadeh Ali , Reidenberg Joy S.
Published: June 2021

The elephant proboscis (i.e., trunk), a unique muscular organ formed by the combined narial and upper lip region, is a complex muscular hydrostat adapted for a multitude of functions, including grasping, holding, and transporting objects, siphoning, holding, and spraying water, and communication. The proboscis has previously been described as containing tens of thousands of muscle fascicles, each working individually to help bend, curve, and curl the trunk in all directions at any point along its length. Although the muscular system of the proboscis has previously been described in some depth, the unique nervous innervation of the proboscis has been given minimal attention. Here, the previously frozen head of an adult female African elephant (Loxodonta africana) is dissected to investigate the anatomy of the neuromuscular anatomy of the proboscis in more depth. In addition to the complexity of interlacing muscle fascicles and the integrated connective tissues, we identified the proboscideal nerve, a large, 2.5 to 3 centimeter thick nerve that is unique to elephants. The proboscideal nerve is made up of the combined distal nerve fibers of the infraorbital branch of the maxillary nerve (CN V3) and zygomatic branch of the facial nerve (CN VII). Specifically, as the infraorbital branch of the maxillary nerve (carrying sensory innervation) exits the cranium through the infraorbital foramen, the zygomatic branch of the facial nerve (carrying motor innervation) runs along the ventral border of the zygomatic arch and joins the maxillary nerve rostrodorsal to the masseter muscle to form the proboscideal nerve. The proboscideal nerve then travels down the length of the proboscis transmitting small sensory and motor nerve fibers to all muscle fascicles and surrounding skin and tissue. Given this anatomy, fine‐tuned neuromuscular control and coordination of the individual nerve fibers are necessary to control the diverse movements that occur throughout the length of the proboscis. This specialized evolutionary adaptation helped shape a functionally innovative organ that grew, modified, and persisted throughout the course of proboscidean evolution.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Radiographic Analysis of Phalanx Shapes
Authors: Neiswonger Madison Paige , Hartnell Layne S. , Bender Emily M. , Zdilla Matthew J.
Published: June 2021

Understanding differences in the characteristics of human phalanxes is important for varied disciplines including anthropology, forensics, and surgery. For the most part, analysis of the phalanxes has been limited to “traditional” morphometric measurements such as length. Therefore, this preliminary study sought to utilize geometric morphometric methods in the analysis of phalanxes. After exclusions, a total of 1830 phalanxes (618 proximal, 610 middle, and 602 distal) from digits 2–4 were analyzed from PA radiographs. A set of four landmark points were applied to each phalanx. Canonical variate analysis (CVA) was performed among phalanx rows (i.e., proximal, middle, and distal). Additionally, CVA was utilized to identify shape variance of phalanxes, within‐row, according to digit number (i.e., 2nd, 3rd, 4th, 5th phalanxes within a given row). The results of this research suggest that geometric morphometric analysis of phalanxes may aid in the differentiation and identification of phalanxes.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differentiation of neuronal population in cytoarchitecture of spinal cord using Modified Thionin Staining
Authors: Potaliya Pushpa , Ghatak Surajit
Published: June 2021

Background

Numerous staining techniques for neuronal tissue have an inherent unreliability even after elaborate procedures. A non‐methylated thiazin dye, thionin, has been extensively used in the past for neuronal studies. However procedure is not suitable for paraffin embedded tissue. In this study a modification of thionin stain was done for paraffin embedded neuronal tissue.

Aim

For better differential staining of neuronal tissue in spinal cord with modified thionin staining.

Material and method

The formalin fixed material was stained by conventional H & E, thionin and a modified thionin method to evaluate staining results. For this spinal cord sections were treated and stained with acid fuschin, oxalic acid and thionin.

Result

The procedure imparted a distinct differential coloration of myelin and neurons. Myelinated fibres appear pinkish whereas nerve cell bodies and glial nuclei were stained blue.

Conclusion

The modified thionin method was found to be a reliable mean to provide satisfactory results with neuronal tissue in spinal cord.

Support or Funding Information

Nil

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cell Based Therapy Reduces Secondary Damage and Increases Extent of Microglial Activation Following Cortical Injury
Authors: Orczykowski Mary E , Calderazzo Samantha M , Shobin Eli , Pessina Monica A , Oblak Adrian L , Finklestein Seth P , Kramer Brian C , Mortazavi Farzad , Rosene Douglas L , Moore Tara L
Published: June 2021

Cortical injury elicits long‐term cytotoxic and cytoprotective mechanisms within the brain and the balance of these pathways can determine the functional outcome for the individual. Cytotoxicity is exacerbated by production of reactive oxygen species, accumulation of iron, and peroxidation of cell membranes and myelin. There are currently no neurorestorative treatments to aid in balancing the cytotoxic and cytoprotective mechanisms following cortical injury. Cell based therapies are an emerging treatment that may function in immunomodulation, reduction of secondary damage, and reorganization of surviving structures. We previously evaluated human umbilical tissue‐derived cells (hUTC) in our non‐human primate model of cortical injury restricted to the hand area of primary motor cortex. Systemic hUTC treatment resulted in significantly greater recovery of fine motor function compared to vehicle controls. Here we investigate the hypothesis that hUTC treatment reduces oxidative damage and iron accumulation and increases the extent of the microglial response to cortical injury. To test this, brain sections from these monkeys were processed using immunohistochemistry to quantify oxidative damage (4‐HNE) and activated microglia (LN3), and Prussian Blue to quantify iron. hUTC treated subjects exhibited significantly reduced oxidative damage in the sublesional white matter and iron accumulation in the perilesional area as well as a significant increase in activated microglia along downstream white matter. Increased perilesional iron accumulation was associated with greater perilesional oxidative damage and larger reconstructed lesion volume. These findings support the hypothesis that systemic hUTC administered 24 hours after cortical damage decreases the cytotoxic response while increasing the extent of microglial activation.

Support or Funding Information

This study was supported by a contract from Advanced Technologies and Regenerative Medicine (ATRM), LLC. [RR# 101115‐PR] who provided the cell therapy product and the vehicle control and by the National Institutes of Health [NIH‐NINDS R21NS081261].

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
IN UTERO EXPOSURE TO CAFFEINE REDUCES DNA METHYLATION IN HUMAN MESENCHYMAL STEM CELLS
Authors: Pregmon Emily , Seat Mara , Poulsen Ryan , Wendler Christopher Charles
Published: June 2021

There is growing evidence that alterations in the intrauterine environment can have lasting effects on an individual's health. By disrupting prenatal development, environmental factors can lead to the fetal programming of adult disease, including cardiovascular disease. One substance that fetuses are frequently exposed to is caffeine, which is a non‐selective adenosine receptor antagonist. Caffeine consumption during the first trimester of pregnancy is reported by 60% of pregnant women. Human studies have shown correlations between caffeine use during pregnancy and low birth weight, childhood obesity, and neurodevelopmental deficits in children. Although, all do not accept some of these deleterious effects, it is concerning enough that the United States and many Nordic countries recommend that pregnant women consume less than 200 mg/day, or about 2 cups of coffee a day.

Our lab has demonstrated that in utero caffeine exposure in mice produces defects in cardiac gene expression, morphology, and function in adulthood. Furthermore, we have shown that in utero caffeine exposure alters DNA methylation patterns in adult hearts. Based on these data, we hypothesize that in utero caffeine exposure leads to altered DNA methylation that is maintained into adulthood leading to altered gene expression and reduced heart function in adults.

It is difficult to study the effects of caffeine on human development at the molecular level. Therefore, we have chosen to examine human mesenchymal stem cells (MSCs) obtained from umbilical cords of infants with known caffeine exposure during development. These cells were obtained from Dr. Kristen Boyle of The Healthy Start Project at the University of Colorado School of Medicine. We obtained a subset of these cell lines that were divided into two groups based on caffeine exposure during pregnancy.

Initially, we isolated RNA and DNA from these MSC lines to compare gene expression and DNA methylation between low and high caffeine exposure groups. A global DNA methylation assay revealed that the high caffeine group had a 29.98% decrease in global DNA methylation compared to the low caffeine group (p ≤ 0.01, N = 3). Furthermore, qPCR revealed a 3.5 fold decrease in DNA methyltransferase 3a (DNMT3a) expression in the high caffeine exposure group. DNMT3a is one of three enzymes, including DNMT1 and DNMT3b, responsible for DNA methylation during embryonic development.

In summary, our initial results indicate a correlation between high caffeine exposure and reduced global DNA methylation within human MSCs obtained from umbilical cords with known caffeine exposure. In addition, we observed altered gene expression in MSCs exposed to high caffeine levels in utero.

Support or Funding Information

Children's Miracle Networkat the University of Florida

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Lethal Replication Roadblock Imposed by a Catalytically Inactive DNA Polymerase
Authors: Hernandez Alfredo J , Lee Seung‐Joo , Chan Seungwoo , Lee Jaehun , Loparo Joseph , Richardson Charles C
Published: June 2021

Bacteriophage T7 encodes its own DNA polymerase, the product of gene 5. In isolation, gene product 5 (gp5) is a DNA polymerase of low processivity, however, it becomes highly processive upon formation of a complex with E. coli thioredoxin. In the course of investigating the transfer of primers from T7 DNA primase to T7 DNA polymerase, produced a gp5 variant lacking polymerase activity. Expression of a gp5 variant where the metal binding site in the polymerase domain was inactivated was found to be highly toxic to E. coli. This toxicity was strictly dependent on the presence of a functional E. coli trxA allele and T7 RNA polymerase‐driven expression. The closely related Klenow fragment of DNA polymerase I containing an engineered gp5 thioredoxin‐binding domain did not show any toxicity. Purified polymerase‐inactive gp5 drastically inhibited the E. coli replisome in vitro, but only in the presence of thioredoxin. Mutation of two critical metal binding site residues promote the formation of long‐lived protein‐DNA complexes that may block the displacement of the E. coli replication machinery. On the other hand, the inactive gp5 mutant only partially inhibits the T7 replisome, T7 DNA polymerase, or Klenow fragment. We envision that this T7 gp5 allele, in combination with a functional copy of TrxA, can be used a genetic building block in the design of synthetic biology approaches as a cell‐growth shutoff mechanism or a timed protein antibiotic.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.