bluuebunny/crossref_metadata_2025 huggingface dataset:
Ibuprofen Treatment Results in Gender‐Specific Proteasome Dysfunction in Murine Kidneys
Authors: Georgieva Anna Y , Tiwari Shuchita , Gomes Aldrin V
Published: June 2021

Non‐steroidal anti‐inflammatory drugs (NSAIDs) are some of the most commonly consumed drugs currently available. In the United States alone, more than 30 billion over the counter NSAIDs are consumed annually, and more than 600 different products contain NSAIDs. These drugs are popular among all generations due to their ability to relieve pain, inflammation, and fevers. The enzymes cyclooxygenase 1 and 2 (COX‐1 and COX‐2) synthesize prostaglandins, which are associated with the pain, swelling and fevers accompanying tissue damage or infection. NSAIDs lessen these effects by inhibiting COX‐1 and/or COX‐2 and reducing the synthesis of prostaglandins. However, previous studies show that there is a significant correlation between chronic NSAID use and cardiovascular disease, with a 1.3–3.36‐fold increase for risk of stroke. Diclofenac, another common NSAID, has been linked to a >95% population decline of the wild Oriental white‐backed vulture due to renal failure. Previously published data from our lab shows that mice treated with Diclofenac have proteasome dysfunction in heart, liver, and kidney tissues. The 26S proteasome is an ATP‐dependent multi‐catalytic protease, which is responsible for degrading 60–80% of intracellular proteins in all tissues. The proteasome degrades poly‐ubiquitinated proteins through the ubiquitin‐proteasome system (UPS). We hypothesize that proteasome dysfunction occurs in murine kidneys when the commonly used NSAID, ibuprofen, is administered to mice. Male and female eight‐week old mice were treated with ibuprofen (100mg/kg/day) for seven days, and then their kidneys were harvested. The activity of the three catalytic sites on the 26S proteasome, β1, β2, and β5, were measured in control vs ibuprofen kidney samples using biochemical activity assays. To determine protein expression levels of proteasome subunits and poly‐ubiquitinated proteins, western blots were carried out using the respective antibodies. Both male and female samples showed a significant decrease in β1 (caspase‐like) activity. Interestingly, female samples had a significant decrease in β2 (trypsin‐like) activity, while males did not, but males show a significant decrease in β5 (chymotrypsin‐like) activity, while females show no significant difference. We found no difference in free ubiquitin or poly‐ubiquitinated proteins in both males or females, as well as no changes in proteasome subunit expression in both males or females. Our results suggest that ibuprofen treatment causes proteasome dysfunction in murine kidneys in a gender dependent manner.

Support or Funding Information

Research was funded by 5P42ES004966 NIH grant and AHA16GRNT31350040 grant to Aldrin Gomes. A.Y.G. was supported by PREP@UCD, NIH 5R25GM116690‐03 to UC Davis.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activation of G Protein‐coupled Estrogen Receptor Prevents High Salt‐induced Hypertension
Authors: Aceves Jeffrey O. , Gohar Eman Y. , Daugherty Elizabeth , Pollock David M.
Published: June 2021

Estrogen protects premenopausal women against cardiovascular and renal diseases. G protein‐coupled estrogen receptor (GPER) is a membrane bound protein that prompts protective effects throughout the cardiovascular and renal systems. Accumulating evidence suggests that GPER activation contributes to the maintenance of cardiovascular and renal health in women and may even be a potential therapeutic approach for post‐menopausal hypertension. The goal of the current study was to test the hypothesis that GPER activation reduces the elevation in blood pressure in ovariectomized (OVX) rats in response to high salt (HS) diet. In our experiment, female Sprague Dawley rats were implanted with telemetry transmitters, subjected to bilateral ovariectomy, and treated with vehicle or G1, a GPER agonist (400 μg/kg/day, osmotic minipump). Two weeks later, rats were placed into metabolic cages to obtain 24‐hour urine samples under normal salt (NS = 0.49% NaCl), and HS (4% NaCl) diets. Vehicle treated rats had significantly higher mean arterial blood pressures after OVX and HS challenge compared to their baseline (113±1 vs. 105±1 mmHg, respectively, n=6, p<0.05). This hypertensive response was prevented by G1 (106±3 vs. 104±2 mmHg, respectively, n=6). Similar effects were observed in both diastolic and systolic blood pressures. Given that the renal endothelin‐1 (ET‐1) plays an important role the regulation of blood pressure and Na+ homeostasis, we tested whether GPER activation regulate the expression of ET‐1 receptors in the kidney. We found that renal inner medulla obtained from HS‐fed OVX rats treated with G1 for 3 weeks elicited a significant reduction in the mRNA expression of ET‐1 receptor subtype A (ETAreceptor) relative to vehicletreated rats (0.62±0.05 vs 1.00±0.13 fold change, respectively, n=5, p<0.05). A separate group of intact female rats were treated with G15, a GPER antagonist (400 μg/kg/day, osmotic minipump). After being challenged with a HS diet, a significant increase in mean arterial pressure was observed compared to baseline (113±1 vs. 109±1 mmHg, respectively, n=6, p<0.05) in rats given G15. These data supports the hypothesis that GPER plays an important role in the maintenance of blood pressure in females fed HS diet.

Support or Funding Information

This project was supported by a STRIDE fellowship from the APS to JOA, AHA 18CDA34110010 and 15POST25090329 to EYG, and P01 HL 069999 and P01 HL136267 to DMP.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assessing Myo‐mechanicals Characteristic of Rectus Abdominis Muscle after Stem Cell Therapy
Authors: Quiroz Sofia , Barbosa Angélica , Floriano Juliano , Graeff Carlos , Marcondes João , Rudge Marilza
Published: June 2021

Hyperglycemia during gestation have the deleterious effect in the skeletal muscle leading to occurrence of urinary incontinence and pelvic floor muscle dysfunctions in women. Induces functional, metabolic and structural changes in skeletal muscle; decreasing the functional capacity of the muscle, currently there is not therapies in diabetic gestational myopathy and the new proposal of muscle regeneration is based in stem cell therapy in biodevice use. To evaluate the functionality of the muscle in the new therapies to investigate the effectiveness of the muscle regeneration by assessing myo‐mechanicals characteristic of rectus abdominis muscle after therapeutic application of biodevice with mesenchymal stem cells in experimental model in diabetic and non‐diabetic pregnant rats. 20 postpartum rats of the Sprague Dawley® specific pathogen free strain, composing IV groups as follows: I. Diabetic group without biodevice (GD), II. Diabetic group with biodevice (GD ‐Bio), III. Non‐diabetic group without biodevice (GND) and IV. Non‐diabetic group with biodevice (GND‐Bio). The study was approved by the Botucatu Medical School Ethics Committee on the Use of Animals (CEUA) No. 1234/2017. After the euthanasia of the animals, the rectus abdominis samples will be removed and immediately mounted on the supports inside the isolated organ chambers of the Myograph equipment (Model 820MS; Danish Myo Technology®), the segments will be attached and connected to isometric transducers, submerged in solution physiological composition of Krebs [(mM): NaCl, 130; NaHCO3, 14.9; dextrose, 5.5; KCl, 4.7; KH2PO4, 1.18; MgSO47H2O, 1.17 e CaCl22H2O, 1.6]. Maintained at 37 ° C and constantly bubbled with 95% O2 / 5% CO2 mixture being fixed between a force transducer and a fixed unit. Application of stabilization protocol and then electrical stimulation. The parcial results show that the treated group has an increase of 3mN more than the untreated group; this indicates a better myo‐mechanicals condition in non‐diabetic rats. The Biodevice treatment is effective for the development of muscle regeneration therapy, being able to increase the contractile capacity of the rectus abdominis muscle in the model of diabetic and non‐diabetic pregnant rats.

Support or Funding Information

This project is funded for Sao Paulo Research Foundation, FAPESP (number grant 2016/01743‐5)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Macrophage BACH1, a heme regulated transcriptional repressor, controls HMOX1 and skeletal muscle regeneration
Authors: Patsalos Andreas , Tzerpos Petros , Halasz Laszlo , Nagy Gergely , Giannakis Nikolas , Pap Attila , Lyroni Konstantina , Koliaraki Vasiliki , Kollias George , Spilianakis Charalambos G. , Nagy Laszlo
Published: June 2021

Skeletal muscle regeneration following injury is a complex process that involves inflammatory components. Macrophage infiltration to damaged tissues upon injuries and their in‐situ differentiation into effector/inflammatory and repair macrophages is indispensable for necrotic debris clearance, and for coordination of tissue repair. Regulatory events controlling the highly dynamic macrophage phenotype switch are not well understood. Here, by using an acute sterile skeletal muscle injury model, and by studying macrophage chromatin accessibility (ATAC‐seq) and gene expression (RNA‐seq) dynamics during inflammation and tissue regeneration, we identified a heme‐ and MARE motif binding transcription factor, BTB Domain And CNC Homolog 1 (BACH1), as a novel regulatory molecule of the macrophage phenotype switch following tissue injury. We show that Bach1 deletion impairs muscle regeneration, alters the dynamics of the macrophage phenotype transition, impacts myoblast proliferation and leads to transcriptional deregulation of critical inflammatory and repair‐related genes. We also find that BACH1 directly binds and regulates distal regulatory elements of these genes, suggesting a role of BACH1 in controlling a spectrum of the inflammatory and tissue repair response genes in macrophages upon tissue injury. As proof of concept, inactivation of heme oxygenase 1 (Hmox1), a known downstream BACH1 target and the most deregulated gene in the Bach1 KO, in macrophages, impairs muscle regeneration by changing the dynamics of the macrophage phenotype switch. Collectively, our data suggest the existence of a regulatory axis, involving BACH1 and HMOX1 that control the phenotype and function of the infiltrating myeloid cells upon tissue damage, shaping the overall tissue repair kinetics.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of Vitamin D Receptor and Local Vitamin D Metabolism at the Different Stages of Skeletal Muscle Growth
Authors: Srikuea Ratchakrit , Hirunsai Muthita , Charoenphandhu Narattaphol
Published: June 2021

Vitamin D metabolism is an important regulatory process for vitamin D3 action on the target cells. Nevertheless, expression of vitamin D receptor (VDR) and its association with the local regulation of vitamin D metabolism in skeletal muscle at the different stages of growth are currently unknown. In this study, hindlimb muscle of male C57BL/6 mice at the different growth stages including development (1‐month‐old), maturation (6‐month‐old), and aging (18‐month‐old) were investigated. Histological features of skeletal muscle were examined to evaluate an alteration of skeletal muscle plasticity across ages. Protein expression levels of VDR and vitamin D metabolizing enzymes (CYP24A1 and CYP27B1) at all ages were analyzed. Serum 25(OH)D3 was quantified to determine its relationship with VDR, CYP24A1, and CYP27B1 protein expression in skeletal muscle. The results revealed a significant increase of muscle fiber cross‐sectional area but not fiber number during developmental to maturation stage of skeletal muscle growth. In contrast, aging muscle presented a significant increase of centronucleated muscle fibers. These changes of skeletal muscle plasticity in mature and aged muscles were accompanied with the increase of serum 25(OH)D3 and VDR protein expression, respectively. In contrast, CYP24A1 and CYP27B1 protein expression in skeletal muscle were not changed at any age investigated. Collectively, these findings demonstrated the associations of 25(OH)D3 and VDR protein expression levels with the stages of skeletal muscle growth. However, the alterations of circulating 25(OH)D3 and VDR protein expression did not affect local vitamin D metabolism in skeletal muscle.

Support or Funding Information

This work was supported by grants(to RS) from the Thailand Research Fund and Office of the Higher Education Commission (MRG6080090) and the Faculty of Science, Mahidol University.

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 Loss of Endogenous Neuregulin‐4 Increases Intestinal Epithelial Permeability and Apoptosis
Authors: Bernard Jessica Kathleen , Schumacher Michael A , Frey Mark R
Published: June 2021

Growth factor receptors such as the ErbB receptor tyrosine kinases can preserve intestinal barrier integrity in the face of inflammation by activating anti‐apoptotic pathways and stabilizing epithelial tight junction components. Activation of ErbB4 with pharmacological administration of its specific ligand neuregulin‐4 (NRG4) promotes epithelial cell survival by anti‐apoptotic pathways (Src and PI3K/Akt) and reduces intestinal inflammation. However, it is unclear whether ErbB4 is an essential regulator of intestinal permeability and if endogenous NRG4 is required for the response to intestinal inflammation. In this study, we investigated the role of endogenous NRG4 in intestinal barrier function in vivo.

METHODS

Baseline intestinal permeability in 11–17 week old wild type and NRG4 knockout mice was determined by oral gavage with FITC‐dextran (4kDa) followed by analysis of blood collected by cardiac puncture 4 h later. Barrier recovery from bacterial antigen challenge was determined 24 h after FITC‐dextran gavage and LPS‐induced injury. Ileal epithelial apoptosis was assessed by TUNEL and cleaved caspase‐3 staining of formalin‐fixed paraffin embedded tissue.

RESULTS

NRG4−/−mice had compromised barrier function compared to wild type littermates at baseline, as shown by an increase in FITC‐dextran leakage to the blood (1.4‐fold increase). Furthermore, NRG4 deletion reduced the ability to recover barrier 24 hours after LPS‐induced injury (2.4‐fold increase, p<0.05). NRG4−/− mice had exaggerated ileal epithelial cell apoptosis 24 h after LPS as well, measured by TUNEL (32 positive cells per 100 crypts (KO) vs 14.58 positive cells per 100 crypts (WT), p<0.05) and cleaved caspase‐3 staining.

CONCLUSION

Endogenous NRG4 promotes integrity of the intestinal epithelial barrier. Further investigation of the role of NRG4 within the intestinal mucosa may lead to more effective strategies for targeting barrier dysfunction seen in diseases such as inflammatory bowel disease or necrotizing enterocolitis.

Support or Funding Information

Supported by NIH award DK095004 and a Senior Research Award from The Crohn's and Colitis Foundation.

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 Signaling by Vitamin E Nicotinate
Authors: Suzuki Yuichiro Justin , Marcocci Lucia , Shults Nataliia V.
Published: June 2021

Vitamin E nicotinate is an ester of two vitamins, tocopherol (vitamin E) and niacin (vitamin B3), in which niacin is linked to the hydroxyl group of active vitamin E. This vitamin E ester can be chemically synthesized and has been used for supplementation. However, whether vitamin E nicotinate is formed in the biological system is unclear. In rats, vitamin E nicotinate was found in the heart and its level was determined to be 30‐times lower in heart failure. The diet used in these experiments contained vitamin E acetate and niacin separately, but not in the form of vitamin E nicotinate. Thus, vitamin E and niacin are esterified to form vitamin E nicotinate in the biological system. Since other forms of vitamin E were not found to be decreased, we propose that the vitamin E nicotinate structure elicits specific biologic functions, independently of serving as a source of active antioxidant. Consistent with this idea, we found that the treatment of cultured human cells with vitamin E nicotinate, but not vitamin E plus niacin, promoted the formation of anandamide within 10 min. Vitamin E nicotinate also activated MAP kinase signaling. These results suggest that vitamin E nicotinate can endogenously be formed in the biological system and the vitamin E nicotinate structure elicits cell signaling.

Support or Funding Information

NIH

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Magnesium levels in relation to the rate of preterm birth: Data from ecological, observational and intervention studies
Authors: Zhang Yijia , Xun Pengcheng , Lu Liping , Shechter Michael , Rosanoff Andrea , He Ka
Published: June 2021

Objective

Experimental studies suggest that magnesium intake may affect the gestation length due to its ability of maintaining uterine stability. However, little is known about the association between magnesium levels/supplementation and the rate of preterm birth.

Research Design and Methods

In addition to ecological data linking soil magnesium content to the geographic variation of preterm birth rate in the US, the relevant epidemiological and clinical studies published until November 2018 in peer‐reviewed journals were retrieved from PubMed, Google Scholar and the related reference lists. The identified studies were assessed systematically and the overall relative risk (RR) and corresponding confidence intervals (CIs) for risk of preterm birth based on magnesium supplementation were estimated by random‐effects models.

Results

The ecological study revealed an inverse correlation between magnesium content in soil and the rates of preterm birth across the US (r=−0.68, P<0.001). Of the six eligible randomized controlled trials, including 3,333 pregnant women aged 20 to 35 years old with 386 preterm infants, the pooled RR (95% CI) were 0.58 (95% CI: 0.35, 0.96) for those in the magnesium supplementation group as compared with the mothers in the control group. Consistently, findings from 11 observational studies generally support an inverse association between serum magnesium levels and the rate of preterm birth, though the data were not able to be quantitatively combined with meta‐analysis.

Conclusions

Accumulated evidence from ecological, observational, and intervention studies consistently supports that magnesium may be beneficial in reduction of preterm birth rate.

Support or Funding Information

This work was partially supported by a grant from the NIH (R01ES021735).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Se deficiency exacerbated AFB1‐induced splenic injury in chicks is related to regulate six selenoproteins and redox/inflammation/apoptotic signaling
Authors: Zhao Ling , Feng Yue , Deng Jiang , Sun Lvhui
Published: June 2021

This study was to explore the mechanism of Se‐mediated protection against AFB1‐induced splenic injury in broilers. Day‐old male chicks (n = 30/group) were fed a Se‐deficient, corn and soy–based diet (BD, 36 μg Se/kg) or the BD + 1.0 mg AFB1/kg, 0.3 mg Se/kg, or 1.0 mg AFB1/kg with 0.3 mg Se/kg for 3 wk. The Se was added as 2‐hydroxy‐4‐methylselenobutanoic acid. Dietary AFB1 induced the poor growth performance and spleen injury, decreased (P < 0.05) body weight, feed intake, feed conversion, and serum interleukin‐1β, increased (P < 0.05) spleen index and serum interleukin‐6, and decreased the splenic lymphocyte number and white pulp region and histiocyte proliferation in Se adequate groups. Notably, Se deficiency exacerbated (P < 0.05) these AFB1‐induced alterations. Moreover, Se deficiency reduced (P < 0.05) splenic glutathione peroxidases (GPX) and glutathione‐S transferases activities and glutathione content in AFB1 administrated groups. Furthermore, Se deficiency also up‐regulated (P < 0.05) the antimicrobial (beta defensin 1 and 2) and apoptotic (Caspase3) genes but down‐regulated (P < 0.05) inflammatory (E3 ubiquitin‐protein ligase CBL‐B) genes in AFB1 supplementation groups. Additionally, Se deficiency down‐regulated (P < 0.05) Gpx3, thioredoxin reductase 1, GPX4 and selenoprotein (SELENO) S and up‐regulated (P < 0.05) Selenot and Selenou in spleen in AFB1 administrated groups. In conclusion, dietary Se deficiency aggravated AFB1‐induced splenic injury in chicks, potentially via the regulation of the 6 selenoproteins and oxidative stress, inflammatory and apoptotic signaling.

Support or Funding Information

Supported in part by Chinese Natural Science Foundation Projects 31772636 and 31501987 and the Fundamental Research Funds for the Central Universities Project 2662018PY030.

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 stocking density and dietary antimicrobial inclusion on biochemical and enzymical variables of male broilers grown to 35 days of age
Authors: Olanrewaju Hammed A , Purswell Joseph L , Collier Stephanie D , Branton Scott L
Published: June 2021

Significant expansion in conventional and antibiotic‐free (ABF) broiler production is expected to continue as demand for poultry products continues to increase and feed prices remain stable. Much concern nowadays is expressed about stocking density as it relates to the well‐being and welfare of broilers. Changes in the levels of various plasma biochemistry and enzyme activity biomarkers are indicators of internal organs' (heart. liver, kidney) health and overall systemic homeostasis. This study investigated the effects of recommended stocking densities and dietary antimicrobial inclusion of male broilers grown to 35 days of age on selected hematobiochemical and enzymical variables. A total of 1952 1‐d‐ old Ross x Ross 708 male chicks were randomly distributed into 32 pens based on 4 assigned stocking density treatments. Conventional (AGP+) and antibiotic free (ABF) diets were equally assigned to each pen with feed and water provided ad libitum. The treatments consisted of 4 densities (27, 29, 33, 39 kg/m2) and 2 diets (AGP+, ABF) arranged in a 4 × 2 factorial. Blood samples were collected from the brachial wing vein of 3 birds per pen on d 15, 28, and 35. The collected blood samples were centrifuged to separate the plasma and used to determine the concentration of biochemical parameters and enzyme activities using an ACE‐AXCEL automatic analyzer. Results show that only UA was affected by stocking density. However, in comparison to broilers fed the ABF diet, broilers with AGP+ had significant effects on ALB, ALP, AMYL, CK, GGT, LDL, TBILI, and TP. Also, age was showed to have significant contributor effects on most selected variables. The results are in broad agreement with those reported in the literature and contribute to our knowledge of blood metabolites and homeostatic variation in developing male broilers. In conclusion, stocking density up to 39 kg/m2 with appropriate environmental management in the presence or absence of antibiotic in the diets may be suitable for both poultry integrators and contract growers to enhance broilers production efficiency without compromising the welfare of broilers grown to 35 days of age.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
‘It all begins with the outside of the body, with a person wrapped in their own skin’: How to make surface anatomy a central part of a curriculum
Authors: Tunstall Richard
Published: June 2021

The majority of patient examinations and interventions within a healthcare setting rely on the use of surface anatomy; why, therefore, do so many of the courses used to train healthcare practitioners place little emphasis on ensuring proficiency, or indeed mastery, in the surface landmarking and localisation relevant to safe practice? Despite the regularity with which it is used in practice, surface anatomy is often treated as a second‐class relation to gross anatomy, as an add‐on component that students will figure out for themselves via logical deduction. To further add to the issue, many current texts present outdated descriptions of surface anatomy and fail to recognise that there is a range of normal positions within which structures may sit.

Arguably, surface anatomy should form one of the core components of any clinical anatomy and medical imaging curriculum for healthcare practitioners and all graduating practitioners should be able to consistently locate and visualise structures relevant to their practice via the skin. This presentation will discuss a range of techniques, methods and resources that have be used to enable surface anatomy to become an explicit and central part of undergraduate and postgraduate training, and will highlight the challenges and outcomes of implementing such an approach.

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 Comprehensive Anatomical Characterization and Radiographic Study of Stage III Testicular Cancer in a 31‐Year‐Old Male Patient
Authors: Mas Jose Luis , Talarico Ernest Francis , Jones Jonathan A.
Published: June 2021

The purpose of this investigation was to characterize an unusual case of stage III testicular germ cell tumor (TGCT) in a 31‐year‐old male with metastases to nodes, bone, viscera and brain, and to understand all possible routes of metastatic disease. Testicular cancer (TC) has an increasing incidence worldwide, and its etiology, risk factors and pathogenesis are not completely understood. Medical records were reviewed, and the cadaveric specimen evaluated by physical examination and gross dissection. Paraffin embedded tissue sections of the primary tumor were stained with Hematoxylin and Eosin (H&E) for histological study. To examine metastatic spread, pre‐ and post‐mortem digital radiologic image acquisition was done using x‐ray films, and high‐resolution CT Scans and MRI Scans. Image analysis, multi‐planar reformatting, and three‐dimensional (3‐D) reconstruction were done on radiographic series. Dissection showed masses bilaterally from the apex through the lung base; masses on the internal thoracic wall, and hepatomegaly and splenomegaly with multiple tumor masses. Testicular parenchyma was composed of primitive germ cells that formed glomeruloid or embryonal‐like structures, as well as areas with a micro‐cystic histologic pattern and areas of fibrous dysplasia. Medical imaging 3‐D video radiographic dissection was notable for a 38.45 mm diameter, mid‐brain tumor; extreme hepatomegaly with numerous tumors, lung tumors, a large penetrating tumor of the left ilium, and multiple tumors throughout both lungs and the thoracolumbar spine (T5‐S1). This study provides insight into the histology and metastatic spread of TGCT that is essential for clinicians to understand in the evaluation and treatment of TC patients.

Support or Funding Information

Methodist Hospitals Southlake Campus (Merrillville, IN)

Rocco Prosthetic and Orthotic Center (Cincinnati, OH)

Indiana University School of Medicine‐Northwest (Gary, IN)

BodyViz (Clive, IA)

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 method for determination of tracheobronchial airway geometries from four different strains of mice
Authors: Hoeng Julia , Foong Clement , Oldham Michael J. , Lucci Francesco , Cockram Steve , Luke Stephen , Yeo Demetrius , Chua Joanne , Peitsch Manuel C. , Kuczaj Arkadiusz
Published: June 2021

Accurate lung morphometry is fundamental for predicting aerosol dosimetry. Currently, lung morphometry is only available for 2–3 species of mice (B6C3F1, BALB/c, and A/J). Based upon in‐situ prepared silicone rubber mouse lung casts, a complete process, including their micro‐CT scanning, segmentation, and automated algorithmic processing enabling the determination of airways geometries was developed for four strains of mice (BALB/c, A/J, ApoE−/− and C57BL/6).

Silicone rubber lung casts were prepared in‐situ from 20 ApoE−/− and C57BL/6 mice. The cured mouse lung casts were manually inspected for casting quality and manual morphometry measurements were performed (tracheobronchial generations 1–6) on selected lung casts prior to high resolution micro‐CT scanning. Micro‐CT scanning of existing in‐situ lung casts from BALB/c and A/J mice were also performed. Micro‐CT Images were then segmented to reconstruct a 3D model of the individual lung casts. A skeleton of each processed lung cast was automatically created by shrinking the 3D model of each airway to its centerline. Algorithms were developed for automatic detection of possible skeleton exceptions like closed loops, trifurcations and isolated nodes to be subsequently manually resolved. Finally, the skeleton was automatically measured extracting major airway morphometry characteristics (e.g. airway generation number, length, diameter, bifurcations angles, and angle to gravity).

The automated measurement procedure was tested/verified by comparing its measurements of airway length, diameter and bifurcation angles with previous manual morphometry measurements from lung casts of identical BALB/c & A/J mice and the automated measurements of lung casts of two BALB/c mice. Not surprisingly, tracheobronchial airway diameters for ApoE−/− and C57BL/6 mice were similar, but were significantly different from the other murine strains examined. It is anticipated that these anatomical differences will result in different aerosol deposition as predicted by various dosimetry codes (NCRP, ICRP, and Multiple‐Path Particle Dosimetry Model).

Support or Funding Information

PMI R&D, Philip Morris Products S.A., Neuchâtel, Switzerland PMI Research Laboratories Pte. Ltd., Singapore Altria Client Services LLC, Richmond, USASynopsys, Exeter, United Kingdom

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Arterial Variation in the Upper Extremity Observed During Cadaveric Dissection
Authors: Polcaro Lauren , Donohue James , Daly Daniel , Tan Yun
Published: June 2021

Background

The brachial artery (BR) is the continuation of the axillary artery, beginning at the lower border of the teres major muscle. It terminates distal to elbow and divides into the ulnar and radial arteries and supplies the forearm and hand.

During routine dissection, anomalies of the BR were found bilaterally in an 81‐year old male and unilaterally in an 89‐year old female. The bilateral variations were not identical. In the left arm, the BR divided into the ulnar and radial arteries 3.32 cm after the inferior border of the teres major muscle. High bifurcation of the BR has an incidence rate of 8%1. The radial artery branched medially from the BR following a course deep and medial to the median nerve, following a unique path as it descended to the forearm. The ulnar artery originated laterally from the BR, coursing deep to the median nerve and medial to the musculocutaneous nerve to descend to the forearm, giving off the common interosseous artery and anterior and posterior ulnar recurrent arteries below the elbow. In the right arm, the radial artery was observed originating from the axillary artery before the subscapular artery, a branch of the axillary artery. A high origin of the radial artery has an incidence rate of 15%1. The radial artery traveled through the two heads of the median nerve and traveled with it to cross the elbow into the forearm. The BR continued down the arm and became the ulnar artery below the elbow. The 89‐year old female showed a unilateral variation called a superficial ulnar artery (SUA). The BR gave rise to a SUA at the level of the inferior border of the teres major. The SUA passed between the two heads of the median nerve and traveled anteriolaterally to the medial epicondyle of the humerus. At the level of the neck of the radius, the BR divided into a deep ulnar artery and a radial artery. Once the SUA entered the medial aspect of the forearm, it ran laterally to the flexor carpi ulnaris muscle and travelled with the ulnar nerve. The SUA and ulnar nerve pass through Guyon's canal together to enter the hand. SUA are quite rare, with an incidence rate of 0.7% to 9.4%2,3. Surgeons must be aware of these variations of the BR as they may complicate procedures.

Support or Funding Information

Center for Anatomical Science and Education, Department of Surgery, Saint Louis University School of Medicine

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 Praise of Selflessness: Students Expectations of an Anatomy Laboratory Experience
Authors: Kramer Beverley , Hutchinson Erin , Hartmann Carol
Published: June 2021

Body donation is viewed as a selfless act on the part of the donors and their families which benefits the training of Health Sciences students for the future good of society. Most anatomists view the use of human bodies, which is made possible by donation, as the cornerstone for acquiring anatomical knowledge. In this context, our study investigated undergraduate health sciences students' expectations of the anatomy laboratory experience at a South African University.

Over 600 of the 1381 students enrolled in anatomy courses for the degrees of medicine, dentistry and allied health sciences at the University of the Witwatersrand, Johannesburg, South Africa during 2016 and 2017, completed a voluntary survey (Human Research Ethics Committee (Medical) Clearance no. M160219). The survey included an open‐ended question asking students about their expectations of the anatomy laboratory experience. Thematic analysis was used to identify major and minor themes emanating from the student responses. The major themes identified from the 464 responses to the question were: an expectation to learn and better understand the anatomy of the human body, the value of visualizing structures linked to practical learning and hands on experience, as well as the authenticity of dealing with human remains. Minor themes which emerged were the importance of the knowledge gained for future clinical careers as well as the acquisition of academic and personal skills. Additionally, students acknowledged the sacrifice of the body donor and their families in their contribution to the students' learning of anatomy.

The laboratory experience and the use of human bodies was valued by the students for supporting and deepening their learning of anatomy and was viewed as enriching. Some students perceived the experience as an essential aspect in the preparation for their future careers as health sciences professionals.

Support or Funding Information

No funding was received for this project

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evolutionary significance of morphological integration of the mandibular symphysis and corpus in extant hominoids
Authors: Pitirri M. Kathleen
Published: June 2021

Morphological integration refers to coordinated variation among traits that are closely related in development and/or function. Patterns of integration can offer important insight into the relationship between phenotypic traits, providing a framework to address questions of evolutionary significance. Although integrative features of the hominoid cranium have been the focus of numerous analyses, studies of integration in the hominoid mandible have received relatively less attention. Since mandibular fragments are among the most commonly preserved aspects in the hominoid fossil record, understanding patterns of morphological integration in extant hominoids has important implications for interpretations of hominoid evolution. Here, we aim to identify the pattern of integration between the shape of the mandibular corpus and symphysis in extant hominoids. To address this, we conducted a Procrutes‐based geometric morphometric analysis to quantify and analyze covariation between the outlines of the mandibular corpus and symphysis in Pongo pygmaeus, Pongo abelii, Gorilla beringei, Gorilla gorilla, Pan troglodytes and Pan paniscus. Corpus shape was quantified using three landmarks and 60 semilandmarks between M1 and M2, and symphysis shape was quantified using five landmarks and 55 semilandmarks in a sample of 116 adult specimens. The relationship between corpus shape and symphysis shape was assessed using two‐block partial least squares analysis. Phylogenetic patterns of shape variation were assessed using evolutionary covariance matrices. Results of this analysis show that relatively taller, narrower symphyses with well defined superior and inferior transverse tori covary with taller, narrower corpora, while relatively shorter, wider corpora covary with wider, narrower symphyses with larger inferior transverse tori. These results also reveal taxonomic differences in patterns of integration between the corpus and the symphysis that may reflect phylogenetically significant shifts in development and/or function of the hominoid mandible.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of an Improved Method for Genetic Fate Mapping of Brain Microglia
Authors: McKinsey Gabriel L. , Chee Elin , Santander Nicolas , Arnold Thomas D.
Published: June 2021

As the resident macrophage cells of the brain, microglia are crucial for the phagocytosis of infectious agents, apoptotic cells and synapses. Developmentally, microglia originate from the embryonic yolk sac and serve important roles in the sculpting of neonatal neural circuits. In the context of severe brain injury or infection, bone‐marrow derived macrophages invade neural tissue, making it difficult to distinguish between invading macrophages and resident microglia. Although bone‐marrow derived macrophages can adopt some features of endogenous microglia, they are not able to fully recapitulate all of their properties, suggesting that there may be important functional differences between these two populations of cells. To distinguish between resident microglia and circulating myelogenous macrophages, we generated a P2ry12‐CreER mouse line. P2RY12 is a microglial‐specific nucleotide sensing GPCR that is important for microglial response to tissue damage. The P2ry12‐CreER line was generated by CRISPR‐facilitated homologous recombination and insertion of a 2A‐CreER sequence into the 3’ end of the P2RY12 coding sequence. Using immunofluorescent labeling and flow cytometry, we determined whether this recombination was specific to parenchymal microglia, as compared to meningeal, choroid plexus or perivascular macrophages. We also describe preliminary experiments that involve transcriptional profiling of P2ry12‐CreER recombined cells, using a Cre‐dependent Rpl22‐HA mouse line. Finally, using mouse models of neonatal and adult stroke, we used the P2ry12‐CreER line to fate map true brain microglia in the context of brain injury. Altogether, these results demonstrate the utility of this novel mouse line for genetic recombination, gene expression and fate‐mapping studies of brain microglia.

Support or Funding Information

NIH K08NS096192

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assessing the Role of Ndc80 Kinetochore Complex in Accurate Chromosome Segregation
Authors: Rodriguez‐Lopez Xavier Luis , Amin Mohammed , Wang Yu , Yao Yiyi , Varma Dileep
Published: June 2021

Mitosis is required to grow and develop from a single cell to a multicellular organism. However, this process can often lead to cancer when its regulation is lost. The Highly Express in Cancer 1 (Hec1) protein is one of the four subunits of the Nuclear Division Cycle (Ndc80) complex present in mitosis. The function of this complex is required for robust kinetochore‐microtubule (kMT) attachment and is regulated by Aurora B Kinase phosphorylation of Hec1. Given that Hec1 protein binds directly to spindle microtubules, it certainly plays a critical role in chromosome alignment and segregation. To assess how Hec1 leads to defects in cell division cycle in human cells and induce cancer, we expressed a non‐phosphorylatable version of Hec1 (Hec1‐9A) in MDA‐MB‐436 (breast cancer) cells. We observed how mitosis was modified by the Hec1‐9A mutant by employing immunofluorescence microscopy to mark the chromosomes and the mitotic spindle in our Hec1‐9A positive and control cells. The Hec1‐9A satisfied the genome surveillance mechanism, thus, allowing cells that presented erroneous chromosome alignment and segregation to exit mitosis. Further studies will be aimed to investigate whether spindle checkpoint is defective at erroneous kMT attachments and whether Hec1 plays a role in other steps of mitosis.

Support or Funding Information

Funded by NIH‐NIGMS #2R25GM096955

Funded by NIH #R00CA178188

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
FLG loss of function mutations R501X and R2447X in Puerto Rico: Association of genetic ancestry with Atopic Dermatitis and Ichthyosis Vulgaris
Authors: Ramirez Aponte Edwin G , Martinez Cruzado Juan C
Published: June 2021

In Puerto Rico, we do not have many studies of Loss of function (LoF) mutations but using the data from the 1000 Genome Project we identified several polymorphisms that predicted to eliminate protein function. In this research, we work with two single nucleotide polymorphisms that produce premature stop codons in the FLG gene. They are known as R501X and R2447X. Both are associated to skin conditions such as ichthyosis vulgaris and atopic dermatitis; they also have been associated to asthma and allergies. The 1000 Genome Project data is based on the analysis of 104 Puerto Ricans only, and we wanted to test if the frequencies in it were accurate for the Puerto Rican population. We genotyped 736 individuals for the mutation R501X and 292 for R2447X using real time PCR. From the 736 individuals, we found 21 heterozygotes for the SNP R501X, this mutation in the FLG gene having an allele frequency of 0.0142. This allele frequency compared to the one previously reported, shows a difference of 0.0114 suggesting that the mutation in Puerto Rico is not rare. This mutation has been identified mainly in 3 geographical regions of Puerto Rico with high European ancestry. We started working with the mutation R2447X and genotyped 292 individuals; and 3 heterozygotes were found. From this samples the mutation shows an allelic frequency of 0.005; and it appears to be localized in the east region of the island. Now, we expect to genotype the same individuals for both mutations and compare their distribution, prevalence and relation between both.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gene Expression of DNA Repair Proteins in Colon Cancer Tissue
Authors: Odufuwa Kuburat Temitope , Elder Rhoderick , Povey Andrew
Published: June 2021

Cancer is a disease that arises from the disruption of cellular and genetic functions, either intrinsically or extrinsically. DNA repair genes play a critical role in maintaining the integrity of the genome and the dysregulation of the key DNA damage repair genes has been implicated in the development, progression and chemotherapeutic resistance of different cancers. Colorectal cancer (or colorectal adenocarcinoma) is one of the most common malignant neoplasms and a leading cause of cancer‐related deaths worldwide. This work will compare the level of gene expression of the DNA repair genes NEIL1, NEIL2, NEIL3, ERCC1, MLH1, OGG1, and NTHL1 with the housekeeping gene GAPDH in colon tumour tissue and matched normal colon tissue. Various molecular biology tools and techniques were employed in this research, including RNA extraction and purification, complementary DNA (cDNA) synthesis and qRT‐PCR analysis. Gene expression levels were determined using the comparative Ct method. For the twelve tumour tissues investigated, there is a reproducible and characteristic pattern of expression of the genes, with trends of expression observed between the Neil group of genes; Neil3 and Ercc1, and between Neil3, Ercc1 and Mlh1. The results showed that across the tumour tissues, the overexpression of one of the Neil genes marks the suppression of the others. Similarly, the results further demonstrated that there is a special relationship between Neil3 and Ercc1, with both genes antagonistic to each other. The expression patterns of these genes are indicators of the level of genetic instability and could serve as biomarkers for cancer diagnosis.

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