bluuebunny/crossref_metadata_2025 huggingface dataset:
Rostrocaudal changes in the protein expression of NMDA receptor subunits within the rostral ventrolateral medulla of sedentary and active rats (875.12)
Authors: Subramanian Madhan , Mueller Patrick
Published: June 2021

Recently we found enhanced sympathoexcitatory responses to glutamate in the RVLM following sedentary conditions. We have also reported increased dendritic branching in more rostral compared to caudal regions within the RVLM of sedentary rats. The mechanisms by which physical inactivity increases glutamatergic neurotransmission and produces structural changes in more rostral regions of the RVLM is unclear. The NMDA subtype of glutamate receptor has been associated with neuroplasticity in other brain regions of sedentary versus active rats. We hypothesized that sedentary conditions would alter protein expression of NMDA receptor subunits in a rostrocaudal manner within the RVLM. To test this hypothesis, male Sprague Dawley rats were allowed to remain sedentary (n=4) or exercise voluntarily via running wheels (n=4) for 14 weeks. Animals were then sacrificed and the RVLM cryosectioned (80 μm) and micropunched bilaterally. Sections were stained using cresyl violet to identify the rostrocaudal boundary of the RVLM. Based on their location relative to the caudal pole of the facial nucleus (FN), punches were pooled into four regions: caudal ‐480 µm (FN‐480), caudal ‐240 µm (FN‐240), rostral +240 µm (FN+240) and rostral +480 µm (FN+480). Preliminary western blot results showed that within the RVLM of sedentary rats, NR1 protein expression normalized to actin was significantly higher at FN‐480 (2.6 ± 0.2) compared to FN+240 (1.6 ± 0.2, p < 0.005). Interestingly, within the caudal regions of the active rats, NR1 protein expression was significantly lower at FN‐480 (1.4 ± 0.2) compared to FN‐240 (2.3 ± 0.2, p < 0.005). Changes were not observed for NR2A protein expression within both groups. Rostrocaudal differences observed in sedentary rats were not consistent with our structural findings. However, it is important to further investigate these novel findings, including examination of other receptor subunits which could relate to the structural neuroplasticity observed in sedentary rats.

Grant Funding Source: R01HL096787
17β‐Estradiol replacement to ovariectomized rats ameliorates depression‐like behavior (876.2)
Authors: Lu Jiangqiang , Xu Yongjun , Sheng Hui , Tang Zhiping , Ni Xin
Published: June 2021

Administration of estrogens elicits antidepressant‐like effects in female rats. However, the mechanism of this effect remains largely unknown. Inflammation and indoleamine‐2,3‐dioxygenase (IDO) have been implicated to play pivotal roles in the depression‐like behavior. The objective of the present study was to investigate whether estrogen deficiency‐induced depression‐like behavior is associated with inflammation and IDO activation in brain. It has been found that ovariectomy induced depression‐like behaviors in rats, while 17β‐estradiol (E2) treatment for 12‐week ameliorated depression‐like behaviors in ovariectomized (OVX) rats. Ovariectomy resulted in a decrease in 5‐HT and BDNF levels and an increase in levels of IDO, proinflammatory cytokines IFN‐γ and IL‐6, TLR‐4 and phosphorylated NF‐κB (p65 subunit) in hippocampus but not in prefrontal cortex. These effects could be restored by E2‐replacement. Serum E2 level was inversely correlated with the levels of IDO, IFN‐γ and IL‐6, while positively correlated with 5‐HT in hippocampus. Our results suggested that estrogen inhibit inflammation and activation of IDO and maintains 5‐HT and BDNF level in hippocampus, thereby ameliorating depression‐like behavior.

Grant Funding Source: Natural Science Fundation of China No.31371197
Blood oxygen content and affinity in developing embryos of snapping turtle Chelydra serpentina (879.8)
Authors: Sartori Marina , Kohl Zachary , Taylor Edwin , Abe Augusto , Crossley Dane
Published: June 2021

In egg‐laying amniotic vertebrates gas exchange takes place over the chorioallantoic membrane (CAM). Oxygen diffuses from air in contact with the shell into blood perfusing the membrane where it combines reversibly with hemoglobin. The capacity of the blood to transport oxygen is determined by the functional characteristics of the hemoglobin. In this study we investigated changes in oxygen content and affinity during development in embryos of snapping turtle (Chelydra serpentina). Blood samples were taken from embryos at 50, 70 and 90% of incubation. Samples withdrawn from the CAM artery had statistically similar mean oxygen contents for all stages (combined mean ± SEM; 0.94 ± 0.13 mL O2/100 mL of blood). Mean CAM venous blood oxygen content was 2.74 ± 0.44 at 50%, 3.25 ± 0.26 at 70% and increased to 4.97 ± 0.47 mL O2/100 mL at 90% of development. Consequently, the arteriovenous difference increased by 72% late in incubation. Oxygen equilibrium curves were the same at 2% and 6% CO2 at 50% incubation but showed an increased affinity in 2% CO2 and a rightward shift in 6% CO2 at 70 and 90% of development. These results show that embryonic turtles increase oxygen content and affinity and show a Bohr shift late in development likely favoring hemoglobin loading at the CAM and unloading at the tissues, ensuring adequate O2 supply for the increased demand of later stages of development.

Grant Funding Source: Supported by NSF CAREER IBN IOS‐0845741 (DAC) and FAPESP 2012/06938‐8 (EWT); 2013/05677‐9 (MRS)
Moderate endurance exercise training does not promote cardiac fibrosis (881.10)
Authors: Billman George , Cagnoli Kristen , Csepe Thomas , Li Ning , Fedorov Vadim
Published: June 2021

It has been recently proposed that prolonged high intensity exercise training induces myocardial damage that results in cardiac fibrosis which, in turn, could precipitate adverse cardiac events including sudden cardiac death. In contrast, moderate endurance exercise has been shown to protect against lethal arrhythmias. It is, therefore, possible that moderate exercise may produce physiologically beneficial adaptions without inducing myocardial fibrosis. To test this hypothesis, mixed‐breed dogs were randomly assigned to either a 10‐wk exercise training (ET, n = 15; progressively increasing treadmill running) or an equivalent sedentary period (SP, n = 10). Fibrosis was histologically quantified in each cardiac chamber using Masson’s Trichrome staining of paraffin‐embedded sections. The ET group exhibited lower heart rate and higher heart rate variability (high frequency component of R‐R interval variability and the standard deviation of R‐R; indices of cardiac vagal regulation) both at baseline and during a submaximal exercise test following the 10‐wk training program, while these variables were not altered in the SP animals. Despite this confirmation that a trained state had been achieved, fibrosis did not vary between the ET and SP groups for any cardiac chamber (Right atrium: SP, 12.1± 2.9 vs. ET, 11.4 ± 2.7; Left atrium: SP, 7.5 ± 4.2 vs. ET, 7.8 ± 2.0; Right ventricle: SP, 8.1 ± 2.3 vs. ET, 9.1 ± 1.6; Left ventricle: SP, 7.2 ± 1.7 vs. ET, 7.6 ± 1.0 %). These data demonstrate that moderate endurance exercise training can produce physiological adaptations without the induction of cardiac fibrosis.
Systemic blood pressure predicts disease severity of idiopathic pulmonary arterial hypertension (884.13)
Authors: Sun Xing‐Guo
Published: June 2021

Introduction: NYHA class and objective measurements obtained during cardiac catheterization (CATH) and incremental cardiopulmonary exercise testing (CPET) are all used to assess the severity of idiopathic pulmonary arterial hypertension (IPAH). Methods: We retrospectively analyzed the first visiting data from 86 IPAH patients without known systemic hypertension evaluated. After signed consent forms, CATH and CPET data were obtained without intervening changes in therapy. We correlated % predicted peak O2 uptake (%pkVO2) and NYHA class with systolic blood pressure at rest (SBPr) and peak exercise (SBPp) during CPET and the following resting CATH measures: mean right atrial pressure (mRAP), mean pulmonary artery pressure (mPAP), total pulmonary vascular resistance (TPVR), SV, and CO. Data are reported as mean±SD. Results: The mean age was 44±14 yr, NYHA class 2.8±0.6, %pkVO2 48±14%, and SBPp was 160±28 mmHg. Correlations from highest to lowest for %pkVO2 were: SBPp=0.51 (p<0.0001), SBPr=0.48 (p<0.0001), SV=0.36 (p<0.01), TPVR=‐0.34 (p<0.01), CO=0.27 (p<0.05), mPAP=‐0.26 (p<0.05), and mRAP=‐0.07 (p>0.05). Correlations for NYHA class were: SV=‐0.29 (p<0.05 and SBPp=‐0.25 (p<0.05) only. Conclusion: In IPAH patients, easily non‐invasive measured resting systemic systolic blood pressure during CATH and CPET correlate better with NYHA, and especially by %pkVO2, than more commonly‐employed invasive measures obtained during CATH.

Grant Funding Source: Chinese National High Technology Research and Development Program (863 Program) Project2012AA021009
PGC‐1α gene transfer rescues dystrophic muscle from advanced disease progression (884.18)
Authors: Hollinger Katrin , Barton Elisabeth , Selsby Joshua
Published: June 2021

Duchenne muscular dystrophy (DMD) is a progressive, fatal, muscle wasting disease caused by a dystrophin deficiency. The benefits of exercise for DMD patients are unclear and advanced disease progression may limit exercise capacity. We found previously that gene transfer of the exercise mimetic, PGC‐1α, increased abundance of utrophin as well as increased mitochondrial biogenesis using prevention and rescue paradigms. It is well accepted that increased abundance of the dystrophin‐related protein, utrophin, serves as an adequate substitute for dystrophin in dystrophic skeletal muscle. Our purpose in this investigation was to determine the extent to which PGC‐1α gene transfer would rescue dystrophic muscle following prolonged disease progression. One year old male mdx mice from our colony were injected in one hind limb with a virus driving expression of PGC‐1α while the contralateral limb was injected with empty capsid. Following three months of treatment we found that virus caused a 5‐fold increase in PGC‐1α expression in the gastrocnemius. Muscle function was measured in the soleus, in vitro. Muscle weight, tetanic force, and cross sectional area were similar between groups. Soleus muscles over‐expressing PGC‐1α increased specific tension by approximately 60% (p<0.05) and were more resistant to fatigue when stimulated over the course of 10 minutes (p<0.05). Force produced during the final contraction was 60% greater in treated soleus muscles than control (p<0.05). These data indicate that PGC‐1α pathway activation may interrupt the disease process even if initiated within the context of advanced disease. Supported in part by CIAG and Physiological Assessment Core of the Wellstone Muscular Dystrophy Cooperative Center at the University of Pennsylvania.

Grant Funding Source: CIAG
Activation of mechanosensitive ion channel TRPV4 promotes normalization of tumor vasculature and improves cancer therapy (893.12)
Authors: Thoppil Roslin , Adapala Ravi Kumar , Cappelli Holly , Ghosh Kaustabh , Dudley Andrew , Paruchuri Sailaja , Keshamouni Venkateshwar , Klagsbrun Michael , Meszaros J. Gary , Chilian William , Ingber Donald , Thodeti charles
Published: June 2021

The impaired development of tumor vessels, attributed to their highly permeable, tortuous, and hyper‐dilatory properties make it a barrier for the efficient delivery of anti‐cancer agents. Although anti‐VEGF therapies have shown to induce transient normalization of the tumor vasculature and increase the efficacy of cancer drugs, they are met with limited success clinically. Here, we demonstrate that a mechanosenstive ion channel TRPV4 regulates tumor angiogenesis and tumor vessel maturation. Tumor xenograft experiments revealed that, compared to tumors in wild type (WT) mice, those in TRPV4 null mice exhibit greater vascular density and immaturity (increased vessel diameter and reduced pericyte coverage). Endothelial cells derived from the tumor (TEC) showed reduced TRPV4 expression and activity associated with high basal Rho activity as well as aberrant mechanosensitivity and abnormal angiogenesis. Activation of TRPV4 with a small‐molecule activator, GSK1016790A, or TRPV4 overexpression suppressed the endogenously high rho activity, normalized the aberrant capillary formation and abnormal migration exhibited by TEC in vitro. Finally, GSK1016790A in combination with anti‐cancer drug Cisplatin, but not alone, reduced tumor growth in WT mice by normalizing the tumor vessels. Together, these studies demonstrate that TRPV4 channels are critical regulators of tumor angiogenesis, and represent a potential novel therapeutic target for tumor vascular normalization and improving the efficacy of anti‐cancer drugs.
KCa3.1 in the early stages of ischemic stroke (893.19)
Authors: Chen Yi‐Je , Yuen Natalie , Wallace Breanna , Wulff Heike , O'Donnell Martha
Published: June 2021

In the early hours of ischemic stroke, a luminal Na‐K‐Cl cotransporter (NKCC) at the blood‐brain barrier (BBB) works with an abluminal Na/K pump to transport Na+ (with water following) from blood to brain. Ischemic factors (hypoxia, aglycemia and vasopressin) increase BBB NKCC activity in a manner dependent on elevation of intracellular Ca2+. Here, we identified the presence of the calcium‐activated KCa3.1 channel on cultured bovine BBB endothelial cells (CMEC). In airway and gastrointestinal epithelia, inhibition of KCa3.1 channels slows Cl‐ efflux and increases intracellular Cl‐ ([Cl‐]i). Since NKCC activity is highly sensitive to inhibition by [Cl‐]i increasing [Cl‐]i in BBB endothelial cells should reduce Na+ secretion into the brain during ischemic stroke. We therefore hypothesized that pharmacological blockade of KCa3.1 will reduce or prevent brain edema. The KCa3.1 blocker TRAM‐34 reduced Rb86 efflux in a dose‐dependent manner and inhibited vasopressin‐induced rises in CMEC [Ca2+]i MR spectroscopy and imaging studies in rat middle cerebral artery occlusion showed that pre‐treatment with TRAM‐34 (40mg/kg, IP) significantly slowed brain Na+ uptake and cytotoxic edema at a time when the BBB is still largely intact. Our findings suggest that KCa3.1 blockade constitutes an attractive approach for reducing edema in the early stages of ischemic stroke or during brain surgeries.

Grant Funding Source: Supported by AHA & NIHGM
CX3CR1 deficiency alters response to L. reuteri treatment of DSS‐induced colitis in mice (902.10)
Authors: Ahl David , Roos Stefan , Phillipson Mia , Holm Lena
Published: June 2021

Preliminary results from our laboratory suggest that two strains of the probiotic bacteria Lactobacillus reuteri ameliorate DSS‐induced colitis in mice. The chemokine receptor CX3CR1 is expressed by circulating monocytes and by intestinal lamina propria resident cells believed to be macrophages and/or dendritic cells. Mice deficient in CX3CR1 (CX3CR1GFP/GFP) develop an aggravated DSS‐induced colitis, suggesting a role of CX3CR1 in the maintenance of colonic homeostasis. The aim of the study was to investigate if the protective mechanism of L. reuteri R2LC or ATCC PTA 4659 on DSS‐induced colitis involve CX3CR1. Colitis was induced by 3% DSS in drinking water, administrated for 8 days to C57Bl/6 (wt) and CX3CR1GFP/GFP mice. Some of the mice were given L. reuteri R2LC or ATCC PTA 4659 by gavage once daily for 15 days in addition to DSS for the last 8 days. Disease activity index (DAI) was calculated from clinical symptoms and measurements of the pro‐inflammatory cytokines IL1‐β, IL‐6, TNF‐α and mKC were also performed in colonic tissue samples. Immunohistochemical (IHC) staining of colonic tissue sections was used to characterize the population of macrophages and dendritic cells. In the CX3CR1GFP/GFP mice, DAI increased slightly more by DSS than in the wt, and the L. reuteri strains did not significantly reduce DAI, in contrast to significant reductions in wt mice. The levels of all proinflammatory cytokines were significantly higher in DSS treated CX3CR1GFP/GFP mice than in wt. Treatment with L. reuteri significantly reduced most cytokines in wt and CX3CR1GFP/GFP mice. Preliminary data from IHC stainings show a phenotypically skewed population of lamina propria immune cells in CX3CR1GFP/GFP mice. These results indicate that the protective role of the two different strains of L. reuteri on DSS‐induced colitis is slightly reduced in CX3CR1GFP/GFP mice, compared to wt mice. Thus, the mechanism by which L.reuteri bacterial strains ameliorate colitis might involve CX3CR1.
Role of PARP‐1 in the modulation of neutrophil function: relevance for inflammatory bowel disease (902.5)
Authors: Larmonier Claire , Shehab Kareem , McFadden Rita‐Marie , Ghishan Fayez , Kiela Pawel
Published: June 2021

Aberrant immune response has been proposed as the underlying cause of Inflammatory Bowel Diseases. Disease progression and relapse are attributed to a hyperactive adaptive immune response to the bowel luminal contents accompanied by a disproportionate and persistent neutrophil (PMN) influx. Poly(ADP‐ribose) polymerases (PARPs), including its most abundant isoform PARP1, catalyzes the transfer of ADP‐ribose units from NAD+ to a number of target proteins. In vivo studies have demonstrated that genetic ablation or pharmacological inhibition of PARP1 ameliorates the experimental colitis pathophysiological changes. Considering the essential role of PARP1 in the regulation of inflammation, we propose to evaluate the effect of Parp1 deletion on neutrophil function in a model of Dextran Sulfate Sodium (DSS)‐induced colitis. In comparison to WT mice, Parp1‐/‐ were protected from the effects of DSS, with significantly lower weight loss, and reduced proinflammatory mediators, including metalloproteinases and chemoattractants. The regulation of neutrophil‐related gene in the colon was also studied by microarray. Similarly, Parp1‐/‐ neutrophils demonstrated an impaired chemotaxis in vitro and reduced motility in vivo in a model of aseptic peritonitis. In light of these current findings, we postulate that the pathogenic role of PARP1 may be mediated in part by the regulation of neutrophil function.

Grant Funding Source: Supported by NIH Grant 1K01DK099268‐01
Unraveling the anatomy of the cardiac skeleton of the heart (913.6)
Authors: du Plessis Maira , Blaak Christa , Tubbs R Shane , Loukas Marios
Published: June 2021

The fibrous skeleton of the heart is an integral and significant structure, with both supportive and functional significance. Yet, its anatomy has not received much emphasis in literature. This fibrous skeleton has many vital functions within the heart, supporting the valves as well as the myocardium, and insulating the atrioventricular bundle as it courses towards the apex. It is a useful structure in cardiac surgery, providing support for repair and replacement of the heart valves. The paper reviews and correlates the anatomy, embryology and clinical significance of this structure. Dissection of the cardiac skeleton was performed on formalin fixed cadavers. It was found that the aortic root (the portion of the left ventricular outflow tract, supporting the aortic leaflets) which is typically described in terms of an “annulus”, displayed triple semi‐lunar leaflet attachment configuration. Unlike the atrioventricular annuli, the aortic root provides much more complex support for the aortic leaflets than the planar attachments of the mitral and tricuspid leaflets, creating a new understanding of the components of the cardiac skeleton. It has been reported that fibrous support of the cardiac skeleton is not uniform across all the valves. Others have shown it to be concentrated more centrally, seen in the posterior part of the right coronary and non‐coronary cusps. The membranous septum share by these cusps is continuous with the right fibrous trigone, the remaining aortic and pulmonary cusps have muscular anchors. Although less frequent than myocardial, pathology can arise in the fibrous elements of the heart, either during embryological development, or later in life. It is clear that this structure needs to be studied more extensively.
First report of a “latissimo‐cleidocostal” muscle variant on the anterolateral thoracic wall (915.12)
Authors: Miller Joseph , Trelease Robert , Sun Zhengzheng
Published: June 2021

During dissection of the upper limb in a gross anatomy course for first year dental students, we identified an unusual, supernumerary muscle in the right torso of a 91 year‐old, Caucasian male cadaver. This consisted of a substantial muscle bundle continuous with the antero‐inferior border of the latissimus dorsi, bearing a lengthy, strap‐like tendon that passed superomedially, deep to the pectoralis minor, to insert broadly on the neck of the clavicle and first costal cartilage. Although this muscle bore some partial resemblance to reported anterior supracostal muscle variants on the anterior thoracic skeleton, which are typically innervated by intercostal nerves, it was unique in its: 1) continuity with the latissimus dorsi, 2) bifid insertion on both clavicle and first rib, and 3) innervation by branches of the thoracodorsal nerve. Furthermore, there was no resemblance to previously described axillary arch muscles and other rare variations of latissimus dorsi muscle slips and insertions. Based on its demonstrated insertions and innervation, we name this unusual variant a “latissimo‐cleidocostal muscle”. This recognition should now serve as a working hypothesis, to be tested with future observation and study, that this muscle is distinct from currently recognized supracostal or other latissimus dorsi variants.
Quantifying adaptive evolution: three‐dimensional phenotypes provide new insight into mechanisms of rapid evolutionary change in threespine stickleback (918.28)
Authors: Jamniczky Heather
Published: June 2021

How do evolutionary transitions occur following environmental change? Threespine stickleback (Gasterosteus aculeatus) are among the most well‐studied natural examples of rapid evolutionary change. They provide an ideal model with which to examine the means by which many parallel examples of adaptive phenotypic modification can occur over very short periods of time. Here we discuss the results of two studies employing a three‐dimensional (3D) approach to phenotypic quantification, combining the techniques of micro‐computed tomography and geometric morphometrics with the tools of molecular ecology to gain new insight into the mechanisms driving adaptive change. First, we tested the hypothesis that two freshwater populations originating from the same putative ancestral population have diverged to more closely resemble each other than they do their recent common ancestor. We demonstrate that sticklebacks establishing new populations in Alaskan freshwater lakes follow similar phenotypic trajectories. Second, we used a quantitative trait locus analysis of a marine‐freshwater cross from British Columbia to test the hypothesis that a small number of loci of large effect are responsible for marine‐freshwater phenotypic divergence. We recover both previously identified QTL as well as a new, previously unreported loci with the potential for wide‐ranging effects on skeletal development. We discuss the framework of an integrative research program aimed at elucidating the means by which key characteristics of the phenotype may allow organisms to adapt rapidly to changing environments.

Grant Funding Source: Supported by the Natural Sciences and Engineering Research Council of Canada
Investigating the function of the caudofemoralis longus muscle and musculoskeletal plasticity in American alligators (Alligator mississippiensis) (919.9)
Authors: Joneson J , Medina A , Owerkowicz T , Elsey R
Published: June 2021

The CFL ‐ a major retractor/medial rotator of the thigh ‐ is substantial in reptiles with a sprawling gait and long tail. The CFL originates on the transverse processes and chevrons of the caudal vertebrae and inserts on the fourth trochanter of the femur. Evolutionary changes to the hindlimb/tail module in archosaurs have been ascribed to a reduced role of the CFL in terrestrial locomotion, but no alteration of CFL function has been attempted. To discern its role in locomotion, and investigate muscular plasticity, we used bilateral tenotomy to deactivate the CFL in juvenile alligators (Mb=180±30g, SVL=213±28mm). After surgery, we found no significant differences between experimental animals and sham‐operated controls (n=6 each) in average speed, stride length or hip height during walking, when filmed on a level grid trackway. Eight months post‐surgery, CFL wet mass and fiber length were significantly reduced, by 23% and 13% respectively, in experimental animals compared to controls (n=12 per group). This suggests that the contribution of the CFL to hindlimb stance/movement is limited, or compensatory mechanisms are present in the hip during slow and steady locomotion. Further study of muscle fiber phenotype and bone growth/remodeling following tenotomy will provide insight into the nature of plasticity associated with disuse atrophy, and help elucidate the role of musculoskeletal strain in evolutionary transformations of the hindlimb and tail locomotor modules in archosaurs.

Grant Funding Source: Supported by: Research Initiative for Scientific Enhancement (RISE), National Institutes of Health
Consequences of the loss of collagen XXIV in bone (922.7)
Authors: Gordon Marion , Jeon Jae Yoon , Zhou Peihong , Hahn Rita , Svoboda Kathy , Ramirez Francesco
Published: June 2021

Fibrillar collagens give integrity and tensile strength to many organs in the body. Collagen XXIV is a non‐traditional collagen in the fibrillar collagen family. To determine what its function might be in bone, collagen XXIV knock out (KO) mice were analyzed. Since femurs of null mice at 3 and 5 months of age were found to be osteoporotic compared to WT mice, our goal was to quantify mRNAs involved in regulation of bone deposition and resorption, and determine whether the loss of collagen XXIV affected their expression. Femur RNA was extracted from WT and KO mice at 7 days, 1 month, 2 months, 3 months, 6 months, 1 year and 1.5 years of age. RNA was converted to cDNA, and real time qPCR was performed using GAPDH as a control. In WT mice, collagen XXIV mRNA was high at 7 d and 1 month of age. At 2 months it was present at less than half of what it was at 1 month. By 3 months its levels were greatly attenuated. This trend continued for all ages up to 1.5 yrs. KO mice showed minor amounts of collagen XXIV mRNA at 7d, 1 month and 2 months of age, with essentially no transcribed product by 3 months and beyond. Expression levels of RANKL, Runx2, and Lrp5 were elevated in the KO mice. For RANKL, values were 1.7 to 2 fold higher in the KO mouse femurs at all ages except at 2 months, where it was 3.2 times higher. Runx2 mRNA levels were always ~2 to 5 fold higher, except at 1 month of age, where it was ~10 times higher. Lrp5 was ~1.6 to 3.5 times higher in KO mouse femurs than in WT femurs. In summary, seven day old and 1 month old WT mice had the highest expression levels of collagen XXIV mRNA. By 2 months of age, expression was halved, and by 3 months it became negligible. The higher expression of RANKL in KO mice may be responsible for the observed osteoporosis at 3 and 5 months of age. Higher levels of Lrp5 and Runx2 may represent a compensatory mechanism to stimulate bone growth.
Hexane fraction of Costus afer Ker Gawl leaf inhibited mitochondrial permeability transition, F1F0 ATPase and scavenged nitric oxide and hydrogen peroxide (957.1)
Authors: Anyasor Godswill , Onajobi Funmilayo , Osilesi Odutola , Adebawo Olugbenga
Published: June 2021

Recent implication of mitochondria in the etiology of inflammatory diseases especially rheumatoid arthritis led to the evaluation of in vitro and in vivo inhibitory effects of an indigenous medicinal plant, Costus afer leaf hexane fraction (CAHFL) on mitochondrial permeability transition (MPT), F1F0 ATPase, nitric oxide (NO•) and hydrogen peroxide (H2O2) using spectrophotometric methods. Results showed that 10, 20, 40 and 60 µg/ml CAHFL significantly (P<0.05) inhibited Ca2+ induced MPT in a concentration dependent manner by 85.66%, 86.93%, 87.47% and 90.47% respectively when compared with untreated mitochondria in vitro. Spermine potentiated CAHFL MPT inhibition by 88.20%, 90.20%, 92.12% and 93.51% respectively. The In vivo study showed that 50, 100 and 250 mg/kg body weight CAHFL significantly (P<0.05) inhibited MPT in arthritic rats by 52.22%, 61.11% and 65.0% respectively in a dose dependent manner compared with control group (untreated) in 7 days. Concentrations of 10, 20, 40, 60 and 80 µg/ml CAHFL significantly (P<0.05) reduced F1F0 ATPase activity in a concentration dependent manner with 3.93 ± 0.01, 3.88 ± 0.04, 3.63 ± 0.07, 3.51 ± 0.05 and 3.49 ± 0.04 mg/ml phosphate (Pi) liberated respectively compared with 10 µg/ml 2,4‐dinitrophenol (6.60 ± 0.03 mg/ml Pi.). CAHFL scavenged NO• and H2O2 in a concentration dependent manner with an IC50¬¬ of 1.77 mg/ml and 0.33 mg/ml respectively. These data indicated that CAHFL possess the ability to protect mitochondria from damage which may account for its anti‐inflammatory action, further strengthening the claim that CAHLF could be a candidate for anti‐inflammatory drug development.

Grant Funding Source: Babcock University research grant [BU/RIC/003]
Genesis of antibiotic resistance V: transport of antibiotic medication across United States ‐ Mexico border regulated by federal agencies (986.5)
Authors: Kannan Subburaj , Subburaj Prabavathy
Published: June 2021

It is a popular perception that antibiotics could be purchased freely from Mexico and taken in to the United States with ease. Unrestricted flow of antibiotics and the misuse of the antibiotics by the general public on both sides of the United States ‐ Mexico border regions have been at the forefront when discussing the underlying mechanisms for the emergence of “Antibiotic Resistance” in bacterial pathogens triggered by infectious diseases. To dispel the popular misconception, a systematic inquiry has been made at the Port of Entry in Eagle Pass, Texas ‐ United States ‐ Mexico border inspection station. As per the counsel of the Customs and Border Patrol, with the supervisory regulation form FDA ‐ Washington D.C, the prescription medicines that are allowed to pass through the port of entry must be for personal use only. It must have the Doctor’s written prescription for a three month period maximum. Next, the drug must be on the list of the medications approved by The United States Food and Drug Administration which available on the www.FDA.gov. This information was affirmed by the Station Chief of Operation, FDA, stationed at the Eagle Pass ‐ United States ‐ Mexico ‐ Port of Entry. If the antibiotic medication exceeds allowed quantity, it will be referred to the Import Section of the FDA stationed elsewhere for further processing without entry in to the United States. However, the trepidation over factors such as rate of inflow of the travelers, the time of consumption to screen and evaluate each visitor is a serious and significant concern. Further details of issues and concerns shall be discussed in Experimental Biology of 2014

Grant Funding Source: Professional Development Funds from SWTJC, Eagle Pass, Texas
Methylation of hippo pathway genes in bone, kidney, breast and prostate cancer cells (991.1)
Authors: Alokail Majed , Alenad Amal
Published: June 2021

Salvador‐Warts‐Hippo pathway, regulates tissue growth in a wide variety of organisms and plays fundamental roles in cancer. DNA hypermethylation being widely used as a feature of many cancers. We identified differential methylation patterns in bone, kidney, breast and prostate cancer cell lines. Methylation analysis of CpG islands revealed by methylation specific PCR. The preliminary finding shows that there are number of genes were differentially methylated in investigated cancer cell lines. Methylation of this pathway vary between these cells. Methylation frequency was detected in number of cancer cells. In MST1 and MST2 analysis, 1 out of 3 bone cancer cells showed methylation of the MST1 and MST2 and 3 out of 6 breast cancer cells showed methylation of MST1 and MST2. In case of prostate cancer cells, 1 out of 4 these cell lines showed methylation of the MST1 without detection of any methylation in MST2. In SAV1, 3 kidney and breast cells showed methylation was detected compared with 2 of prostate cells and non of the bone cancer cells showed any methylation status. In LATS1 and LATS2 many kidney cell lines observed methylation patter for both LATS1 and LATS2, however, only 2 breast cells showed methylation of LAST1 compared with 1 of bone cells and non of prostate cancer cells showed any methylation. In conclusion, hippo pathway observed epigenetic variations in different cancer cell lines within the same cancer type.

Grant Funding Source: King Saud University
Concordance of Affymetrix GeneChip® Human Transcriptome Array 2.0 and USB® VeriQuest™ real‐time PCR data (LB207)
Authors: Ujvari Andrea , Chamnongpol Sangpen , Kourennaia Olga , Myers Kim , Bressler Vincent , Hubbard Jason , Schweitzer Anthony , Kubu Chris
Published: June 2021

Global gene expression analysis using Affymetrix GeneChip Human Transcriptome Array (HTA) 2.0 combined with Affymetrix Transcriptome Analysis Console (TAC) Software empowers scientists not only to gain information about RNA expression at the gene‐level, but also about RNA alternative splicing at the exon‐level. Real‐time PCR is routinely used to verify microarray results due to its high sensitivity and wide dynamic range, and was used in this study to validate Affymetrix GeneChip HTA 2.0 data. Gene‐level expression data for the classic MAQC A and MAQC B samples (used by the MicroArray Quality Control Consortium) were validated with commercially available primers and design software. Excellent fold‐change correlation of microarray and qPCR data were observed using USB VeriQuest Probe real‐time PCR (R=0.96, slope=0.70). For alternative splicing validation of Affymetrix GeneChip HTA 2.0, total RNA from three different tissues (liver, muscle and brain) were used. Exon level information provided by TAC Software was utilized to design custom PCR primers for USB VeriQuest SYBR Green real‐time PCR. All alternative splicing events selected for validation were confirmed by real‐time PCR. We show excellent inter‐platform concordance between Affymetrix microarray and VeriQuest real‐time PCR data.

Grant Funding Source: Supported by Affymetrix, Inc.
Exposition of Müller AO classification of tibial fractures using cadaveric bone models (LB23)
Authors: Abdullatif Victor , Kirakosian Arman , Nusse Gloria
Published: June 2021

The Müller AO classification system of bone fractures has substantially served osteology education and medicine for many decades. The present study aims to provide a physical representation using castings of cadaveric tibial bones. These models will illustrate the fracture classifications in accordance with said system. Castings will have simulated fractures using surgical and mechanical‐force methods. Proximal, diaphysieal, distal, and malleolar fractures will be showcased. Cadavers are supplied by the University of California San Francisco’s Willed Body Program and San Francisco State University’s Human Anatomy Laboratory.

Grant Funding Source: none
Ethnomedicinal survey of folkloric plants used in managing breast cancers by the traditional medical practitioners of North‐east Nigeria (LB240)
Authors: Mohammed Zainab , Hamza Hajja , Daja Aliyu , Gidado Abubakar , Hussaini Isa
Published: June 2021

Cancer is a major leading cause of death in the world today. Breast cancer is the most common cancer in women in 140 countries. The widespread use of medicinal plants is not limited to the developing countries, there’s a renewed interest by the public and scientific organisations concerning plants and cure for diseases. The increasing cost of modern health care has made the role of ethnomedicine a very important one. A majority of the Nigerian population use traditional medicine for relief from various ailments largely due to inadequate medical facilities, high poverty rate and many other socio‐ economic/ socio‐ cultural factors. Local herbalists claim the possession of remedies useful in curing different cancer types. However, such knowledge is secretive and kept within close family circle.

This work is aimed at providing an inventory of North East Nigerian plants with anti‐ breast cancer potentials for future verification and scientific validation. Semi‐structured questionnaire and oral interview were used to obtain relevant ethnomedicinal data. Twenty five different plant species were identified to possess possible anti‐ breast cancer potentials. Plant names, family, part used, relevant pharmacologic activities and pictures of plants were documented.
Single serve frozen meals at lunch yield greater 24‐hour energy intake reduction compared to shakes (LB306)
Authors: Reimers Kristin , Jacobson Mary , Ward Tabra , Meyer Michael
Published: June 2021

Providing 蠄 450 kcal is an unrecognized attribute of most single serve frozen meals (SSFM), including those not marketed for weight management. Preliminary research shows energy compensation does not occur at other eating occasions when self‐selected lunch meals are replaced with portion‐controlled meals. Meal replacement is an evidence‐based strategy for weight management, with most evidence based on shakes as the intervention. While effective, shakes do not offer a traditional meal experience. This study examined efficacy of single serving frozen meals (SSFM) vs. shakes as meal replacements by assessing participants’ average 3‐day energy intake and satiety scores under 3 conditions: baseline, SSFM and shake. Participants (n = 23, 74% female, BMI 31.58 ± 2.95) consumed study food exclusively for 3 consecutive days/week, consisting of self‐selected buffet meals at the test facility and snacks to consume away from the facility. After the baseline 3‐d ad‐libitum condition, participants replaced their self‐selected lunch with a 280 kcal shake or SSFM in a random, counterbalanced manner. Participants completed 10 cm satiety rating scales 3 hours after lunch, and researchers weighed foods before and after consumption to calculate energy consumed. Mean ± SD hunger and desire to eat scores were lower 3 hours after SSFM vs. shake (5.64 ± 2.17 vs. 6.60 ± 1.87 and 5.67 ± 2.42 vs. 6.71 ± 2.11, respectively, p < 0.01). 24‐hour energy intakes were less during both SSFM and shake interventions vs. baseline (1900 ± 453, 1970 ± 431, and 2400 ± 596 kcal, respectively, p < 0.01). Additionally, compared to baseline, SSFM as a lunch meal replacement resulted in greater 24‐hour energy reduction than replacing lunch with a shake (‐500 ± 426 vs. ‐430 ± 388 kcal, respectively, p < 0.01). These results support the use of single serve frozen meals as a meal replacement for weight loss.
Endothelial nitric oxide synthase (eNOS) and soluble guanylyl cyclase (sGC) expression in the ovarian artery during the estrous cycle in overfed or underfed sheep treated with arginine (Arg). (LB324)
Authors: Redmer Dale , Bass Casie , Kaminski Samantha , Grazul‐Bilska Anna
Published: June 2021

The aim of this study was to determine if Arg supplementation to control, overfed, and underfed ewes impacts protein and mRNA expression of eNOS and sGC in the ovarian artery. Ewes were categorized by weight and randomly assigned to one of three different nutrition groups: control (C; 2.14 Mcal/kg; n=37), overfed (O; 2xC Mcal/kg; n=37), and underfed (U; 60% of C Mcal/kg; n=37) beginning 60 days prior to Arg treatment and were randomly assigned to one of two treatments; Arg (L‐Arg‐HCl; 155 µmol Arg/kg BW) or saline (~10 ml) i.v. beginning on day 0 of the first estrous cycle until tissue collection at the late‐luteal phase of the first estrous cycle, or early or mid‐luteal phases of the second estrous cycle. Ovarian vascular pedicle was fixed and imaged, and RNA was extracted from the ovarian artery (OA). Compared to saline controls, Arg increased mRNA expression of eNOS nearly 300‐fold and sGC approximately 150‐fold in OA for O group during the mid‐luteal phase (P<0.0001; n=5 for each group). Arg (n=3) increased eNOS 7‐fold (P<0.06) compared to saline (n=7) in U during the early luteal phase. No other Arg effects were observed in either other diet groups or on other luteal phases. eNOS protein was localized in blood vessel endothelium of the ovarian pedicle, whereas sGC‐beta protein was expressed in both vascular smooth muscle and endothelium of blood vessels. During the early or mid‐luteal phase when the corpus luteum is actively growing or fully functional, Arg can dramatically influence the NO system in OA by increasing mRNA expression of eNOS and sGC in U or O, and suggests that Arg supplementation may alter vascular function and blood flow to the ovary in compromised animals.

Supported by USDA‐AFRI grant 2011‐67016‐30174, and Hatch Projects ND01754 and ND01748 to ATGB and DAR.
Inhibition effect of Vitamine E δ‐tocotrienol on PhIP/DSS‐induced colon carcinogenesis in CYP1A‐humanized mice (LB325)
Authors: Chen Yu‐Kuo , Liu Anna , Lee Mao‐Jung , Wang Hong , Cheung Connie , Yang Chung S.
Published: June 2021

Our previous research indicated that combination of 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP), the most abundant heterocyclic amine produced during the cooking of meats and fish, with dextran sodium sulfate (DSS) rapidly induced colon carcinogenesis in transgenic CYP1A‐humanized mice (hCYP1A‐mice). Herein, we modulated the dosage of DSS (0.8‐1.5%) to establish a suitable model for cancer prevention research and to evaluate the inhibition effect of δ‐tocotrienol (δ‐T3) on colon carcinogenesis. The treatment with 200 mg/kg PhIP, followed by 1% DSS in the drinking water for 7 days moderately induced colon carcinogenesis in hCYP1A‐mice with a tumor multiplicity of 4.17 ± 0.58 at week 8. Tumor multiplicity was reduced in 0.025% and 0.5% δ‐T3 treatment groups by 49% (P < 0.05) and 58% (P < 0.01), respectively. Supplementation with δ‐T3 also increased the respective levels of tocotrienol in the serum and colon tissues. These results demonstrate the inhibitory effect of δ‐T3 against colon carcinogenesis, and the inhibitory activity may be due mainly to the actions of δ‐T3.

Grant Funding Source: NSC‐102‐2221‐E‐020‐021
Pre‐antiretroviral therapy (ART) selenium status predicts HIV clinical but not virologic failure post‐ART (LB326)
Authors: Shivakoti Rupak , Gupte Nikhil , Yang Wei‐Teng , Mwelase Noluthando , Kanyama Cecilia , Sugandhavesa Patcharaphan , Semba Richard , Christian Parul , Campbell Thomas , Gupta Amita , NWCS 319 and PEARLS study team
Published: June 2021

Selenium status, with important roles in oxidative stress and immunity, has a non‐linear U‐shaped association with all‐cause mortality in US populations; in HIV‐infected persons, low selenium is associated with increased viral load. Factors associated with selenium deficiency and whether selenium is independently associated with HIV ART outcomes in HIV‐infected persons is unknown. We performed a multi‐country case‐cohort where cases were HIV‐1 infected adults with either clinical failure (incident WHO stage 3, 4 or death by 96 weeks) or virologic failure (HIV RNA levels 蠅 1000 copies/mL for 2 successive visits at 蠅16 weeks). Median baseline selenium levels were 82.04 µg/L (IQR: 57.28 ‐ 99.89) and selenium deficiency prevalence was 53% varying widely by country from 0% (US) to 100% (Zimbabwe). Independent risk factors for baseline selenium deficiency (<85 µg/mL) were country, previous TB (adjusted odds ratio (aOR) 7.57, 95% CI 1.46‐39.21), low albumin (aOR 3.61, CI 1.13‐11.52), and log C‐reactive protein (aOR 2.74, CI 1.25‐6.02). Adjusted models showed a non‐linear U‐shaped association between selenium and clinical failure but no associations of selenium with HIV virologic failure. In summary, lower and higher levels of selenium pre‐ART were associated with increased hazards of clinical failure post‐ART. Selenium supplementation of HIV‐infected subjects initiating ART should be approached with caution.
Ingestion of leucine + glycine with glucose produces a partially additive effect on plasma glucose despite a less than additive effect on serum insulin (LB427)
Authors: Nuttall Frank , Iverson Jennifer , Gannon Mary
Published: June 2021

Leucine (Leu) is well recognized to stimulate insulin secretion and to reduce the glucose rise when ingested with glucose. Glycine (Gly) reduces the glucose rise, but only weakly stimulates insulin secretion. We have shown that both were particularly potent in their ability to attenuate the glucose response to 25 g glucose. The current study was designed to determine whether the glucose‐attenuating effects would be even greater if Gly and Leu were ingested together with glucose.

On 4 separate days, 9 healthy subjects were studied. Test meals were given at 0800 hr. Subjects received water only on day 1, followed by 25 g glucose only, Leu + Gly (1 mmol/kg fat free body mass each), or Leu + Gly with 25 g glucose, on subsequent days in random order. Serum/plasma glucose, insulin and glucagon were measured frequently thereafter for 2.5 hrs. Net areas under the curve were calculated.

The glucose area response decreased by 77% (p<0.02) after ingestion of Leu + Gly + glucose compared to ingestion of glucose alone. The insulin area response increased by only 24% (p<0.05). When compared to our previously published data on the individual amino acids, the decrease in glucose response with Leu + Gly + glucose was partially additive. The increase in insulin response was less than additive. The glucagon response was considerably less than additive.

Thus, there is a mechanistic interaction between Leu and Gly when ingested with glucose that results in a markedly attenuated glucose response. This occurred in the presence of an attenuated insulin response. Changes in glucagon response could not explain the results. Overall, the way in which ingested amino acids interact with each other and affect glucose metabolism is complex and not well understood.

Grant Funding Source: Supported in part by Funds from the Department of Veterans Affairs
Parental perceptions of Rhode Island EFNEP (LB464)
Authors: van Asch Patrick , Gorman Kathleen , Melanson Kathleen , Sebelia Linda , Tovar Alison
Published: June 2021

Parental perceptions of Rhode Island EFNEP Patrick van Asch, Kathleen Gorman, Kathleen Melanson, Linda Sebelia, Alison Tovar Nutrition and Food Sciences, University of Rhode Island Childhood obesity in the United States has tripled over the last thirty years. In Rhode Island (RI), 19% of children ages 2 – 4 are overweight and 16% are obese, which is higher than the national average (16% and 12% respectively). The Expanded Food and Nutrition Education Program (EFNEP) targets low‐income households and provides knowledge and skills to improve nutritional well‐being. This project aimed to assess parental perceptions of the RI‐EFNEP curriculum and how it addresses childhood obesogenic behaviors (sugar‐sweetened beverage consumption, physical activity, sleep duration, and screen‐time) using individual face‐to‐face semi‐structured interviews. Sixty‐minute interviews were conducted at local community centers with caregivers (n=22) of 2 – 8 year old children who had completed the program. A translator was present for Spanish speaking participants (n=14). Content analysis was utilized to analyze themes coded in Microsoft NVivo 10. Overall, caregivers reported a positive experience with the program and felt empowered to change their families’ health as a result (e.g. consuming more whole grains). In addition to learning about nutrition, they reported being able to improve health behaviors (e.g. increasing physical activity). Suggestions for improving the curriculum included more instruction in label reading, physical activity, media literacy, food demonstrations, portion control, and tips to get their children to be healthier. Although not part of the curriculum, caregivers reported learning little about sleep duration, screen‐time, and physical activity with regards to their children’s health. Since RI‐EFNEP targets at‐risk, low‐income households, the results from this project can be used to tailor the curriculum to focus more on childhood obesity prevention.

Grant Funding Source: Supported by the Rhode Island Foundation