bluuebunny/crossref_metadata_2025_split huggingface dataset:
Exploring the Developmental Thermal Biology of an Abundant Native Amphibian in Southern California, the Baja California Chorus Frog
Authors: Korito Lindsey , Manzanares Samantha , Bucsky Julie , Mueller Casey A. 
Published: June 2021

The Baja California chorus frog (Pseudacris hypochondriaca), a member of the Pacific chorus frog complex, is probably the most abundant native amphibian in southern California. The persistence of this species when many other amphibians are in decline provides a unique opportunity to examine the developmental and physiological attributes of a successful species. Developing stages are often particularly susceptible to environmental influences, and understanding physiological function during this time can provide insight into the drivers of species abundance and distributions. The focus of our research is to examine how the developmental physiology of the Baja California chorus frog is influenced by, and responds to, temperature. We collected egg clutches from local ponds within 24 h of fertilization. The clutches were separated and incubated at 10°C, 15°C and 20°C. Oxygen consumption rate (Vo2) was measured throughout development at each temperature. Embryos and tadpoles were fixed throughout development in 4% paraformaldehyde in phosphate buffered saline, then transferred to 70% EtOH, removed from their egg (if not already hatched) and photographed under a dissecting microscope. The yolk and embryo/tadpole were separated, weighed, dried at 65°C for 24 h, and reweighed. The stage and length of each embryo/tadpole was determined from photographs. Average time to 50% hatch and hatch window duration was determined across clutches. Development rate was strongly affected by temperature, with warmer temperatures reducing time to hatch. Temperature also influenced the duration of hatching, with the hatch window ranging from a few days at warmer temperatures to over a week at 10°C. Preliminary results indicate Vo2 was higher and more variable in embryos incubated at 20°C, but there was a limited temperature effect on growth, with length, dry mass and yolk dry mass similar between temperatures throughout development. These results indicate the conversion of yolk into tissue occurred at the same rate across temperatures, and the energy allocated to growth was not altered. However, because more oxygen was consumed at 20°C, the cost of development was higher than at 15°C. Individuals incubated in warmer temperatures may be “wasting some oxygen”, but this does not appear to confer negative physiological consequences. We will further explore morphological and physiological responses to temperature in embryos and tadpoles of this species to understand if robust thermal biology contributes to the success of the species.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Variation in atmospheric oxygen levels affects adult insect wing morphology and flight performance
Authors: Lyn Sheena L. , Christensen Jacob , VandenBrooks John , Schaefer Charles , Parker Greg 
Published: June 2021

Objective and HypothesesChanges in atmospheric oxygen over geologic time have been hypothesized to drive several evolutionary events in the history of insects. However, the insect fossil record is composed of mostly isolated wings, which presents challenges in interpreting the data and testing these hypotheses. Therefore, our objective was to understand how insect wing morphology varies with atmospheric oxygen as a critical first step in addressing these hypotheses. Given that wing veins contain tracheae which have been shown to vary with rearing oxygen elsewhere in the body, we hypothesized that wing vein diameters should be inversely correlated with rearing oxygen and that these changes would affect flight performance of these insects.MethodsDrosophila melanogaster and Blatella germanica were reared from hatching to adults under three different oxygen concentrations: 12% (hypoxia), 21% (normoxia) and 31% (hyperoxia). The wings were then dissected and imaged using a mechanical stage mounted on an inverted microscope. Wing area, wing vein diameters, wing vein lengths, and tracheal diameters were all measured using ImageJ. In a second experiment, flies were again reared in three different oxygen concentrations and then flown in all three oxygen levels to test the effect of these changes in wing morphology on flight performance.ResultsIn both Drosophila and Blatella, mass specific wing area, wing vein diameters, and tracheal diameters were all inversely correlated with rearing oxygen. In terms of flight performance, flies reared in 31% oxygen flew the best. However, overall flies flew the best in the oxygen level they were reared under.ConclusionsThese results demonstrate that rearing oxygen impacts wing morphology in several groups of insects and that these changes in morphology directly impact flight performance. These changes suggest that wing morphology in the fossil record can be used to interpret evolutionary changes. Additionally, this suggests that wing vein diameter may be a potential proxy for atmospheric oxygen over geologic time.
Early Neuronal Inflammation May Be a Contributing Factor to Changes in Epidermal Innervation Associated with Neuropathy in Prediabetic Mice
Authors: Reller Abigail M. , Umbaugh David S. , Schaller Megan L. , Maciejewski J. Claire , Wooten Joshua S. , Guilford Brianne L. 
Published: June 2021

Neuropathy, a crippling diabetic complication, was initially purported to result from lack of glycemic control. However, recent research has linked neuropathy to obesity in individuals without diabetes. Furthermore, evidence indicates mice show signs of neuropathy after high fat feeding. However, the pathogenesis of neuropathy induced by a high fat diet is still unknown.PURPOSEThe purpose of this study was to determine if early neuronal inflammation is a contributing factor to changes in epidermal innervation and mechanical hypersensitivity associated with neuropathy in prediabetic mice fed a high fat diet.METHODSMale C57Bl/6 mice were randomized to a standard diet (Std, 15% kcal from fat) or high fat diet (HF, 54% kcal from fat) for 2, 4, or 8 weeks (n = 11–12 mice per group). Inflammatory mediators (IL‐1α, IL‐1β, IL‐2, IL‐3, IL‐4, IL‐5, IL‐6, IL‐10, IL‐12p70, IL‐17, MCP‐1, IFN‐γ, TNF‐α, MIP‐1α, GMCSF, RANTES) were quantified in the lumbar dorsal root ganglia (DRG) using Multiplex ELISA. Additionally, qRT‐PCR was performed to measure mRNA expression of genes associated with inflammation (IL‐1β, IL‐6, IL‐10, MCP‐1, TNF‐α, RANTES) in epididymal adipose. Mechanical sensitivity associated with neuropathy was determined using the von Frey test and measured every other week, beginning the week before the dietary intervention. In order to quantify intraepidermal nerve fiber (IENF) density and a specific population of peptidergic nociceptive nerve fibers (TrkA) in the hindpaw footpad skin, fluorescence immunohistochemistry was performed.RESULTSCompared to Std, HF had greater bodyweight, fasting blood glucose, and insulin after 8 wks. After 2 wks, IL‐1α was higher in HF compared to Std. After 4 wks, IL‐5, IENF, and TrkA nerve fiber density were higher in HF compared to Std. There were no significant differences in hindpaw sensitivity or adipose mRNA expression of IL‐1β, IL‐10, IL‐6, TNF‐α, RANTES, or MCP‐1 after 2, 4, or 8 wks.CONCLUSIONIn mice fed a high fat diet for 4 wks, increased IENF and TrkA nerve fiber density was accompanied by an increase in the inflammatory mediators IL‐1α and IL‐5 in the lumbar DRG despite lack of increased inflammatory gene expression in the adipose. This suggests a high fat diet causes an early increase in neuronal inflammation and increased nociceptive nerve fiber density that may be responsible for mechanical hypersensitivity, a sign associated with “painful” neuropathy in prediabetic miceSupport or Funding InformationSupported by Southern Illinois University Edwardsville Seed Grants for Transitional and Exploratory ProjectsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Increases in insulin signaling following electrical pulse stimulation are blunted in myotubes derived from severely obese individuals with or without type 2 diabetes
Authors: LIM SEONGKYUN , CHAVES ALEC , ZHENG DONGHAI , HOUMARD JOSEPH 
Published: June 2021

IntroductionExercise training has been shown to be effective in improving skeletal muscle insulin sensitivity, but individuals with severe obesity and/or type 2 diabetes (T2D) may not respond as robustly as lean individuals. Electrical pulse stimulation (EPS) applied to primary human skeletal muscle cell cultures (myotubes) has been used as an in vitro exercise model to understand responses to contractile activity alone.PurposeThe purpose of this study was to examine the effect of antecedent EPS on insulin signaling in myotubes from lean and severely obese individuals, with and without type 2 diabetes.MethodsHuman muscle cells were cultured from muscle biopsies obtained from lean (BMI: 21.7± 0.75 kg/m2), obese (BMI: 39.1± 0.99 kg/m2), and diabetic (BMI: 40.7 ± 9.44 kg/m2) individuals. Following differentiation, mature myotubes were stimulated with electrical pulses, set at 11.5 V, 1 Hz and 2 ms for 24 h. Following EPS, myotubes were treated with 100nM insulin for 10 minutes, after which the cell lysate was harvested for western blot analysis to measure the phosphorylation status of AKT (pAKT).ResultsInsulin stimulation increased pAKT levels in all three groups (p = 0.04, Mean ± SEM, 2.86 ± 0.43, 2.1 ± 0.33, 1.85 ± 0.12; fold change over control in lean, obese, and T2D respectively). However, in response to EPS, only the lean group increased insulin‐stimulated pAKT further (p=0.03, 2.86 ± 0.43 vs 4.07 ± 0.11, fold increase over control).ConclusionOur data show that EPS increases insulin‐stimulated pAKT levels in myotubes from lean individuals, but this response is absent in myotubes from individuals with severe obesity with or without T2D, indicating that there is an inherent defect in their cellular response to electrical/contractile activity.Support or Funding InformationNational Institutes of Health (DK56112, JAH).
Changes in Hepatic Expression of Hyaluronan Synthases (HAS2 and HAS3) After Treatment with Steptozotocin in Channel Catfish
Authors: Dougherty Megan , Schmidtberger Abigail , Nevarez Ericka , Kobayashi Yasuhiro 
Published: June 2021

Streptozotocin (STZ) is commonly used experimentally to induce hyperglycemia in rodents. As an alternative model organism for investigating human obesity, channel catfish show a genetic predisposition toward an accelerated growth rate resulting in a phenotype similar to obese humans. In our previous studies, STZ treatment resulted in hypoglycemia and abnormal liver morphology in channel catfish, indicating that STZ treatment possibly induces liver damage. The mechanisms involved in the STZ‐induced hypoglycemic and tissue damage have never been examined. Another noted marker for liver damage stems from hyaluronan synthases (HAS), which have been associated with tissue damage in both humans and rodents. We hypothesized the expression of HAS mRNA would change in response to liver damage induced by STZ in channel catfish. Juvenile channel catfish were treated with 0, 3.6, 36, 180, or 360 mg STZ/kg body weight (n=4 fish/treatment) on day 0. Liver samples were collected on day 7. Hepatic expression of HAS2 and HAS3 mRNA was examined, using real‐time PCR. Expression of HAS2 and HAS3 mRNA decreased in response to an increased dose of STZ (p < 0.05 and p < 0.10, respectively). These results suggest STZ induces severe liver damage that down regulates the expression of HAS2 and HAS3 mRNA. However, the effects of STZ on gene expression appear to be global, and the tissue damage induced by STZ is not reversible by changes in the amount or activity of HAS.Support or Funding InformationThis research was funded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS‐1238831). It was also supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20 GM103418.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Perivascular Adipocytes Store Norepinephrine by Vesicular Transport
Authors: Ahmad Maleeha , Darios Emma S. , Ayala‐Lopez Nadia , Burnett Robert , Watts Stephanie W. 
Published: June 2021

Perivascular adipose tissue (PVAT) exerts an anti‐contractile effect on blood vessels in healthy individuals, a mechanism lost in obesity‐related hypertension. To understand this process, we must first investigate the role of the functional adrenergic system residing within PVAT. We have shown that PVAT contains functional norepinephrine (NE), and that PVAT adipocytes have NE transporters, similar to sympathetic neurons. We hypothesize that NE is stored in vesicles within PVAT adipocytes, and that the vesicular monoamine transporter (VMAT) is used to do so. High‐performance liquid chromatography showed the presence of NE in normal male Sprague Dawley rat superior mesenteric artery (SMPVAT; 503.60 ± 34.70 ng/g tissue) and mesenteric (MPVAT; 148.20 ± 15.00 ng/g tissue) PVATs (N=6). Using immunofluorescence, we tested for VMAT1 and VMAT2 in these PVATs (N=6), and both transporters were present surrounding the lipid of the adipocytes. Immunofluorescence of MPVAT (N=4) and SMPVAT (N=4) revealed the co‐localization of NE and VMAT2 surrounding the lipid of PVAT adipocytes, and especially concentrated around the nucleus. We developed a protocol imaging live primary rat MPVAT adipocytes to capture real‐time uptake of Mini202, a fluorescent VMAT probe functioning as a false neurotransmitter (N=4). Cells were stained with 500 μM Mini202 (see data table). Addition of Rose Bengal (100 μM), a potent VMAT inhibitor, significantly reduced Mini202 signal. Addition of plasma membrane transporter inhibitors, nisoxetine (NE transporter, 10 mM) and corticosterone (organic cation 3 transporter, 10 mM) also significantly reduced Mini202 signal. Interestingly, immunofluorescence results support that neither VMAT1 (N=4) nor VMAT2 (N=4) is present in retroperitoneal adipocytes, suggesting that perivascular adipocytes are unique in their ability to store NE. This study supports a new function of perivascular adipocytes in storing amines and provides a basis for understanding NE storage by PVAT in obesity‐related hypertension.Support or Funding InformationFunding provided by NIH NHLBI PO1HL70687 and the APS UGREFThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inactivation of the Pyruvate Dehydrogenase Complex Results in a Decrease in Butyrate Oxidation and Enhancement of Cellular Proliferation
Authors: Simon Emily , Park Bohye , Spilman Kasey , Donohoe Dallas R. 
Published: June 2021

Bacteria in the colon ferment dietary fiber producing the metabolite butyrate, which has been shown to have protective effects against colorectal cancer. Butyrate has two main functions in colonic epithelial cells (colonocytes); metabolically, as the primary energy source, and in gene expression, as a histone deacetylase (HDAC) inhibitor. These two functions are not independent of each other. Cancerous colonocytes oxidize butyrate at a lower capacity than non‐cancerous colonocytes which impacts its role in the inhibition of HDACs. Previously, we found that dichloroacetate, which inhibits pyruvate dehydrogenase kinase 2 (PDK2) resulted in an increased oxidation of butyrate in colorectal cancer (CRC) cells that was associated with diminished pyruvate dehydrogenase complex (PDC) phosphorylation. Increased phosphorylation of this complex inhibits the activity and is a common feature in cancer cells. Therefore, we hypothesized that increased phosphorylation of PDC in CRC cells mediates the metabolic shift away from utilizing butyrate, which is a predominant feature in non‐transformed colonocytes. Here, we report that targeted knockdown of PDK2 by RNA interference showed a significant increase in butyrate oxidation by the CRC cell. This effect was associated with a general decrease in PDH phosphorylation and lactate production, while also having increased levels of acetyl‐CoA. In addition, knockdown of pyruvate dehydrogenase phosphatase (PDPC1) did not significantly alter butyrate oxidation in CRC cells. We next tested whether the oxidation of other short‐chain fatty acids (SCFAs) (acetate and propionate) was also regulated by PDK2 and the phosphorylation of PDH. Interestingly, the oxidation of these two SCFAs was not significantly affected by PDK knockdown. Therefore, this suggests that the phosphorylation of PDH by PDK2 is an important event in regulating the CRC cell's metabolic shift away from the oxidation of butyrate and toward glycolysis.Support or Funding InformationUTK Start‐up FundsThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activation of Hippo Signaling by rhBMP‐2 Suppresses the Proliferation of Human Colorectal Cancer Cell
Authors: Liu Yu Chuan , Kim Soo Mi 
Published: June 2021

Despite that the use of recombinant human bone morphogenetic protein (rhBMP)‐2 has been debated for a decade due to its oncogenic characteristics. However, the underlying molecular mechanism and safety issues of rhBMP‐2 usage remain poorly understood. In this study, we investigated the effect of rhBMP‐2 and its signaling pathways involved in colorectal cancer cell (CRC) using HT‐29 and HT‐116 cells. RhBMP‐2 significantly inhibited proliferation of CRC cells in dose‐dependent by MTT assay. Cell cycle arrest in G1 phase was induced at 24h after rhBMP‐2 treatment. RhBMP‐2 also stimulated Smad4, p53 and p21 levels, and reduced cyclin D1, cyclin‐dependent kinase (CDK) 4 and CDK6 activities. On the other hand, rhBMP‐2 treatment resulted in reduced protein expression levels of poly (ADP‐ribose) polymerase (PARP) whereas those of cleaved PARP and cleaved caspase‐9 were significantly increased in CRC cells. In addition, rhBMP‐2 increased RASSF1, MST1, MST2, Mob1, LATS1, Sav1, and p‐YAP protein levels. Therefore, our results indicate that rhBMP‐2 suppresses CRC cell proliferation which is mediated via the hippo signaling pathway. Therefore, targeting BMP‐2 may constitute a potential therapeutic strategy for CRC.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Live Cell Fluorescence Imaging Reveals Intercellular Calcium Waves and Chloride Channel Activation During Rotavirus Infection
Authors: Chang‐Graham Alexandra , Perry Jacob , Engevik Melinda , Danhof Heather , Hyser Joseph 
Published: June 2021

BackgroundRotavirus (RV) causes severe diarrheal disease in children worldwide. During infection, RV increases cytosolic calcium in intestinal epithelial cells through release of endoplasmic reticulum calcium and activation of the store‐operated calcium entry (SOCE) pathway. Increased cytosolic calcium is critical for RV replication and activation of secretory pathways in GI epithelium, yet the molecular details of these processes remain poorly defined. Thus, characterizing RV‐induced calcium signaling at the molecular level is needed to understand RV pathogenesis. We conducted live cell fluorescence imaging in cell lines and human intestinal enteroids (HIEs) as a biologically relevant model of the human small intestine to measure calcium signaling dynamics and chloride channel activation during RV infection.Methods & ResultsWe first engineered African green monkey kidney MA104 cells and HIEs to stably express GFP‐based genetically encoded calcium indicators to measure calcium dynamics throughout RV infection using live cell epifluorescence microscopy. We found that RV significantly increases the number and magnitude of calcium transients, which were attenuated with SOCE blockers. Furthermore, we observed that single RV‐infected cells triggered long‐distance intercellular calcium waves (ICWs) that encompassed surrounding uninfected cells in both MA104 cells and both duodenum and jejunum HIEs. Treatment with the ectoNTPase apyrase or purinergic receptor inhibitors blocked ICWs and reduced RV replication, mucin release, and serotonin secretion. These results indicate that extracellular ATP/ADP signaling elicits ICWs and is an important mediator of calcium dynamics during RV infection. Finally, we created HEK293 cell lines stably expressing halide‐sensitive YFP, a chloride channel biosensor, and either the calcium‐activated chloride channel anoctamin 1 (Ano1) or cystic fibrosis transmembrane conductance regulator (CFTR). We found that RV activated both mouse Ano1 and CFTR, which were attenuated with channel‐specific blockers, and suggests that multiple chloride channels can be activated during RV infection.ConclusionsOverall, our results support that RV causes dynamic calcium signaling during infection that is mediated by SOCE and purinergic signaling. Both pathways are potent stimulators of chloride secretion, thus they are potential therapeutic targets for developing life‐saving anti‐diarrheal drugs. Furthermore, we are the first to show that viruses can exploit purinergic signaling to generate ICWs that encompass uninfected cells and thereby potentially amplify the pathophysiological signaling important for diarrhea.Support or Funding InformationBCM Seed Funding, NIH F30 DK112563‐01This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
WW Domain‐containing Oxidoreductase Regulates Keratinocyte Growth for Maintaining Epidermal Homeostasis
Authors: Chou Ying‐Tsen , Hsu Li‐Jin 
Published: June 2021

Human fragile WWOX gene encodes a candidate tumor suppressor WW domain‐containing oxidoreductase. Previous studies have shown that ectopically overexpressed WWOX induces apoptosis in cancer cells and suppresses tumor growth both in vitro and in vivo. Functional suppression of WWOX by antisense mRNA, dominant negatives, and small interfering RNA protects cells from apoptosis by tumor necrosis factor, anticancer drugs, UV light, and ectopic p53 in vitro. Interestingly, substantial evidence revealed that progression of various cancers to a pre‐metastatic stage positively correlates with upregulation of WWOX protein expression, suggesting that WWOX may act more than a tumor suppressor. In normal human epidermis, WWOX is abundant in proliferating epithelial cells of the skin, and its expression is increased in intensity toward the superficial layers. In this study, we determined a reduced number of Ki67‐positive proliferating keratinocytes in the epidermis of our newly developed Wwox gene knockout (Wwox−/−) mice. A marked reduction in the basal (K5‐positive) and suprabasal (K10‐positive) keratinocyte numbers was examined in the keratinized squamous epithelium of the Wwox−/− mouse skin. Apparent decreases in Wwox−/− epidermal and dermal thickness were found. To further verify whether WWOX regulates keratinocyte growth, we detected significantly decreased cell proliferation in both ex vivo Wwox−/− primary keratinocyte cultures and WWOX‐knockdown HaCaT cells as compared to the controls. No significant increase in the number of TUNEL‐positive cells was observed in the Wwox−/− epidermis. Moreover, our recent data showed that keratinocyte differentiation was slightly enhanced in WWOX‐knockdown HaCaT cells. Together, our results clearly demonstrated a crucial role of WWOX in regulating normal keratinocyte proliferation for maintaining skin homeostasis.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.