bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of voluntary running with defined distances on body adiposity and its associated inflammation in mice fed a high‐fat diet
Authors: Yan Lin , Sundaram Sneha 
Published: June 2021

Sedentary lifestyle contributes to obesity. This study examined the effect of quantitative voluntary running on body adiposity and its associated inflammation in mice fed a high‐fat diet. Male C57BL/6 mice were assigned into six groups and fed the AIN93G (sedentary) or a high‐fat diet (sedentary, unrestricted running, or with 25%, 50%, or 75% restriction in running activity) for 12 weeks. The average daily running distance was 8.3, 6.3, 4.2, and 2.1 km for the unrestricted, 25%, 50%, and 75% restricted runners, respectively. The high‐fat diet increased body fat mass by 46% compared to the AIN93G diet in sedentary mice. Running reduced the fat mass in a dose dependent manner; there was no difference between sedentariness and running at 2.1 km/d. The high‐fat diet significantly increased blood glucose, plasma insulin, HOMA‐IR, and plasma and adipose concentrations of leptin and monocyte chemotactic protein‐1 (MCP‐1) in sedentary mice. Running, despite continued consumption of the high‐diet, reduced these variables in dose‐dependent manners. Running at 8.3 (unrestricted) and 6.3 km/d (25% restricted) had the greatest but similar reductions in, whereas running at 2.1 km/d did not affect (except MCP‐1), these measurements. Running, regardless of daily distance, significantly reduced MCP‐1 in both plasma and adipose tissues. In conclusion, voluntary running at 6.3 km/d is optimal to reduce body adiposity and its associated inflammation and metabolic disturbance in C57BL/6 mice. Results that running at 2.1 km/d reduced MCP‐1 without affecting body fat mass show the benefits of moderate amount of exercise in improving low‐grade inflammation. It suggests that different mechanisms, rather than reducing body adiposity, are responsible for this improvement.Support or Funding InformationThe U.S. Department of Agriculture, ARS, research project 5450‐51000‐045‐00D.
Weight loss due to low‐fat feeding in female mice with obesity induced by high‐fat feeding reduces both body fat and bone status
Authors: Sawin Emily A , Stroup Bridget M , Murali Sangita G , Ney Denise M 
Published: June 2021

BackgroundGlycomacropeptide (GMP) is a 64 amino acid (AA) glycophosphopeptide released from κ‐casein during cheese making that promotes satiety, reduces body fat, increases bone mass and infers prebiotic and anti‐inflammatory effects.ObjectiveTo determine if GMP, when fed as the primary protein source, increases fat oxidation with weight loss and improves body composition and bone status after weight loss compared to casein.MethodsWeanling female C57Bl6 mice were fed a high fat casein diet (60% en) for 28 wks and then crossed over to high and low fat diets (HF 60% en; LF 13% en) with casein or GMP for another 10 wks. We assessed body weight, food intake and body composition by DXA. Fatty acid oxidation was measured in liver, muscle, and fat tissues using 14C‐palmitate. Plasma concentrations of amino acids, hormones and cytokines were determined. Bone biomechanical performance was assessed with the 3‐point loading test.ResultsMice that were fed the low fat diets after crossover had significantly reduced body mass, body fat, and energy intake compared with the mice that were fed the high fat diets. After weight loss, female mice fed the low fat diets had significantly smaller femoral cross‐sectional area and femoral bone mineral density compared to mice that were crossed over to the high fat diets.ConclusionCrossing over to low fat diets after weight gain on a high fat diet improves body composition by reducing overall body mass and body fat through reduction in energy intake. The reduction in body mass coincides with a reduction in bone status as femoral cross‐sectional area and bone mineral density are reduced.Support or Funding InformationSupported by USDA Hatch WIS01790 and a Wisconsin Alumni Research Foundation Accelerator Grant.
Minimum Selenium Requirements Based on Muscle and Kidney Glutathione Peroxidase Activity in the Turkey
Authors: Taylor Rachel M , Bourget Victor G , Sunde Roger A 
Published: June 2021

Liver and blood glutathione peroxidase (GPX) activities have been used extensively as biomarkers of selenium (Se) status in laboratory and livestock animals. In livestock species used for meat, selenoenzyme activity in the muscle should be considered when setting Se requirements. We recently reported that the Se requirement in rapidly growing young turkey poults is 0.3 μg Se/g diet, based on plasma and liver GPX activity (PLOS ONE 11:e0151665), vs. the published requirement of 0.2 μg Se/g diet (NRC 1994). To examine Se requirements based on breast and thigh muscle, cardiac muscle, and kidney (a tissue with high GPX activity), we fed day‐old male poults (n=50) 10 graded levels of Se (0–1.0 μg/g) for 28 d, and measured GPX1 and GPX4 activity, and transcript expression of all 24 turkey selenoproteins in each tissue. Se response curves and plateau breakpoints were plotted to determine extent of deficiency and minimum Se requirements for each biomarker. The minimum dietary Se requirement for growth was 0.05 μg Se/g. Poults receiving 0 μg Se/g diet had 1 and 8% of plateau GPX1 activity in kidney and heart, respectively, and minimum Se requirements were 0.30 and 0.34 μg Se/g diet, respectively. Breast and thigh GPX1 activity Se response curves were hyperbolic but did not reach well defined plateaus, as GPX1 activity was significantly higher in poults fed 1.0 μg Se/g vs. 0.4 μg Se/g. GPX4 activities in Se‐deficient poults were 9, 22, 8, and 27% of GPX4 plateau activities in kidney, heart, breast and thigh, respectively, and minimum Se requirements were 0.25, 0.31, 0.32 and 0.30 μg Se/g diet, respectively. qPCR of total RNA from each tissue showed that 2–6 transcripts out of 24 total selenoprotein transcripts were significantly downregulated by Se deficiency to 31–73% of Se‐adequate plateaus, with GPX1 and SELH transcripts downregulated in every tissue studied; minimum Se requirements based on transcript expression were 0.08–0.19 μg Se/g diet, and no selenoprotein transcript was up‐ or downregulated significantly by supernutritional Se. This study shows that the turkey minimum dietary Se requirement is 0.3 μg Se/g diet based on heart and kidney GPX activity and on breast and thigh GPX4 activity, just as determined previously based on gizzard, liver and plasma GPX activities. The hyperbolic Se response curves for breast and thigh GPX1 activity suggest that 0.4 μg Se/g diet may be warranted for the 0–4 wk old rapidly growing poult.Support or Funding InformationUSDA NIFA (Hatch 233618 and 1004389), and by the Wisconsin Alumni Foundation Selenium Nutrition Research Fund (No. 2046295).
Oncogenic activity of MAN2A1‐FER fusion in liver cancer and other human malignancies
Authors: Luo Jianhua , Chen Zhanghui , Monga Satdarshan , Michalopoulos George , Yu Yan P 
Published: June 2021

Oncogenic fusion gene is one of the fundamental mechanisms driving the progression of human cancers. MAN2A1‐FER, a fusion gene between the mannosidase domain of MAN2A1 and tyrosine kinase domain of FER, was found in 6 different types of human malignancies. MAN2A1‐FER fusion translocated FER domain from cytoplasm to Golgi apparatus, and led to phosphorylation of N‐terminus of EGFR and activation of EGFR signaling pathway. Expression of MAN2A1‐FER generated dramatic increase of growth and invasion of cancers, while removal of MAN2A1‐FER through knockout generated significant lower level of growth and metastasis. The presence of MAN2A1‐FER increased the sensitivity of human cancers to FER kinase inhibitor crizotinib or EGFR kinase inhibitor canertinib. Hydrodynamic tail‐vein injection of MAN2A1‐FER gene resulted in rapid development of liver cancer in mice with somatic Pten deletion. Taken together, we concluded that MAN2A1‐FER fusion gene is one of the key drivers for human cancer development.Support or Funding InformationNCI, DOD and UPCI
Lipidomic Profiling Identifies Cytochrome P450 as A Therapeutic Target for Colitis‐Associated Colorectal Cancer
Authors: Wang Weicang , Yang Jun , Song Mingyue , Wan Debin , Yang Haixia , Sanidad Katherine , Hammock Bruce D. , Xiao Hang , Zhang Guodong 
Published: June 2021

Eicosanoids and associated lipid mediators play central roles in regulating inflammation and carcinogenesis. While most previous studies of lipid mediators in colorectal cancer have only analyzed single or limited number of lipid mediators, few systematic studies have been carried out. Here we used a LC‐MS/MS‐based lipidomics, which can analyze >100 bioactive lipid mediators produced by cyclooxygenase (COX), lipoxygenase (LOX), and cytochrome P450 (CYP) enzymes, to profile azoxymethane (AOM)/dextran sulfate sodium (DSS)‐induced colorectal cancer in mice. Lipidomics showed that CYP‐derived fatty acid epoxides were the only lipid mediators which were increased in both plasma and colon tissues of AOM/DSS‐treated mice. RT‐PCR and western blotting validated that the expressions of CYP were increased, while the expression of soluble epoxide hydrolase (the dominant enzyme in degrading fatty acid epoxides) was reduced, in colon tissues of AOM/DSS‐treated mice. To study the functional roles of CYP enzymes, we treated mice with SKF‐525A and clotrimazole, which are two different selective inhibitors of CYP enzymes involved in biosynthesis of fatty acid epoxides, and found that CYP inhibitors significantly suppressed AOM/DSS‐induced colorectal cancer in mice. To elucidate the specific lipid mediator involved in the pro‐colorectal cancer effects of CYP, we treated mice with epoxyoctadecenoic acids (EpOMEs) which are metabolites of linoleic acid produced by CYP, and found that EpOMEs worsened DSS‐induced colitis. Together, these results suggest that the previously unappreciated CYP pathway plays critical roles in promoting progression of colorectal cancer through formation of pro‐inflammatory EpOMEs, and CYP enzymes could be novel therapeutic targets of colitis‐associated colorectal cancer.Support or Funding InformationUSDA NIFA 2016‐67017‐24423, NIEHS R01 ES002710 and Superfund Research Program P42 ES04699
Protein Kinase C Epsilon Peptide Inhibitor Exerts Cardioprotective Effects in Myocardial Ischemia/Reperfusion Injury
Authors: McIntyre Anahi , Adekayode Christine , Heron Joseph , James Ifeanyi , Woodley John , Dittakavi Tejaswi , Benjamin Israel , Liu Stephanie , Chen Qian , Barsotti Robert , Young Lindon 
Published: June 2021

During myocardial ischemia/reperfusion (I/R), the generation of reactive oxygen species (ROS) contributes to post‐reperfusion cardiac injury and contractile dysfunction. Activation of protein kinase C epsilon (PKC ɛ) during I/R has been shown to increase ROS release, in part, by its stimulation of increased uncoupled endothelial nitric oxide synthase activity. We hypothesize that using a cell permeable PKC ɛ peptide inhibitor (PKC ɛ‐) (N‐myr‐EAVSLKPT, MW=1054 g/mol, 10μM or 20μM) will improve post‐reperfused cardiac function and attenuate infarct size compared to untreated controls in isolated perfused rat hearts subjected to I(30 min)/R(90 min). Male Sprague‐Dawley rats (275–325 g) were anesthetized with sodium pentobarbital (60 mg/kg) and anticoagulated with heparin 1000 units IP. PKC ɛ‐was dissolved in Krebs' buffer and infused during the first 10 min of reperfusion. PKC ɛ‐treated hearts exhibited significant improvement in post‐reperfused cardiac function at 90 min in the maximal rate of left ventricular developed pressure (+dP/dtmax): 56±5%; n=6 (10μM) and 46±3%; n=4 (20μM) compared to untreated controls (n=6) which only recovered to 32±5% of baseline values for +dP/dtmax respectively (p<0.05). Furthermore, PKC ɛ‐treated hearts showed significant reduction in infarct size of 27±2% (10μM) and 28 ±2% (20μM) compared to untreated control I/R hearts, 40±3% (p<0.05). The results suggest that PKC ɛ‐is effective in improving cardiac function and reducing infarct size and is a putative treatment that could aid in clinical myocardial infarction/organ transplantation patient recovery.Support or Funding InformationThis study was supported by the Center for Chronic Disorders of Aging, the Division of Research and the Department of Bio‐Medical Sciences at Philadelphia College of Osteopathic Medicine.
Chronic Dehydration Induced Changes in Mouse Supraoptic Nucleus (SON): Contribution to Chronic Kidney Diseases (CKD)?
Authors: Song Zhilin , Roncal Carlos , Milagres Tamara , Lanaspa‐Garcia Miguel , Andres‐Hernando Ana , Johnson Richard 
Published: June 2021

Chronic recurrent dehydration (WAN) can cause CKD, maybe through a fructokinase (KHK) dependent way. VP is a peptide hormone that is synthesized in the hypothalamic SON and paraventricular nucleus (PVN), and is subsequently transported to and stored in posterior pituitary (PP), from where it is released into systemic circulation. VP may mediate some types of CKD through currently unclear mechanisms. Previous studies have demonstrated that acute dehydration (24 hr water deprivation) may activate a local polyol pathway: aldose reductase (AR)‐fructose‐KHK, leading to elevated VP synthesis and secretion, thus worsening CKD. Current studies evaluated the effects of chronic dehydration on the AR‐KHK axis in mouse SON. Mice were subject to heat stress for 30 min at every hour for seven hours during day with limited water access at night only, for five weeks. Mice were sacrificed and SONs and PPs were extracted out and assayed by rtPCR and ELISA for mRNA expression and hormone content, respectively. Chronic dehydration induced significant increase in mRNA expression of the following markers: AR, KHK‐A, KHK‐C, VP, transient receptor potential vanilloid I (TRPV1, the osmo‐sensor), and oxytocin, indicating activation of the AR‐KHK axis and increased VP synthesis /secretion. Further analysis revealed that VP storage in PP was slightly reduced in WT mice by chronic dehydration but was significantly reduced in KHK A/C double knockout mice. KHK‐KO did not significantly affect VP storage in PP of control mice. These results suggest that chronic dehydration may lead to CKD, at least partially, through elevated VP synthesis and secretion driven by activation of the AR‐fructose‐KHK pathway. Future studies will further clarify the underlying mechanisms.Support or Funding InformationDOD PR130106;NIDDK DK048520;AHA12GRNT11940024
Sex Differences in the Utilization of Essential and Non‐Essential Amino Acids in Lepidoptera
Authors: Davidowitz Goggy , McCue Marshall , Levin Eran 
Published: June 2021

The different reproductive strategies of males and females have selected for differences in function and behavior that may translate into differences in resource use. Here we studied sexual differences in the utilization of essential and non‐essential amino acids by the Carolina sphinx moth (Manduca sexta: Sphingidae). We labeled two essential amino acids (leucine and phenylalanine) and one non‐essential amino acid (glycine) with an isotopically enriched first carbon (13C). These isotopically enriched amino acids were added either to the larval diet or to the adult nectar. We determined the allocation of these amino acids using a stable carbon isotope analyzer of the 13CO2 of the moth's breath as well as allocation to flight muscle and to the spermatophore. On day one after eclosion from the pupae, unfed male moths oxidized larval‐derived amino acids in higher ratios than unfed females, but this trend reversed after four days of starvation: females oxidized more amino acids than males. Males oxidized nectar‐derived essential amino acids more than females, whereas there was no difference between males and females in the oxidation of non‐essential amino acids. Males allocated significantly more nectar‐derived essential and non‐essential amino acids to flight muscle than did females. Last, males allocated relatively few nectar‐derived amino acids to their first spermataphore compared to a four‐fold increase to their second and third spermatophores. We conclude that males and females have fundamentally different allocation strategies of amino acids and that these allocation strategies differ for essential, compared to non‐essential, amino acids. We suggest that these differences in amino acid allocation strategies reflect the need of the males to maintain their ability to fly and find mates, a lower priority for females.Support or Funding InformationNSF grant IOS‐1053318 to GD
Diet Composition Modulates Intestinal Hydrolytic Enzymes in White‐Footed Mice (Peromyscus leucopus)
Authors: Wang Luping , Karasov William 
Published: June 2021

Animals can digestively accommodate to diets with varying substrate levels by routinely possessing plenty of spare digestive capacity or by modulating digestive capacity to match changes in dietary substrate level. There are few studies of this in wild mammals, though in laboratory mice and rats levels of intestinal disaccharidases and peptidases are known to change in correlation with dietary carbohydrate and protein levels, respectively. We maintained white‐footed mice for 12–81 days on diets enriched in either starch (diet C), protein (diet P) or lipid (diet L). There was no effect of either diet, or time on the diet, on their body mass when tissues were harvested. There were no significant effects of body mass on small intestinal mass or length, and no significant effects of diet on small intestinal mass or length. Diet treatment affected the summed activities of maltase, sucrase and aminopeptidase‐N. Post‐hoc Tukey comparisons showed that in all cases summed activity was lowest for diet L, with no differences between diets C and P. Activities of all three enzymes tended to be highest most often in midintestine, and whatever factor(s) reduced enzyme activities on the L diet apparently did so in all intestinal positions. This is only the second study in a wild small mammal, to our knowledge, testing for digestive modulation to diet composition. Like the other study, we found evidence of modulation, but the pattern differed from that observed in laboratory mice and rats in that the expected correlations with dietary carbohydrate and protein were not apparent. Maltasisc activity was relatively high in white‐footed mice, which suggests that they may routinely possess plenty of spare digestive capacity.Support or Funding InformationSupported by NSF 1354893 to WHK.
Genetic Background Influences Patterning and Implementation of Stereotypy in the Early Postnatal Mammary Ductal Tree
Authors: Hadsell Darryl , Firozi Urvashi , Hadsell Louise , Cox Timother C , Lee Adrian 
Published: June 2021

The extent to which ductal branch pattern is controlled by hard‐wired stereotypical mechanisms has been a long standing question in biology. In the mammary gland, prevailing wisdom has held that ductal patterning is stochastic. However, a detailed analysis of mammary ductal patterning using appropriate 3‐dimensional approaches has not been done. Prior published work from our group demonstrates that genetic background has a profound effect on mammary ductal patterning. The hypothesis for these experiments was that ductal patterning and the implementation of stereotypical branching behaviors during early postnatal development differs with genetic background. To test this hypothesis, 12 measures of ductal geometry and two of stereotypy were compared among 3D reconstructions derived from whole‐mounted mammary glands that were stained with the epithelial marker E‐cadherin and imaged by optical projection tomography. Samples were taken on postnatal day (PN) 17 from females of the C57BL/6J (N=5), PWK/PhJ (N=3), and KK/HlJ (N=3) strains. Comparison of graph size, defined as total number of branches, and total ductal length (mm) revealed that overall development was the greatest (P<0.05) in KK/HlJ (56.33±1.76, and 31.57±0.357, respectively), intermediate in C57BL/6J (50.33±6.16, and 23.67±.798, respectively), and least in the PWK/PhJ (41.60±3.12, and 20.51±1.39, respectively). The average local ductal bifurcation angles were also larger (P<0.05) in KK/HlJ than PWK/PhJ (107.01±0.67° and 95.45±3.39°, respectively). Lastly, as a measure of stereotypy, graph discordance was lowest (P<0.05) in KK/HlJ, intermediate in C67BL/6J, and highest in PWK/PhJ (0.47±0.013, 0.54±0.03, and 0.61±0.02, respectively). These results highlight the utility of 3D imaging methodologies in understanding mammary ductal development and support the conclusion that differences in ductal development among differing genetic backgrounds may also be linked with differential implementation of stereotypical patterning.Support or Funding InformationThis work supported by a grant from the DOD number DAMD‐0017.
Studies on HIV‐1 RNA‐protein interactions.
Authors: Hadler Nicole , Stockdale Eryn , Wise Kenneth , O'Carroll Ina 
Published: June 2021

In the absence of a cure and a vaccine, the human immunodeficiency virus (HIV‐1) continues to affect millions of people worldwide. HIV is a retrovirus, a unique family of viruses that replicate via a DNA intermediate. The newly replicated HIV RNA must be spliced in order for it to exit the nucleus of the host cell and be packaged into new viral particles. Normally, unspliced or partially spliced HIV RNA's are trapped within the nucleus and degraded. To combat this, HIV has evolved an accessory protein, Rev. The protein Rev binds to the RRE (located on the unspliced or partially spliced RNA) in order to create a complex that allows for the unspliced HIV RNA genome to be exported out of the nucleus.The secondary structure of the HIV‐1 Rev‐Response Element contains five unique stems. The protein Rev binds to sites situated on stems I and II. We have used electrophoretic mobility shift assays (EMSAs) to assess the significance of the various stems, Stems I, III, IV and V, in the functionality of the Rev‐RRE complex. In this assay, electrophoresis was used to resolve the Rev‐RRE complexes after titrating the mutants with Rev protein. RRE mutant RNAs were obtained in vitro and purified. Free RNAs and protein‐bound RNAs were resolved on a gel; the effectiveness of RNA mutants to bind Rev provide insight into the role of the RNA structure in Rev binding.Support or Funding InformationThis project is supported by the Defense Threat Reduction Agency and the US Naval Academy.
Extraction of Protein Thermodynamic Parameters from High‐Throughput Differential Scanning Fluorimetry Assays
Authors: Wright Thai , Stewart Jamie , Konkolewicz Dominik , Page Richard C 
Published: June 2021

Thermodynamic properties of protein unfolding have been extensively studied; however, the methods used have typically required significant preparation time and high protein concentrations. Here, we present a facile, simple, and high‐throughput differential scanning fluorimetry (DSF) method that enables a complete set of thermodynamic parameters of protein unfolding to be extracted. This method assumes a two‐state, reversible protein unfolding mechanism and provides the capacity to quickly analyze the biophysical mechanisms of changes in protein stability and to more thoroughly characterize the effect of additives, inhibitors, or pH. We show the utility of the DSF method by analyzing the thermal denaturation of lysozyme, carbonic anhydrase, chymotrypsin, horseradish peroxidase, and cellulase. Compared to similar biophysical analyses by circular dichroism, DSF provides greater than 24‐fold reduction in experimental time. This study opens to door to rapid characterization of protein stability on low concentration protein samples.Support or Funding InformationThe authors acknowledge financial support from the US National Science Foundation (Award No. MCB 1552113 to RCP) and institutional support from Miami University through the Robert H. and Nancy J. Blayney Professorship (to RCP).