bluuebunny/crossref_metadata_2025_split huggingface dataset:
Northeastern Montana range pastures maintaining distinct American bison herds yield different trace metal availability and intestinal parasite infections in the American Bison on the Fort Peck Indian Reservation
Authors: Rhodes Ryon Sun , Snyder Anna , Topliff Christina , Kelling Clayton , Coon Steven D. 
Published: April 2020

BackgroundThe Fort Peck Reservation tribes maintain two distinct bison herds on different pastures in Montana which includes a cultural herd (purebred) and a business herd (contains beef genes). The different pastures housing the herds have different Animal Unit Months (AMU) ratings (a measurement of forage/acre per adult animal) with the cultural herd pasture having a higher rating. With the limited grazing opportunities, it has been discovered that many of these bison have serious health problems related to malnutrition which can cause lower birth rates. Previous studies indicate metals such as copper (Cu) and zinc (Zn) are necessary for proper health and birthrates. In addition, poor calving rates in apparently healthy bison with adequate protein in diet might simply be from the different metal deficiencies.HypothesisDecreased absorption of Cu and Zn are due to increased absorption of selenium and molybdenum. These deficiencies along with parasitic infections are causing health problems both bison herds on the Fort Peck Reservation.MethodsWe successively isolated liver and fecal samples from euthanized bison to test for toxic and essential trace minerals, and to probe for present parasites in the gastrointestinal tracts. Ova were counted using light microscopes from intestinal fecal matter obtained from the bison.ResultsMineral imbalances were detected as elevated molybdenum (Mo) in both herds. Elevated Mo can deplete liver copper (Cu). Deficient levels of Cu, manganese (Mn) and Zn were evident in both herds. On average cultural animals had Cu levels on of 7.75 ppm, well below the suggested range of 75 ppm while the business herd liver Cu content was averaged to be even lower at 4.33 ppm. Additionally, Mn was found to be deficient in the cultural herd (2.1 ppm where 3.0 ppm is normal) but not in the business herd. Zn was deficient in both herds at 28 ppm where 100 ppm is normal. Also, we determined that bison in both herds were either free of internal parasites or had low burdens of infestation. Seven business herd and 15 cultural herd fecal present in the cultural herd, none in the business herd. We believe health problems from malnutrition contribute to decreased weights and birthrates of the bison on the Fort Peck Indian Reservation samples were analyzed. The business herd had samples that exhibited undetectable ova or parasites per gram of feces while the cultural herd samples had low levels of Strongyle type ova (0.4–7.78 ova/gram of feces). Three of the cultural herd samples in additionally had Trichuris sp. (0.4 ova/gram), Nematodirus (0.2–0.67 ova/gram), and/or Coccidia (0.6 ova/gram).ConclusionsMalabsorption of nutrients is caused by increased absorption of selenium and molybdenum but not infections from parasites in the Bison digestive tract.Support or Funding InformationFunded by the USDA
ARL15 Regulates CNNM2‐dependent Mg2+ Transport by Modulating its N‐linked Glycosylation
Authors: de Baaij Jeroen H.F. , Zolotarov Yevgen , Ma Chao , Franken Gijs , Tremblay Michel L. , Hoenderop Joost 
Published: April 2020

BackgroundA large Genome‐wide association study identified that ARL15, a small GTP‐binding protein, is associated with urinary Mg2+ excretion. Within the kidney, ARL15 is highly expressed in the thick ascending limb (TAL) and distal convoluted tubule (DCT), where Mg2+ reabsorption is tightly regulated. However, the exact function of ARL15 and the mechanism by which ARL15 regulates renal Mg2+ handling are still unknown.MethodTo identify protein‐interaction between ARL15 and Cyclin M (CNNM) proteins, proximity‐dependent biotin identification (BioID) and co‐immunoprecipitation were performed. Immunohistochemistry were used to investigate co‐localization in mouse kidney and human embryonic kidney (HEK293) cells. Furthermore, cell surface biotinylation and 25Mg2+ uptake assays were used to assess cell surface expression of CNNM2 and Mg2+ transport activity. The glycosylation pattern of CNNMs was determined by far lectin Western blot and glycosidase assays.ResultsWe identified members of the CNNM family as direct interaction partners of ARL15 by BioID. Immunoprecipitation with truncated CNNM2 proteins indicated that ARL15 interacts with CNNM2 at its carboxyl‐(C)‐terminal conserved CBS domain. CNNM2 and ARL15 co‐localize in the basolateral membrane of DCT. Interestingly, overexpression of ARL15 in HEK293 cells showed subcellular localization in the Golgi‐apparatus and resulted in an increased N‐glycosylation of CNNM proteins. This ARL15‐mediated glycosylation was Mg2+‐sensitive and encompassed hybrid and complex glycosylation. The functional consequences of ARL15‐dependent glycosylation were examined by 25Mg2+ uptake experiments. ARL15 increased 25Mg2+ uptake via CNNM2 by increasing its cell surface expression.ConclusionARL15 increases CNNM2 plasma membrane expression by regulating its N‐glycosylation pattern. Altogether, our results establish ARL15 as novel regulatory mechanism of Mg2+ transport within the DCT.Support or Funding InformationThis work was financially supported by grants from the Netherlands Organization for Scientific Research NWO Veni 016.186.012 and VICI 016.130.668).
Comparison and Analysis of Degree of Newborn Neurocranial Bone Development in Two Prdm16 Mutant Mouse Models
Authors: Leslie Erin , Moskal Russell , Jones Jennifer , Bjork Bryan 
Published: April 2020

This study is part of a larger project examining the role of Prdm16, a transcriptional cofactor involved in TGF‐β signaling pathways, on craniofacial development. Prdm16 is expressed in various tissues, including cranial neural crest and the brain. Two, Prdm16 mutant mouse models, cleft secondary palate‐1 on a C57Bl6/J background strain (csp1) and Prdm16 conditional gene trap on an FVB/NJ background strain (cGT), display non‐syndromic clefting of the secondary palate and micrognathia due to loss of Prdm16 function. Our earlier pilot project on adult cGT and csp1 mice revealed that cGT heterozygous mutants have a relatively narrower skulls compared to their wild type (wt) littermates and that csp1 heterozygous mutants have relatively wider skulls than their wt counterparts. Homozygous mutants of both strains die shortly after birth. The aim of this study is to quantitatively assess variation in bony neurocranial development in newborn het and mut csp1 and cGT mice and contrast this with the wt morphology to determine if variation in degree of bone development in these strains is apparent at the newborn stage.Crania of newborn heterozygous mutant, homozygous mutant, and wt csp1 and cGT mice were microCT scanned and the resulting 2D images were volume‐merged to create a 3D model for each specimen. The neurocranial bones from each 3D model were segmented into separate images for analysis. For each specimen, we calculated the surface area and volume of each neurocranial bone and statistically compared these values among the genotype groupings.Smaller bone surface areas and volumes were interpreted as indicating less bone development. Homozygous mutant mice of both strains had significantly less developed neurocranial bones than their wt counterparts. The heterozygous mutant mice in both strains displayed substantial variation in degree of bone development. The calvarial bones were especially under‐developed in homozygous mutant csp1 and cGT mice, but the occipital and alisphenoid bones were well‐developed in most specimens.Our results suggest that reduced Prdm16 expression in newborn homozygous mutant csp1 and cGT mice correlates with overall underdevelopment of the neurocranium compared to wt mice. Bone development patterns in the heterozygous mutant mice are less apparent and require additional analysis. These findings complement our previous research on adult csp1 and cGT mice and additional insight on the downstream effects of Prdm16 expression on craniofacial development.Support or Funding InformationNIH/NIDCR (R15DE023982) and Midwestern University Core Facility
Insights into the Effect of Genetic Variation in CYP2C9 (*11) on Drug Binding.
Authors: Davie Dylan S. , Parikh Sonia , Shah Manish B. 
Published: April 2020

Cytochrome P450s (CYP) are a family of membrane associated heme‐containing enzymes involved in xenobiotic metabolism with recent relevance in pharmacogenomics. The human CYP2C9 enzyme metabolizes up to 15–20% of drugs that include warfarin, losartan, ibuprofen, and tolbutamide. It is highly polymorphic with several genetic variations associated with significantly altered activity of the enzyme. The CYP2C9*11 allele occurs predominantly in African Americans but also in Caucasian and other ethnic populations with a frequency of around 0.05. The *11 genetic variation represents a substitution of amino acid residue at 335 from arginine to tryptophan (R335W) that has demonstrated decreased catalytic activity towards various substrates including S‐warfarin. The R335W was created in CYP2C9 wild‐type (WT) construct using site‐directed mutagenesis and co‐expressed in E. Coli alongside chaperone proteins (GroEL/ES). Generation of purified CYP2C9*11 protein enabled catalytic studies and crystallization in complex with the drug substrate losartan. The enzymatic assays revealed marked reduction in losartan turn‐over by CYP2C9*11 compared to the WT enzyme. Additionally, comparison of the amino acid sequence revealed that R335 is conserved across the CYP2C subfamily of enzymes. The R335 is located on the distal J‐J’ loop and interacts with multiple amino acid side chains on the J and J’ helices. The computational analysis suggests that the substitution with hydrophobic and bulkier tryptophan located on the surface region may result in destabilizing interactions with several helices including those located near the active site heme, affecting protein stability and function. Overall, these observations establish the effect of distal variation prompting further evaluation by structural and biophysical studies.Support or Funding InformationThe research was funded by the start‐up funds from the Albany College of Pharmacy and Health Sciences.
Fatty Acid Desaturase Gene‐Induced Omega‐3 Deficiency in Amerindian‐Ancestry Hispanic Populations
Authors: Blomquist Sarah , Coletta Dawn , Mandarino Lawrence J. , Hallmark Brian , Yang Chaojie , Rich Stephen , Manichaikul Ani W. , Chilton Floyd 
Published: April 2020

Hispanic populations in the US have increased risk of obesity, elevated circulating triglycerides, nonalcoholic fatty liver disease, and diabetes. Our previous studies suggest that ancestry‐related differences in the frequency of variants in the fatty acid desaturase (FADS) gene cluster in the context of the modern Western diet can lead to inadequate circulating and tissues levels of n‐3 long‐chain (LC‐; ≥ 20 carbons) polyunsaturated fatty acids (PUFA); these deficiencies in turn have the capacity to increase metabolic disease risk. Specifically, we and others have demonstrated Amerindian (AI)‐Ancestry populations have high frequencies of FADS gene variants that may lead to less efficient n‐3 LC‐PUFA biosynthesis. To test this hypothesis, concentrations of fatty acids, including LC‐PUFAs in plasma phospholipids were analyzed in 1,102 Hispanic American participants from Multi‐Ethnic Study of Atherosclerosis (MESA) cohort, which represent six Hispanic subgroups based on self‐reported region of origin: Central America, South America, Mexico, Cuba, Dominican Republic, and Puerto Rico. These data revealed a striking inverse relationship between genome‐wide AI‐ancestry and levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Additional analyses revealed that the genotype at FADS SNP rs174537, for which the frequency of the T allele increases with AI‐ancestry, could primarily explain the associations. There also was a strong association between the number of T alleles (additive model) at rs174537 and circulating triglycerides (18.5 mg/dL per T allele; P=2.0 x 10−5) as well as an important marker of vascular inflammation sICAM‐1 (b=12.7 ng/mL, P=0.02).We also genotyped the FADS SNP, rs174537, in a separate adult Arizona Hispanic cohort (Arizona Insulin Resistance [AIR] registry) and compared the FADS genotypic frequencies with other racial/ethnic groups. Importantly, the TT genotype associated with limited LC‐PUFA biosynthesis ranges from <1% in African‐Ancestry populations to 40.2% in AI‐Ancestry Hispanics in the AIR Registry, with ~11 % in European‐Ancestry populations. These data demonstrate that high AI‐ancestry populations have a dramatically higher frequency of the TT genotype versus African and European populations. Importantly similar to MESA, there also was a significant association between the number of T alleles and plasma triglycerides (p = 0.0036; GG = 125±8.18 mg/dl, GT = 135±5.63 mg/dl, TT = 143±5.58 mg/dl) again suggesting the AI‐associated TT genotype places this population at higher risk of hypertriglyceridemia. Together, these data reveal that FADS gene*dietary PUFA interactions give rise to n‐3 LC‐PUFA deficiencies and may enhance metabolic and inflammatory diseases in a large proportion of individuals in AI‐Ancestry Hispanic populations.Support or Funding InformationFunding source: NIH R01 AT008621
Inhibition of Sphingosine Kinase 2 Attenuated Cisplatin‐induced Nephrotoxicity in Mice
Authors: Xie Dengpiao Li , Hu Gaizun , Wang Weili , Li Pin-Lan , Chen Chaoling , Li Ningjun 
Published: April 2020

Sphingosine‐1‐phosphate (S1P) is a bioactive sphingolipid metabolite and shown to participate in various kidney diseases. S1P is produced by sphingosine kinases (Sphks), which phosphorylate sphingosine into S1P. It has been demonstrated that knockout of Sphk2 gene protects the kidneys in unilateral ureteral obstruction model. The present study tested the hypothesis that inhibition of Sphk2 protects the kidneys against cisplatin‐induced nephrotoxicity. Male C57BL6 mice were divided into three groups: vehicle (Ctrl), cisplatin + vehicle (Cis) and cisplatin + ABC294640, a Sphk2 inhibitor (Cis + in). Cisplatin (10 mg/Kg) was given by weekly i.p. injection and Sphk2 inhibitor (35 mg/Kg) gavage every other day for 4 weeks. Plasma creatinine levels were significantly higher in Cis group than that in Ctrl, whereas it was significantly lower in Cis+in group than that in Cis (mean: 0.17 ± 0.02, 0.38 ± 0.05, and 0.20 ± 0.02 mg/dL in Ctrl, Cis and Cis+in group, respectively, p<0.05). Western blot analysis showed that the levels of fibrotic protein alpha‐smooth muscle actin (α‐SMA) in the kidneys were significantly increased in Cis group and that this increase in α‐SMA was blocked in Cis+in group. The relative α‐SMA protein levels were 1.0 ± 0.04, 1.8 ± 0.11 and 0.8 ± 0.19 in Ctrl, Cis and Cis+in group, respectively (p<0.05). The increased level of Kidney Injury Molecule‐1 was also significantly reduced in Cis+in group compared with that in Cis group. These results suggest that Sphk2 pathway mediates cisplatin‐induced kidney damage. Manipulating Sphk2 may be used as a therapeutic strategy in the management of cisplatin‐induced nephrotoxicity.Support or Funding InformationNIH grants R01DK54927, R01DK107991, R01HL145163
ADAM17 impairs arterial fluid shear stress mechanosensing in aged mice
Authors: Tian Yanna , Davila Alec , Clermont Andrew , Balogh Marta Bagine , Czikora Istvan , Bagi Zsolt 
Published: April 2020

Junctional Adhesion Molecule A (F11R/JAM‐A), which can be shed by a disintegrin and metalloproteinase ADAM17, has been implicated in arterial mechanotransduction. We hypothesized that in aging ADAM17 activation impairs wall shear stress mechanosensing via an augmented F11R/JAM‐A shedding. Using videomicroscopy we found a reduced wall shear stress (WSS)‐induced dilation in isolated, pressurized gracilis muscle arteries in 30‐month old (aged) mice, when compared to 12‐week old (young) controls. Dilations to acetylcholine or to the NO donor, sodium nitroprusside were not different between the two groups. We found that ADAM17 expression was significantly upregulated in arteries in aged mice, which was associated with reduced endothelial cell junction localized F11R/JAM‐A. After activation of ADAM17 with phorbol‐myristat‐acetate, WSS‐induced dilation was reduced in young mice, whereas ADAM17 inhibition by TIMP3 augmented WSS‐induced dilation in aged mice. Cultured HUVECs exposed to increasing levels of WSS (0 to 16 dyne/cm2) progressively changed their orientation to the direction of flow. In the presence of recombinant ADAM17 or after siRNA‐mediated silencing of F11R/JAM‐A, endothelial cells failed to efficiently orient with the direction of flow. Gene transfer of ADAM17 cleavage resistant, mutated JAM‐A(V227Y) resulted in a normal endothelial orientation to flow even after ADAM17 activation. Collectively, our results uncover a novel role for ADAM17 in impairing F11R/JAM‐A‐dependent WSS mechanosensing in arteries of aged mice.Support or Funding InformationNational Institute of Aging (R01AG054651)
Loss of CLC‐5 reduces endocytic capacity and alters cholesterol distribution in the proximal tubule
Authors: Shipman Katherine , Long Kimberly R. , Rbaibi Youssef , Raja Sripriya , Baty Catherine J. , Weisz Ora A. 
Published: April 2020

Dent disease is a progressive X‐linked disorder affecting the kidney proximal tubule (PT) caused by loss of function of the Cl−/H+ exchanger, CLC‐5. Early symptoms include low molecular weight (LMW) proteinuria resulting from inefficient reabsorption of filtered proteins by the PT. However, the role of CLC‐5 in this process in unclear. Knockout of Clc‐5 in mice recapitulates the LMW proteinuria observed in human disease and causes decreased PT levels of megalin and cubilin, the receptors that bind to and recover filtered proteins. How loss of CLC‐5 leads to reduced receptor expression remains unknown. We used a previously developed opossum kidney (OK) cell culture model that recapitulates morphologic and functional features of the PT in vivo to study the role of Clc‐5 in the endocytic pathway. In Clc‐5 siRNA knockdown (KD) and CRISPR/Cas9 mediated Clc‐5 knockout (KO) cells, there was a significant decrease in endocytic capacity that was rescued by heterologous expression of wild‐type human CLC‐5. Additionally, we used CRISPR/Cas9 technology to generate Clc‐5 knockout mice and confirmed LMW proteinuria in the KO. Heterozygous females also have reduced PT albumin uptake and megalin expression. Previous gene expression studies in Clcn5 KO mice suggest there are alterations in cholesterol and lipid metabolic enzymes, and elevated cholesterol levels have been shown to alter recycling endosome organization and impair fast recycling. We hypothesize that loss of CLC‐5 disrupts cholesterol homeostasis, resulting in reduced recycling and concomitantly increased degradation of megalin. Consistent with this, we observed an accumulation and redistribution of cholesterol in Clc‐5 KD OK cells. We are currently working to quantify cholesterol in Clc‐5 KO OK cells. In female mice with only one functional copy of Clc‐5, we observed a dramatic a change in the distribution of cholesterol in the PT when compared to WT mice. Current studies are focused on quantifying the trafficking kinetics and distribution of megalin in the PT in the absence of CLC‐5 and determining whether altered cholesterol metabolism contributes to the reduced endocytic capacity of the PT and resulting LMW proteinuria in Dent disease.Support or Funding InformationNIH F31 DK121394, R01 DK101484, R01 DK100357, P30 DK079307, and ASN Pre‐Doctoral Fellowship
Implementing Evidence‐Based Teaching Practices: Support for Educators through the Promoting Active Learning & Mentoring Research Coordination Network (PALM Network)
Authors: Johnson Kathryn M.S. , Stieben Margaret E. , Wick Sue 
Published: April 2020

The Promoting Active Learning & Mentoring Research Coordination Network (PALM Network) provides one‐to‐one year‐long mentored support to biology faculty and postdoctoral fellows seeking to implement evidence‐based, effective active learning strategies into their classrooms. These PALM fellows are paired with mentors, recognized for their experience in education reform, typically faculty at another academic institution. Mentors assist fellows as they design, implement, and assess teaching innovations in their own classroom, and fellows visit the mentor’s institution to observe evidence‐based practices in the mentor’s classroom. In addition to individual mentoring, PALM fellows meet regularly, virtually and in‐person, with other fellows and mentors, providing a diverse and broad network of support for their continued development as educators. Support and assistance from professional societies provides an operational framework, promotion, and recognition to the Network. To date, 28 fellow‐mentor pairs have participated in the PALM Network. Here we present the findings of the mid‐point evaluation of the 5‐year NSF‐funded network, using a mixed‐methods assessment approach. Fellows (n=13) and mentors (n=11) completed online surveys upon acceptance or after one year of participation in the PALM Network. At the time of acceptance into the PALM Network, self‐reported confidence of the network member’s ability to use backward design to develop active learning activities and assessments of student learning was dependent upon whether the respondent was a fellow or a mentor (p ≤ 0.046). For those surveyed after one year in the PALM Network, self‐reported confidence in these abilities were not different between fellows and mentors (p ≥ 0.122), indicating the abilities of the fellows shifted towards that of the mentors. Grounded‐theory analysis of semi‐structured in‐person interviews with mentors and fellows (n=8, representing 7 mentor‐fellow pairs) participating in the PALM Network for at least 6 months, but less than 2 years, revealed PALM Network members are enthusiastic, talented, and ambitious educators and researchers, dedicated to implementing education reform. PALM fellows value the one‐to‐one mentoring and the support of a broad network. PALM Network fellow‐mentor collaborations result in successful implementation of transformational pedagogical projects. Although implementing change at the fellow’s home institution was sometimes difficult, the mentors and other PALM Network members provided valuable advice and support for the fellows to facilitate their work. Both fellows and mentors highly recommend the PALM Network. Therefore, the PALM Network provides effective support for educators implementing evidence‐based best practices in their classrooms.Support or Funding InformationThis work was funded by NSF‐RCN‐UBE #1624200, including a sub‐award to American Physiological Society.