bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hormesis ultraviolet radiation and its effects on Drosophila melanogaster performance
Authors: Berry Raymond , Lopez-Martinez Giancarlo 
Published: April 2020

Hormesis can be protective as a multifarious response that results in upregulation of genes and organismal performance. The protective effects of hormesis in animal models includes improved survival, mating, extended longevity, and improved performance at old age (i.e. mating); amongst others. The sun is a source of energy used by terrestrial life in many trophic levels allowing for exposure to irradiation (UVR), a natural environmental stressor that increases production of reactive oxygen species (ROS) .This type of environmental stress is damaging by attacking lipids, proteins and DNA. Exposure to prolonged UVR conditions early in life (pupae) leads to changes in performance later in life. This response can be classified as preconditioning, pretreatment, conditioning, or cross tolerance. Previous studies scantily provide appropriate levels of exposure needed for UVR hormesis that would increase production of ROS and antioxidant enzymes. The age of the individual at the time of treatment can have dramatic effects on hormesis with low dose UVR effects that induce protection, an extension of normal life span and improved organismal performance shown in other insects. The vinegar fly, Drosophila melanogaster, was exposed to UVA at different intensities. Vials containing pupae, separated from food and kept in same vial were exposed to 0, 30, 45 or 60 minutes of ultraviolet radiation A (nm = 365). Different rates of UVA exposure (mW/m2), produced by the distance from the source, show slight differences in treatment survival/adult emergence and flight ability. Low doses of UVR lead to protective and beneficial responses, while exposures to high doses are damaging and detrimental. Ultimately, data such as these can be refined to develop a model of dose response that can be used to estimate hormesis, or the biphasic response.Support or Funding InformationNIH RISE 5R25GM061222‐19
Effects of Cannabidiol on RAW 264.7 Macrophage Viability and Inflammatory Markers
Authors: Danchine Veronika Denise , Caldari Cristina 
Published: April 2020

Cannabidiol (CBD) is a non‐psychoactive derivative from the marijuana plant. There has been an explosion of readily available commercial products infused with and marketing CBD contents, and yet there is a paucity of information regarding the specific effects of CBD on immunity. Our laboratory is interested in examining the effects of CBD on innate immunity, using the immortalized murine macrophage cell line, RAW 264.7. The objective of this research was to determine the effects of CBD on the proliferation, activation, and inflammatory cytokine production in lipopolysaccharide (LPS)‐activated murine macrophages. Additionally, we wanted to determine if maintaining the murine macrophages in serum‐free media during CBD incubation would have an effect on cell viability. RAW 264.7 cells were incubated with either 0.2 or 20 μM of CBD for 24 hours in serum‐free media (SFM) or complete media (CM), followed by a 6 hour, 0.01 μg/ml LPS challenge in SFM. Cell viability was determined via cell counts with Trypan blue exclusion and MTS assay. Macrophages cultured in CM tended to have higher percentages of live cells (Trypan blue exclusion method) compared to those cultured in SFM (54.7+5.4% vs 31.8+5.4% live; ANOVA, p=0.07). Additionally, culturing macrophages in CM also resulted in significantly higher viability (MTS assay) compared to macrophages in SFM (0.58+0.06 vs 0.29+0.06 490nm absorbance; ANOVA, p=0.03). There was no significant effect of CBD concentration (ANOVA; p=0.4) or CBD concentration*media (ANOVA; p=0.2) type on cell viability. These results suggest that concentrations of 0.2 and 20 μM CBD have no detrimental effects on murine macrophage viability and proliferation and that CBD pre‐incubation of macrophages should be performed in complete media. Additionally, data will be presented on the effects of CBD on macrophage activation, including nitric oxide (NO) and interleukin (IL)‐6 production.Support or Funding InformationDenison University Biology Department and Denison Lisska Center for Scholarly Engagement
Creation of a Novel Absorbent Bandage for Pressure Ulcer Treatment
Authors: Dunlap Jared N. , Fischer Kristin , Wolyniak Michael J. 
Published: April 2020

Research was undertaken to explore the possibility of using solid, microporous materials in a bandage that when applied to a pressure ulcer that achieves the following criteria:

‐contains a microporous material as the secondary absorbent that withholds moisture under bodily conditions.
‐sustains an ideal moisture level on the wound bed to create an ideal healing environment.
‐expect an absorbance capacity for water from the microporous material under body‐like conditions over 25%.
‐accounts for multiple different secretion types in and around wounds.
‐integrates this adsorbent into a comfortable gauze bandage for easy for use by doctors and nurses.
‐prevents infection and bacterial growth in this bandage by integrating a heat‐stable antibiotic into the bandage.The goal of this research was to understand if these goals were feasible, then design a bandage containing a microporous material that could prevent pressure ulcer progression and infection, and further test the used components for effectiveness and unintended reactions. Comparative literature research was conducted on several heat stable antibiotics and on multiple microporous materials including kaolin clays, silica gel, and zeolite. It was determined that synthetic zeolite (4 Angstrom Molecular Sieve) was the ideal candidate to use in the bandage. A highly effective, heat stable antibiotic was also chosen to integrate into the bandage: Trimethoprim‐Sulfamethoxazole. The combination of the two materials yields the potential for the long‐term application of this bandage. However, when acting as a Hemostatic Agent, zeolite has been found to cause an exothermic reaction when it contacts blood. In order to continue my construction and study of the bandage, this interaction needs to be further understood in relation to pressure ulcers. Bovine serum, whole blood, and ratios of fractionated blood was tested in tissue wells with zeolite at physiological temp (37□) for any significant exothermic reaction amounting to a temperature change over 5□ from physiological temp. No significant temperature change was noted with Fetal Bovine Serum and will be used as a control. Another goal includes testing upon the zeolite mechanism of action if such a temperature change occurs with blood ratios found in pressure ulcers, in order to potentially mitigate the effects when implemented in a bandage. Another goal is testing to study effectiveness of antibiotics when integrated and subjected to heat and vacuum with zeolite. The end goal is finally designing and constructing a prototype bandage.Support or Funding Information‐Hampden‐Sydney College Undergraduate Research Council.
How does GDP‐fucose get into the Endoplasmic Reticulum for O‐fucosylation of EGF Repeats and TSRs?
Authors: Yuan Youxi , Haltiwanger Robert S. 
Published: April 2020

Fucose is a common monosaccharide found in glycan chains of all mammalian cells. Fucose is transferred from GDP‐fucose to glycoproteins by fucosyltransferases, resulting in terminal fucosylation of N‐linked glycans or mucin‐type O‐glycans, or direct modification of serine or threonine residues by Protein O‐fucosyltransferases (POFUT) 1 or 2. Defects in fucosylation can lead to developmental disorders or cancer. In particular, POFUT1 adds O‐fucose to Epidermal Growth Factor‐like (EGF) repeats of Notch receptors, while POFUT2 adds O‐fucose to Thrombospondin type 1 Repeats (TSRs) found in multiple cell‐surface and secreted proteins, including the A Disintegrin And Metalloproteinase with ThromboSpondin motifs (ADAMTS) family of extracellular proteases. Elimination of Pofut1 or Pofut2 results in early embryonic lethality in mice, indicating these two fucosylation pathways play essential roles in mouse development. Both POFUT1 and POFUT2 are located in the endoplasmic reticulum (ER), while fucosyltransferases involved in terminal fucosyaltion are localized in the trans‐Golgi. GDP‐fucose is synthesized in the cytoplasm either through a de novo or salvage pathway, and the GDP‐fucose is then transported into the lumen of the ER or Golgi for use by the fucosyltransferases. Currently two GDP‐fucose transporters, SLC35C1 and SLC35C2, have been identified, both of which are localized to the Golgi. It is unknown how GDP‐fucose is transported into the ER where Notch receptors and ADAMTS proteins are fucosylated. In order to uncover where the GDP‐fucose for POFUT1 and POFUT2 comes from, we have generated CRISPR‐Cas9 mutant cell lines in which fucosylation is impaired in two ways: 1. Inactivating GDP‐fucose biosynthetic enzymes from the salvage or de novo pathways; 2. Inactivating the GDP‐fucose transporters. HEK293T cells have proven to be an excellent model for fucosylation studies. All the fucose biosynthesis genes are normally expressed, and impaired fucosylation can be expected in knockout mutant cell lines. Using CRISPR‐Cas9 technology, the following genes have been knocked out: GMDS and TSTA3 of the de novo pathway, FUK and GFPP of the salvage pathway, and the GDP‐fucose transporters SLC35C1 and SLC35C2. O‐Fucosylation of EGF repeats and TSRs and terminal fucosylation of N‐linked glycans is being analyzed using Mass Spectrometry and lectin binding. Notch function is also being evaluated. These results should reveal whether a preferred biosynthetic pathway or transporter for different forms of fucosylation exists.Support or Funding InformationSupported by NIH grant GM061126.
Elucidating the Mechanism of DNA Repair in Bdelloid Rotifers by Using RNAi Knockdown and CRISPR/Cas9 Editing
Authors: Gardner Storm Devin , Schurko Andrew Michael 
Published: April 2020

Genomic DNA regularly becomes damaged by numerous ways, including desiccation, radiation exposure, and during recombination in cell division. Fortunately, cells have DNA repair pathways that seek out and correct damage. The bdelloid rotifer Adineta vaga is an excellent model organism to study DNA repair because it has a highly efficient DNA repair system. Bdelloids can recover from desiccation and return to life by repairing DNA damage endured in the desiccated state. Bdelloids have also been shown to recover from DNA damage caused by massive doses of ionizing radiation. The mechanisms of this miraculous DNA repair in these aquatic invertebrates are unknown. This project explores using RNA interference (RNAi) and CRISPR/Cas9 genome editing as tools to investigate the function of DNA repair genes, MSH6 and MLH1, in bdelloid rotifers. When target genes are knocked down or inactivated, DNA repair in bdelloids should be compromised. For RNAi, two methods were explored. For the “feeding method,” bdelloids were fed strains of E. coli that express double stranded RNA (dsRNA) copies of MLH1 and MSH6. For the “soaking method,” dsRNA copies of MLH1 and MSH6 were synthesized, and bdelloids were transfected with these molecules. The expectation was that ingested or transfected dsRNA would induce RNAi in bdelloid rotifers. Real‐time PCR followed to quantify expression of the target gene and determine if the RNAi knockdown was successful. For the feeding method, an MLH1 knockdown was observed 6–12 hours after feeding, but the effect was absent 24 hours after feeding. For the soaking method, an MSH6 knockdown was only present when rotifers were transfected with 0.2 μg dsRNA and incubated for 48 hours. As an alternative tool for studying gene function, CRISPR/Cas9 genome editing was done to mutate DNA repair genes. For utilizing CRISPR/Cas9, a single guide RNA that targets MLH1 was synthesized and used in a Cas9 in vitro cleavage assay to determine its ability to cut the target sequence. The guide RNA was then combined with Cas9, and these ribonucleoprotein complexes were electroporated into bdelloids. Mutation detection using T7 endonuclease I was then done to identify alterations to the gene sequence. Gene knockdowns using RNAi and gene mutations using CRISPR/Cas9 will provide a reverse genetics approach to identify genes that have a role in bdelloid DNA repair. This could enhance our understanding of similar mechanisms in traditional model systems, which could have implications for biomedical research.Support or Funding InformationThis work was supported by grants from the Hendrix College Odyssey Program and the Hugh Stevens Moseley Undergraduate Research Awards.
Renomedullary Interstitial Cell Endothelin A Receptors Regulated Blood Pressure and Renal Function
Authors: Hu Chunyan , Lakshmipathi Jayalakshmi , Stuart Deborah , Peti-Peterdi Janos , Kohan Donald E. 
Published: April 2020

Renomedullary interstitial cells (RMIC) may modulate blood pressure (BP) and renal function through release of natriuretic factors and through direct effects on medullary blood flow (RMIC encircle vasa recta). Endothelin‐1 (ET‐1) modulates RMIC vasoactive factor release and contraction via ETA receptors (ETAR) in vitro. Consequently, we tested the hypothesis that knockout of ETAR in RMIC regulates renal function and BP in vivo. We bred ETARf/f mice containing loxP‐flanked exons 6–8 of the EDNRA gene with tamoxifen‐inducible tenascin‐C‐CreER2 knockin mice. Tenascin‐C‐CreER2:ETARf/f mice were injected with tamoxifen (160 mg/kg IP) for 5 consecutive days at 3 months of age to obtain RMIC ETAR KO. Littermates of the same genotype without injection served as controls. EDNRA gene recombination occurred in inner medulla, but not cortex. Inner medulla ETAR mRNA content fell by 80%, while ETBR mRNA levels were unaffected. ETRA KO did not affect BP in mice on normal (0.3% Na+) or low (0.03% Na+) salt diets. In contrast, ETAR KO reduced salt‐sensitive systolic BP elevation in mice on a high (3.2% Na+) salt diet (ETAR KO 127±3 vs. control 134±2 mmHg, n=13, 15, p<0.05). Urine volume and urinary Na+ and K+ excretion were higher in high‐salt diet‐treated ETAR KO mice compared with controls during the first 3 days of high salt intake. Urinary ET‐1, nitrite/nitrate, and prostaglandin E2 excretion were increased in high salt diet‐treated ETAR KO mice vs. controls. No significant differences were found in GFR, vasopressin, aldosterone, or renin levels between high salt fed ETAR KO and control mice. Inner medullary protein and mRNA levels of total NKCC2 and ENaC‐α and ‐β were decreased, and inner medullary cyclooxygenase‐2 (COX‐2), but not COX‐1, protein levels were increased, in high salt fed ETAR KO vs. control mice (n=8). Exogenous ET‐1 did not detectably alter RMIC intracellular Ca2+ concentration or cause RMIC contraction as assessed by ex vivo inner medulla confocal imaging. We conclude that disruption of RMIC ETAR reduces salt‐sensitive hypertension and this is associated with reductions in NKCC2 and ENaC together with elevations in several autocrine and paracrine natriuretic factors. These effects of RMIC ETAR, at least based on ex vivo studies, may not involve RMIC contraction. While further study is required, our data suggest a novel regulatory pathway for maintaining water/electrolyte homeostasis.Support or Funding InformationNHLBI P01 HL136267
Coronary Vascular Cell Stiffness Influences Coronary Blood Flow
Authors: Montgomery Kathlyene R. , McCallinhart Patricia E. , Trask Aaron J. 
Published: April 2020

Our laboratory has previously shown that type 2 diabetic (T2DM) coronary resistance microvessels (CRMs) undergo inward hypertrophic remodeling associated with reduced stiffness. Reduced T2DM CRM tissue stiffness is associated with decreased elastic modulus of coronary vascular smooth muscle cells (VSMCs). The goal of this study was to test the hypothesis that reducing or augmenting coronary vascular cell stiffness will increase or decrease coronary blood flow, respectively. Coronary blood flow was measured from hearts isolated from 16‐week‐old control Db/db and diabetic db/db mice and perfused on a Langendorff system at a constant pressure of 80 mmHg. Latrunculin B (1 μM final concentration; n=2 per group) was infused to depolymerize actin and reduce vascular cell stiffness and jasplakinolide (0.2 μM final concentration; n=4 per group) was infused to increase actin polymerization and increase vascular cell stiffness. After drug infusions, hearts were immediately transferred to an adjacent Langendorff apparatus and were perfusion fixed with 4% paraformaldehyde for 2 mins at 1 mL/min. Hearts were then placed in 4% PFA for 24–48 hours and were stored in 70% EtOH until embedding in paraffin and sectioning. Sections were evaluated for F‐ and G‐actin by immunofluorescence. In pooled Db/db and db/db analyses, Latrunculin B caused an increase in coronary blood flow over baseline (34.1 ± 15.0% above baseline, n=7), and jasplakinolide caused a 26.5 ± 4.1% reduction in coronary blood flow from baseline (n=8). Immunohistochemical analysis confirmed a decreased trend in F/G actin in the coronary microvessels of latrunculin B‐perfused hearts (1.00 vs. 0.82 ± 0.08, n=1–4 per group), while F/G actin tended to be increased in the CRMs of jasplakinolide‐perfused hearts (1.00 ± 0.00 vs. 1.50 ± 0.16, p=0.12, n=2–6 per group). Importantly, the F/G actin ratio in jasplakinolide‐treated hearts was not different measured in the myocardium 50–100 μm away from a CRM (1.00 ± 0.06 in vehicle‐perfused hearts vs. 1.01 ± 0.10 in jasplakinolide‐perfused hearts, p=0.96, n=2–6 per group), demonstrating that the drug was efficacious only in the peri‐vascular region. These data support the hypothesis that coronary blood flow increases as vascular cell stiffness is decreased and coronary blood flow decreases as vascular cell stiffness is increased. Collectively, these data suggest that coronary vascular cell stiffness can modulate coronary blood flow.Support or Funding InformationKRM was supported by an American Heart Association Summer Undergraduate Research Fellowship (17UFEL33420025). NIH R00HL116769, R21EB026518, and S10OD023438 to AJT.
Mentorship In Converting Research Into A CURE: The Mentoring The Integration Of Research Into The Curriculum (MIRIC) Initiative
Authors: Wolyniak Michael , Barton Lance F. , Resendes Karen K. 
Published: April 2020

The American Association for the Advancement of Science (AAAS) Vision and Change document has spurred several initiatives designed to improve the way in which undergraduates learn science. Often, these initiatives have been disseminated as one‐time workshops that generate awareness of and interest in developing authentic research experiences for undergraduate STEM classrooms. Conversely, whole “standardized” curricula have been developed and made available for adoption. However, successfully generating the sustainable change necessary to bring real reform to undergraduate science education should benefit both students and faculty scholarship. To create sustainable change, long‐term faculty development initiatives focused on mentorship are needed so that instructors can develop and implement sustainable curricula with local relevance. Experienced instructors seasoned in developing and implementing course‐based undergraduate research experiences (CUREs) based on their own scholarship and local resources can convey their experiences to mentees interested in using these pedagogical techniques as the centerpiece of their own teaching. The Council on Undergraduate Research (CUR) Biology Division has created the Mentorship for Integrating Research Into the Classroom (MIRIC) program to provide a means for members with an interest in developing improved and sustainable active learning techniques to gain experience in this style of teaching through close, long‐term interaction with a veteran teaching mentor. Developed from the former ASCB Mentorship in Active Learning and Teaching (MALT) program, MIRIC focuses on the development of instructors who wish to develop a dynamic CURE. Current and future life science instructors pair themselves up with seasoned veterans of CURE development and work with them and their students over the course of a semester or longer to develop a CURE that will allow the mentee to bring authentic research into his or her classes. In our pilot studies, we collected qualitative data based on participant interviews that suggest that MIRIC mentorships have made positive gains in promoting sustainable active learning techniques among participants. Since then, we have surveyed the first cohort on both the success of their CURE development process as well as the benefits of the mentoring relationship on both the mentors and mentees using validated questions derived from Mathur (2012) and Pamuk and Thompson (2009).Support or Funding Information‐The Council on Undergraduate Research (CUR)
Cux1 regulates cilia length in polycystic kidney disease
Authors: Holthaus Conner , Kumar Emmanuel , Rumschlag Matthew , Wee Zhi Nee , Pasillas Melissa , Heuvel Greg Vanden 
Published: April 2020

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is one of the most common inherited disorders affecting the kidney. Renal cyst development in ADPKD results from mutations in the PKD1 or PKD2 genes, which encode the proteins polycstin1 (PC1) and polycystin2 (PC2), respectively. PC1 and PC2 proteins are localized to primary cilia where they are proposed to form a receptor channel complex that detects flow transmitting a calcium‐mediated signal. Primary cilia are critical to the pathogenesis of ADPKD, which is one of many ciliopathies that exhibit renal cystic disease. Cux1, a murine homolog of the Drosophila gene Cut, is a cell cycle dependent transcriptional repressor that regulates expression of the cyclin kinase inhibitor p27. Cux1 is highly and ectopically expressed in mice carrying a collecting duct specific mutation of Pkd1 (Pkd1CD) and in human ADPKD cells where it promotes cell proliferation. Mice carrying mutations in both Cux1 and Pkd1 have reduced cystic disease and an increased life span. A role for Cux1 in regulating genes involved in cilia assembly and function has recently been identified in the Galapagos cormorant, however, the role of Cux1 in cilia in the mammalian kidney is not known. To begin to determine whether Cux1 regulates ciliogenesis we evaluated cilia morphology and the expression of the ciliary protein, OFD1 (oral‐facial‐digital‐1), identified as a target of Cux1 in the Galapagos cormorant. Cilia analysis was performed on kidneys isolated from wild type, Cux1 transgenic, Pkd1CD, and Pkd1CD/Cux1 double mutant mice. Cilia morphology was assessed by immunofluorescence labeling of alpha‐tubulin and the collecting duct marker dolichos biflorus agglutinin (DBA) to identify cells in which Pkd1 was deleted. Cilia in Pkd1/Cux1 double knockout kidneys were significantly shorter than cilia in the Pkd1CD kidneys, consistent with previous studies showing that decreased cilia length corresponds to decreased cystic disease. OFD1 is an inhibitor of ciliogenesis and OFD1 expression in the various mouse models demonstrate that OFD1 expression corresponds to Cux1 expression. OFD1 protein levels were the lowest in the kidneys of mice constitutively expressing Cux1 and were highest in mice with deletions of Cux1. Taken together, our results reveal a novel role for Cux1 in regulating ciliogenesis in the kidney and suggest that the reduced cystic disease in the Pkd1/Cux1 double mutant mice results from reduced cilia length.Support or Funding InformationAmerican Association of Anatomy FGAP, Western Michigan University Homer Stryker MD School of Medicine