bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of hepatic cytochrome P450 oxidoreductase in bone development (921.1)
Authors: Kellogg Dean , Tang Kejian , Roman Linda , Fajardo Roberto , Masters Bettie Sue , Panda Satya 
Published: June 2021

Cytochrome P450 oxidoreductase (POR) is the only known electron donor for Cytochromes P450 (CYPs), heme oxygenase (HO), and dehydrocholesterol reductase in the endoplasmic reticulum (ER). Hepatic CYPs are essential for the metabolism of cholesterol, steroids, sex hormones, and xenobiotics, such as drugs and toxins. Thus, alterations in POR activity will have multiple effects on the physiological activity of multitudinous metabolites. Human mutations in POR have been reported and present as altered steroidogenic profiles, severe craniofacial and bone defects resembling Antley‐Bixler syndrome (ABS). Liver‐specific POR null (LCN) mice were examined to study endo‐ and xeno‐biotic metabolism. Surprisingly, previous studies demonstrated only minor changes in plasma and intestinal lipid content of LCN mice, suggesting that lipid homeostasis is maintained despite significant hepatic steatosis. Here, we show for the first time that the deletion of POR from the liver leads to decreased bone volume in both cortical and trabecular compartments, as measured by micro computed tomography analysis, but no notable skull defects. In bone‐specific POR null mice, however, we have previously observed defects in skull morphology and trabecular bone density. This difference suggests that the action of hepatic CYP‐derived circulating metabolites is critical for normal bone development and is distinct from bone‐ specific CYP activity. There are several potentially important metabolites that could affect bone homeostasis and require hepatic CYP hydroxylation such as vitamin D, retinoic acid and steroid hormones. These data highlight the importance of both systemic and local POR activity in healthy bone development.Grant Funding Source: Supported by NIH grant GM081568 to BSSM and SPP
Analysis of yeast SUMO ligase MMS21‐dependent genome maintenance pathway (925.4)
Authors: Liang Jason , Lee Nancy , Albuquerque Claudio , Putnam Christopher , Kolodner Richard , Zhou Huilin 
Published: June 2021

Genome instability often characterizes many cancers and is indicative of high frequency of mutations and genome rearrangements. Several pathways, including the DNA damage checkpoint, play a role in maintaining the stability of the genome by suppressing gross chromosomal rearrangements (GCRs). Additionally, eukaryotic genomes contain numerous repetitive sequences, segmental duplications and regions of copy number variation, which influence the outcomes of gross chromosomal rearrangements.Using a genetic assay in Saccharomyces cerevisiae to screen for suppressors of GCRs, we recently reported that Mms21‐dependent sumoylation specifically suppress GCRs mediated by homeologous sequences. These types of duplication‐mediated GCRs are attributed to Rad52‐dependent non‐allelic homologous recombination (NAHR). We will present findings that show unexpectedly, the DNA damage checkpoint to be required for the formation of duplication‐mediated GCRs in the absence of Mms21‐dependent sumoylation. The canonical DNA damage checkpoint genes MEC1, RAD24, MEC3, RAD9 and RAD53 are required for formation of duplication‐mediated GCRs in the mms21 mutant, while TEL1, MRC1 and CHK1 are minimally involved. Among the checkpoint genes, mutation of RAD9 strongly suppresses GCRs formed in the mms21 mutant and this suppression is independent of either SIZ1 or SIZ2, which encode two other SUMO ligases in yeast. We will also present findings on the role of DNA damage checkpoint in regulating the SUMO proteome. These results show that the outcomes of GCRs are dynamic and are governed by a fine balance of distinct genome maintenance pathways.Grant Funding Source: GM080469, T32CA009523 ,LICR
Determining the genetic basis of equine deafness via candidate gene studies (928.3)
Authors: Innamorati Katherine , Lipsett Koren 
Published: June 2021

White fur or coat pigmentation and the presence of blue eyes have been linked to cochleosaccular deafness in multiple mammalian species. At this time, no hereditary explanation has been presented for this phenotype as it appears in horses. A candidate gene study was established to determine the genetic basis for this disorder in a pedigree of Spanish Mustangs. KIT, a receptor tyrosine kinase implicated in multiple mammalian deafness studies, was sequenced in both the affected Spanish Mustangs and related members of the pedigree. A mutation (KI16 + 1037A) in the 5’ splice junction of exon 17 known to cause the sabino coat pattern appeared to be unlinked from the deafness phenotype. Sabino coloration is distinguished as a white coat, ranging from minimal white splashes to an entirely white mammal. Future research will focus on the sequencing of additional candidate genes affecting melanocyte migration and melanin production, including EDNRB, MITF, PMEL17, and SOX10.Grant Funding Source: Supported by HHMI
Transcriptional changes in gamma‐secretase conditional knockout mice (934.5)
Authors: Arbella Jennifer , Yu Gang , Dries Daniel 
Published: June 2021

gamma‐secretase is an intramembrane protease composed of four transmembrane proteins: presenilin, nicastrin, Pen‐2, and Aph‐1. Several studies have examined the role of gamma‐secretase in neurons but none have done so in non‐neuronal cells. Therefore, our lab developed a conditional knockout mouse in which gamma‐secretase was conditionally deleted from the myelinating cells of the brain. Here, we use quantitative PCR to confirm the specificity of the conditional deletion and to probe a series of putative genes for their altered expression in conditional knockout mice. Moreover, we begin to look at transcriptional changes in the conditional knockout animals using using RNA‐seq.Grant Funding Source: Supported by National Institute of Neurological Disorders and Stroke grant 1R01NS079796
Live cell monitoring of gene modulation provides a better understanding of gene function (946.12)
Authors: Weldon Don , Williams Yuko , Koong Victor , Huryn‐Selvar Karyn 
Published: June 2021

One of the most common methods for understanding gene function within a live cell is modulating the gene through knockdown or over‐expression systems. These experiments provide information regarding the role a particular gene plays within a given biological process. Unfortunately, determining the modulation in gene expression is currently performed at the population level as in the case of qPCR and or is ultimately an endpoint determination as in the case of RNA FISH.We propose a new method for determining the RNA expression levels of target genes using a nanoparticle technology which can be used in live cells without the need for transfection reagents. Live cell gene expression detection affords the ability to monitor changes in expression levels over time. More data can be collected over the course of a single experiment at the cellular level while monitoring gene expression in a dynamic manner as opposed to studying many samples at varied time points in the case of RT‐PCR or FISH.Here we present data illustrating the benefits of detecting gene expression in live cells while modulating both Survivin and ER alpha genes. U2OS cells which have an ER alpha knock‐in under the control of an inducible promoter were monitored using the nanoparticle detection method prior to and during the induction process enabling the monitoring of the gene modulation. Knockdown experiments were also performed using Survivin as a target gene to illustrate down regulation of genes and the ability to monitor the loss of function as well.Monitoring the modulation of genes within live cells can assist in determination of gene function by allowing for the collection of many more data points within a single experiment without the need for duplicate samples at various time points. Furthermore, as this method is non toxic the live cells can be further characterized or used in subsequent experiments providing even more data from the same cells.
Quantifying the two populations of dual oriented P6 in nontypable Haemophilus influenzaeand Pal in Escherichia coli (948.6)
Authors: Shaw Juliana , Schmidt Rachel , MacPherson Victoria , Pichichero Michael , Michel Lea 
Published: June 2021

Nontypable Haemophilus influenzae (NTHi) causes otitis media and other respiratory diseases in people of all ages, conferring the need for a vaccine against this bacterium. P6 is a lipoprotein and a vaccine candidate for protection against NTHi. Recent studies show that P6 is inserted into the outer membrane of NTHi in two distinct orientations‐ facing out toward the extracellular space and in toward the periplasm (Michel et al., J of Bacteriology 195: 3252). To further investigate the two orientations of P6, we utilized a cell‐impermeable biotinylating reagent (NHS‐LC‐LC‐biotin) and streptavidin‐agarose beads to separate the outward and inward facing P6. We then used standard protein detection methods to quantify the two populations. Results showed that the great majority of P6 faces in toward the periplasmic space of NTHi. Similar methods were used to quantify the two populations of Pal, P6’s homologue in Escherichia coli. Our results showed that, like P6, Pal is mostly oriented facing in toward the periplasmic space of the bacterial cell. We propose that, in light of our results, there may be more dual oriented lipoproteins in biology.Grant Funding Source: This study was supported by NIH NIDCD RO1 08671 (to MEP) and the Rochester Institute of Technology.
Overexpression of the novel gene CIA7 in Chlamydomonas reinhardtii (995.3)
Authors: Pedraza Ricardo , Ynalvez Ruby 
Published: June 2021

Heavy metal pollution has become a significant environmental and public health problem that the world faces. C. reinhardtii is an excellent model organism that has often been used in studies of heavy metal tolerance. This alga contains the novel gene CIA7 that possibly encodes a protein that may play a role in metal binding. This gene was isolated from inorganic carbon acquisition mutants. By cloning and overexpressing CIA7 in C. reinhardtii, it can be determined if CIA7 is critical in C. reinhardtii’s metal toxicity tolerance. This gene of interest was first extracted from the alga and then amplified using PCR. Vectors pENTR™/D‐TOPO® (Invitrogen) and psL72 plasmids were used for cloning CIA7 in E.coli. C.reinhardtii cells were then transformed through electroporation. Growth analysis between wild type C.reinhardtii D66 and transformants at different metal concentrations were done to determine the effect of CIA7 on C.reinhardtii under metal stress. If a link between this protein and heavy metal tolerance is identified, this could potentially lay the foundation for its use in heavy metal pollution.Grant Funding Source: Supported by: Norman Hackerman Advanced Research Program
Role of lysophosphatidic acid in traumatic brain injury: anti‐LPA antibodies are neuroprotective after experimental TBI (999.3)
Authors: Wojciak Jonathan , Sabbadini Roger , Crack Peter , Morganti‐Kossmann Cristina , Zhang Moses , Pebay Alice , Conquest Alison , Morris Andrew 
Published: June 2021

Lysophosphatidic acid (LPA) is a bioactive phospholipid with a potentially causative role in neurotrauma. LPA receptors are increased after human brain injury and LPA levels are upregulated in the CSF and blood of traumatic brain injury (TBI) patients. Further, LPA levels correlated with initial injury severity using GCS, GOSE and ISS clinical assessments. Blocking LPA signaling with the LPA‐directed monoclonal antibody, B3/Lpathomab, was neuroprotective in a mouse controlled cortical impact model of TBI. B3 significantly reduced lesion volume measured both by standard histological assessments and MRI. Diminished tissue damage coincided with improvement in functional outcomes. B3 was anti‐inflammatory by limiting cerebral mRNA expression of pro‐inflammatory cytokines such as interleukin‐1b. These data strongly suggest that LPA dysregulation/upregulation is causal to injury progression following TBI, and that interventions to reduce LPA may have a significant impact in mitigating early and late phases of neurotrauma. We propose that anti‐LPA mAbs could satisfy a critical unmet need by limiting the initial neuronal damage and inflammatory processes while subsequently stimulating regenerative processes to optimize long‐term functional outcomes after neurotrauma. Importantly, LPA may also be a novel biomarker for TBI.Grant Funding Source: Lpath, Inc.
The Mechanism for Tau Tangles (LB102)
Authors: Ambessie Sallem , Shire Sequoia , Simpson Nick , Yenew Mihret , Dix Randy , Slawson Chad 
Published: June 2021

Alzheimer’s disease (AD) affects more than 5.2 million people in the US. The disease is characterized by the presence of “Tau Tangles”, which is the aggregation of the protein Tau. The Tau protein is expressed in the nervous system and helps stabilize microtubules, which help establish cell shape and serve as tracks for neuronal transport. The Tau protein is regulated through phosphorylation and in patients with AD, the Tau protein is hyperphosphorylated, which can lead to the formation of “Tau Tangles”. These tangles are made up of free floating Tau and can interfere in the neuron communication because of a structural change in the protein due to the phosphorylation. In a normal mature neuron, tubulin is present in over tenfold excess of the Tau protein, and almost all Tau protein is bound to microtubules. In AD, the ratio between free floating Tau and microtubule bound Tau increases. In neurons affected by AD, hyperphosphorylated and glycosylated Tau disengages from microtubules and disrupts microtubule organization. Tau kinases phosphorylate Tau; however the regulation of Tau phosphorylation is still unknown. The Olathe North SMART Team (Students Modeling A Research Topic) modeled the Tau kinase using 3D printing technology.Grant Funding Source: Supported by grants from the NIH‐SEPA and NIH‐CTSA.
Getting “Rit‐R” Iron in the Cell: The Role of RitR in Reducing Iron Transport into Streptococcus pneumoniae. (LB111)
Authors: Allbee B. , Anderson N. , Blank D. , Bolgert J. , Bothe A. , Britt N. , Geiser E. , Hammer T. , Janecek E. , Kalmer I. , Lawniczak J. , Lesiecki M. , Levy J. , Marshall M. , Meaux N. , Ruzicka M. , Temmer J. , Tiffany K. , Butt A. , Dyke S. , Vandenberg B. , Wankowski J. , Zeitlow E. , Tiffany Karen , Silvaggi Nicholas 
Published: June 2021

According to the World Health Organization, pneumonia is the leading cause of death in children worldwide, and infection of lung tissue by Streptococcus pneumonia causes the bulk of bacterial pneumonia cases in children. The atypical response regulator, RitR (Repressor of iron transport Regulator), helps S. pneumoniae survive in the hostile oxidizing conditions in the lungs. RitR has two domains, a DNA‐binding domain (DBD) and an aspartate‐less receiver domain (REC). In its “inactive” form, these domains are docked (i.e., close together) and the DBD is unable to bind DNA. In its “active” form, the domains are undocked; the DBD is freed from the REC. The “active” form dimerizes and can bind to DNA to turn off the iron transport genes. To study the changes that occur when RitR is activated, the Cedarburg High School SMART (Students Modeling A Research Topic) Team used 3D printing technology to model inactive RitR and a hypothetical active RitR dimer. RitR helps the bacterium survive in the oxygen‐rich environment in lungs by stopping iron transport into the bacterial cell. If iron is transported into the cell, oxygen forms reactive oxygen species that damage and kill cells. S. pneumonia cells without RitR are unable to infect lung tissue, so RitR is a potential target for drug design.Grant Funding Source: Supported by a grant from NIH‐CTSA.
Somatic mutant alleles of POLE found in human cancers suppress proofreading and replication fidelity in vitro (LB121)
Authors: Henninger Erin , LeCompte Kim , McBride Christine , Bunnell Bruce , Pursell Zachary 
Published: June 2021

While it is now appreciated that tumor cells contain thousands of mutations, identifying their source(s) has remained problematic. Whole genome sequencing from colorectal and endometrial cancers recently identified mutations in DNA polymerase (Pol) epsilon, one of three main eukaryotic nuclear replication Pols. These mutations largely reside in the exonuclease domain, are associated with a hypermutated phenotype, and are predicted to promote tumorigenesis. However, the effects of these somatic mutations on enzyme function have not yet been studied. To address this, we introduced several of these mutations in recombinant human Pol ε and measured their effects on replication fidelity in vitro. While the mutants retained wild type DNA synthesis activity, 3'‐5' exonuclease proofreading was suppressed. Surprisingly, this suppression was variable, ranging from ~2‐fold to almost complete ablation, and resulted in increases in frameshifts and base pair substitutions in an in vitro forward mutation assay. Mutants also had increased ability to bypass abasic sites. Exonuclease inactivation of one POLE allele in human cells resulted in decreased sensitivity to H2O2 treatment. These results suggest that Pol ε mutant alleles may promote viability and mutagenesis, and possibly tumorigenesis, in part through bypass of oxidative stress‐induced DNA lesions.Grant Funding Source: NIH RR020152
The Effect of Artesunate on Myeloperoxidase Activity of Human Polymorphonuclear Neutrophil. (LB134)
Authors: MINARI JOSEPH 
Published: June 2021

Myeloperoxidase is the most abundant enzyme found in the polymorphonuclear neutrophil and is known to play a central role in the host defense system of the leukocyte. The enzyme has been reported to interact with some drugs to generate free radical which inhibits its activity. This study investigated the effects of artesunate on the activity of the enzyme and the subsequent effect on the host immune system. In investigating the effects of the drugs on myeloperoxidase, the influence of concentration, pH, partition ratio estimation and kinetics of inhibition were studied.This study showed that artesunate is concentration‐dependent inhibitor of myeloperoxidase with an IC50 of 0.078mM. Partition ratio estimation showed that 60 enzymatic turnover cycles are required for complete inhibition of myeloperoxidase in the presence of artesunate. The influence of pH on the effect of artesunate on the enzyme showed least activity of myeloperoxidase at physiological pH. The kinetic inhibition studies showed that artesunate caused a competitive inhibition with an increase in the Km value from 0.12mM to 0.26mM and no effect on the Vmax value. The Ki value was estimated to be 2.5mM. The results obtained from this study show that artesunate is a potent inhibitor of myeloperoxidase and it is capable of inactivating the enzyme.It is considered that the inhibition of myeloperoxidase in the presence of artesunate as revealed in this study may partly explain the impairment of polymorphonuclear neutrophil and consequent reduction of the strength of the host defense system against secondary infections.
Case Report: Contribution of the Radial and Ulnar Arteries on the Blood Supply of the Thumb (LB22)
Authors: Valentin Natalia , Del Rosario Cristina , Veras Wilson 
Published: June 2021

Understanding the vascular patterns of the hand and its variations is important for surgeons, radiologists and anatomists due to the constant innovations on microsurgical techniques for the traumatized hand. Even though the thumb contributes 40 percent of the overall function of the hand, its irrigation is sometimes neglected. The principal irrigation of the thumb is normally provided by the princeps pollicis artery. It has been described that this artery usually arises from the radial artery or in other occasions can arise directly from the deep palmar arch, or from a common trunk with the radilis indicis artery. However, anatomical variations to this description have been found in previous studies. In order to determine the origin of the princeps pollicis artery, 12 hands were dissected out of 6 adults formalin preserved cadavers from the Gross Anatomy Laboratory of the Universidad Central del Caribe during the fall semester of 2012‐2013. In the present study we have found 58.3% of the specimens with the princeps pollicis artery arising from the first dorsal metacarpal artery. In 85.7% of these cases there is a persistent anastomosis between the princeps pollicis and the superficial palmar arch. Additionally, the irrigation of the thumb was found as normally described in 25% of the specimens. The remaining 16.6% of the specimens were discarded. These results allow us to suggest further investigation in this topic to provide significant statistics in terms of the incidence of this anatomical variation in the population.Grant Funding Source: Supported by: This publication was possible by grants from the National Center for Research Resources (5 G12 RR 003035‐27) and the National Institute on Minority Health and Health Disparities (8G12 MD 007583‐27) from the National Institutes of Health
Effects of two popular weight loss programs on changes in body composition and visceral adipose tissue in women (LB296)
Authors: Coletta A , Mardock M , Lockard B , Byrd M , Simbo S , Jagim A , Kresta J , Baetge C , Jung Y , Koozehchian M , Khanna D , Kyul H , Oliver J , Greenwood M , Rasmussen C , Kreider R 
Published: June 2021

The purpose of this study was to determine if different types of exercise and diet interventions promote greater changes in visceral adipose tissue (VAT). 54 sedentary women (34±8 yrs, BMI 35±6) were randomized to participate in the Curves (C) or Weight Watchers (W) weight loss programs for 16‐wks. Participants in the C program followed a 1,200 kcal/d diet for 1‐wk and 1,500 kcal/d diet for 3 wks (30:45 CHO:PRO). Subjects then ingested 2,000 kcals/d for 2‐wks (45:30 CHO:PRO) and repeated this diet while participating in the Curves with Zumba program 3‐d‐wk. Remaining subjects followed the W point‐based diet program, received weekly counseling, and were encouraged to exercise. DEXA body composition with VAT was determined at 0 &amp; 16 wks. MANOVA revealed a significant time (p<0.001) and group x time (p=0.035) effect. Both groups lost a similar amount of weight (W ‐6.3±4, C ‐4.8±4 kg, p=0.17), fat mass (W ‐3.1±6, C ‐5.9±6 kg, p=0.13), and VAT mass (W ‐98±468, C ‐240±445 g, p=0.26). Subjects in the C group experienced greater gains in FFM (W ‐3.3±5.9. C +1.2±4.1 kg, p=0.002) and tended to lose more body fat (W ‐1.1±8, C ‐4.5±5.5 %, p=0.07). Changes in VAT mass significantly correlated with changes in weight (r=0.38), fat mass (r=0.73), FFM (r=‐0.62), and body fat (r=0.64). Results indicate that different types of diets can differentially affect changes in body composition and VAT.Grant Funding Source: Curves International, Inc. (Waco, TX)