bluuebunny/crossref_metadata_2025_split huggingface dataset:
CTNNA1 is Expressed in U937 Cells and is Regulated by miR9
Authors: Patel Nikhil , Rabquer Bradley James 
Published: June 2021

Rheumatoid arthritis (RA), a chronic, auto‐immune, inflammatory disorder characterized by progressive joint destruction, affects 0.3–1% of people worldwide and is a major cause of disabilities. Monocyte (MN) trafficking into inflamed joints is critical to the pathogenesis of RA. Micro RNAs (miRNAs) are approximately 22‐nucleotide long single‐stranded, noncoding RNA which bind to mRNA, thus making them act as gene expression regulators. miRNAs bind to complementary sites on the 3′‐end of target mRNAs and either block their translation or enhance their degradation, resulting in downregulation of certain genes. Previous studies in our lab have identified a gene of interest called CTNNA1, as prior research has suggested miR9 to be a suitable binding match. CTNNA1 plays a role in several pathological processes, including muscle contraction and tumor metastasis. It's hypothesized that miR9 binds to CTNNA1 and that miR9 will mediate its degradation.Methods and ResultsU937 monocytes were cultured, mRNA was isolated, and cDNA was prepared. During this, the DNA sequence of CTNNA1 in humans was analyzed so primers could be designed to bind to the predicted 3′ UTR binding sites of miR9. PCR was then performed to yield the isolated, predicted binding site, and then base pair size was confirmed via electrophoresis gels. The gene product was then cut with restriction enzymes Sac I and Sal I, ligated into the pmirGLO vector, and was transformed into E. coli cells and plated for incubation. Selected E. coli colonies expressing the gene were grown and incubated in overnight cultures.ConclusionCTNNA1 is expressed by U937 cells and it's 3′ UTR has been cloned into pmirGLO for mir9 binding assessment.Support or Funding InformationAlbion College Biology Department and the Foundation for Undergraduate Research, Scholarly, and Creative ActivityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The IMPC: A Global Scientific Infrastructure for Understanding the Role of Genes in Complex Traits
Authors: Muñoz‐Fuentes Violeta , Meehan Terrence F. , Lloyd KC Kent , Mallon Ann‐Marie , Smedley Damian , Parkinson Helen 
Published: June 2021

The International Mouse Phenotyping Consortium (IMPC) is building a functional catalogue of the mammalian genome by producing and phenotyping a knockout mouse strain for every protein‐coding gene. To date, over 4,000 knockout mouse lines, many for poorly understood genes, have been characterized and made available to the research community in a coordinated effort involving more than a dozen global research centers and dedicated publicly‐available online resources. Using a standardized adult phenotyping pipeline, centers measure each mouse for more than 250 phenotypic parameters including metabolite content in blood, hearing capacity, and skeletal malformations. In addition, over 1,000 embryonic lethal mouse lines are analyzed in a specialized embryonic development pipeline that uses high‐resolution 3D imaging. All data is quality controlled and analyzed by a dedicated informatics consortium and all abnormal phenotypes automatically compared to clinical features of human disease populations to identify robust mouse models of disease and inform efforts in precision medicine. We will present our discoveries into the wide‐ranging sexual dimorphism of phenotypic traits, the latest findings in metabolism, hearing and aging and provide examples of how researchers are using IMPC resources to identify causative alleles within GWAS loci. The plethora of new genetic disease models as well as the basic and translational knowledge that has arisen from our analysis has recently earned the IMPC recognition by the G7 for being the first global research infrastructure for the life sciences.Support or Funding InformationNIH Common Fund:U54H G006370, U54H G006364This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of Coding and Noncoding Variants for Human CKD Using Novel Strategies
Authors: Prokop Jeremy W. , Yeo Nan C. , Ottmann Christian , Link Brian A. , Mendenhall Eric M. , Freedman Barry I. , Lazar Jozef , Jacob Howard J. 
Published: June 2021

Interpreting the mechanism of genetic variants is one of the greatest challenges impeding the analysis of the rapidly increasing volume of patients' genomic data, particularly for rare variants that do not reach GWAS significance due to low population frequencies. There are 153 GWAS hits for altered kidney function due to commonly inherited genetic variants, yet minimal knowledge of the mechanism of these variations is known at the current time. Utilizing a computational strategy to filter 3,921 genetic variants that are found within linkage disequilibrium of the GWAS hits we have nominated 4 damaging protein coding variants and more importantly 30 noncoding variants predicted to alter transcription factor based gene regulation. One of these noncoding variants, rs17319721 associated with risk of chronic kidney disease (CKD), is found near the SHROOM3 gene and is thought to play a functional role in podocytes of the kidney. In this study we show this variant falls upstream of a novel transcriptional start site that results in a shorter SHROOM3 isoform lacking the PDZ domain. This variant disrupts binding of TCF7L2 in podocytes and the single variant (rs17319721) alters transcription levels of SHROOM3 as determined by variant mutation into the genome using CRISPR/Cas9 technology.To begin deciphering the role of rare variants within human genomes we performed a full assessment of the ExAC database for >60,000 sequenced human exomes, revealing several coding variants in SHROOM3 that are likely to alter function, including a coding variant (P1244L) we show to have association to CKD in humans. We determined that P1244L attenuates the interaction of SHROOM3 with 14‐3‐3 binding and alters signaling through the Hippo pathway, a known mediator of CKD. Animal studies show a loss of function in zebrafish kidney due to the variant, resulting in increased glomerular permeability. These data demonstrate multiple new SHROOM3‐dependent genetic and molecular mechanisms that regulate CKD and demonstrate a novel analytical pipeline with broad application to systematically characterizing risk mechanisms of CKD and other complex diseases.Support or Funding InformationSupport for this study was from NIH‐K01ES025435 (to JWP), NIH‐R01HL069321 (to HJJ), NIH‐R01EY014167 (to BL), the Collaborative Research Centre 1093 through the Deutsche Forschungsgemeinschaft (DFG, to CO), and HudsonAlpha Institute for Biotechnology.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In the absence of weight gain, survival rates of DF508‐CF female mice are increased by genistein diet.
Authors: Al‐Nakkash Layla , Fairbourn Nathan , Mylavarapu Charisma , Dbeis Ammer , Lord Ryan , Chandrashekar Archana , Bowman Taylor , Banayat Tatum , Hodges Craig 
Published: June 2021

Mice homozygous for the most common disease‐associated mutation, DF508, are characterized with severe intestinal disease and require constant laxative treatment for survival. This pathology mimics the intestinal obstruction (meconium ileus) seen in some CF patients. Genistein is a naturally occurring isoflavone found in soy. Patch clamp studies have demonstrated that genistein increases the open probability of DF508 CFTR to wild‐type (WT) levels. We evaluated whether dietary genistein alone would eliminate the need for laxatives for survival of the DF508 CF mouse, thereby improving mortality rates. At age 21 days, we maintained male and female DF508 mice on three diet regimens for 45 days; normal diet, normal diet + Colyte or 600 mg genistein/kg diet, 600G. Survival rates and body weight data were determined. At the completion of the diet study, tissues were extracted, immediately frozen at −80ºC until use and evaluated later for protein expression and histological evaluation. Survival rates for each diet group were as follows; males fed normal diet = 38% (8/21 mice), males fed normal diet + Colyte = 83% (35/42 mice), males fed 600G = 60% (9/15 mice) and females fed normal diet = 47% (9/19 mice), females fed normal diet + Colyte 71% (27/38 mice), females fed 600G = 87% (13/15 mice). Body weight at the end of the diet study was greater in males fed 600G (21.96±0.68 g, n=14) versus those males on Colyte (18.86±0.64 g, n=8). There was no change in overall weight gain in the female groups. Jejunum villi length, crypt depth and wall thickness was comparable between diet groups. Total protein expression of jejunum Na+/K+‐ATPase and NKCC1 was comparable between groups. Total expression of jejunum GLUT2 and GLUT5 were unchanged, however SGLT1 expression was significantly increased 2‐fold in females fed 600G. We are currently exploring whether liver protein expression and serum metabolic markers are modified to better elucidate the mechanism of action of 600G to improve weight in males and to improve survival rate in females. We conclude that feeding DF508 female mice 600G genistein‐diet abolished the dependence upon laxatives for survival. These studies could have implications in reducing the sex‐dependent differences in the age of survival of CF female patients.Support or Funding InformationSupported by Soy Health Research Program and MWU intramural funds (LA).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
High Fat Diet Impacts Gut Microbiome Differently in Male and Female Sprague Dawley Rats
Authors: Gonzalez Agnes P. , Rodriguez Christine D. , Brofen‐Quiñones Samuel , Sambolin‐Escobales Lubriel , Cruz Myrella , Appleyard Caroline B. , Chompre Gladys 
Published: June 2021

Obesity has increased during the last couple of decades and is strongly related to cardiovascular disease, hyperlipidemia, diabetes, and neurological disorders, such as depression, which tend to be more prevalent in women. A high fat diet (HFD) is associated with negative effects on the gut microbiome, including increased Firmicutes/Bacteriodetes ratio (F/B ratio) which may affect the integrity of the intestinal barrier. Gut microbiome transplants from male to female rats have been shown to decrease diabetes type 1 in females, suggesting sex differences in the gut microbiome. Previous studies in our lab have shown that Sprague Dawley male rats fed with a HFD showed a depressive‐like behavior and also a decrease of tight junction proteins in the gut suggesting gut permeability. However, it is not clear whether the effects of a HFD on the microbiome are based on gender differences.AIMTo ascertain whether the administration of a HFD will affect differently the gut microbiome in males and females.METHODSThere were four groups of Sprague Dawley rats (n=12/group): Female Low‐Fat Diet (FLFD), Female High Fat Diet (FHFD), Male Low‐Fat Diet (MLFD), Male High Fat Diet (MHFD). Female and male postnatal rats were fed with a LFD (10% fat) for 6 weeks. Then, half of each group were transferred to a HFD (60% fat) for 8 weeks. We measured absolute weight daily, and rats were tested for depressive like syndrome in a porsolt swim test one day before sacrifice. Fat and feces were collected from all animals to assess differences in the gut microbiome.RESULTSRats showed no differences in absolute weight nor in percent weight gain in either group, however, MHFD showed an increase of epidydimal fat tissue compared to MLFD. FHFD group showed a significant increase in immobility compared with the FLFD group suggesting depressive like syndrome (p<0.05). Measurement of depressive like syndrome in males was limited by their size and did not reach significance. In gut microbiome, MHFD showed a significant increase in the F/B ratio compared with the MLFD, however, in females there were no significant differences. Females on a high‐fat diet had a significant increase in Proteobacteria compared to FLFD (p<0.05). In contrast MHFD showed a significant increase in Verrucomicrobia compared with the MLFD (p<0.05).CONCLUSIONSA HFD affects the gut microbiome of males and females differently. The significance of this study is that sex differences may be a potential factor in how a HFD might contribute towards various pathophysiological conditions via differing effects on the gut microbiome.Support or Funding InformationSupported by NIGMS/INBRE P20 GM103475‐14 and RCMI G12 MD007579This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metabolic plasticity through pregnancy with normal pregestational BMI
Authors: Delgadillo‐Velazquez Jaime , Patiño‐Gonzalez Nayeli , Nambo‐Venegas Rafael , Millán‐Cortés Alexis , Vadillo‐Ortega Felipe , Palacios‐Gonzalez Berenice 
Published: June 2021

During normal pregnancy, metabolic changes occur causing weight gain and insulin resistance to provide the fetus with substrates for growth and development. Hence, a metabolic plasticity must exist to allow the mother to adapt correctly upon the pregnancy metabolic overload, preventing adverse outcomes like gestational diabetes. The aim of the study was to analyze the metabolic plasticity during pregnancy in women with normal pregestational BMI. Glucose, triglycerides and total cholesterol were determined, and indirect calorimetry was performed during fasting and after the consumption of a high‐carbohydrate meal. Fasting triglycerides and total cholesterol were significantly higher during the third trimester. Postprandial glucose concentrations were significantly lower at second trimester and higher in third trimester compared to first trimester. There was a significant increase in fasting respiratory quotient (RQ) during the last trimester compared to the first and second trimester. Regarding ΔRQ, there was a significant decrease during the last trimester compared to the first trimester. In conclusion, at the end of pregnancy there is a lower metabolic flexibility, associated with the presence of physiological insulin resistance.Support or Funding InformationThis study was funded by CONACYT grant FOSISS 2016 # 273110This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Partial Sleep Deprivation on Lipid Profile
Authors: Covassin Naima , Bukartyk Jan , Cundrle Ivan , Singh Prachi , Calvin Andrew D. , St Louis Erik K. , Somers Virend K. 
Published: June 2021

BackgroundIncreasing evidence suggests that insufficient sleep is associated with increased morbidity and mortality. Studies linking short sleep with metabolic derangements such as obesity, insulin resistance and dyslipidemia, are especially persuasive. Nevertheless, the vast majority of these data have been obtained from observational studies. We sought to evaluate the effects of 9 days of partial sleep deprivation compared to normal sleep on lipid profile in healthy subjects undergoing an in‐laboratory, experimental protocol.MethodsEight healthy, nonobese, normolipidemic subjects (4 males, age 25.4±6.8 years, body mass index 25.2±3.9 kg/m2) completed a randomized, controlled, crossover, 16‐day in‐patient study. Each sequence consisted of a 4‐day acclimation period, followed by 9 days of experimental partial sleep deprivation (4 hours of sleep opportunity; 00:30 AM – 4:30 AM); or normal sleep (9 hours of sleep opportunity; 10:00 PM – 7:00 AM), and a subsequent 3‐day recovery segment. Subjects consumed a controlled, weight maintenance diet (15% protein, 55% carbohydrates, and 30% fat) during the study. Fasting blood samples for measures of total cholesterol, high‐density lipoprotein (HDL) cholesterol, low‐density lipoprotein (LDL) cholesterol, and triglycerides were obtained in both study sequences on Days 4, 7, 10, 13, and 16.ResultsCompared to normal sleep, experimental partial sleep deprivation did not cause significant changes in total cholesterol (partial sleep deprivation vs normal sleep: 160.1±21.8 mg/dL vs 164.9±23.7 mg/dL, P=0.724), HDL cholesterol (54.1±14.7 mg/dL vs 52±12.8 mg/dL, P=0.75), LDL cholesterol (86.5±18.9 mg/dL vs 89.5±19.7 mg/dL, P=0.329), or triglycerides (97.7±26.6 mg/dL vs 96.6±38.3 mg/dL, P=0.633). Analysis of time course of lipid values across study days did not reveal appreciable patterns (all P>0.295). Likewise, body weight was similar during both study sequences (partial sleep deprivation vs normal sleep: 69.6±14.2 kg vs 69.2±13.5 kg, P=0.459).ConclusionsExperimental sleep restriction does not alter circulating lipids or weight in healthy subjects. Aberrant metabolic effects of insufficient sleep may be mediated by dietary intake and occur when access to food is unrestricted, or may emerge only after longer exposure to sleep curtailment.Support or Funding InformationAmerican Heart Association grant 16SDG27250156; National Institutes of Health R01 grants HL114676 and HL114024; European Union project FNUSA‐ICRC no. CZ.1.05/1.1.00/02.0123.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sagittal Craniosynostosis and Related Cranial Anomalies: Genetic Insights
Authors: Olatunde Josh , Frolov Andrey , Tersigni‐Tarrant MariaTeresa , Martin John 
Published: June 2021

BackgroundSagittal craniosynostosis (scaphocephaly) is a pathologic condition that results in premature fusion of the sagittal suture restricting the transverse growth of the skull leading to elevated intracranial pressure and consequent neurodevelopmental delay. It can occur most commonly as an isolated event resulting in nonsyndromic craniosynostosis, or it can occur less commonly in conjunction with other anomalies resulting in syndromic craniosynostosis. Although in past years close attention was paid by researchers regarding the mechanisms that govern development of craniosynostosis, much more remains to be learned about this cranial anomaly, in particular, the genetic mechanisms associated with scaphocephaly. The clearer understanding of the latter not only could shed more light on the respective mechanisms but could also help to improve and predict better outcomes of the respective corrective clinical procedures.ResultsThe present study reports a unique case of a male cadaver 56 years of age with sagittal craniosynostosis with a 0.56 cranial vault index as measured from respective radiographic images. Upon closer examination of the individual's head it was concluded that he underwent, most likely early in his life, a sagittal strip synostectomy with replacement. The additional maxillofacial anomalies included edentulous, mandibular prognathism with class 3 malocclusion and 15 mm of negative overjet, and exostosis of the hard palate. The latter was preliminary characterized as torus palatinus. One of the most interesting features of the present case was the abnormal appearance of the replacement bone strip and its vertical displacement that were accompanied by a presence of abnormal dura and venous sinuses with no brain indentations on the dural surface of the strip. Histologic analyses of cranial soft and hard tissues should help to clarify a nature of the respective pathologies. In order to better understand molecular mechanism(s) responsible for the development of sagittal craniosynostosis with the particular maxillofacial anomalies, the gene coding region (exome) of DNA extracted from the cadaveric tissue was screened for putative mutations employing Illumina Next Generation Sequencing Platform. This will help to unveil new genetic variations associated with the aberrant skull development in humans and optimize clinical approaches for its correction.ConclusionThe current case most likely represents syndromic sagittal craniosynostosis where surgical intervention was mainly unsuccessful resulting in abnormal development of the replacement bone strip and intracranial tissue.Support or Funding InformationThis study was supported by the Center for Anatomical Science and Education, Department of Surgery, Saint Louis University School of Medicine.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
New in vivo microRNA biotechnology reveals specific roles for the miR‐200 family in craniofacial development
Authors: Sweat Mason E. , Yu Wenjie , Eliason Steven , Sweat Yan Yan , Cao Huojun , Hong Liu , Amendt Brad A. 
Published: June 2021

The molecular regulators of stem cell differentiation are a hot topic and the proteins involved in this process have been well studied, but the function of microRNA (miR) remains elusive.ObjectivesTo investigate the role of the miR‐200 family in the regulation of craniofacial development and dental epithelial stem cells (DESCs). Previously, we found that the miR‐200 family was highly expressed in the differentiated cell types of the lower incisor but was absent from the stem cell niche, suggesting it may contribute towards differentiation.MethodsWe constructed a plasmid‐based miR inhibitor system (PMIS) and used it to inhibit the miR‐200 family in mice. One inhibitor line specifically targets the seed sequence specific for miR‐200a, and ‐141, while the other is specific for the seed sequence found in miR‐200b,‐200c and ‐429, and crossing the lines allows the functional inhibition of the entire miR‐200 family.ResultsDouble inhibitor mice are neonatal lethal and have poorly differentiated lower incisors. The teeth are smaller and they lack cells expressing cell differentiation markers (DSP and amelogenin) of mature odontoblasts and ameloblasts. Several transcription factor regulators of stemness which are direct targets of the miR‐200 family, including Sox2, are ectopically expressed in normally differentiated cells. A proliferation cell labeling BrdU assay demonstrated that the majority of dental epithelial cells are proliferating in double inhibitor mouse incisors.ConclusionsThis study is among the first to use targeted inhibition to study specific miRs and we found that the miR‐200 family is required for the appropriate differentiation of DESCs and the family functions by reducing the expression of transcription factors promoting stemness outside of the stem cell niche compartment of the lower incisor. Overall, craniofacial development is affected due to a lack of cell differentiation.Support or Funding InformationUniversity of Iowa College of DentistryThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.