bluuebunny/crossref_metadata_2025_split huggingface dataset:
Diet Significantly Influences the Interleukin‐6 Response to Mouse Models of Peritoneal Sepsis
Authors: Klug Mikahla A. , Patrick‐Eisenberg Austin , Thomas Blake , Rose Christian , Houghton Jeffrey , Gigliotti Joseph C. 
Published: May 2022

IntroductionSepsis remains to be a tremendous health burden in the United States, affecting 1.7 million Americans and causing over 250,000 deaths each year. Previous work has shown high‐calorie diets to significantly alter the immune response and outcomes in animal models of sepsis; however, the diets used model overt metabolic dysfunction and not dietary patterns observed in a “typical” American. We developed a novel Americanized diet (AD) that incorporates several of the nutritional inadequacies reported in Americans and set out to determine how consumption of the AD influences immunological outcomes of peritoneal sepsis in mice.MethodologyMale C57Bl/6 mice were used in accordance to protocols approved by the Institutional Animal Care and Use Committee at LUCOM and compliant with the Guide for the Care and Use of Laboratory Animals. Mice received ad libitum access to experimental diets [chow, AD, or a commercially available high‐calorie Western diet (WD)] for 8‐weeks and peritonitis was induced with cecal‐ligation and puncture (CLP) or intraperitoneal injection of a fecal slurry (fecal peritonitis, “FP”). For both CLP and FP, blood was collected 24‐hours later for quantification of circulating interleukin‐6 (IL6). Another group of mice were fed their assigned diets and the spleens cultured ex vivo in DMEM media with lipopolysaccharide (LPS, 10ug/mL) or saline for 3 hours. Tissue and supernatant were collected and frozen until assayed for IL6 and inflammatory gene expression using a commercially available PCR‐microarray. All data were analyzed using general linear model procedures in SPSS with Tukey’s post hoc test and group averages were considered statistically significant with a P‐value less than 0.05.ResultsMice consuming AD and WD had ~3‐fold greater circulating IL6 following CLP (P=0.04) and FP (P<0.05) as compared to mice consuming chow. LPS stimulation of splenic tissue ex vivo caused a significant increase in secreted IL6 protein (P<0.001). Mice fed WD had the greatest IL6 concentration (425 pg/mL), which was greater than mice fed chow (163 pg/mL, P=0.04) but not mice fed AD (360 pg/mL, P=0.9). PCR‐array analysis confirmed the upregulation of IL6, with mice fed WD having double (P=0.03) the mRNA expression of splenic IL6 after LPS as compared to mice fed AD and chow. CCL12 expression was similarly upregulated in mice fed WD as compared to mice fed AD and chow (P=0.005).ConclusionOur data support the hypothesis that diet significantly influences the severity of animal models of sepsis. Consumption of a nutritionally inadequate AD caused a similar increase in circulating IL6 as compared to mice consuming a high calorie WD. However, the source of the additional circulating IL6 is unknown, ex vivo spleen stimulation studies with LPS did not report an increase in IL6 production in mice fed the AD. Future studies will determine if and how diet influences IL6 production in other tissues.
Adipose‐derived human soluble (pro)renin receptor causes resistance to Losartan treatment in high‐fat diet male and female mice
Authors: Nichols Kellea , Arthur Gertrude , Hinds Terry , Yiannikouris Frederique 
Published: May 2022

Obesity, affecting more that 37% of the US, contributes to hypertension. Despite the use of one or more anti‐hypertensive treatments, 48% of the hypertensive population remains with resistant hypertension, which prompts the development for new therapeutic targets. We demonstrated that obesity increased the expression of prorenin receptor (PRR) in the adipose tissue and elevated plasma soluble PRR (sPRR). In addition, the infusion of mouse sPRR increased blood pressure in male mice fed high fat‐diet (HF); indicating that adipose‐derived sPRR could increase circulating sPRR and contribute to hypertension. However, there is a critical gap in the functional role of human sPRR in obesity‐hypertension. In this study, we aim to define whether adipose‐derived human sPRR contributes to obesity‐hypertension.Human sPRR‐Myc‐tag transgenic mice were bred with mice expressing adiponectin/Cre to selectively express human sPRR in adipocytes (adi‐HsPRR). Adi‐HsPRR and control littermate (CTL) male and female mice were fed HF‐diet for 20 weeks (N=8‐15/group). Body weight was assessed weekly and body composition monthly. Blood pressure was measured by telemetry after 15 weeks of diet.Adipose‐derived human sPRR did not significantly elevate body weight or fat mass (Male: CTL. 18.3±1.0g; adi‐HsPRR. 17.5±0.8g. Female: CTL. 15.6±1.5g; adi‐HsPRR. 11.9±1.3g; p>0.05). Systolic blood pressure (SBP) significantly increased in HF‐fed male and female mice. However, adipose‐derived human sPRR did not further elevate SBP (24h SBP. Male: CTL. 136.0±1.7 mmHg; adi‐HsPRR: 133.4±1.5 mmHg; Female: CTL. 131.9±2.8 mmHg; adi‐HsPRR: 130.6±3.1 mmHg; p>0.05). Surprisingly, the anti‐hypertensive effect of losartan (Los) to lower blood pressure was significantly reduced in adi‐HsPRR male and female mice (Male: CTL. ΔSBP: ‐12.1±1.5 ΔmmHg; adi‐HsPRR: ‐7.8±0.6 ΔmmHg; Female: CTL. ΔSBP: ‐13.4±1.1 ΔmmHg; adi‐HsPRR: ‐5.7±2.3 ΔmmHg; p<0.05). In 3T3‐L1 cells, sPRR significantly increased phosphorylation of ERK1/2, which was not completely blunted by Los indicating that human sPRR could act as a partial agonist of AT1R or activate ERK1/2 independently of AT1R. Our data suggests that adipose‐derived sPRR does not stimulate AT1R‐mediated contractility, instead impairs Los efficacy.
Metabolic Syndrome and SIRT1 Mutation Impair Ca2+Channels in Coronary Smooth Muscle Cells of Ossabaw Miniature Swine
Authors: Thamba Aish , Reed John , Strobel John , Byrd James , Alloosh Mouhamad , Coutts Alex , Sturek Michael 
Published: May 2022

BackgroundNormal metabolic health and sirtuin1 (SIRT1) by its deacetylase activity maintain voltage‐gated Ca2+ channel (VGCC) function that promotes gene expression to maintain contractile coronary smooth muscle (CSM) phenotype. MetS impairs VGCC function and sarcoplasmic reticulum (SR) Ca2+ release, contributing to dedifferentiation of CSM to proliferative and osteogenic phenotypes.HypothesisMetS and impaired SIRT1 will impair VGCC and SR Ca2+ release channels in CSM.MethodsUsing CRISPR/Cas9 methodology a point mutation (SIRT1L100P) was made in Ossabaw miniature swine to mimic the naturally occurring mutation in humans and decrease SIRT1 activity, thereby resulting in hyperacetylation (Ac) of Ca2+transporters and impaired function. Four groups of pigs were used to analyze genotype and diet interactions: wild type lean, SIRT1 lean, wild type MetS, SIRT1 MetS. Pigs were age 4 months at the start and fed normal chow (lean) or atherogenic diet (MetS) for 7 months. CSM cells were enzymatically dispersed and Ca2+ measured with fura‐2. Depolarization with 80 mM K+ assessed Ca2+ influx through VGCC and the peak Ca2+ response to 5 mM caffeine to open SR Ca2+ release channels assessed the caffeine‐sensitive SR Ca2+ store.ResultsTwo‐way ANOVA showed SIRT1 mutation (p=0.02) and MetS diet (p<0.0001) significantly decreased VGCC function independently, but not additively or synergistically. Interaction of SIRT1 genotype and MetS diet was significant (p<0.0001). No significant effect of genotype or diet was observed on SR calcium store release.ConclusionSIRT1L100P mutation is likely to contribute to coronary atherosclerosis and SIRT1 activators may be effective therapies.
Novel Functional FOXC1 Variants in Familial and Sporadic Atrial Septal Defect
Authors: Chen Huan‐Xin , Hou Hai‐Tao , Wang Xiu‐Li , Yang Qin , He Guo‐Wei 
Published: May 2022

ObjectiveAtrial septal defect (ASD) is one of the most common forms of congenital heart disease. Genetic defects play important roles in the pathogenesis of ASD but are complex and unclear. This study tries to discover the genetic pathogenesis in familial and sporadic ASD patients.MethodsIn Stage I, in a family with 30 members whole exome sequencing was performed in 4 ASD patients and the identified genetic variants were screened in the rest of the family members. In Stage II, 335 unrelated sporadic and isolated ASD patients were enrolled to test whether the genetic variants were involved in these patients. In Stage III, biological functions of the variants were elucidated at the cellular level and the CRISPR/Cas9 was used to construct variant‐specific mutant mice in order to observe the abnormality of the heart structure.ResultsA novel heterozygous missense variant, FOXC1 c.518G>A (p.R173H) was identified in the ASD family by whole exome sequencing. Subsequently, other 4 heterozygous variants in FOXC1, including 2 novel missense variants: c.556‐558delAAG (p.K186del) and c.559G>A (p.D187N) were identified in unrelated 335 sporadic ASD patients. In dual‐luciferase reporter assay, the transcriptional activity of R173H decreased to almost 30% of the wild‐type. In the Foxc1 R173H site‐specific mutant mice, the atrial septum became very thin in tissue slices of the heart that was similar to the echocardiographic finding in the above family members.ConclusionVariants in FOXC1 likely cause cardiac anomaly, particularly ASD. These findings provide a new insight into genetic mechanisms and counseling of familial and sporadic ASD.
The Cognitive Load of Learning Anatomy in a Virtual World
Authors: Touliopoulos Evangelea , Ives Rowan , Chan Sachi , Lohit Simran , Reis Isabella R. , Sinha Sakshi , Nehru Amit , DeYoung Veronica , Mitchell Josh P. , Brewer‐Deluce Danielle , Sonnadara Ranil , Wainman Bruce C. 
Published: May 2022

IntroductionCognitive load refers to the amount of working memory that is being used in a task, like memorizing the anatomical landmarks on distinct boney specimens. Critically, cognitive load may be compromised when the load imposed by the environment and the content to be learned together exceeds a student’s capacity. Previous research shows that stereoscopic materials delivered in virtual reality (VR) can be more mentally taxing compared to desktop (i.e., two dimensional) delivery but may be similar to that encountered in real life. There is no data on the cognitive load of autostereoscopic displays. Given the increased reliance upon digital media for teaching in learning in anatomy classrooms, it is prudent to better understand the cognitive load imposed on learners across a variety of modalities employed.MethodsCognitive load will be compared across three different learning modalities: immersive virtual reality (VR, displayed on the Oculus Quest 2TM), autostereoscopic (displayed on the AlioscopyTM screen), and an identical printed, physical model. During a four‐minute learning phase, undergraduate students, with no prior formal anatomy education, will learn 10 anatomical landmarks on a displayed bony model (calcaneus, zygomatic bone, or hemipelvis) in each of the three modalities. A Stroop test will be administered as a secondary task throughout the learning phase to evaluate cognitive load. Stroop test reaction time, and accuracy of the participants' responses to the Stroop test will be recorded. After the learning phase, an untimed, recognition‐based test will be administered wherein participants will be asked to recall the ten landmarks learned with the aid of a 3D‐printed bone identical to the one used in the learning phase. Performance will be evaluated based on landmarks correctly identified and the results will be correlated with cognitive load measured in each learning modality.ResultsWe hypothesize that the cognitive load will be highest for VR when compared to the cognitive load on the AlioscopyTM and physical model modalities which would manifest as lower reaction times and/or accuracy on the Stroop test. Further, we hypothesize that cognitive load will inversely correlate with the recognition test performance.ConclusionThe results of this study will allow educators and students to make informed decisions when deciding which learning modalities should be used for anatomy education or any other education that requires nominative learning on complex objects. Understanding which modalities minimize cognitive load and improve learning will help improve outcomes and allow for more efficient anatomical education.
Mathematical Modeling of Protracted HCMV Replication using Genome Substrates and Protein Temporal Profiles
Authors: Monti Christopher E. , Mokry Rebekah , Schumacher Megan , Dash Ranjan , Terhune Scott 
Published: May 2022

RationaleClinical manifestations of human cytomegalovirus (HCMV) infection, a major cause of morbidity and mortality in the immunocompromised, are associated with the lytic replication cycle. Lytic replication occurs over 96 hrs and is marked by viral DNA (vDNA) synthesis and expression of viral proteins, the latter of which can be generalized into distinct temporal classifications. Given the complexity of the lytic replication cycle, mathematical modeling can be used to probe this elaborate system. Recently, mathematical modeling has been used to study the early lytic replication cycle and the transition between lytic and latent cycles. To date, there appears to be few modeling studies focusing on the late phases of the lytic replication cycle.MethodvDNA replication kinetics were probed using qPCR assays with absolute quantification over varying multiplicities of infection (MOIs) in fibroblasts. An empiric model of vDNA replication was generated and parameterized using these data and then compared to qPCR data obtained post hoc. The vDNA model was used to drive a model of late viral infection that incorporated protein temporal class expression and culminated in virion production. This model was parameterized using data from immunoblots of representative proteins from the late temporal profile and viral titering data at three different MOIs. Predictions from this model were then compared to post hoc high‐temporal‐resolution, fluorescence‐based concentration measurements of one of the representative proteins over a 96‐hr replication cycle.ResultsA MOI‐dependent model of vDNA and late viral replication was developed from the experimental data. The model of late viral replication is mechanistic. It predicts feedback inhibition of late viral protein production and an increase in late viral protein degradation following an increase in vDNA. Both the vDNA and late replication models yielded 3 primary predictions: (1) presence of saturation kinetics at high MOIs, (2) inefficient replication at low MOIs, and (3) a range of MOIs in which vDNA replication and virus production are maximized. Saturation kinetics at high MOIs likely represents physical limitations of cellular machinery. Inefficient replication at low MOIs likely indicates a minimum MOI required to sustain infection. Identification of a maximal range of MOIs likely represents a set of conditions most favorable to HCMV lytic replication where extraneous viral material is minimized, while both the number of cells and the amount of viral material delivered to each cell is maximized.ConclusionA mathematical model of HCMV vDNA synthesis and late protein expression was generated and predicts a range of MOIs where maximal replication occurs, which can be extended to predict protein dynamics during infection.
Tau Mediated Coupling Interactions between Pol III Core DNA Synthesis and DnaB Helicase Unwinding
Authors: Welikala Malisha U. , Perera Himasha , Behrmann Megan , Trakselis Michael 
Published: May 2022

DNA replication is a vital process ensuring the passing down of intact genetic information over generations. DNA replication in the model organism, Escherichia coli (E. coli), has been extensively studied, providing significant insights into the various enzyme mechanisms. However, while the prime functions of individual components of the replisome have been deciphered, the more complex interactions between the components is an active area of study to understand replisome coupling. At the core of the replisome, the DnaB helicase unwinds the double strand DNA which is followed by rapid synthesis of nascent strand by Polymerase III core (Pol III core). It is proposed that the Tau subunit of the clamp loader complex acts as the physical link between both DnaB and Pol III core, leading to a coupling phenomenon between unwinding and synthesis.Using a reconstituted E. coli replisome in vitro, unwinding of DNA substrates with variable lengths and assemblies can be monitored through electrophoresis to better understand the kinetics of coupling. Mutations have been introduced in the exterior surface of DnaB, disrupting its charged interactions with the excluded strand, and promoting more constricted conformation that result in faster unwinding. The unwinding kinetics of wild‐type versus mutant DnaB enzymes on different length substrates are being examined. Further experiments include Pol III synthesis alone or with proposed coupling from the tau‐subunit and can simultaneously examine unwinding and synthesis. Mutations that affect DnaB conformation or disrupt Pol III‐tau interactions will be utilized to assess their relative influences in maintaining a coupled replisome. Conducting in vitro experiments will provide mechanistic insight into the nature of interactions between these replisome components in a coordinated system.Coupling interactions are also being investigated in vivo using precise CRISPR‐Cas9 genome editing. Coupling mutations have been introduced in the dnaX gene encoding the tau‐subunit. These mutant strains exhibit a reduced growth rate when compared to wild‐type strains, indicating that disrupted coupling leads to reduced doubling times. These strains are being further characterized to determine the genomic and cellular consequences of helicase‐polymerase decoupling in vivo. Therefore, a combination of in vitro and in vivotechniques are being used to better understand the importance of coupling during DNA replication to maintain a stable genome.
Inhibition of Wnt/β‐catenin Signaling Pathway on Melanogenesis‐ Insights from Zebrafish (Danio rerio)
Authors: Silva Praneeth T. , Atukorallaya Devi S. A. 
Published: May 2022

IntroductionVertebrate color patterns are composed of specialized pigment‐producing cells, the chromatophores. Melanin is the main pigment type that colors the skin and skin appendages in mammals. Melanin is produced by specialized cells called melanocytes. Melanocytes have embryonic neural crest cells origin and their development and functions are ubiquitous across vertebrates, residing in dermal and epidermal skin layers. Studies of melanogenesis have provided experimentalists with a valuable system for understanding pattern formation in the developing embryo. In addition, studying these mechanisms may have important consequences for the clinical sciences. Alcohol is a widely abused teratogenic substance. Fetal alcohol spectrum disorder (FASD) is described as birth defects associated with prenatal alcohol exposure (PAE). PAE is likely one of the leading causes of birth defects but pathophysiology of FASD associated body pigmentation disorders are needed to be investigated further. Zebrafish (Danio rerio) (ZF) is identified as a good model to study FASD and melanocyte biology. Wnt signaling pathway is a conserved signal transduction pathway which is important in embryonic development and it regulates many aspects of melanogenesis. The current study was designed to investigate the effect of Alcohol‐Wnt signaling pathway activity on ZF melanogenesis.Materials and MethodsWild‐type (WT) ZF embryos were treated with Wnt pathway inhibitor (LiCl) and combined chemical treatment (LiCl and 1% alcohol) at 10 hours post fertilization (hpf). The differential effects of above substances on early development, differentiation, patterning of melanocytes were investigated using cell biology and imaging techniques.ResultsMelanocytes of LiCl exposed samples showed defects in migration, arrangement and survival. Ectopic melanocytes and developmental aberrations of eye pigmentation were noticed in both chemical treatments. Significant variation of the melanocyte cell size, shape and body pigmentation patterning was observed among the LiCl and combined treated samples compared to the control.ConclusionEmbryonic LiCl and alcohol exposures affect the development and patterning of the melanocytes in ZF. The molecular mechanisms underlying the above phenotypes will be investigated further to understand the FASD associated pigmentation defects.
Thoracopelvic Covariation Across Ontogeny in Humans
Authors: Zaleski Sarah 
Published: May 2022

Clarifying human thoracopelvic covariation patterns can contribute to an understanding of body shape evolution. Previous research has shown that although adult human males and females share a common thoracopelvic covariation pattern, males tend to exhibit mediolaterally wide thoraces and narrow pelves, and females tend to exhibit mediolaterally narrow thoraces and wide pelves (2020 Am J Phys Anthropol 173(3) 514‐534). The current study tests for sex differences in thoracopelvic covariation across ontogeny.I analyzed decedent computed tomography data from the Center for Forensic Imaging at the University of New Mexico with a sample of 44 females and 48 males, aged 0‐21 years. I used SlicerMorph to collect three‐dimensional landmark coordinate data representing thoracic and pelvic shapes as well as R geomorph and Morpho for geometric morphometric analyses. After separate Generalized Procrustes Analyses on the pelvis and thorax, I analyzed thoracopelvic covariation with two‐block partial least squares analysis. To compare covariation magnitudes, I tested effect sizes of sex‐specific two‐block partial least squares. I assessed sex‐specific covariation patterns by projecting the male data into the female two‐block partial least squares analysis and then comparing the slopes of sex‐specific data via major axis regressions and the smatr R package.Results from the two‐block partial least squares analyses indicate significant covariation between the thorax and pelvis in females and males (p=0.001). The first latent variables explain 94.47% and 94.40% of the total covariation in the female and male samples, respectively. In both sexes, as the thorax becomes less pyramidal and more barrel‐shaped with development, the pelvis becomes more anteroposteriorly elongated. The sex‐specific two‐block partial least squares analyses do not exhibit significantly different magnitudes of thoracopelvic covariation. Slope comparison between sexes does not indicate significantly different patterns of thoracopelvic covariation.Previous research suggests that adult thoracopelvic covariation is driven by sex differences. The current study shows that subadult thoracopelvic covariation tracks with growth and development in both sexes, which is expected in the ontogenetic sample. Subadult females and males do not exhibit significantly different magnitudes nor patterns of thoracopelvic covariation. Processes such as sagittal balance of the upper body over the pelvis may influence thoracopelvic covariation.