bluuebunny/crossref_metadata_2025_split huggingface dataset:
Integration of Histology and Pathology in a Senior Elective Course Curriculum
Authors: Cui Dongmei , Daley William 
Published: June 2021

The Anat. 653 Review of Histology was a traditional basic science course offered at the University of Mississippi Medical Center, which provided a microscopic review of specimens of the tissue and organ systems to the M4 students who intend to pursue specialty training in pathology and other fields. A newly modified course not only provides a microscopic review of histology, but also involves integrated clinical cases, pathological images and discussion sessions of both basic science and clinical information. Three‐dimensional integrations include (1) integration of reading materials of both histology and pathology; (2) integration of microscopic review of histological and pathological images; (3) integration of basic science and clinical faculty in small group discussion sessions, comparing the normal cells, structures and tissues with the altered abnormal tissues and associated diseases. The students have an opportunity to consult and discuss with faculty members from both the Anatomy Department and the Pathology Department. They also have the opportunity to give a presentation of selected clinical cases during the discussion sessions. The integrated Histology and Pathology review course will be much better at preparing senior year medical students, especially those who intend to pursue specialty training in Pathology in their upcoming residency than the traditional review course.Grant Funding Source: University of Mississippi Medical Center
Teaching surface anatomy through near‐peer instruction
Authors: Carp Shayna J , Metzger Keith , Rennie William 
Published: June 2021

Study ObjectivesUnderstanding of surface anatomy is crucial for clinical care, including physical examination, trauma management and surgery. This study evaluates the utility of a series of modules focused on surface anatomy in aiding student understanding of anatomical landmarks, as well as their ability to apply knowledge. Modules were developed in the context of larger lab sessions by an MS2, and guided by faculty, presented to MS1s. This provided opportunities for professional development of a medical student and for faculty members to mentor.MethodsA series of modules relating surface anatomy to clinical scenarios was developed by an MS2, guided by faculty members. These were presented to MS1s at Hofstra North Shore‐LIJ SOM on a weekly basis. Each of the stations was developed around a specific goal and students were guided through activities using Socratic questioning. Effectiveness of the modules was evaluated through exam questions related to surface anatomy, and compared to the same questions from a cohort the previous year that did not have the modules.ResultsPreliminary results indicate that students who received the surface anatomy modules performed better than those who did not on questions related to surface anatomy. It is yet to be determined whether the intervention group is able to more broadly apply new knowledge by utilizing general principles of surface anatomy.
Amphibian muscle development and homologies: broader developmental and evolutionary implications, with notes on metamorphosis and terminal additions
Authors: Ziermann Janine , Diogo Rui 
Published: June 2021

There has been a lot of controversy about the homologies of amphibian muscles. There is hardly any study comparing the development and adult configuration of neotenic and non‐neotenic salamanders and of frogs with direct and indirect development. Here we report our findings and explore their broader evolutionary implications, e.g. for the understanding of metamorphosis. For instance, we discuss the controversial homologies of the depressor mandibulae, levator hyoidei and branchiohyoideus, which are crucial for the understanding of amphibian biology and of the origin of amniote muscles. In three investigated salamander species, including a neotenic one, at least part of the larval depressor mandibulae derives from the larval levator hyoidei; in the two non‐neotenic salamanders some fibers of the larval depressor mandibulae become part the larval branchiohyoideus. The confusion about the homology and evolution of these and other muscles comes partly from the fact that many larval muscles disappear during metamorphosis. This contradicts the commonly accepted views that 1) during ontogeny there is a tendency to differentiate more muscles, and 2) that changes of development during evolution often refer to terminal additions. Our recent studies in modern humans have provided similar data and also contradicted these two latter views; therefore we also discuss the implications of our results for human evolution.Grant Funding Source: start‐up package from Howard Univ. College of Medicine
Germ cell nuclear factor (Gcnf/Nr6a1) plays a novel role in neural crest cell induction
Authors: Achilleos Annita , Dennis Jennie , Bhatt Shachi , Sakai Daisuke , Trainor Paul 
Published: June 2021

The neural crest (NC) is a multipotent, migratory cell population that is a hallmark of vertebrate evolution and gives rise to a diverse array of tissues including peripheral nervous system and craniofacial skeleton. Formation of the NC encompasses several steps including induction and specification, epithelial to mesenchymal transition (EMT), migration and differentiation into diverse cell types. Previous work in aquatic and avian systems revealed that Wnt, BMP and FGF signaling pathways drive NC formation. However, knockout mouse models failed to recapitulate a role for these pathways in mammalian NC induction. Our approach uses a mouse mutant in germ cell nuclear factor (Gcnf)/Nr6a1, to study mammalian NC formation. Our analysis of Gcnf−/− embryos revealed a total absence of NC posterior to the first branchial arch and a great anterior reduction in cranial NC. Both phenotypes result from a failure of neural progenitors to differentiate into NC and undergo EMT. Along with our global gene expression and protein interaction analysis, we show that Gcnf regulates two key processes during NC formation: stem cell maintenance and EMT. Therefore, we hypothesize that Gcnf acts as a bimodal switch (i) repressing pluripotency genes and activating NC‐specific genes and (ii) regulating EMT during NC delamination. This is the first example of a total loss of NC in a specific domain of the neuraxis in a mammalian system.Grant Funding Source: AAA Postdoctoral Fellowship
A simple PCR‐based strategy for estimating species‐specific contributions in chimeras and xenografts
Authors: Ealba Erin Lynn , Schneider Richard Allen 
Published: June 2021

Various tissue‐engineering strategies for repair and regeneration involve using cells from different species in chimeras or xenografts. Yet a major challenge is distinguishing between donor‐versus host‐effects on gene expression. This study provides a simple molecular strategy to quantify species‐specific contributions in chimeras and xenografts using species‐specific primers for ribosomal protein L19 (RPL19). cDNA from quail, duck, chicken, mouse, and human was mixed in pairs as a dilution series, species‐specific RPL19 expression was measured, a linear regression was calculated, and the ratios of donor and host cells were estimated. To test the accuracy of the approach, quail cells were transplanted into transgenic‐GFP chick and resulting chimeras were analyzed with species‐specific primers. Fluorescent‐activated cell sorting validated the RPL19 strategy. To apply the RPL19 strategy, Runx2 expression was measured in quail‐duck chimeras. Elevated Runx2 levels correlated with higher percentages of donor cells. Finally, RPL19 primers were also shown to discriminate human‐mouse samples. Thus, this study offers a straightforward and universal strategy that enables chimeras and xenografts to be screened efficiently and rapidly prior to obtaining critical information at the molecular level. Supported by K08 DE021705 and T32 DE007306 from NIDCR to ELE.; R01 DE016402 from the NIDCR to RAS.Grant Funding Source: NIDCR
Function of CDK8 Submodule and TRIM28 in Transcriptional Regulation at HSPA1B
Authors: Bunch Heeyoun , Calderwood Stuart 
Published: June 2021

Recently, it has been reported that a majority of induced or developmental genes possesses Pol II paused at the promoter proximal site about 50–100 bp from the transcription start site (TSS). HSP70 has been well characterized for the promoter proximal pausing before induction that releases the paused Pol II for processive elongation.In order to understand the promoter proximal pausing of Pol II in HSP70 transcriptional regulation, we investigated the function of CDK8 submodule (CDK8, Med12, Med13, and Cyclin C) at HSPA1B using an in vitro transcription system. The results show that CDK8 submodule activates transcription at HSPA1B and that Pol II is stably paused at around +70 at HSPA1B. The activating function of CDK8 submodule is dependent on the timing of its availability to the transcription machinery. In spite of their function as activators, CDK8 submodule and P‐TEFb are unable to relieve Pol II from the pausing site as individual. We also screened a transcription factor(s) that possibly regulates the Pol II promoter proximal pause via oligo pull‐down assays followed by mass spectrometry (MS). TRIM28 was identified to bind near the Pol II pausing site of the HSPA1B non‐template DNA. In vivo knock‐down of TRIM28 increases HSPA1B mRNA expression under the unstressed condition. TRIM28 depletion also alters the expression profile of other known pausing genes, proposing the function of TRIM28 in the Pol II promoter proximal pausing.
Regulation of CFTR gene expression by recruitment of an intestinal transcription factor network.
Authors: Kerschner Jenny , Harris Ann 
Published: June 2021

We recently identified members of a transcription factor (TF) network that interact with a critical cis‐regulatory element for the cystic fibrosis transmembrane conductance regulator (CFTR) gene. This element, located in intron 11 of the gene, 100 kb distal to the promoter with which it interacts, contains an intestine‐selective enhancer, and associates with enhancer signature proteins and tissue‐specific TFs important for regulating CFTR expression. Chromatin immunoprecipitation identified FOXA1, FOXA2, HNF1, and CDX2 as trans‐interacting factors at this and other CFTR cis‐regulatory elements. Moreover, siRNA‐mediated knockdown of CDX2 or FOXA1/A2 causes a significant reduction in endogenous CFTR transcription in intestinal cells, suggesting that these factors are important for maintaining high levels of CFTR expression in these cells. More recently we showed that local changes in chromatin structure and nucleosome positioning are triggered by the interaction of members of the TF network. Our data suggest this group of TFs may play a more global role in regulating intestinal expression of the gene. The mechanism by which they regulate CFTR expression and influence chromatin environment is under analysis.Funding: Cystic Fibrosis Foundation; National Institutes of Health (R01HL094585 and R01HD68901); and Northwestern University Cellular and Molecular Basis of Disease Training Grant
Unraveling the architecture of the A1 complex of the Nanoarchaeum equitans A1A0 ATP synthase
Authors: Mohanty Soumya , Hogue Christopher Warren Victor 
Published: June 2021

ATP synthases are multimeric motors which are indispensible for cellular energetics. The archaeal ATP synthases are the oldest but are relatively understudied. Nanoarchaeum equitans is a nanosized parasitic hyperthermophile with several ancestral traits. Genomic annotation shows that the typical archaeal ATPase subunits are missing, suggesting that the N.equitans ATP synthase is either compromised or constructed partially. The annotated regulatory B subunit is shortened compared to the B subunits of other archaeal ATP synthases, which led to the speculation whether the A and B subunits are capable of forming a functional complex. This study aims at understanding the architecture and functionality of the catalytic A1 complex. The recombinant N. equitans ATPase subunits were purified, reconstituted in‐vitro and characterized using dynamic light scattering, size exclusion chromatography and Blue Native PAGE and isothermal titration calorimetry. Results show the stoichiometric association of A and the annotated smaller B subunits in the presence of nucleotides to form higher order complexes. Therefore our results suggest that the N.equitans ATP synthase might be a minimalistic, yet functional form which would provide a vital connection to the understanding of the evolution of ATP synthases.Supported by Mechanobiology Institute, National University of Singapore (Research Grant R714–009‐007–271)
Weight loss and role of fat free mass in obese patients
Authors: Ippoliti Flora , Canitano Nicoletta , Businaro Rita 
Published: June 2021

Study objectiveWe evaluated body composition analysis, to monitor obese patients in the early stages of immune‐metabolic dysregulation. The expanded activated adipocytes induce a low‐grade chronic inflammation involving an altered body composition of Fat Free Mass (FFM),mostly muscle and bone tissue leading to the onset of Medically Unexplained Symptoms (MUS).Materials and methodsWe enrolled 109 patients 79 females and 30 males all free of disease but with MUS and problems of excess weight.We obtained parameters known as prognostic indicators for inflammatory processes by using BIA‐ACC bioimpedance device (Biotekna‐Italy), performed at the first medical examination, (time T0), and after 6 weeks of proteic, low‐caloric diet (1200 calories).ResultsAll patients decreased the BMI but 3 subgroups showed: a)75.23% loss of Fat Mass but also FFM b)17.43% loss of FM with preservation or even increase FFM c)7.34% loss of FFM alone. The main components with a cumulative variance of 94% were only 3 which shows that the analyzed data are statistically highly consistent. The 1st principal component is the FFM correlated closely with BMR, TBW and TBCa, to a lesser extent with Gly, TBK and TBprotein.The 2nd component is AT, FM, AAT with particular relevance to the visceral fat.The 3rd component is the stress with inflammation.ConclusionThe number of parameters provided by the analysis of body composition after appropriate low‐calorie diet, have allowed us to evaluate, also in the presence of a decrease in BMI, the real changes in FM and FFM in relation to all the other variables and the essential role of the preservation of FFM during weight loss.
Characterization of mutations in small heat shock proteins that influence substrate binding
Authors: McMenimen Kathryn , Caminiti Courtney , Liu Pei , Kamalova Aichurok , Weber Caroline 
Published: June 2021

The small heat shock proteins (sHsp), ubiquitous cellular homeostasis machinery, are oligomeric chaperone proteins. sHsps play an important role in maintaining cell function and survival under stress conditions such as high temperatures. By binding to non‐native proteins in an ATP‐independent manner, sHsps effectively prevent harmful aggregation of denatured proteins, promote proper protein folding, or facilitate protein degradation. Defects in sHsps can result in abnormal accumulation of proteins or aberrant protein folding, which lead to many pathologies such as cataract, tumor and neurodegenerative diseases including Parkinson's disease and Alzheimer's disease.Structural studies have demonstrated that sHsps contain a highly conserved α‐crystallin domain flanked by variable N‐terminal and C‐terminal regions. They usually exist in large oligomers and are capable of adopting different oligomeric states in response to different conditions. Therefore, because of their dynamic structures, the specific mechanisms of how sHsp bind to substrate and function are still not well understood. Nonetheless, it is suggested that the variable N‐terminal region of the protein contributes to substrate binding and oligomerization.We have made several mutations in the sHsps to characterize regions that influence oligomerization and make contacts with substrate proteins. Our studies suggest that the hydrophobic regions of the N‐terminal domain are important for substrate‐binding. Additionally, mutations in the a‐crystallin domain also contribute to the dynamics of protein oligomerization and sHsp‐client interactions.
Quantitative Profiling of Cysteine S‐nitrosylation Susceptibility in Cancer
Authors: Zhou Yani , Convertier Shalise , Weerapana Eranthie 
Published: June 2021

We aimed to quantitatively measure the susceptibility of cysteine S‐nitrosylation (SNO), the transfer of a NO moiety to cysteine (Cys) on protein. SNO represents a form of redox modulation and plays an important role in diverse cellular processes. Recent developments of chemical methods to detect SNO have made it possible to identify SNO‐Cys sites. However, these methods do not monitor the extent of Cys to undergo nitrosylation in a complex proteome.We utilized activity‐based protein profiling to identify nitrosylated Cys and to measure the susceptibility of Cys SNO in cell lysates after treatment with NO donor. A gel based dose‐dependent experiment revealed that Cys residues on different proteins demonstrated diverse susceptibilities to SNO. Isotopic tandem orthogonal proteolysis‐activity‐based protein profiling (isoTOP‐ABPP) method was applied to further quantitatively monitor the SNO susceptibility. Several novel Cys SNO targets were identified and verified by overexpression in mammalian cells. One of these is the serine/threonine‐protein kinase 1 (AKT1) which plays a critical role in many different signaling pathways, and might contribute to tumor progression. Our study showed that one Cys on AKT1 is highly susceptible to SNO. We are exploring structural motifs that govern this selectivity, and further investigate the role of nitrosylation in modulating AKT1 activity in vivo.
Investigating the targeting and regulation of Ded1, a DEAD‐box ATPase, in returning repressed mRNAs to translation
Authors: Pollard Beth , Hilliker Angela 
Published: June 2021

Translation regulation is important, especially during stress, in neurons, and in early development, where many mRNAs are stored in translationally repressed states. These repressed mRNAs can be returned to translation, but this process is poorly understood. We have previously shown evidence that the RNA helicase, Ded1, performs two sequential functions to return a repressed mRNA back to translation in an ATP‐dependent manner. First, Ded1 promotes the formation of a “pre‐48S mRNP” that includes the initiation factors eIF4F and Pab1. Then, Ded1 hydrolyzes ATP, allowing the mRNA to become translated. We hypothesize that Ded1 acts as an ATP‐dependent switch to return mRNAs back to the translating pool.We are investigating how Ded1 forms this pre‐48S intermediate. Ded1's ability to form this repressed mRNP in vitro depends on the presence of a polyA tail. Additionally, Ded1 increases the recruitment of Pab1 to the m7G cap. We are currently investigating the relationship between Pab1 and Ded1 in formation of this repression complex.
The role of mTORC1 and mTORC2 in UVB‐induced
Authors: Lewis Martha , Shantz Lisa 
Published: June 2021

Mammalian target of rapamycin (mTOR) plays an important role in cell proliferation and survival. mTOR has been shown to exist in at least two complexes, mTORC1 and mTORC2. mTORC1 is inhibited by rapamycin, contains the mTOR catalytic subunit as well as the proteins RAPTOR and mLST8. It shows that mTORC1 phosphorylates p70 S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E binding protein 1 (4EBP1). mTORC2 is known to be the kinase responsible for AKT phosphorylation on Serine 473 (S473). Based on previous research, we hypothesize that mTOR is a useful target in the prevention and treatment of nonmelanoma skin cancer. The goal of this project is to expand those results using wild‐type (WT) and RICTOR‐null (Rictor−/−) mouse embryo fibroblasts (MEFs). Since phosphorylation of AKT on Threonine 308 (T308) has been reported to be facilitated by S473 phosphorylation,Aimis to determine the phosphorylation state of AKT T308 in WT and Rictor−/− cells, both with and without UVB treatment and to examine the effect of Torin‐2, which inhibits both complex on proliferation in WT and Rictor−/− MEFs.RESULTSWestern blot analysis demonstrated that UVB exposure induces AKT phosphorylation at T308 in both cells suggesting that the absence of mTORC2 signaling does not prevent T308 phosphorylation in response to UVB. MTS analysis showed that mTORC1 inhibition with rapamycin had no significant effect on cell proliferation. Treatment of WT and Rictor−/− cells with Torin‐2 decreased proliferation in a dose‐dependent manner. Since TORC2 activity is absent in MEFs lacking Rictor, this result suggests the presence of a rapamycin‐insensitive mTORC1 activity that may play a role in cell proliferation.
Adenovirus 36 as a Risk Factor for Clinical Overweight/Obesity
Authors: Voss Jameson , Burnett Daniel , Olsen Cara , Haverkos Harry , Atkinson Richard 
Published: June 2021

We evaluated Adenovirus 36 (Ad36) serostatus and follow‐up body mass index (BMI), waist circumference (WC), and clinical records among Air Force (AF) recruits.Male enlistees (n=500) were followed from time of entry into AF beginning in 1995 to 2012 or separation, for 2915 person‐years. Numerically randomized selection occurred within four BMI quartiles (ranging from 20–30 kg/m2), stratified by year of entry. The exposure, Ad36 antibody, was assessed by ELISA from banked serum.The primary outcome, follow‐up BMI, was similar by serostatus. This contrasted with secondary outcomes. The hazard of clinical overweight/obesity showed interaction (p=0.007) between baseline BMI and serostatus. At a baseline BMI of 22.5, survey weighted cox proportional hazards showed the seropositive had a 4.0 fold increased hazard of diagnosis of clinical overweight/obesity (p=0.003) as compared with seronegative, controlling for baseline BMI. The maximum WC was higher among seropositive.Ad36 serostatus is associated with increased hazard of overweight/obesity diagnosis among lean, male, AF recruits.The opinions are those of authors, not the DoD or USUHS. IDCRP funded the study and Obetech, LLC provided lab testing.
Food access, social safety nets, and social stigma among low‐income families in Massachusetts
Authors: Ghavami Sarvnaz Modarresi , Peterman Jerusha Nelson , Anliker Jean 
Published: June 2021

Social safety nets like SNAP, WIC, and fuel assistance can alleviate food insecurity and improve household resources. They may not prevent health consequences of food insecurity unless they promote purchase and consumption of healthful foods. Social stigma around use of benefits can prevent vulnerable families from utilizing benefits, which may further complicate food insecurity. We analyzed focus group transcripts (6 focus groups, 21 participants) with low‐income, largely food insecure individuals in Massachusetts to assess use of social safety nets and related social stigma. We used directed content analysis to examine pre‐identified themes and to identify new themes. Pre‐ identified themes included the use of SNAP and WIC to extend resources and stigmatization of the use of social safety nets by the members of the community. New themes were utilization of fuel assistance to free up resources, saving SNAP for special and healthy foods, stigmatization of low cost foods by children, stigma within the community for resource allocation of benefit users, and self‐stigmatization. Different stigma associated with the use of social benefits and purchase of low cost foods might prevent low‐income individuals from utilizing. In this context, allocation of certain benefits, such as SNAP, to purchase low quality foods might lead to health complication in the future.Funding source: USDA‐CSREES #H994