bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of Growth Supplements on the Differentiation of Adipose Derived Stem Cells from Burned and Non‐Burned Human Skin
Authors: Mangum Lauren Heard , Wrice Nicole , Larson David , Natesan Shan , Christy Robert 
Published: June 2021

Severe traumatic injuries, such as large total body surface area burns, limit the availability of autologous cells for wound repair. Previous experiments from our lab have demonstrated that adipose derived stem cells (ASCs) from burned debrided skin are viable, display an immunophenotype indicative of stemness, and are capable of tri‐lineage differentiation. Traditional tissue culture utilizes undefined, animal‐derived components that present numerous complications for the utilization of ASCs in human medicine. Accordingly, there is a need for defined, xeno‐free cell culture media. Human platelet lysate (hPL) is one potential media supplement for the replacement of fetal bovine serum in stem cell culture. In this study, burned human ASCs (BH‐ASCs) and ASCs derived from abdominoplasty (HAP‐ASCs) were compared in regard to the ability to proliferate and differentiate in media supplemented with traditional 10% FBS, 10% hPL, or grown in MesenPRO RS™. ASCs were obtained from the stromal vascular fractions of subcutaneous adipose tissue from burned and non‐burned skin. Cells were cultured in DMEM with 10% FBS or 10% hPL, or in MesenPRO RS™. Cells were induced to differentiate towards an adipogenic or osteogenic lineage at an early passage utilizing traditional differentiation media supplemented with FBS or 10% hPL. DMEM or αMEM was used for adipogenic and osteogenic lineages, respectively, with the addition of standard induction agents. Adipogenic differentiation was assessed after 14 days by Oil red O staining, triglyceride content, and expression of markers of adipogenesis, including PPARγ, leptin, and adiponectin. Osteogenic differentiation was assessed by alizarin red S staining and expression of markers of ostogenesis, including BMP‐2 and RUNX2. Results indicate that adipogenic and osteogenic differentiation appears to be influenced by both the ASC source (burned vs. non‐burned skin) and by the supplement in which they are grown and differentiated. Use of MesenPRO RS™ and supplementation with hPL appears to enhance differentiation towards both lineages over standard FBS. Prior experiments in our labs have indicated that both burn and non‐burn ACSs are capable of trilineage differentiation, however there is a need to characterize these cells, particularly the BH‐ASCs in xeno‐free media, with regard to their stemness and immunosuppresive capabilites if they are to be utilized in any capacity for regenerative medicine. Ongoing research efforts in our laboratory seek to address these issues to better define the utility of BH‐ASCs as a burn wound therapeutic.Support or Funding InformationThis work was possible due to the generous funding from the US Army Medical Research and Material Command and ORISE
Transcriptomic Analysis of the Right and Left Ventricle in Normoxia and Hypoxia (a Model of Pulmonary Arterial Hypertension)
Authors: Gorr Matthew W , Sriram Krishna , Insel Paul A 
Published: June 2021

The pathobiology of pulmonary arterial hypertension (PAH) involves the remodeling of the pulmonary artery within the lung characterized by vasoconstriction, hypertrophy, fibrosis and thrombosis. These factors lead to increased pulmonary vascular resistance (PVR), which, in turn, can produce progressive remodeling of the right ventricle (RV) and eventual right heart failure. Current therapy for PAH emphasizes approaches that decrease PVR, but does not work for some patients and does not specifically target the RV. This study tested the hypothesis that the RV contains a unique subset of genes expressed during remodeling that may serve as a platform to identify and develop novel treatment strategies. To test this hypothesis, we subjected Sprague Dawley rats to hypoxia (10% O2, H‐rats) for two weeks, or to normoxia (N‐rats) as a control (n=3–4/treatment). H‐rats had a significant, 2.19‐fold increase in the Fulton Index (right/[left + septum] ventricular weight): 0.458 ± 0.036 (H‐rats), 0.210 ± 0.020 (N‐rats); mean ± standard deviation, Student's t‐test p<0.0001), indicating significant RV remodeling. To examine mRNA expression, we utilized RNA sequencing (RNA‐seq) of the RV and the left ventricle (LV). Using differential expression analysis, we identified 2,430 genes with significantly (False discovery rate [FDR] <0.1 using multiple testing) altered expression in the RV of H‐rats vs. N‐rats and 504 genes with significantly altered expression in the LV between the two groups of rats. Differentially expressed genes in the RV of H‐rats vs. N‐rats included GPCRs and other widely used drug targets. In normoxia, the RV and LV differentially expressed 299 genes compared to 423 genes under hypoxic conditions. These results indicate that: 1) a subset of genes are differentially expressed in the RV compared to the LV and 2) a large change in the transcriptome occurs with hypoxia‐induced RV remodeling. These findings raise the possibility that treatments for PAH might be directed at gene products selectively expressed in RV remodeling and be able to spare the LV, thus providing a novel approach for treating a currently unmet need for patients with RV hypertrophy and failure.Support or Funding InformationSupported by Department of Defense (W81XWH‐14‐1‐0372) and the National Institutes of Health (5T32HL007444‐33)
Deficiencies in RNS2‐mediated ribosomal RNA turnover cause changes in the Pentose Phosphate Pathway flux and alter cell growth in Arabidopsis
Authors: MacIntosh Gustavo C , Morriss Stephanie , Liu Xiaoyi , Bassham Diane 
Published: June 2021

Enzymes belonging to the RNase T2 family are essential for normal ribosomal RNA turnover in eukaryotes. In Arabidopsis thaliana this function is performed by RNS2. The null mutant rns2‐2 has increased rRNA half‐life and constitutive autophagy. To determine the effect of defective rRNA turnover on cellular homeostasis, we used comparative transcriptome and metabolome analyses of 10 day old wild‐type and rns2‐2 seedlings to identify molecular processes affected in the mutant. Bioinformatics analyses suggested additional phenotypes that were confirmed through direct plant size measurements and microscopy.Few genes were differentially expressed in the rns2‐2 mutant, indicating that control of autophagy in this genotype is mainly achieved at the posttranscriptional level. Among differentially expressed genes we identified transcripts related to carbon flux processes, particularly the pentose phosphate pathway, and metabolite analyses confirmed changes in the levels of PPP intermediates. We also found differentially expressed genes related to cell wall loosening and found a decrease in monosaccharide components of cell wall hemicellulose. As a potential effect of weaker cell walls, rns2‐2 plants are larger than wild‐type controls, due to larger cells and increased water content. We propose that lack of rRNA recycling in rns2‐2 cells triggers a change in carbon flux, which is redirected through the pentose phosphate pathway to produce ribose‐5‐P for de novo nucleoside synthesis. rRNA or ribosome turnover is thus essential to maintain nucleoside homeostasis in Arabidopsis.Support or Funding InformationThis work was supported by a grant from the United States National Science Foundation [MCB‐1051818] to G.C.M. and D.C.B.
3D Integrated Tissue and Organ Printing System to Produce Human Body Parts with Structural Integrity
Authors: Lee Sang Jin 
Published: June 2021

Tissue engineering and regenerative medicine strategies could offer new hope for patients with serious tissue injuries or end‐stage organ failure. Scientists are now applying the principles of cell transplantation, material science, and engineering to create biological substitutes that can restore and maintain normal function in diseased or injured tissues/organs. Among various three‐dimensional (3D) biofabrication technologies developed for the field of tissue engineering, 3D printing technology is one of the most attractive and powerful methods for use in constructing a structure that mimics native tissues, and thus many researchers are focusing on development of new printing technologies and applications. The hypothesis driving development of 3D bioprinting is that by precisely placing cells in relation to each other, an environment that encourages physiologically relevant cues both to and from these cells can be created, resulting in a tissue construct with functionality. Thus, we have developed 3D integrated tissue and organ printing (ITOP) system to allow construction of biological substitutes that mimic the structure and function of native tissues or organs. This system makes it possible to precisely place various cell types and biomaterials in a single 3D architecture. Our proof of concept experiments demonstrates the capabilities of our ITOP by fabricating structures of various tissue constructs of appropriate shape and size needed for clinical use. This novel organ printing system provides a leap forward in our ability to fabricate tissue constructs that could be used in translational applications.Support or Funding InformationThis work was supported, in part, by grants from the Armed Forces Institute of Regenerative Medicine (W81XWH‐08‐2‐0032), the Telemedicine and Advanced Technology Research Center at the US Army Medical Research and Material Command (W81XWH‐07‐1‐0718) and the Defense Threat Reduction Agency (N66001‐13‐C‐2027).
Characterizing the Role of Yeast Cyclin Pcl1 in the Establishment of Pheromone Receptor Polarity
Authors: Pai Chih Yu (Chloe) , Sukumar Madhushalini , Stone David 
Published: June 2021

Chemotaxis (directed cell movement) and chemotropism (directed cell growth) are vital mechanisms essential to a variety of biological processes, including cell development, cancer metastasis, angiogenesis, and axon guidance. Both chemotaxis and chemotropism depend on the ability of the cell to interpret shallow and complex chemical gradients. One of the best‐studied models of eukaryotic directional sensing is the mating response of Saccharomyces cerevisiae (budding yeast). Yeast exist as two haploid mating types, MATa and MATα, that can sense pheromone gradients, chemotrop toward the closest mating partner, and eventually fuse at their tips to form diploid cells. In vegetative cells, the G protein coupled receptor (GPCR) that binds pheromone and its associated heterotrimeric G protein are uniformly distributed on the plasma membrane. Upon receptor activation, Gβγ is released from Gα and Gβ is rapidly phosphorylated on multiple residues. Free Gβγ signals through a MAP kinase cascade to trigger cell‐cycle arrest and changes in gene expression. Pheromone‐activated receptors are globally internalized and then reappear as polarized crescents that mark the chemotropic growth site, where the cell ultimately forms its mating projection. Previous studies have shown that the receptor crescents are visible before detectable actin polarization and that the receptors can polarize without actin‐dependent directed secretion [1]. On the contrary, inhibiting receptor internalization confers a moderate defect in gradient sensing [2]. A key question raised by these observations is how does the yeast cell interpret the external signals and establish the chemotropic growth site? From a directed genetic screen, we found that Pcl1 is critical for the establishment of receptor polarity. Pcl1 is a cyclin required by the cyclin dependent kinase Pho85, which has been implicated in polarization during budding. It is known that Gβ phosphorylation is crucial to receptor polarization, and Gβ is predicted to be a substrate of Pho85‐Pcl1 [3]. Additionally, our data indicated a genetic interaction between Gβ and Pcl1. Here, we show that Pcl1 localizes to the mating projection during the mating response. Moreover, Pcl1 and Gβ interacted directly on the plasma membrane of pheromone‐treated cells, as indicated by a Bimolecular fluorescence complementation assay. Lastly, Pho85 inactivation also appears to affect receptor polarity. Taken together, these results suggest that Pho85‐Pcl1 regulates polarization of the receptor by phosphorylating Gβ.Support or Funding InformationNational Science Foundation
Gulliver Prototype Development and Deployment
Authors: Williams Claire , Berdy Brittany , Epstein Slava 
Published: June 2021

The current objective of our project is to develop a technology, coined Gulliver, aimed at enhancing the study of microorganisms. This device will expand upon established in situ technology to cultivate and isolate microorganisms in one step. Our long term goal is for this device is to not only isolate and culture in one step, but also to eventually equip our device with nanosensors to allow in situ monitoring of bacterial growth and activity, and transmit this data directly to the researcher, allowing an unprecedented view into the microbial world. The Gulliver design consists of a flat mounting device with a 4 mm hole in the center to act as the growth chamber. On one side of the device, a polycarbonate membrane with 0.03μm pores is attached, allowing for diffusion of nutrients and growth signals from the environment. On the reverse side, the same membrane is affixed, however it contains an approximately 1 μm entry aperture, to allow entry of a single microbial cell into the growth chamber. Once the microorganism enters the pore, the entrance is blocked. As the organism continues to grow and divide, it is provided with nutrients from the environment through the diffusion pores, and a pure colony will develop inside the device. After a period of in situ incubation, the device is removed from the environment, the growing biomass retrieved from the growth chamber, “domesticated” and grown on a petri dish for further study. Currently, testing is underway using fluorescently labeled bacterial species to prove the concept of autonomous cultivation and isolation in on step. Devices will be incubated in mixed cultures of fluorescently labeled bacteria, and growth will be collected from each chamber. Fluorescence will be used to ensure a pure culture is obtained. Thus far, we have demonstrated an ability to reliably produce 1 μm pores in polycarbonate membrane. Furthermore, we have designed a prototype of the device consisting of the polycarbonate membrane adhered to a silicone or PTFE base. The prototype is capable of withstanding incubation in an aqueous environment, without material degradation, for up to 3 months, providing an extended period of time for environmental study of microorganisms. The overall goal of this project is to gain a fuller picture of the microbial world, allowing researchers to access novel microorganisms, study the important environmental processes they perform, and utilize the valuable bioactive compounds they produce.Support or Funding InformationThis particular segment of the project is supported by the NSF, the Alpha Fund Grant, and an Northeastern Undergraduate Research and Creative Endeavors Award.
PIP4Kγ as a potential target for Huntington's disease
Authors: Giridharan Sai Srinivas Panapakkam , Al‐Ramahi Ismael , Hasegawa Junya , Safren Nathanial , Patnaik Samarjit , Gee Amanda , Titus Steve , Zheng Wei , Ferrer Marc , Southall Noel , Barmada Sami , Botas Juan , Marugan Juan , Weisman Lois S 
Published: June 2021

Phosphoinositide lipids provide spatial and temporal regulation of cell signaling and membrane traffic. The PI5P‐4‐kinase family is one of the less studied phosphoinositide lipid kinases. These enzymes catalyze the conversion of phosphatidylinositol 5‐phosphate to phosphatidylinositol 4,5‐bis phosphate. A recent study by Vicinanza et al., a suggested that depletion of the PI5P‐4‐kinase, PIP4Kγ promotes a non‐canonical autophagy pathway. Moreover, this depletion reduced the presence of huntingtin‐exon1‐polyglutamine aggregates. Expression of exon1 of huntingtin with an extended polyglutamine tract is a model for huntingtin protein aggregates. To further test the hypothesis that PIP4Kγ is a potential target to reduce huntingtin aggregates, we tested the depletion of PIP4Kγ in cell culture models that express full length mutant huntingtin (mHtt). We determined that depletion of PIP4Kγ, but not PIP4Kα or PIP4Kβ, lowers the levels of mHtt in patient fibroblasts and in immortalized HdhQ111 mouse striatal neurons. We show that PIP4Kγ‐dependent depletion of mHtt requires lysosomal function because the depletion is blocked by the lysosomal inhibitor, bafilomycin A1. Moreover, we show that depletion of PIP4Kγ increases autophagic flux, as measured by an increase in LC3‐II levels by western blot analysis and an increase in LC3‐II puncta by microscopy. Our studies suggest that PIP4kγ should be further pursued as a target for Huntington's disease.Support or Funding InformationThis work was supported by AHA postdoctoral fellowship 14POST20480137 (Sai Srinivas Panapakkam Giridharan), NIH grants R01GM050403 and NS064015 (Lois S Weisman) and by HSS funding 1R03MH084839 (Juan Marugan)
Validation of the Dietary Screening Tool in a Middle‐Aged Appalachian Population
Authors: Marra Melissa Ventura , Thuppal Sowmyanarayanan V , Bailey Regan L , Johnson Elizabeth 
Published: June 2021

The Dietary Screening Tool (DST) was developed and validated to assess nutritional risk in older adults. The purpose of this study was to assess its use in a middle‐aged sample of adults (45–64 years) in Appalachia (n=87) and evaluate its validity with multiple 24‐hour dietary recalls and selected biomarkers. Adults identified by the DST as at nutrition risk (<60 of 100 points) were significantly more likely to have had lower overall diet quality as assessed by the Healthy Eating Index (64 vs 50, p<0.001) when compared to those not at risk or potential risk. Dietary intakes of vitamin A, vitamin C, vitamin B2, vitamin B6, niacin, vitamin K and potassium were also lower among those considered at risk, despite having significantly higher energy intakes (1653 ± 106 vs 2032 ± 76 kcal/day, p=0.004). The diets of those at risk were higher in added sugars (33 g vs 23 g, P=0.005) and lower in dietary fiber (10 g vs 12 g, p<0.001) than those not at risk. The at risk group had significantly lower serum levels of 13‐cis‐β carotene (40 vs 60 nmol/L, p<0.02), trans‐ β‐carotene (395 vs 711 nmol/L, p<0.001), β‐Cryptoxanthin (137 vs 198 nmol/L, p<0.01), cis‐Lutein (90 vs 137 nmol/L, p<0.02), and trans‐lutein (292 vs 424 nmol/L, p=0.01). The DST was able to identify middle‐aged adults at nutritional risk.Support or Funding InformationUSDA Northeast Regional Project NE1439, WVU Health Disparities Mountains of Excellence Flash Funding and the WVU ADVANCE Sponsorship Program.
Developmental Changes in Resting Gamma Power from Age Three Months to Five Years Are Modulated by Infant Diet
Authors: Pivik R T , Andres A , Cleves M A , Tennal K B , Gu Y , Badger T M 
Published: June 2021

Gamma band activity (30–50 Hz) is a significant EEG component related to intelligence, memory and language processes, but there is limited information regarding the early development of this activity and none considering how infant diet may influence this development. The present study examined changes in gamma power from 3 months through 5 years in 497 healthy children who were breast‐fed (BF: 168, 84 boys), or fed milk formula (MF: 169, 89 boys), or soy formula (SF: 160, 90 boys) as infants. Recordings of resting EEG (5 min; eyes open) were obtained quarterly during infancy and then yearly. We used ANOVAs adjusted for relevant growth and environmental factors (gender, head circumference, gestation, birth and visit weight, SES and mother's IQ) to compare group, age, and group by age changes in gamma EEG power from 44 homologous sites distributed over frontal, central, temporal, parietal and occipital scalp regions. Across sites and groups, gamma power was lowest at 3 months (all p < .001), and across age gamma power was consistently higher at frontal than other sites (all p < .05). From 3 to 6 months gamma power increased dramatically at all sites for all groups [all p < .001; mean increase >100% for all sites, greatest for central‐parietal regions (187% mean increase across groups)]. After 6 months, gamma power generally increased across sites until a time after which significant (p < .05) decreases became evident relative to preceding peak levels. The ages after which decreases became evident varied within and across brain regions, with site‐specific time points of change ranging from 1 to 4 years and occurring later (after 3–4 years) in anterior frontal than other regions. Temporal sites differed from others in showing sharp decreases both early (after 9 months or 1 year) and later (after 3 years). Group variations in site‐related time points of change were most evident for frontal and occipital regions. Infant diet‐related effects were evident at 3 months [BF < MF, SF (p ≤ .001) and were generally sustained across age‐‐ particularly at parietal‐occipital sites where BF < MF differences (p < .05) were most prominent. These findings provide new information regarding the regional development of EEG gamma activity from infancy to the onset of middle childhood, and show that this development is modified by infant diet. Previous reports of negative associations between resting gamma and measures of language skills and intelligence suggest potential developmental advantages for breastfed infants.Support or Funding InformationFunded by USDA‐ARS Project 6026‐51000‐010‐05S