bluuebunny/crossref_metadata_2025_split huggingface dataset:
Migration of Jurkat cells in response to CypA
Authors: Jurjus Rosalyn A , Pushkarsky Tatiana , Bukrinsky Michael 
Published: June 2021

Cyclophilins (CyPs) are ubiquitously distributed proteins, identified initially as the main binding target for cyclosporine A (CsA). A potential contribution of cyclophilin in inflammatory diseases is suggested from several studies. The activities of secreted cyclophilins are related to interactions with CD147, a transmembrane glycoprotein that plays a major role in lymphocyte migration and activation and is a binding partner to the β1 integrin. Our hypothesis is that the CypA/CD147/β1 interaction is important in mediating cellular migration in inflammation. Using Jurkat cells as our model, we first started by optimizing Jurkat cells adhesion, spreading and migration.In timelapse migration studies, Jurkat cells replated on fibronectin‐collagen I and poly‐lysine showed an increase in velocity by 3 folds when CypA was added to the media. The velocity of these cells significantly decreased when CD147 antibody (Ab) was added, compared to the corresponding isotype control. However, some element of CypA activation remained in the presence of CD147 Ab and it was not concentration dependent. Adding the CD147 Ab without prior CypA exposure did not have a significant effect. This suggests that the CypA‐CD147 activation might involve the β1 integrin. A β1 integrin blocking Ab combined with CD147 Ab is expected to completely prevent the CypA activation and the cell migration.Grant Funding Source: R56AI081152
Identification of genes essential for Francisella invasion of non‐phagocytic cells
Authors: Lo Karen Y , Nano Francis E , Guttman Julian A 
Published: June 2021

The intracellular bacterium Francisella tularensis subspecies tularensis (F. tularensis) is the causative agent of tularemia, a disease that can result in 30–35% mortality if left untreated. F. tularensis host cell invasion is crucial for their pathogenesis as without invasion disease does not occur. Upon host entry, the bacteria colonize both macrophages and epithelial cells. We hypothesize F. tularensis possesses unique virulence factors specific for the invasion of epithelial cells. To identify F. tularensis genes involved in this process, we utilized both in vitro and in vivo experimental approaches using a biosafety level 2 bacterial surrogate, F. tularensis subspecies novicida (F. novicida). A F. novicida mutant library was screened for bacteria with reduced invasion and intracellular growth using gentamycin‐based survival assays in hepatocytes in vitro. Select candidate mutants were tested in mice for their ability to colonize murine livers. We discovered that bacteria containing mutations in at least 6 previously uncharacterized genes had significantly reduced bacterial loads, resulting in mice that survived for extended durations as compared to wild‐type infections. Our findings demonstrate that novel bacterial proteins are crucial for Francisella virulence and these findings open a new door to the discovery of novel targets for the development of therapeutics and prophylactics.Grant Funding Source : NSERC and CIHR
The spectrin cytoskeleton is required for efficient intestinal bacterial infections
Authors: Guttman Julian Andrew , Ruetz Tyson Joel 
Published: June 2021

Crucial aspects of bacterial pathogenesis such as the invasion of host cells and cell‐to‐cell transmission require the hijacking of host cell cytoskeletal systems. Although much work has examined the role of the actin cytoskeleton during numerous bacterial infections, the involvement of the spectrin cytoskeleton has remained overlooked. The spectrin cytoskeleton is required for many cellular processes including cell motility, organization of the plasma membrane and epithelial cell polarization. To determine whether this cytoskeletal system influenced microbial infections we tested the hypothesis that bacterial pathogens control the spectrin cytoskeleton as part of their disease processes. To test this hypothesis we utilized cell biological techniques including the immunolocalization of spectrin cytoskeletal components and RNA interference, and coupled these analyses with microbiological techniques such as wild‐type and mutant bacterial infections, invasion assays and colonization experiments. We found that the spectrin cytoskeletal network was integral for Salmonella, Listeria, Shigella and pathogenic E. coli infections. We showed that host cell invasion and the generation of bacterial motility structures during infections required spectrin cytoskeletal components and knockdown of specific spectrin‐associated proteins haulted Listeria, Shigella and Salmonella internalization as well as pathogenic E. coli attachment to host cells, thus impeding the infections. These findings demonstrate that the spectrin cytoskeleton is essential for bacterial pathogenesis and could be used as a target for future therapeutics. This work was funded through NSERC and the CIHR.Grant Funding Source: CIHR and NSERC
Alpha II‐Spectrin Interacting Partners in the Inner Ear Serve as Pointers to the Protein Composition of the Striated Organelle
Authors: Chidavaenzi Robstein Lovejoy , Lysakowski Anna 
Published: June 2021

Inner ear hair cells sense sound and head movements via apically‐placed stereocilia. These stereocilia are comprised of paracrystalline arrays of parallel actin filaments that pack more densely at the base to form stereociliar rootlets. The later, generally described in the literature as anchored in the cuticular plate, have been observed by our lab to extend well into the cell body where they associate with an underlying cytoskeletal structure called the striated organelle (SO). Made up of alternating thin (~10nm) and thick (~35nm) filaments, the function of this organelle remains unknown. To ascertain its function, we are attempting to determine its protein composition. So far, we have been using α‐II‐spectrin, an actin crosslinking and molecular scaffold protein, as a biological marker for the SO since we localized it there with confocal fluorescent and EM immunogold studies (Vranceanu et al., submitted). Further employing α‐II‐spectrin as bait, we pulled down interacting partners in vestibular endorgan tissue using co‐immunoprecipitation and liquid chromatography mass spectroscopy. Here we identify some of these interacting proteins and present evidence that they also localize to the SO. Based on this putative protein composition, we can now speculate as to probable function for the organelle in inner ear electro‐mechanotransduction.Grant Funding Source: American Hearing Research Foundation
Is BMP‐2 a component of the particulate matrix that is responsible for epithelial/mesenchymal transformation during heart development
Authors: Abd-Elhamid Tarek H. , Conway Marianne L. , Sinning Allan R. 
Published: June 2021

Early cardiac development involves an epithelial/mesenchymal transformation (EMT) of endothelial cells in the atrioventricular (AV) canal and outflow tract (OT) regions of the heart. It has been shown that the myocardium secrets factors, collectively called particulate matrix that induce this process. Previous reports from our laboratory showed that, only myocardial conditioned media (MyoCM) or EDTA extract can replace the myocardium and drive mesenchyme formation. Reports from other laboratories suggest that bone morphogenetic protein 2 (BMP‐2) has a role in EMT. So, we asked whether BMP‐2 is a part of the particulate matrix. Initial experiments using immunostaining of stage 15–16 chick embryo sections with anti BMP‐2 antibody revealed that BMP‐2 signals were detected within the myocardium of the cushion forming regions and in a group of cells in the endocardial layer of these regions in the heart, but not in the extracellular matrix. To confirm these results, whole heart lysate, EDTA extract from stage 15–16 embryos and soybean agglutinin (SBA) binding proteins from MyoCM will be examined for the presence of BMP‐2 by western blotting analysis.Grant Funding Source : none
A successful method for engaging students in active learning using prosected cadavers
Authors: Khirallah Samar , Walker James J 
Published: June 2021

Over the past several years, we have taught a graduate level gross anatomy course using prosected cadavers. The laboratory component of the course constituted instructor‐led demonstrations of the anatomical structures. To create a more engaging and active learning environment, we developed a new teaching strategy that allowed students to construct their own anatomical knowledge through cooperative discovery‐ learning exercises. In the new method, instructors tagged structures on the cadavers prior to the laboratory sessions and then students worked in small groups to identify the tagged structures. We then compared the two methods by assessing both students’ learning and attitude. Four traditional demonstration sessions were alternated with 4 active learning sessions. Each lab session was followed by a short practical identification quiz. A survey was administered to evaluate students’ attitudes. Quiz scores were significantly higher following active learning sessions (mean score 54 vs 38) and continued to improve as students became more effective learners. The survey clearly showed a unanimous preference of the improved design. Structured exercises that emphasized kinesthetic manipulation of the anatomical specimens in conjunction with a cooperative learning environment advanced the learning process significantly and provided the motivation for advanced self‐learning.Grant Funding Source: none
Getting crafty in the classroom: Reinforcing students’ knowledge of muscular attachments and positions with fabric muscles
Authors: Bee Mary Tracy , Montante James M , Grabowski Greg , Forbes William C 
Published: June 2021

Helping students to visualize the origin, insertion, position and relationships of muscles is a challenge. We present a visual, interactive exercise to reinforce student understanding in this area. Students were asked to place precut two‐dimensional felt models of the muscles on a life sized skeleton or disarticulated bones matching the correct bony origins and insertions.In order to assess the impact of this approach on student learning, the students were split into two groups. The first group spent 15 minutes utilizing the felt leg muscles while the second group spent 15 minutes reading their books to study the muscular attachments of the leg muscles. A pre‐test and a post‐test covering the material were administered to both groups. Students who worked with the felt muscles showed a greater improvement relative to the students who studied independently (p<0.05). The groups switched activities when studying the foot muscles and similar results were found. Student satisfaction was assessed with a survey administered after each unit. The majority of students indicated that the activity was fun and felt that the activity was an effective way to learn the material. We advocate the use of this type of activity to engage the students and to provide an opportunity for the students to reinforce their understanding of muscular attachments.
Beneficial Aspect of Cadaver Dissection – Medical Students Perspective
Authors: Kraszpulska Barbara 
Published: June 2021

The aim of the study was to assess the beneficial aspect of cadaver dissection in the process of learning anatomy. The objectives of the study were: to determine medical students’ perspectives regarding if and how cadaver dissection was beneficial, and to determine the degree of respect toward the donors.An anonymous survey was completed by 103 first‐year medical students (42 female; 61 male) at WSU at the end of the anatomy course (involving complete cadaver dissection). Ninety five percent of the students found the dissection to be beneficial. They listed as the most valuable aspects of doing dissection: seeing 3D orientation of anatomical structures (53%), helping with assigned material (40%), and understanding relationships in the human body (21%). All students (100%) declared appropriate respect and appreciation for the donors, although less than half (48% male; 34% female) would consider donating their own body. Medical students themselves were generally in favor of organs donations ‐ 90% male and 74% female. Ninety‐five percent of students disagreed with the idea of completely replacing cadaver dissection with computer‐assisted program.These results suggest that medical students regard cadaver dissection as an effective time learning experience, which reinforces lecture and textbook material and provides a better understanding of the complicated relationships in the human body.
Accurate Accountability of Anatomical Donors and Material Permits Quality Control for the Medical School and Maintains Public Trust
Authors: McGillicuddy Michael P. , Caceres Noel , Marquez Samuel 
Published: June 2021

SUNY Downstate recently purchased a state‐of‐the‐art accountability program system from SRS Computing (SRSC) for tracking donors in our school. The program was used originally for the Cremation Industry. The SRSC program offers a robust managing system that permits the direct application for medical school education. It allows the tracking of donors from the moment of receipt by assigning a number in the form of a bar code that is synchronized to the Case Management Tracking System, which then monitors the donors throughout the duration of the course. The donor is logged at the time of arrival to Downstate and is constantly monitored each & every step the donor is being used. If pro‐sections are taken from the donor for the purposes of medical research, that section is labeled with a bar code tag permitting tracking of the specimen until its return to the donor at the end of the study. This system allows Downstate to return the trust given to the school from each of the donor families that all Donors & Donor materials are returned for their intended disposition. The system allows selected data to be readily available for review such as date of death and age including the donor's family request for the disposal of cremains at the end of the study or course. In the past there have been allegations of body parts and materials being sold for profit, this tracking system ensures the location of any and all donors and donor materials in real time.Grant Funding Source: None
Crystallization of Escherichia coli 70S Ribosome in complex with tRNAs
Authors: Young Brandon , Steitz Thomas A. , Innis Axel 
Published: June 2021

The prokaryotic ribosome is responsible for translation, the process of creating polypeptides from amino acids. Specifically, ribosomes translate the codons in mRNAs into polypeptide chains. The prokaryotic ribosome is made up of a large subunit and a small subunit, which are known as the 50S (Svedberg units) and 30S subunits, respectively. The larger subunit catalyzes peptide bond formation within the peptidyl transferase center, while the smaller subunit mediates decoding of the mRNA. Together they make up the 70S prokaryotic ribosome. Studies have shown that certain nascent polypeptides can bind to the exit tunnel, resulting in inhibition of peptide bond formation which is referred to as ribosome stalling. Through the use of X‐Ray crystallography we will crystallize the 70S ribosome in complex with tRNAs and a nascent chain within the exit tunnel producing a structure. Although, no structure has been produced yet, we have been able to narrow down conditions that will optimize the growth of these crystals. This optimization will allow for production of the structure which will result in us being able to further explore the process of nascent chain mediated‐translational arrest in the Escherichia coli (E. coli) 70S ribosome.This research was supported in part by a grant from the Howard Hughes Medical Institute.
Improvement of defective Cystic fibrosis airway epithelial repair after CFTR rescue
Authors: Trinh Nguyen Thu Ngan , Adam Damien , Bardou Olivier , Anik Privé , Maillé Émilie , Lingée Sarah , Ferraro Pasquale , Desrosiers Martin , Coraux Christelle , Brochiero Emmanuelle 
Published: June 2021

Airway damage and remodelling is an important component of the progression of lung cystic fibrosis (CF) pathology. Our histological analyses of bronchial tissues from CF patients revealed several ultrastructural anomalies, including epithelial desquamation and tissue remodelling. Although some areas were in the process of epithelial regeneration, the repair mechanisms seemed obviously insufficient in maintaining epithelial integrity in CF airways. Our aim was thus to study the CFTR function in airway repair processes. Wound‐healing assays were performed to study the early phase of epithelial repair, in particular cell migration, in human CF and non‐CF airway cells. Our experiments showed that airway cells from CF patients repaired slower than non‐CF cells. In addition, CFTR inhibition or silencing in non‐CF primary airway cell monolayers partially inhibited wound‐closure. Interestingly, wt‐CFTR transduction in CF airway cell lines or CFTR correction with VRT‐325 in CFBE‐ΔF508 cells and primary CF bronchial monolayers significantly improved wound‐healing. Altogether our results demonstrated that functional CFTR play a critical role in wound‐repair, and CFTR correction may represent a novel strategy to promote the airway repair processes in CF.Project funded by CFC (Cystic Fibrosis Canada).
MITOCHONDRIA‐TARGETED TREATMENT ATTENUATES VASCULAR OXIDATIVE STRESS, ENDOTHELIAL DYSFUNCTION AND HYPERTENSION
Authors: Dikalov Sergey , Nazarewicz Rafal , Bikineyeva Alfiya , Dikalova Anna 
Published: June 2021

Mitochondrial reactive oxygen species (ROS) have been recently implicated in the development and progression of cardiovascular diseases. We have suggested that mitochondria targeted antioxidants can be beneficial for treatment of endothelial dysfunction, hypetension other parthological conditions. In this work we have studied specific molecular mechanisms responsible for stimulation of mitochondrial O2• and its downstream targets using cultured human aortic endothelial cells and angiotensin II‐induced hypertension. Depletion of cytoplasmic Nox2 isoform of NADPH oxidase inhibited stimulation of mitochondrial ROS by angiotensin II. Meanwhile, treatment with the mitochondria targeted superoxide dismutase mimetic mitoTEMPO or SOD2 overexpression inhibited production of mitochondrial O2•, reduced activity of redox dependent c‐Src and cytoplasmic NADPH oxidase, restored vascular NO in angiotensin II stimulated cells and reduced blood pressure in mice with angiotensin II and DOCA‐salt induced hypertension. We have suggested that mitochondrial ROS stimulates cytoplasmic oxidative stress via release of H2O2. Indeed, scavenging of mitochondrial H2O2 with mitochondria targeted glutathione peroxidase mimetic mitoEbselen inhibited production of cytoplasmic O2• by NADPH oxidase and reduced blood pressure in angiotensin II induced hypertension. Production of angiotensin II‐induced mitochondrial ROS was inhibited by specific PKCε peptide antagonist, ATP‐sensitive potassium channel blocker 5‐hydroxydecanoate, inhibitors of complex I rotenone, inhibitors of complex II malonate or malate. These studies show that mitochondrial ROS play an important role in feed‐forward stimulation of oxidative stress by activating mitochondrial reverse electron transfer which can be treated with mitochondria‐targeted antioxidants.This work has been funded by National Institutes of Health HL094469.
Effects of storage conditions on soil molecular biology for forensic science purposes
Authors: Larson Sabreena A , Patel Niraj , Freeman Victoria , Hustedt Jason , Drijber Rhae , Carter David , Bailey Cheryl 
Published: June 2021

The object of this project is to identify changes in soil microbial community DNA and Fatty Acid profiles caused by storage, using capillary electrophoresis single‐strand conformation polymorphism (CE‐SSCP) and fatty acid methyl ester (FAME) analysis.There were six different storage treatments tested (−80°C, −20°C, 4°C, freeze dried, air dried, oven dried) on soil samples collected from 4 different locations in Nebraska. Samples were taken at a depth of 0.0 cm to 5.0 cm from three biological replicates at each collection site. After collection, soils were sieved and subsamples were placed into each storage treatment for five weeks then analyzed, with the exception of one subsample (fresh) which had DNA and fatty acids extracted within 36 hours of collection. Additional fresh samples were collected and processed two weeks after initial collection and seasonally.The CE‐SSCP and FAME analysis will illustrate the microbial community and its diversity for an individual soil sample through the number of peaks, molecular size of peaks and relative peak heights. Using statistical analysis it can be determined which storage treatments altered the soil microbial community profile when compared to the fresh subsample. Results will be presented to show the effectiveness of these two methods to detect small variations in the soil microbial community. Funded by National Institute of Justice (25‐6228‐0159‐001).
Association of Berberine into hydrogel as drug delivery system
Authors: Souza Camila R , Lunardi Claure N 
Published: June 2021

Hydrogels are hydrophilic polymer networks having the capacity to absorb water, ranging from about twenty to thousand times their dry weight. Berberine (BS) is a natural product, a quaternary ammonium salt from the group of isoquinoline alkaloids found in medicinal plants as Berberis Vulgaris. It has been reported that BS has some activity against and has already been used to treat dysentery, hypertension, inflammation, and liver disease in China and Japan. In this work, berberine was used as a model drug to study the its association with different types of hydrogel of Polyvinyl alcohol (PVA 10%); gellan gum (GG 2%) and gellan gum‐PVA crosslinked with cysteine (cys) (GG2%PVA2%2%cys) preparations. Parameters such as fraction of retained water (Wf); hydration percentage (%H); Swelling (DSw) and time course profile (t=100%) (TC) were evaluated.The results showed that the GG2%PVA2%cis was able to retain more water and swelling than the other preparations. The time course of release of the BS to the medium was greater for GG2%PVA2%cys making it a candidate to drug delivery studies in biological assays. Supported by FAPDF, CAPES, CNPq, FINATEC and DPP‐UnB.
Case‐Control Study of Eating Behaviors of Young Adults with and without Diet‐Related Chronic Health Condition (DRCHCs)
Authors: Quick Virginia , McWilliams Rita , Byrd-Bredbenner Carol 
Published: June 2021

Young adults with DRCHCs (i.e.,type 1 diabetes, celiac disease, cystic fibrosis, inflammatory bowel disease, irritable bowel syndrome) must adhere to dietary restrictions to control their diseases. These restrictions may put them at risk for disturbed eating. This study aimed to determine if young adults (18–26 years) with and without DRCHCs differed with regard to disturbed eating behaviors (Binge Eating; Inappropriate Compensatory Behaviors; Eating, Weight, and Shape Concerns; and Restraint scales from Eating Disorders Examination Questionnaire; Emotional and Disinhibited Eating from Three‐Factor Eating Questionnaire; and Night Eating Questionnaire). Each DRCHC participant (cases=166) was matched with 4 healthy participants (controls=664) based on gender and BMI (±0.50 BMI units). Conditional logistic regression analyses indicated cases were twice as likely to have been diagnosed by a healthcare provider with an eating disorder (p=0.08, OR=1.99, CI90 [1.03–3.83]). Cases were significantly more likely to use excessive exercise to manage their weight (p=0.04, OR=1.41, CI95 [1.02–1.94]) and more likely to misuse medicine (e.g., laxatives) to control their weight (p=0.04, OR=1.14, CI95 [1.00–1.29]) than controls. Findings indicate that healthcare professionals should screen DRCHC patients for disturbed eating behaviors and eating disorders.Funding: Kappa Omicron Nu FellowshipSociety Topic Area: ASN‐5002 Nutrition interventions for risk factor modification in chronic disease