bluuebunny/crossref_metadata_2025 huggingface dataset:
A prospective, randomized, placebo‐controlled trial of high‐dose vitamin D for prevention of vitamin D insufficiency in early stage chronic kidney disease
Authors: Alvarez Jessica A , Law Jennie , Coakley Kathryn , Wright Breanne , Hao Li , Ziegler Thomas R , Tangpricha Vin
Published: June 2021

Objective

Vitamin D insufficiency [25‐hydroxyvitamin D [25(OH)D] <30 ng/ml] is common in individuals with chronic kidney disease (CKD), even in the earlier stages. We aimed to determine if high‐dose vitamin D3 could prevent vitamin D insufficiency in early stage CKD (stage 2 and 3).

Methods

: This was a prospective, randomized, controlled trial of vitamin D3 (cholecalciferol 50,000 IU per wk for 12 wks followed by 50,000 IU every other wk for the remainder of the yr compared to placebo, ClinicalTrials.gov # NCT00781417). Serum 25(OH)D and PTH were measured at baseline, 12 wks, and 52 wks.

Results

48 subjects were enrolled. At baseline, 57% of patients were vitamin D insufficient. In the vitamin D group, insufficiency rates decreased to 20% and 14% at 12 and 52 wks, respectively, and were lower (P<0.05) than the placebo group (77% and 47% at 12 and 52 wks, respectively). Mean serum 25(OH)D increased from 26.7 ± 6.8 to 42.8 ± 16.9 ng/ml in the D group at 12 wks (P<0.05) and remained elevated at 1 year. PTH decreased in the D group by 12 wks (−16.9 ± 30.3 pg/ml, P<0.05). 25(OH)D or PTH did not change in the placebo group.

Conclusions

Our high‐dose vitamin D3 treatment strategy for 1 yr was sufficient to improve and maintain optimal 25(OH)D concentrations in the majority of subjects with early stage CKD. Further study should assess the impact of early vitamin D therapy on other clinical outcomes.

Grant Funding Source : Emory, Atlanta Veterans Affairs, T32 DK007298‐31
Transcriptional regulation of the Tocopherol Transfer Protein
Authors: Ulatowski Lynn , Dreussi Cara , Manor Danny
Published: June 2021

Vitamin E is a family of neutral plant lipids of which RRR α‐tocopherol (tocopherol) is selectively retained in the body and thus is considered the most biologically active form. Tocopherol's ability to scavenge free radicals defines tocopherol as the major lipid‐soluble antioxidant in humans. Tocopherol's essential function in humans has been recognized for decades, as well as its specific role in protecting cells from oxidative stress‐related damage. Low tocopherol levels were found to associate with increased risk for oxidative stress‐related diseases, like Alzheimer's and cardiovascular disease. Tocopherol transfer protein (TTP) is a cytosolic protein that facilitates intracellular transfer of tocopherol and maintains whole body vitamin E status. Mutations in TTP result in the heritable disease ataxia with vitamin E deficiency (AVED; OMIM #600415), characterized by progressive spinocerebellar ataxia and very low vitamin E levels. Despite the well‐documented role of TTP in maintaining normal tocopherol status, the mechanisms that regulate transcription of the TTPA gene are unknown. Our study found that expression of the TTPA gene in immortalized human hepatocytes is significantly enhanced upon treatment with chemical agonists of the transcription factors PPARδ, RXR, and CREB, as well as by H2O2 and a hypoxia mimetic. We also cloned a 2 Kb proximal region of the human TTP promoter, and used it to drive expression of a reporter construct. Promoter “bashing” experiments revealed distinct sites of transcriptional repression and activation. Finally, we show that single nucleotide polymorphisms (SNPs) have profound impact on the transcriptional activity of the cloned TTP promoter. These findings may explain altered responses to vitamin E supplementation in human clinical trials. Taken together, our results suggest that TTP is indeed transcriptionally regulated and this regulation may have a profound effect on vitamin E status in humans.

Grant Funding Source : T32DK07319 and RO1DK067494
Using the phenylalanine (PHE) stable isotope pulse method to measure intracellular protein breakdown and metabolic shunting in the context of sepsis in the pig
Authors: Ten Have Gabriella A , Engelen Marielle P , Wolfe Robert R , Deutz Nicolaas E
Published: June 2021

To develop an easy applicable clinical method to measure protein metabolism, we tested the PHE stable isotope pulse method (PULSE) and compared the results with a simultaneously performed primed‐continuous infusion protocol (CONTI) during sepsis in pigs with stimulated whole body protein breakdown (WbRa).

12 pigs with Pseudomonas aeroginosa induced hyperdynamic sepsis and 8 controls were studied 17h after start. WbRa CONTI with L‐[13C6]PHE and WbRa‐PULSE with L‐[15N]PHE. Plasma PHE enrichment by LC‐MS, WbRa‐CONTI = I/TTR, compartmental analysis (SAAM II) PULSE. Data (mean (SE)); stats (Prism).

WbRa‐CONTI was 22% higher in septic pigs {924 (23) to 1130 (46) nmol/kg BW/min} (p=0.0032), while WbRa‐PULSE 36% {1722 (66) to 2342 (191); p=0.0164}. Sepsis show a substantial expansion of pool 1 and 2 and metabolic shunting (pool 2><1), while intracellular PB increased 31% {2620 (82) to 3425 (305); p=0.0426}. Ra‐PULSE relates to intracellular PB (p<0.0001) and Ra‐CONTI to Ra‐PULSE (P<0.001).

Changes in Ra‐CONTI reflect intracellular PB with high sensitivity. PULSE in addition shows that sepsis in the pig is related to a substantial increase in metabolic shunting in relation to expansion of the pools (probably intra‐ and extracellular). It is suggested that the CONTI method can be replaced with the easy applicable PULSE method that also provides additional information about intracellular fluxes and breakdown.
Radical hysterectomy teaching module with a three‐dimensional digital model of the female pelvis
Authors: Dunnigan Samantha Kay , Nguyen Hai M , Johnson Marjorie
Published: June 2021

Performing radical hysterectomy requires thorough knowledge of pelvic anatomy, which is complex and difficult to visualize. The goal of this project is to create a teaching tool to enable surgeons worldwide to improve their understanding of pelvic anatomy, leading to less invasive procedures and better patient outcomes. The pelvic model was created using AMIRA, a 3D segmentation and surface‐rendering program. Anatomical areas of interest were labeled on consecutive 2D slices from the Visible Human Project, which were then reassembled to form a 3D model and incorporated into animated videos. An animated instructional program based on radical hysterectomy was created, which includes rotational viewing, real‐life video, clinical narration, correlated imaging, labeling and a glossary. The glossary standardizes anatomic and surgical terminology to help overcome language barriers when training doctors whose native language is not English. Due to the detail, ease of use and versatility, this tool will be an excellent resource for surgeons training to perform radical hysterectomy, allowing for better patient outcomes especially in developing countries where this technology was not previously available.

Grant Funding Source: Ontario Graduate Scholarship
Integration of anatomy, literature research skills and clinical practice through a peer‐teaching and peer‐assessment exercise at pre‐clinical level in an undergraduate medical course
Authors: Filgueira Luis , Eppler Elisabeth
Published: June 2021

The aim of this educational project was to explore the feasibility of enhancing clinical anatomy teaching in an undergraduate medical course by engaging a sub‐group of advanced preclinical students (n=54/186) in providing lecture material as a group presentation (n= 3). Focussing on specific syndromes or diseases, the aims of the educational exercise were to enhance knowledge in clinical anatomy, but also to integrate anatomy with clinical topics. Additional educational aims were to train and enhance literature research, team work, oral communication and presentation skills, as well as to engage students in peer‐teaching and peer‐assessment exercises.

The lectures were recorded for revision purpose and the content tested using a multiple choice exam at the end of semester. The group presentations were assessed by the peers. Additional self and teacher assessment of the presentations was done.

At the end of semester, students’ perception of this educational exercise was assessed through an online questionnaire. In addition, marks of peer, self and teacher assessment, as well as final marks for this and other units were statistically analysed to evaluate for the sub‐group and the whole class whether this educational exercise enhanced the average student outcome.

Study evaluation indicates that the described educational exercise is feasible and valued by the students.
Curricular Integration: Not what we are bringing together, but who
Authors: Hopkins Robin
Published: June 2021

Across the professions (nursing, teaching, engineering, medicine, law, etc.) there is a call for early training to be taught in a way that is more relevant to practice in the field. National reports making recommendations on the future training of professionals emphasize the integration of classroom‐based knowledge with practice‐based knowledge. In medical education the call is for the basic medical sciences, which are typically taught by basic scientists in the university setting during the first two years of medical school, to be integrated with the context of clinical practice. The implementation of integration is presented as curricular change, with the assumption of relatively unproblematic collaboration between basic scientists and physicians to teach medicine in clinically relevant ways. One implication of such reform is that basic scientists, more than clinicians, will be required to change their former ways of knowing, teaching, and assessing performance in the basic sciences. This is not trivial, as it involves not only their sense of competency, but also their very identity as professionals and academics. Drawing from the literature on educational change, in the context of anatomy education, this presentation will introduce a conceptual framework that will help us better think about and investigate the experience of making such a shift in professional competence and identity for anatomists.

Grant Funding Source : UBC Four Year Fellowship
Anatomy Education through Dissection and Peer Teaching: Special Dissections of the Head and Neck
Authors: Koba Alexandra M. , Johnson Laura , Babin Kathryn P.
Published: June 2021

Special dissections of the head and neck were performed in the context of an undergraduate human anatomy course to enhance the human anatomy curriculum through peer teaching dissection and the use of learning aide's created by the students. The purpose of peer teaching of dissection in the undergraduate setting was to present an entirely unique approach where the students themselves can re‐create and advance a hemi and coronal section of the head and neck, along with intricate dissections of the ears, eyes, and oral cavity. The dissections allowed a regional approach for students to think critically and allow different types of learners to complement each other's problem solving skills. Students were able to manipulate and understand how various systems overlap in the head and neck and therefore be able to visualize internal structures. A step‐by‐step dissection manual was created where new dissections and techniques were recorded and often referenced. The use of this study tool and peer teaching of dissections has been a powerful means of advancing the undergraduate anatomy curriculum and learning anatomy as a dynamic basis for future pre‐clinical studies.

Grant Funding Source : Grant‐in‐Aide‐of‐Research, GIAR, Administered by Sigma‐Xi, the Scientiific Research Society, San Francisco State University IRA Grant.
Myocardial Regeneration through Tissue Engineering
Authors: Vunjak-Novakovic Gordana
Published: June 2021

The overall goal of cardiac tissue engineering is to re‐establish the structure and function of infarcted myocardial tissue. Interdisciplinary research at the interface of stem cells and bioengineering holds promise that functional cardiac tissue grafts can be generated in vitro, and used for implantation as well as models to study cardiac development and disease. In all cases, the notion is that the biological potential of cardiovascular cells can be mobilized by providing controllable environments that regulate cell differentiation and assembly by factors present during native development. Key requirements for cardiac regeneration include an appropriate cardiac tissue matrix, electromechanical cell coupling, and functional vascularization. This talk will discuss some recent and ongoing work towards meeting these requirements. We will also discuss the important interactions between the engineered tissue grafts and the inflammatory environment of the infarct bed that largely determine the functional outcomes of tissue engineered heart repair.

Funding: NIH (HL076485, EB002520, RR026244), Helmsley, NYSTEM
Mechanisms of ESCRT‐mediated cargo sorting and degradation
Authors: Audhya Anjon , Schuh Amber , Mayers Jonathan , Fyfe Ian , Edwardson Michael
Published: June 2021

Vesicle‐mediated cargo transport within the endomembrane system requires precise coordination between adaptor molecules, which recognize sorting signals on substrates, and factors that promote changes in membrane architecture. At endosomal compartments, a set of protein complexes collectively known as the ESCRT machinery sequesters transmembrane cargoes that harbor a ubiquitin modification and packages them into vesicles that bud into the endosome lumen. Using atomic force microscopy, we define the architecture of each ESCRT complex in the presence of physiologically relevant lipid bilayers. This analysis indicates that ESCRT‐0, ESCRT‐I, and ESCRT‐II function as heterotetrameric complexes that collectively bind and sequester ubiquitin‐modified cargoes. Additionally, our analysis reveals a mechanism by which ESCRT‐III membrane scission activity is both spatially and temporally regulated during multivesicular endosome biogenesis. A revised model describing ESCRT function during protein sorting in the endocytic pathway will be presented.
Functional genomics to decipher cancer dependencies and mechanisms
Authors: Hahn William C
Published: June 2021

Recent advances in genomics now make it possible to consider enumerating all of the genetic lesions in specific cancers. While these approaches will yield critical information regarding the identify, number, and types of alterations found in human tumors, a complementary approach to decipher the molecular basis of malignant transformation depends upon the application of genome scale tools to annotate the function of genes involved in cancer initiation and progression. Over the past several years, we have developed genome scale RNAi libraries and open reading frame expression libraries that permit a systematic evaluation of genes involved in cancer initiation and maintenance. Using these libraries, we have now performed screens in a panel of human cancer cell lines to systematically identify cancer vulnerabilities. By combining these functional approaches with information derived from mapping the structural abnormalities present in cancer genomes, we have identified several new oncogenes that contribute to cancer development. In pilot studies, we have identified several genes that act as synthetic lethal partners to known oncogenes or mediate resistance to targeted therapies. In recent work, we have expanded the scale of these experiments to genome scale in more than 100 cell lines. Recent insights from these efforts will be discussed. Taken together, these studies suggest that combining forward and reverse genetic approaches with information derived from the cancer genome characterization projects will yield a comprehensive list of cancer vulnerabilities and establish a general approach for the rational identification of oncogenic and co‐dependent pathways in cancer.
Asymmetry and aging of mycobacterial cells: a novel mechanism of diversification
Authors: Fortune Sarah M
Published: June 2021

Cells use both deterministic and stochastic mechanisms to generate cellâ€“to‐cell heterogeneity, which enables the population to better withstand environmental stress. Within a clonal population of mycobacteria there is significan heterogeneity in elongation rate that arises because mycobacteria grow in an unusual, unipolar fashion. Division of the asymmetrically growing mother cell gives rise to daughter cells with different functional properties. Importantly, the physiologically distinct subpopulations of cells that arise through asymmetric growth and division are differentially susceptible to clinically important classes of antibiotics.
TorsinA – unlocking the mysteries of an AAA+ ATPase in the endoplasmic reticulum and nuclear envelope
Authors: Hanson Phyllis I. , Heyden Abigail Vander , Naismith Teresa V.
Published: June 2021

TorsinA is an AAA+ ATPase in the endoplasmic reticulum (ER) and nuclear envelope (NE) lumen that is mutated in dominantly inherited early onset DYT1 dystonia. TorsinA is an essential protein in mice, but its cellular function and the basis for disease are not well understood. We have previously shown that localization and activity of torsinA within the ER and NE are dynamically regulated by an interacting transmembrane protein, LULL1 (TOR1AIP2). To understand how torsinA responds to LULL1, we now identify features in both proteins responsible for controlling torsinA localization. We find that distinct regions mediate binding and torsinA re‐targeting, and establish among other things that the extreme N‐terminus of LULL1 must be present for torsinA to be directed to the NE. The N‐terminus of LULL1 is cytosolic, while its torsinA‐binding domain is lumenal, raising the intriguing possibility that control of torsinA localization involves communication between the cytosol and the ER lumen. When retargeted to the NE, torsinA displaces a subset of linker of the nucleoskeleton and cytoskeleton (LINC) complex proteins and may thereby modify NE structure and nuclear positioning. The disease causative torsinA mutant also responds to LULL1, but only weakly displaces LINC proteins.

These findings identify a mechanism for regulating torsinA localization and activity via LULL1, and suggest that activity at the NE is perturbed by the disease‐causative mutation. Changes in torsinA activity at the NE may contribute to DYT1 dystonia.
Unbiased identification of novel AMPK substrates by chemical genetics
Authors: Brunet Anne
Published: June 2021

The energy‐sensing AMP‐activated protein kinase (AMPK) is activated by low nutrient levels. We showed that AMPK mediates lifespan extension by some dietary restriction regimens in C. elegans. Functions of AMPK other than its role in cellular metabolism are beginning to emerge. To identify novel AMPK substrates in mammalian cells, we used an unbiased chemical genetic screen. We created an analog‐specific version of AMPK that uses a bulky analog of ATP and transfers the phosphate group to specific substrates in cells. Coupled with mass spectrometry, this approach allowed us to identify 30 novel AMPK substrates. Our results indicate that AMPK phosphorylates components of the mitotic apparatus, providing an intriguing link between energy sensing and the completion of mitosis. Our findings indicate that AMPK coordinates nutrient status with mitosis completion, which may be critical for the organism's response to low nutrients during development, or in adult stem and cancer cells. These studies may also help uncover mechanisms by which dietary restriction extends lifespan.
A novel role of a lysosomal acid lipase in the regulation of longevity
Authors: Wang Meng
Published: June 2021

Lipid metabolism plays a key role in human health and longevity. Dysregulation of lipid metabolism has been implicated in many age‐related diseases. As a major site of lipid metabolism, adipose tissue exerts crucial endocrine effects on aging. However, it remains poorly understood how lipid metabolism is coupled to lifespan control. We found a lysosomal acid lipase that promotes lifespan and healthspan in C. elegans. The underlying molecular mechanisms will be discussed in the talk.
Systemic polyhydroxylated fullerene toxicity in fathead minnows (Pimephales promelas Rafinesque, 1820)
Authors: Whitley Elizabeth M. , Jovanovic Boris , Palic Dušan
Published: June 2021

Carbon nanoparticles have the potential to serve as nanopharmaceutical delivery systems for drugs, genes, and immunogens. They are able to both generate and scavenge reactive oxygen species (ROS) and, like some other nanomaterials, may induce autophagy. Controversy exists regarding their potential for toxicity. Systemic administration of hydroxylated fullerene nanoparticles to fish has been reported to result in renal accumulation and nephropathy, as well as impairment of innate immune responses. To better characterize biocompatibility and the potential for adverse systemic effects, we administered the water soluble fullerenol, C60(OH)24, to fathead minnows (Pimephales promelas), a model organism for environmental toxicology studies. Briefly, adult minnows were injected intraperitoneally with 20 micrograms/g, the lowest lethal dose observed in a preliminary study. Fish died or were euthanized moribund within 36 hours post‐injection. Sections of whole fish from control and treatment groups were evaluated using a four‐category scoring system comprising 31 different histologic features. Statistically significant tissue changes were observed in the kidney and sensory system and included renal edema, degeneration of tubular epithelium, pigment accumulation and loss of hematopoietic tissue in the trunk kidney and degeneration or death of ciliated sensory cells in the ear. Investigation into cellular and molecular mechanisms associated with these morphologic changes may provide valuable information to direct development of safe fullerene‐based delivery systems. This work was supported by the Iowa State University Office of the Vice President for Research.
Liver Morphologic Changes Induced by Semicarbazide
Authors: Da Costa Miguel Mendes , Duarte Ana Rute , Capela-Silva Fernando , Farinha Rodrigo , Cabrita António Silvério
Published: June 2021

This study evaluates the liver morphologic changes associated with the ingestion of semicarbazide contaminated food.

Two groups of 10 Wistar rats were studied. Group 1 was submitted to no manipulation and group 2 was submitted to the administration of food contaminated with semicarbazide at a concentration of .6g/Kg. All animals were sacrificed after four weeks and a complete necropsy was performed. The slides of liver stained by HE were evaluated for morphologic lesions.

In group 1 no significant morphologic changes were found. In group 2 some morphologic lesions were found such as sinusoidal congestion, small focal haemorrhages, cellular hyaline infiltration and oedema, macrovesicular steatosis, bile duct hyperplasia and flooding of cell's cytoplasm. Anisocariose was observed in all cases, from which pyknosis and nucleoli increased by angular nuclei were the most evident. The most marked lesions observed in the liver were hyaline infiltration, bile duct hyperplasia and flooding of cell's cytoplasm. Also some inflammatory tissue was found in the organ with the presence of lymphocytes and macrophages.

We may conclude that ingestion of semicarbazide at high doses in short period of time is hepatotoxic.
Elucidating the role of β‐catenin in hepatocellular tumor angiogenesis
Authors: Delgado Evan R , Monga Satdarshan Pal
Published: June 2021

Angiogenesis is a process that plays an important role in tumor growth and progression. Various anti‐angiogenic therapies have been pursued such as targeting Vascular Endothelial Growth Factor (VEGF) with mixed results. Hepatocellular carcinoma (HCC) is the 3rd leading cause of death by cancer, and angiogenesis has been thought to be aiding in the disease process. Since, hyperactive Wnt signaling is implicated in roughly 30% of all HCC cases due to stabilizing mutations in the β‐catenin gene, we hypothesized its important role in regulating angiogenesis through modulation of factors such as VEGF and FGF‐2, shown independently to be targets of β‐catenin pathway. In this study we show for the first time that various factors such as EphrinA1, FGF‐2, VEGF‐A, and Urokinase among others are regulated by β‐catenin in HCC cell lines. We used a novel antisense oligonucleotide known as Peptide Nucleic Acids (PNA) that targets the transcription start site of β‐catenin gene and is efficaciously taken up by the cell. After PNA treatment of HepG2 cells, β‐catenin knockdown was verified by Real Time‐PCR and Western Blot. A Real Time‐PCR array was next used to analyze the effect on factors involved in angiogenesis. We identified several pro‐angiogenic factors that were downregulated upon β‐catenin suppression. Thus, our studies show a role of β‐catenin in angiogenesis, particularly in HCC, where a therapy targeting β‐catenin may not only impact cell viability and proliferation, but also reduce tumor angiogenesis.
MG624, an α7‐nicotinic receptor antagonist, inhibits angiogenesis in human small cell lung cancer
Authors: Dom Aaron M. , Brown Kathleen C. , Lau Jamie K. , Witte Theodore R. , Hardman W. Elaine , Luo Haitao , Chen Yi Charlie , Dasgupta Piyali
Published: June 2021

Small cell lung cancer (SCLC), a highly angiogenic tumor, displays a 90% association with smoking. Nicotine promotes angiogenesis in lung cancer via alpha‐7‐nicotinic acetylcholine receptors (α7nAChRs). Therefore, α7nAChR antagonists should attenuate nicotine‐induced angiogenesis and be useful for the therapy of SCLC. Here we show that the α7nAChR antagonist MG624 suppresses proliferation of human microvascular endothelial cells from the lung (HMEC‐L) in response to nicotine. MG624 displays potent anti‐angiogenic activity in the Matrigel, rat aortic ring and chicken chorioallantoic membrane models. MG624 also inhibited nicotine‐induced angiogenesis in human SCLC tumors xenografted in athymic mice. The administration of MG624 did not cause any discomfort or lethargy in mice. Chromatin immunprecipitation assays show that MG624 decreases the recruitment of Egr‐1 on FGF2 promoter thereby decreasing its transcription and suppressing nicotine‐induced angiogenesis. Our results suggest that anti‐angiogenic agents like MG624 may be useful for the therapy of human SCLC.
Inhibitory effects of Macrolactin A and 7‐O‐succinyl macrolactin A on angiogenesis and cancer cell invasion
Authors: Kang Youra , Park Sumin , Kim Hyun Young , Kim Dong Hee , Kim Jung-Ae
Published: June 2021

The macrolactin compounds are known to have broad range of pharmacological activities such as antibacterial, anti‐viral, neuro‐protective, and antiproliferative activities. In the present study, we investigated anti‐angiogenic activities of macrolactin A (MA) and its derivatives, 7‐O‐malonyl macrolactin A (MMA) and 7‐O‐succinyl macrolactin A (SMA) using in vivo chick chorioallantoic membrane (CAM) assay. MA and SMA showed strong inhibitory activity against not only growth factor (VEGF)‐induced but also inflammatory cytokine (TNF‐α and IL‐8)‐induced angiogenesis. However, MMA did not show any inhibitory effect. In in vitro angiogenesis using human umbilical vein endothelial cells (HUVECs), MA and SMA significantly inhibited VEGF‐induced angiogenic processes, proliferation, tube formation and invasion of HUVCs. We also found that MA and SMA concentration‐dependently inhibited HT‐1080 human fibrosarcoma cell invasion in a matrigel‐coated transwell assay. Taken together, our results suggest that MA and SMA may be valuable drug candidates for treating cancer‐ and chronic inflammatory disease‐associated abnormal angiogenesis as well as cancer invasion.
Pro‐Angiogenic Capacity of MMP‐9 Produced by Different Types of Inflammatory Leukocytes is Determined by the Levels of TIMP‐1 Complexed with the MMP‐9 Proenzyme
Authors: Zajac Ewa , Schweighofer Bernhard , Kupriyanova Tatyana , Casar Berta , Rimann Ivo , Juncker-Jensen Anna , Deryugina Elena , Quigley James P
Published: June 2021

By delivering a potent matrix metalloproteinase, MMP‐9, inflammatory neutrophils and macrophages (mε) play an important role in tumor angiogenesis. We have previously demonstrated that uniquely high angiogenic capacity of neutrophil MMP‐9 (nMMP‐9) is determined by its TIMP‐free status. Herein, we compared biochemically and functionally nMMP‐9 with the MMP‐9 produced by monocytic cells, including monocytes and innate (M0), tumor suppressing (M1), and tumor promoting (M2) macrophages (mε). Angiogenic potential of a specific inflammatory leukocyte type was related to gene and protein expression levels of TIMP‐1 and MMP‐9, ultimately determining how much TIMP‐1 is complexed with secreted MMP‐9. Resembling neutrophils that release TIMP‐free MMP‐9, M2 mε produce MMP‐9 complexed with very low amounts of TIMP‐1. Correspondingly, M2 mε induced relatively high levels of neovascularization, yielding only to neutrophils in their angiogenic potential. Low TIMP‐1/MMP‐9 ratios inversely correlated with the high proteolytic activity of nMMP‐9 and M2 mε MMP‐9 in a substrate cleavage assay. Finally, the lack of TIMP‐1 in nMMP‐9 and low TIMP‐1 burden in M2 mε MMP‐9 were functionally linked to their high capacity to induce new blood vessel formation, thereby highlighting a biochemical mechanism underlying unique angiogenic potential of MMP‐9 delivered by inflammatory neutrophils and pro‐tumorigenic macrophages.
Base Pair: High school teacher professional development creates generational science education impact
Authors: Rockhold Robin William , Sullivan Donna , McDaniel Olga
Published: June 2021

Base Pair, a collaboration between the University of Mississippi Medical Center (UMMC) and a diverse group of secondary schools, uses research‐based, experiential development of teachers to amplify the impact of a biomedical research mentorship program on regional secondary school science education. Initiated in 1991–92, Base Pair brings high school teachers into UMMC for summer training in subjects from use of inquiry labs including DNA isolation, bacterial transformation, protein purification and quantitation, restriction enzyme digestion of DNA, gel electrophoresis, PCR analysis for DNA associated with genetically modified food, and enzyme linked immunosorbent assays to grant‐writing and biotechnology curriculum design. A total of 119 teachers have been trained and 120 teacher‐initiated grants have resulted from Base Pair‐trained teacher applications. As a result, a 74% rate of funding and approximately $438,190 new funds have been generated. A core of teacher trainees who continue regular interaction with UMMC mentors have initiated 4 novel inquiry‐based curricula that have directly impacted over 1,000 students, and a science research course that has been adopted by the state department of education. Core teachers also delivered workshops to over 600 additional teachers in local, regional and national meetings and are authors on 83 published scientific presentations. Mentorship of teachers has increased science education resources, advanced teacher professionalism and extended the impact of UMMC outreach. (Supported by Howard Hughes Medical Institute)
Integrating statistical analysis and experimental design into life science curricula with secondary teachers
Authors: Clase Kari Lynn , Adedokun Omolola , Charles Andrea , Fatum Leslie , Ha Soo , Janowiak Kirk , Parker Loran , Pelaez Nancy , Solis Elvia
Published: June 2021

Preparing students for careers in biological sciences will require proficiency in experimental design, statistical analysis and reasoning that students currently lack. To address this problem, a collaborative project aims to integrate statistical analysis (SA) and experimental design (ED) into the life science curricula at both high school and university levels. The project is developing cohorts of “teacher scientists” who both reform their own curricula and disseminate lessons learned to other secondary teachers through regional workshops. To assess the effectiveness of these workshops, a post‐participation survey was administered.

Responses were obtained from 21 science educators in the first workshop. Preliminary analyses indicate that the workshop was effective at teaching participants about laboratory techniques for teaching statistics concepts concurrently with biology content. Teachers indicated that they were likely to adopt a curriculum that integrates biology and statistics using the techniques from the workshop. Additionally, the teachers were interested in continuing discussions with their peers and university faculty for support integrating ED and SA into high school curricula.

This project was funded by the Howard Hughes Medical Institute (Deviating from the Standard: Integrating Statistical Analysis and Experimental Design into Life Science Education)
CO2 rebreathing: an undergraduate lab to study the chemical control of breathing
Authors: Domnik Nicolle Jasmin , Turcotte Scott E , Yuen Nathaniel Y , Iscoe Steve , Fisher John T
Published: June 2021

Duffin and McAvoy's (1988) modification of the Read (1967) CO2 rebreathing (CO2RB) method is widely used to study chemical control of breathing (ChemoCB). We used a CO2RB laboratory to challenge undergraduate science majors (to investigate the ChemoCB). We collated and analyzed 8 years of data to study the reproducibility of the laboratory and compared the results to the literature. Students (aged 19–21 yrs; IRB approval) performed the technique using a rebreathing bag containing 95% O2 to suppress peripheral chemoreceptor activity. Ventilation was measured following hyperventilation to deplete tissue CO2 stores, which also enhances detection of the central chemoreflex threshold (CCT). Data were analyzed by 3 investigators. Of 37 data sets, 7 were rejected due to technical issues. CO2 sensitivity and CCT were 4.69±3.12 L/min/mmHg (mean±SD) and 43.5±3.9 mmHg respectively. The CCT was normally distributed, whereas sensitivity was skewed to the left. Values agree well with those reported by Duffin & McAvoy (1988). We conclude CO2RB is a robust method for undergraduate investigation of the principles of chemical control of breathing. Funded by Queen's University, NSERC and CIHR.
Applying new technologies to old problems: undergraduate research projects for isolating and characterizing local Daphnia sp. and Chlamydomonas sp
Authors: Daggett Melissa A. F.
Published: June 2021

Independent research projects offer undergraduate students important opportunities to do applied learning in the sciences. This poster describes the methods and results of two such projects performed during the past year at Missouri Western State University in which students isolated local species of Daphnia or Chlamydomonas and carried out a series of student‐directed cell and molecular biology techniques in order to phylogenetically characterize their isolates. Students engaged in these projects reported on the usefulness of being able to learn concepts and techniques in more detail than when presented as part of a regularly schedule academic course.
Use of high fidelity human patient simulation (HPS) in second year medical pharmacology instruction
Authors: Ishac Edward JN , Robinson Susan E , Welch Sandra P
Published: June 2021

This study examined the use of HPS (Laerdal SimMan 3G) in a pharmacology general principles course and its impact on student learning, critical thinking and problem‐solving. Eight simulation cases were developed: six involving the autonomic nervous system and two with pain management. Students were assigned to one of 36 teams consisting of 5–6 members. Each team encountered only one case scenario. Teams were required to take a patient history, perform a physical exam, request laboratory tests, make a diagnosis, administer appropriate pharmacotherapy and monitor treatment outcome. Two different scenarios were performed concurrently and the two teams were then combined for a student led debriefing session that was faculty facilitated. Student survey comments were extremely positive. Results from 195 students included HPS complemented the course objectives (5.0 out of 5.0), promoted higher level, critical thinking (4.9/5.0), was a more effective learning experience than lectures alone (4.9/5.0) and improved student ability to function as a health professional team (4.7/5.0). We conclude that HPS provides an effective learning experience that encourages active learning and problem‐solving skills as a member of a health professional team.
Predicting strategies to improve student perceptions about connections between biology knowledge areas using innovative network analysis software
Authors: Dasgupta Annwesa , Pelaez Nancy
Published: June 2021

Students vary in their knowledge, experience and confidence (KEC) when they enter a course. A goal is to build their understanding by integrating emergent properties of life from molecules to organisms with evidence from experiments. What connections do students see among knowledge areas? What factors describe the clusters perceived by students? A perception inventory was administered in an undergraduate biology course for first year students, covering target course outcomes in potentially overlapping subjects: the physical and chemical basis of life, molecular basis of regulation, plant biology, animal biology, and the experimental and empirical basis of biology. For each outcome, students’ reported KEC were averaged to yield one score per student. Factor Analysis validated the target subject areas identified for the course. A correlation network diagram energized using the Kamada‐Kawai transformation, revealed that students’ prior KEC with biomolecules and molecular representation was unrelated to their KEC for experimental biology and both were unrelated to their KEC for plant physiology. By the end of the course, plant and animal physiology were connected but still separate from the experimental and empirical basis of biology and from molecules. Network diagrams show connections students perceive and guide innovative instructional strategies to help students make new connections.
Physiology with the singing greeting card beeper
Authors: Belusic Gregor , Zupancic Gregor
Published: June 2021

The singing greeting card beeper is a great mechanoelectrical transducer, usable in a number of different physiological experiments at no cost. It is an AC‐coupled sensor with a time constant of ca. 30 ms and a linear response up to at least 5 N, delivering relatively large voltage upon slightest deformation. If implemented as a human finger pulse sensor, it performs equally well as commercial‐grade sensors. It can also serve as a force transducer for sensing limb displacement in testing the reflexes such as the patellar reflex. If attached to a hammer, it can be used to report the moment of mechanical stimulation of the leg muscles. In testing the reaction times, it can serve as a microphone to report the moment of acoustical stimulation. We developed a range of experiments suitable for secondary school and undergraduate level. The implementation of the sensor into the curriculum has been accepted very well on the national as well as on the international scale. The idea was awarded the ADInstruments Macknight award in 2011.

The development of the sensor and experiments has been supported by the European Social Funds, operative program of human resources development 2007–2013.
mTOR Inhibitors Reduce Cerebral Vasospasm in a Subarachnoid Double‐Hemorrhage Canine Model
Authors: zhang Weiguang , chen chunhua , Meng Haiwei , Han Hongbin , Han Jingyan , Liu Shuwei , Zhou Changman
Published: June 2021

mTOR pathway promotes cell proliferation under environmental stress. In the present study, we investigated the role of mTOR pathway in subarachnoid hemorrhage brain injury. Thirty dogs were randomly divided into 5 groups: sham, SAH, SAH+DMSO, SAH + Rapamycin and SAH+AZD8055. An established canine double‐hemorrhage model of SAH was used by injecting autologous arterial blood into the cisterna magna on days 0 and 2. Angiography was performed at days 0 and 7. Clinical behavior, histology, immunohistochemistry, and Western blot of mTOR, P70S6K1, 4E‐BP1 and PCNA in the basilar arteries were examined. Severe angiographic vasospasm was obtained in the SAH and SAH+DMSO dogs compared with in Sham respectively. In the SAH+ RAPA and SAH+ AZD8055 groups, a moderate vasospasm, mTOR, P70S6K1, 4E‐BP1 and PCNA increased in the sample of spastic basilar arteries (p<0.05). Rapamycin and AZD8055 attenuated angiographic vasospasm while improving appetite and activity scores (p<0.05) on days 5 to 7. Rapamycin and AZD8055 significantly reduced the level and expression of mTOR, P70S6K1, 4E‐BP1 and PCNA (p<0.05). In conclusion, mTOR molecular signaling pathway play a significant role in cerebral vasospasm (VSMCs proliferation) following SAH, and mTOR inhibitor has potential to become an attractive strategy to treat vasospasm following SAH.
Hsp60 secretion and migration from cancer cells: a proposal for a multistage pathway
Authors: Campanella Claudia , Cappello Francesco , Bucchieri Fabio , Merendino Anna , Fucarino Alberto , David Sabrina , Pitruzzella Alessandro , Farina Felicia , Burgio Giosalba , Corona Davide , de Macario Everly Conway , Zummo Giovanni , Macario Alberto J.L.
Published: June 2021

Hsp60 can be released by tumor cells to promote growth. Hsp60 exits mitochondria, reaches the blood and distant targets by mechanisms not yet understood. We determined Hsp60 levels and secretion in normal and tumor cell lines in cell lysates, medium, exosomes, membranes, lipid rafts, and other materials, by ultracentrifugation, electrophoresis, Western blotting, ELISA, EM, and enzymatic tests. Exosomes were purified and characterized by EM and by presence of ALIX, Hsp90, Hsp70, ATPase, and AChE. The tumor, not the normal cells secreted Hsp60, in the absence of cell death, involving lipid rafts‐exosome pathways; Hsp60 associated with lipid‐rafts and cell and exosomal membranes. A working model predicts: Hsp60 gets in the cytosol from mitochondria or from synthesis in the cytosol without entering the organelle, arrives at the cell membrane and binds lipid rafts and, through lipid‐raft mediated endocytosis, is included in early endosomes and then in multivesicular bodies and, lastly, in exosomes for secretion. Hsp60 in exosomes may then reach components of the innate immune system and trigger inflammation; endothelial cells to elicit neoangiogenesis; and other tumor cells to block apoptosis and promote cancer survival.
Nexilin is a dynamic protein at Listeria monocytogenes comet tails and enteropathogenic E. coli pedestals
Authors: Law H T , Bonazzi Matteo , Jackson Jeremy , Cossart Pascale , Guttman Julian A.
Published: June 2021

The extracellular pathogen enteropathogenic Escherichia coli (EPEC) and the intracellular bacterium Listeria monocytogenes (Listeria) control the actin cytoskeletal machinery during their infections as part of their disease processes. Through the subversion of this cytoskeletal system, motile actin‐rich structures (EPEC pedestals and Listeria comet tails) are generated. Nexilin, an actin‐binding protein, is thought to facilitate actin‐based cell motility. Consequently, we tested the hypothesis that nexilin is recruited to EPEC pedestals as well as Listeria comet tails and is needed for proper functioning of those structures. Using a combination of immunolocalization, RNA interference and time‐lapse imaging, we demonstrate that nexilin co‐localizes with actin in EPEC pedestals and Listeria comet tails. As stationary EPEC and Listeria becomes motile, nexilin focuses distally in these long actin‐rich structures. When nexilin expression was perturbed, EPEC pedestals remain unaltered, however Listeria comet tails had aberrant morphologies. Our data suggests that nexilin is redistributed to the dissociating end of actin filaments where it may act as a platform for bacterial propulsion when infecting host cells.

Grant Funding Source: CIHR, NSERC, Institut Pasteur, Institut National de la Santé et de la Recherche Médicale, Institut National de la Recherche Agronomique, European Research Council, Howard Hughes Medical Institute
Anatomical Correlation of Musculoskeletal Diagnostic Images
Authors: Russell Byron , Garner Bryan
Published: June 2021

The use of diagnostic ultrasound has become mainstream in the evaluation of musculoskeletal disorders. Correct interpretation of the images is paramount in the evaluation of musculoskeletal conditions. The purpose of this study was to compare the images obtained using diagnostic ultrasound to the actual tissues measured in preserved cadavers. Ten images were collected from a variety of anatomical sites including the hip, knee, plantar foot, Achilles tendon and shoulder. The thickness of tissues was measured on the images followed by dissection and measurement of the tissue thickness on the cadaver. The calculated thickness and the actual thickness were compared. The correlation between the calculated thickness and the measured thickness was r = .72. The results indicate a moderate to good relationship between the calculated and measured tissue thickness.
3‐D Arthroscopy: A New Frontier in Surgical Visualization
Authors: Roach Victoria A , Mistry Manisha R , LeBel Marie-Eve , Wilson Tim D
Published: June 2021

At present, support of 3‐D visualization for laparoscopic surgical procedures is growing. This is likely due to the ability to interpret anatomical structures during the 3‐D visualization previously impossible in 2‐D endoscopic procedures. With the ability to interpret depth cues accurately, surgeons better orient themselves in body cavities with greater ease and fewer movements, ultimately correlating to reduced patient morbidity. Considering the validated success of 3‐D in endoscopy, it is surprising that this technology has not been applied to arthroscopy; the endoscopy of joints. At present, millions of arthroscopic surgeries are performed using primitive 2‐D scope technology on the 3‐D anatomy of the joint cavity, posing a significant conceptual roadblock for novice orthopedic surgeons. This study aims to apply technology used in 3‐D laparoscopes to build a 3‐D arthroscope prototype to enhance operative precision via accurate visualization of the joint. This study also aims to test the efficacy of a prototype for skill acquisition via the Global Ratings Scale for surgical skills, completion time measurements, and instances of perceptual depth errors. It is hypothesized that testing surgical applications of this 3‐D visualization in trainees will result in reduced training time, fewer perceptual errors, increased operative precision and decreased burden of cognitive load in the user.

Grant Funding Source : Departmental
Shear Stress‐Activated Angiopoeitin‐2 Modulates Endothelial Cell Repairs and Vasculogenesis via Wnt/β‐catenin Signaling Pathway
Authors: Yu Fei , Takabe Wakako , Harrison Michael , Li Rongsong , Gu Jie , Chi Neil , Lien Ching-Ling , Hsiai Tzung
Published: June 2021

Fluid shear stress is intimately involved in vascular endothelial cell repairs. It is recognized that pulsatile shear stress up‐regulated Angiopoetin‐2 expression (Ang‐2) in human umbilical vein endothelial cells (HUVEC). We hypothesized that Ang‐2 was implicated in HUVEC repairs and vasculogenesis via Wnt/β‐catenin signaling pathway. Knock‐down of Ang‐2 expression with siRNA resulted in an impaired HUVEC migration and tube formation. Wnt3a, a Wnt activator, up‐regulated Ang‐2 mRNA expression (p < 0.05 vs. no addition, n = 3), whereas DKK‐1, a Wnt inhibitor, inhibited Ang‐2 expression (p < 0.05 vs. PBS treated, n = 3), leading to impaired migration and tubule formation. We further demonstrated that subintestinal vessels (SIVs) were absent in heat shock inducible DKK‐1:gfp zebrafish at 72 hpf. Knock‐down of Ang‐2 with morpholino oligomers in the transgenic zebrafish embryo (TG(flk1:GFP)) at 1‐cell stage resulted in impaired vasculogenesis to the entire vasculature at 48 hpf. Treatment with ionomycine, a Calcium (Ca2+) ionophore, also resulted in both impaired HUVEC repairs and suppression of SIV formation in TG(flk1:GFP) lines at 72 hpf as evidenced by down‐regulation of Ang‐2 mRNA expression (p < 0.05 vs. DMSO treatment, n = 3). Taken together, our findings suggest that shear stress‐activated Ang‐2 is implicated in endothelial cell repairs and vasculogenesis via Wnt/β‐catenin signaling pathway.

Grant Funding Source : NIH