bluuebunny/crossref_metadata_2025_split huggingface dataset:
TIMP‐1 protein but not secretion is increased by shear stress in the skeletal muscle microvasculature
Authors: Uchida Cassandra , Milkiewicz Malgorzata , Fudalewski Tomasz , Gee Eric , Haas Tara L. 
Published: June 2021

Luminal splitting is a form of angiogenesis caused by increased shear stress. A feature of luminal splitting is a lack of extracellular matrix proteolysis correlated with decreased production of matrix metalloproteinase (MMP)‐2. We hypothesized that an MMP inhibitor, tissue inhibitor of matrix metalloproteinase (TIMP)‐1, is upregulated by shear stress. TIMP‐1 mRNA increased in the EDL of prasozin‐treated rats (p<0.05, n=4) as measured by RT‐PCR. Rat skeletal muscle microvascular endothelial cells were exposed to shear stress (12 dyne/cm2). TIMP‐1 mRNA increased in cells after 2 and 24 hours of shear (p<0.05, n=3), and TIMP‐1 protein increased at 2 and 24 but not 4 hours (p<0.05, n=3) as assessed by Western. TIMP‐1 secretion decreased at 2 hours of shear and returned to control levels at 24 hours (p<0.01, n=3) as measured by reverse zymography of concentrated media. Similarly, MMP‐2 activity increased in media at 2 but not 24 hours of shear (p<0.05, n=7) as assessed by an in vitro MMP‐2 activity assay (Anaspec). 2 hours of shear caused no change in TIMP‐1 surface staining (ns, n=3) as assessed by confocal microscopy, but did cause an increase in intracellular TIMP‐1 staining (p<0.05, n=3). Our results show that intracellular TIMP‐1 protein is increased by shear stress, due to increased production and decreased secretion. The function of intracellular TIMP‐1 is unknown.Funded by the Heart and Stroke Foundation of Canada.Grant Funding SourceHeart and Stroke Foundation of Canada
Cooperation between Vezf1 and RhoB during vascular development
Authors: Stuhlmann Heidi , Benjamin Laura , Victor Andrea , Ocaya Pauline 
Published: June 2021

Vascular endothelial zinc finger 1 (Vezf1) encodes a zinc finger transcription factor that is essential for developmental angiogenesis and lymphangiogenesis. Vezf1−/− embryos show vascular remodeling defects, loss of vascular integrity, internal bleeding and embryonic death. Haploinsufficiency is observed in 20% of Vezf1+/− embryos, resulting in lymphatic hypervascularity, edema and hemorrhaging (Kuhnert et al Dev.Biol.2005).Recent studies suggested that Vezf1 might functionally interact with Rho GTPases. We studied cooperation between Vezf1 and RhoB during vascular development by generating embryos with compound Vezf1;RhoB null alleles. Strikingly, loss of RhoB exacerbates the phenotypes associated with loss of one Vezf1 allele. The phenotypes include: 1) enlarged head vessels 2) vascular anomalies at multiple sites in the head, hypervascularity, and hemorrhaging of internal organs, and 3) hypervascular lymphatic phenotype with blood‐filled vessels in the neck. These structures display similarities with human vascular and lymphatic anomalies. Thus, the results suggest that Vezf1 and RhoB cooperate in vivo, and indicate their potential role in human vascular pathologies.
Hypoxia Response and Stroke Susceptibility with Sickle Cell Anemia (SCA)
Authors: Milbauer Liming C , Enenstein Judy A , Kiley Jim , Johnson Darrell , Roney Mark , Solovey Anna , Hebbel Robert P 
Published: June 2021

Individuals with SCA are susceptible to stroke at an early age. Since hypoxia is a hallmark of stroke, we hypothesize that hypoxic responses of some genes may differ constitutively in individuals at risk for sickle‐stroke. Blood outgrowth endothelial cells (BOEC) were isolated from African American donors without SCA (NS) as well as SCA donors at risk (AR) or not at risk (NAR) of early stroke. Risk was assessed as occlusive disease at the Circle of Willis. BOEC were incubated under hypoxic (0.5% O2) conditions and induction of 9 known hypoxia response genes were assessed by qRT‐PCR. Combined data from 30 min and 1h hypoxia, indicated significant differences (p<.05) between groups in fold induction of several genes: AR>NAR (Elk‐1, tPa, HIF1α, HIF1β and Endothelin‐1), AR>NS (ELK‐1, tPa), and NS>NAR (Elk‐1, HIF1α). Slightly significant differences (p<.1) included AR>NAR (Egr‐1, Flk‐1) and AR>NS (Flk‐1, Egr‐1, HIF1α, Endothelin‐1). Under normoxic conditions, most genes did not differ at p<.05 between groups. The 9 genes are known for complex impacts on inflammation, angiogenesis, apoptosis and cerebral ischemia. However, the high hypoxia response of AR compared to NAR BOEC suggests that AR individuals may lack protective mechanisms that NAR have. In conclusion, individuals with SCA at risk for stroke respond to hypoxia with exaggerated expression of several genes.Funding: NIH (HL076540 & HL55552)
TB Meningitis, Reactive Oxygen Species (ROS) and the Pathological Ossification of the Petroccipital Fissure and Cochlear Cannaliculus
Authors: Balboni Armand L , Bergemann Andrew D , Laitman Jeffrey T 
Published: June 2021

Fibrocartilage ossification is a well‐controlled process in which chondrocytes within the tissue stop proliferating and differentiate into hypertrophic chondrocytes before undergoing apoptosis and the induction of tissue mineralization. We previously reported a pathological ossification of the fibrocartilagenous petroccipital fissure (POF) and cochlear cannaliculus (CC) in individuals with TB meningitis (Balboni et al., 2008). However, a mechanism by which such ossification could occur has not been shown. Recent work by Yang et al. (2007) demonstrates that TB meningitis causes a marked increase in the production of ROS in the central nervous system. To test whether an increase in ROS could cause POF/CC ossification we cultured ATDC5 chondrocytes in the presence of control medium(low ROS activity), differentiation medium(high ROS activity), or differentiation medium plus the antioxidant Deferiprone®, and observed them for evidence of hypertrophy. ATDC5 cells were also tested for the expression of the hypertrophic markers mmp13, typeX collagen, and runx2 via RT‐PCR. We show that increased ROS levels induce chondrocyte hypertrophy in vitro. Markers of elevated ROS levels are detected in hypertrophic ATDC5 chondrocytes while Deferiprone® protects cells from oxidative stress and inhibits these markers. These data demonstrate that POF/CC ossification could occur via ROS formed during TB meningitis.Grant Funding SourceFaculty Research Award
Effect of Autologous Platelet‐Rich Plasma on Distraction Osteogenesis of the Mandible in Rabbits.
Authors: Jurjus Abdo R , Rahme Mark , Saghieh Said 
Published: June 2021

Distraction osteogenesis of the jaws has been used in the treatment of pediatric craniofacial deformities. Successful clinical outcome depends on a coordinated multidisciplinary treatment.Platelet rich plasma (PRP) containing growth factors extracted from autologous blood and used to increase the healing potential of bone and soft tissue has been, in addition to bone marrow cells (BMC), recently tested during distraction osteogenesis of long bones in humans. The specific aim of this research is to study the effect of PRP on bone healing after mandibular distraction in rabbits and explore future possible application in humans. Right mandibular distraction was performed in 12 rabbits divided into 2 groups of 6 each. PRP and physiological saline were injected in the callus following distraction in the tested and control group respectively. The rabbits were sacrificed after consolidation period of 45 days and the mandibles were surgically removed. Two rabbits, 1 from each group, were excluded from the study the first due to lack of adequate distraction, the second died on the 30th day of consolidation period. Bone mineral density, radiographic analysis, mechanical properties and histological features of the lengthened bones were assessed on 10 rabbits 5 in each group.The data analysis showed a faster bone remodeling with an increase in the amount of trabeculation but no significant increase in bone mineral density and content in the experimental group in comparison to the control group.The value of this research lies in the potential acceleration of healing after mandibular distraction using PRP which is greatly needed since the mandible is usually subjected to complex functional demands.
Sinusoidal endothelial cell crosstalk with other liver cells
Authors: DeLeve Laurie D. , Wang Xiangdong 
Published: June 2021

Hepatic sinusoidal endothelial cells (SEC) line the microcirculation and have a unique phenotype: open fenestrae clustered in sieve plates and lack of a basement membrane. In normal liver, the SEC phenotype is maintained by paracrine secretion of VEGF by hepatocytes and hepatic stellate cells (HSC) and, downstream, VEGF‐stimulated autocrine production of nitric oxide (NO) by the SEC. VEGF‐stimulated NO production by SEC also promotes HSC quiescence.Capillarization, loss of SEC fenestration and formation of a basement membrane, precedes liver fibrosis and seems to be permissive for the final common step in fibrosis: activation of HSC. If loss of the differentiated SEC phenotype precedes and permits fibrosis, characterizing dysregulation of cell‐cell crosstalk in liver disease becomes important in understanding liver fibrosis. Progressive loss of paracrine secretion of VEGF, loss of SEC VEGF receptors, and decline in SEC NO production occurs in parallel with increasing severity of liver disease in experimental models of steatosis, steatohepatitis and cirrhosis. Further elucidation of the interplay between hepatocytes, HSC and SEC may provide therapeutic strategies to prevent fibrosis.
Expression and characterization of active francisella tularensis FabI enzyme, a potential antibacterial target
Authors: Bohn Kelsey C , Chmielowski Jennifer N , Wen Lisa , Huang Jenq‐Kuen , Timsina Yam N , Chand Phani C , Pathak Ashish K 
Published: June 2021

Francisella tularensis (F. tularensis) has been identified by the United States Centers for Disease Control (CDC) as one of the top six bacteria that could potentially be used as biological weapons. This research focuses on an enzyme of fatty acid biosynthesis in F. tularensis, namely FabI. FabI is an enoyl‐ACP reductase which catalyzes the last reduction step in each elongation cycle of type‐II fatty acid synthesis. The enzyme has been identified as a potential target for antibacterial drug development. In this research, the gene encoding of FabI was custom synthesized and cloned in pET15b expression vector. The enzyme was over‐expressed in E. coli, purified using Ni‐NTA affinity column, and its activity assayed spectrophotometrically. The pure FabI was tested with a set of indole‐2‐carboxylic acid compounds for their ability to inhibit FabI. We have discovered several new compounds with promising activity against F. tularensis FabI. The best result was obtained with indole‐2‐carboxylic acid analog WIUAKP‐031. The compound at 37.5 μM inhibited 98% of the FabI enzyme with IC50 value of 6 μM. The availability of the functional F. tularensis FabI will greatly aid in structural studies of this essential bacterial enzyme and facilitate the identification of small molecule inhibitors of type II fatty acid synthase.Supported in parts by seed grants from University Research Council, Western Illinois University.
Glycosylation of cyclooxygenase‐2 (COX‐2) influences effectiveness of COX‐2 inhibitors
Authors: Graham Kamara Whitney , Sevigny Mary B. , Gabriel Bianca S 
Published: June 2021

Cyclooxygenase‐2 (COX‐2) is an enzyme that catalyzes the rate‐limiting step in the prostanoid synthesis pathway, which plays an important role in a variety of physiological and pathophysiological processes including inflammation and cancer. COX‐2 exists as two major glycoforms of 72 and 74kDa, the latter resulting from an additional oligosaccharide chain at amino acid residue Asn580. The purpose of this study is to determine if this additional glycosylation affects the inhibitory ability of various COX‐2 inhibitors. COS‐1 cells were transiently transfected with either the wild‐type or Asn580‐mutant COX‐2 gene. Subsets of both cell groups were treated with various concentrations of either aspirin, flurbiprofen, ibuprofen, or celecoxib. After addition of the COX‐2 substrate arachidonic acid to inhibitor‐treated and untreated (control) cells, media was collected and subjected to an ELISA which measured levels of the downstream product prostaglandin E2. Results indicate that, at low concentrations, both aspirin and flurbiprofen are more effective at inhibiting the activity of the 72 than the 74 kDa glycoform, whereas ibuprofen and celecoxib are more effective at inhibiting the 74 kDa glycoform. This indicates that glycosylation of COX‐2 at Asn580 influences the efficacy of COX‐2 inhibitors.Research supported by the Dept. of Natural Sciences and Mathematics at Dominican University of California.
Biochemical characterization of Calponin homology domain of human IQGAP1
Authors: Kurella Vinodh Babu , Lecour Louis F , Worthylake David K 
Published: June 2021

Human IQGAP1 is a190 kDa molecular scaffold that contains several domains required for interaction with proteins such as F‐actin, calmodulin, Cdc42/Rac1, β‐catenin and E‐cadherin. Calponin homology domain (CHD) is a ~160 amino acid module located in the N terminal region of IQGAP1. The CHD belongs to the type 3 family of CH domain containing proteins such as actin‐binding proteins, calponin and Vav. IQGAP1 localizes to the cell cortex, binds to and crosslinks actin via its CHD. When this activity occurs at cadherin‐based cell‐cell junctions, adhesion is stabilized. Calmodulin a calcium sensor, negatively regulates the interaction of IQGAP1 with its binding partners. Preliminary calorimetric analysis suggests that Ca2+ /calmodulin but not apo‐calmodulin (calcium fee) binds to the N‐ terminal region of IQGAP1 containing the CHD. The mode of binding is apparently sequential with two calmodulin molecules binding to one IQGAP1 CHD. We have just obtained crystals of this CHD and are currently screening for crystals of a complex between Ca2+/calmodulin and CHD. It is expected that these structures will reveal conformational changes in the CHD necessary for complex formation and why Ca2+ /calmodulin but not apo‐calmodulin binds to the CHD.Louisiana board of regents grant
Intracellular translocation in Aktion: small molecule inhibitor effects on AKT localization
Authors: Strelko Cheryl L. , Roberts Mary F. 
Published: June 2021

Upregulation of the AKT pathway leads to a variety of human cancers. Phosphatidylinositol ether analogs, or PIAs, are small molecules designed to inhibit the Ser/Thr kinase AKT, by binding to its pleckstrin homology (PH) domain and preventing translocation from the cytoplasm to the nucleus. D‐3′‐deoxy‐diC8‐phosphatidylinositol is a soluble PIA that has an IC50 of 40 µM in U937 human lymphoma cells, which predominantly express Akt1. This compound (and not related further deoxygenated inositol lipids or L‐inositol species) does not appear to affect activation and growth of cells that have other Akt isoforms as well as Akt1 (e.g., B cells). We have shown that D‐3′‐deoxy‐diC8PI inhibits phosphorylation of proteins downstream of Akt1. However, near normal levels of phosphorylation of Akt1 strongly indicate that plasma membrane translocation of Akt1 has not been significantly inhibited. A GFP‐Akt1 fusion protein was expressed in U937 and MCF‐7 cell lines, and the movement of Akt1 when treated with D‐3′‐deoxy‐diC8PI was tracked using confocal microscopy. Treatment with D‐3′‐deoxy‐diC8PI did not inhibit plasma membrane or nuclear translocation, indicating cytotoxicity occurs by a different mechanism. Binding of the Akt1 PH domain in the absence and presence of the short‐chain PIA has also been examined to confirm the in vivo results.(This work is supported by N.I.H. GM60418).
Effect of Pepsin Hydrolysis on Amylase Activity and Lack of Amylase Inhibitory Activity of Sweet Potato Protein
Authors: Patterson Katherine W. , Truong Van den , Clare Debra A. , Allen Jonathan C. 
Published: June 2021

The objectives were to determine the effect of pepsin hydrolysis on amylase activity and test for amylase inhibitory activity of Beauregard sweet potato protein. Protein was extracted from sweet potato flesh, peel, and leaf. Samples with or without pepsin digestion at pH 2 were run on SDS PAGE. Proteins resistant to pepsin digestion were found in the flesh, peel, and leaf. Amylase activity and amylase inhibitory activity were determined using starch as substrate, sugar reducing ends as product, and dinitrosalicylic acid as the detection medium. Dinitrosalicylic acid reacts with sugar reducing ends to absorb at 540 nm. No amylase activity was found in the leaf extract. Digestion with pepsin eliminated amylase activity in the flesh and peel extracts. Amylase inhibitors were not apparent in any of the fractions before or after pepsin hydrolysis. Amylase inhibition by sweet potato protein does not appear to be the mechanism for the low glycemic response to Beauregard sweet potatoes. Gastric digestion is likely to eliminate native amylase activity in the sweet potato thereby requiring pancreatic amylase to digest carbohydrate in vivo.Support: N.C. Sweet Potato Commission.Grant Funding SourceN.C. SWEET POTATO COMMISSION
Validation of dual stable isotope method for measuring cholesterol absorption in hamsters
Authors: Ghanam Khadija , Wang Yanwen 
Published: June 2021

Although dual stable isotope method has been used to measuring cholesterol absorption rate in hamsters, this method is adopted from humans and has not been well validated in hamster model. Thus, we have conducted two studies in hamsters to validate this method in terms of optimum time points and effective dose of two cholesterol tracers. After 2 wk of adaptation, male Golden Syrian hamsters were administered orally with 18O‐cholesterol (0.45 mg in study 1 and 0.6 mg in study 2) and intravenously injected with 13C‐cholesterol (0.18 mg in study 1 and 0.24 mg in study 2). Blood samples were collected at 0, 6, 24, 48, 72, 96, and 120 h, respectively following the injection. The enrichment of 18O and 13C in free cholesterol of red blood cells was analyzed using isotope ratio mass spectrometry (IRMS) and used to calculate cholesterol absorption rate. The results showed that both tracers increased gradually in the first 6 or 24h and gradually declined over next 4 days. The tracers at low and high doses behaved similarly over the time. The ratio of 18O/13C reached a plateau at 48 h and stayed constant until 96 h. The percent cholesterol absorption rate did not differ among 48, 72 and 96 h. These results suggest that cholesterol absorption rate can be measured precisely using the dual stable isotope method at 48 or 72h with the injection of lower dose of stable isotope.Supported by Atlantic Innovation Fund, Canada
Maternal MTHFR C677T Genotype & Depressed Mood during Pregnancy Affects SLC6A4 Methylation in Infants at Birth
Authors: Wade Rachel E , Oberlander Tim F , Brain Ursula , Austin Jehannine , Devlin Angela M 
Published: June 2021

In utero and early postnatal exposure to depressed maternal mood may program childhood behaviour via epigenetic processes. Methylenetetrahydrofolate reductase (MTHFR) is an enzyme important for methyl metabolism. A common variant in the MTHFR gene, C677T, is associated with depression. We investigated the effect of maternal MTHFR C677T genotype on maternal mood (Edinburgh Postnatal Depression Scale, EPDS), during pregnancy (n=82 women, all receiving folate supplements) and on promoter methylation of two genes implicated in depression, brain derived neurotrophic factor, BDNF, and sodium‐dependent serotonin transporter, SLC6A4, in the women and their infants at birth. Women with the MTHFR 677TT genotype had greater (P<0.05) mid gestational depressed mood (mean age 24.98±0.51 weeks). Maternal MTHFR 677TT genotype was also associated with lower (P<0.05) maternal and infant SLC6A4 promoter methylation but not BDNF methylation. We also found a negative relationship (P<0.01) between maternal depressed mood (EPDS scores) and maternal SLC6A4 promoter methylation. These findings show that maternal MTHFR C677T genotype and mood influence maternal and infant SLC6A4 promoter methylation and suggests that disrupted maternal methyl metabolism and depression during pregnancy affects gene‐specific DNA methylation patterns, which may have long‐term consequences for childhood behaviour.Grant Funding SourceCIHR
Student Involvement in Local Wellness Policies of Local Education Agencies in Pennsylvania
Authors: Jomaa Lamis Hassan , McDonnell Elaine , Hartman Terryl , Weirich J. Elaine , Orlofsky Charles , Probart Claudia 
Published: June 2021

The purpose of this study is to investigate Student Involvement (SI) in Local Wellness Policies (LWPs) of Pennsylvania Local Education Agencies (LEAs) and to explore predictors of SI. According to the 2004 federal mandate, each LEA participating in a school meal program must develop a LWP and involve a number of stakeholders in the development process. Research supports SI in school‐based interventions to promote healthy eating and physical activity. LWPs from Pennsylvania LEAs (n=721; including 539 public and 182 private LEAs) were abstracted and analyzed using SPSS 15. Logistic regressions were run to examine predictors for involving students on wellness committees, in annual reviews, in school menu planning, as well as assessing student needs and promoting peer‐education. LEAs with comprehensive and rigorous policies were more willing to engage students in LWPs, regardless of the sponsor type (public or private), enrollment, location, or socioeconomic status. LEAs that included students in policy development were more likely to involve students on on‐going wellness committees. In conclusion, LEAs that wrote comprehensive and rigorous policies were more likely to have SI. Demographic characteristics did not appear to be barriers to SI. Involvement of students in drafting wellness policies increases the likelihood of on‐going SI, which may lead to greater student buy‐in for wellness initiatives.Grant Funding SourceRobert Wood Johnson Foundation
A Culturally tailored Nutrition Intervention to Promote Omega 3 fatty acid Consumption from fish Among African‐American Women of Childbearing age: A Pilot Study
Authors: Jones Samira Soyini , Greenwood MRC , Zidenberg‐Cherr Sheri 
Published: June 2021

The prevalence of obesity, type 2 diabetes and cardiovascular disease continues to rise among African‐American women. Consumption of high caloric, non‐nutrient dense food choices and lack of adequate physical activity may increase their risk for these detrimental outcomes. Additionally, studies have shown the importance of omega 3 fatty acids, specifically docosahexanoic acid (DHA) and eicosapentanoic acid (EPA), on reducing the risk of cardiovascular disease, insulin resistance, and improved neurocognitive functioning in adult populations. The aim of this project is to assess the effectiveness of a four week nutrition intervention tailored to the needs of African‐American women on increasing nutrition knowledge and improving dietary behaviors. Preliminary data on fish consumption prior to intervention indicate most women, on average, consumed fish 1‐2 times per month and improved to at least 4 times per month. Increases in knowledge of the health benefits of fish consumption and the risks associated with consumption of mercury‐contaminated fish were observed. Dietary behaviors such as 'limiting food portions' were found to be consistently correlated at baseline to post‐intervention (r =0.368, p=0.027) indicating improvements in this area. Intervention strategies have included use of evidence‐based nutrition education curricula that are community‐based participatory approaches for adults.Grant Funding SourceUC Davis Department of Nutrition