bluuebunny/crossref_metadata_2025_split huggingface dataset:
Involvement of Endothelial Small GTPase RhoA in Angiogenesis In Vitro and In Vivo
Authors: Zahra Fatema Tuz , Sajib Md Sanaullah , Ichiyama Yusuke , Akwii Racheal , Tullar Paul , Kubota Yoshiaki , Mikelis Constantinos M 
Published: June 2021

Angiogenesis, the formation of new blood vessels, is a highly orchestrated process requiring a well‐regulated balance between pro‐angiogenic and anti‐angiogenic factors and corresponding signaling networks. It is an essential process during development and in specific adult conditions such as wound healing and ovulation. Imbalanced angiogenesis is a characteristic of several diseases such as cancer, inflammation, rheumatoid arthritis, osteoarthritis and age‐related macular degeneration. The Rho family of small GTPases plays a central role in a variety of cellular processes involving cytoskeletal rearrangement, cell movement, microtubule dynamics, signal transduction, gene expression which are necessary for vascular development and angiogenesis. Among the small GTPases of the Rho family, RhoA, Rac1 and Cdc42 are the best characterized. Although the role of endothelial Rac1 and Cdc42 in embryonic vascular development and retinal angiogenesis has been studied recently, the role of endothelial RhoA is yet to be explored. In this study, we aim to identify the involvement of endothelial RhoA in angiogenesis in vitro and in vivo. In vitro, RhoA seems to be involved in endothelial cell proliferation, cell migration, invasion and sprouting triggered by important angiogenesis inducers, such as Vascular Endothelial Growth Factor (VEGF) and Sphingosine‐1 Phosphate (S1P). To study the involvement of endothelial RhoA in vascular development in vivo, we generated endothelial‐specific RhoA deficiency through the Tie2‐Cre and Cdh5‐CreERT2 promoters. In vivo deletion of RhoA in endothelial cells during embryogenesis leads to decreased survival of endothelial RhoA‐deficient mice. However, the few mutant survivors do not present morphological or behavioral abnormalities. In line with that, inducible RhoA deficiency in the retinal vessels in different developmental days did not affect the radial growth, the number of filopodia per area and the area with deep vascular plexus. Ongoing experiments aim to clarify the compensation mechanisms of RhoA deficiency in physiological angiogenesis in vivo versus in vitro models, providing a better understanding of the role of endothelial small GTPase RhoA in normal physiology and human diseases.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Study the function of GTPase related genes wasf1 and ect2 in zebrafish vascular development
Authors: Chiu Yan‐Hui , Wu Chang‐Yi 
Published: June 2021

Vascular development is important in vertebrates and regulated by complex signaling pathways. In the process of forming vessels are mainly divided into vasculogenesis and angiogenesis. Zebrafish is a powerful vertebrate model organism to study vascular development. Our previous study in zebrafish identified the transcription factors nr2f1b/islet2 required for blood vessels, mainly focus on the patterning of intersegmental vessels (ISV) and caudal vein plexus (CVP). The further microarray results showed that the expression of several GTPase related genes was regulated by nr2f1b/islet2, and we are interested in how those GTPase related genes contribute to vascular patterning in zebrafish.In this study, we reported two novel GTPase related genes wasf1 and ect2 that investigated their significance for the vascular development. In‐situ hybridization show that ect2 and wasf1 mRNA is expressed in developing vessels. Knockdown of ect2 and wasf1 by morpholino microinjection cause the defect in blood vessel formation. TUNEL assay and AO staining data show that vascular defects do not caused by cell death, but likely due to the impairment of proliferation and/or migration. Consistent with the defect in vascularture, loss of ect2 or wasf1 reduced the expression of the vascular markers. We further perform Western blotting to examine the relations between wasf1/ ect2 and major angiogentic VEGF/ BMP signals. We showed that GTPase related proteins wasf1 and ect2 play an important roles of vascular development mediated by VEGF/ BMP signalings in zebrafish.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Changes in the expression of ERK, JNK and p38 in small intestine related to prolonged intake of commercial sweeteners and their relationship with periodontal state in BALB/c mice
Authors: Sánchez Carlos Manuel Abascal Sherwell , Estrada José Antonio , Contreras Irazú 
Published: June 2021

IntroductionObesity is a public health problem associated with increased consumption of high‐calorie diets, which predisposes the development of non‐communicable diseases such as obesity and diabetes mellitus type II. A common strategy for their prevention is the addition of non‐nutritive sweeteners in diet. The need for evidence on the molecular interactions between sweeteners and mucous membranes after chronic consumption of these compounds has generated hypotheses that establish a relationship between sucralose intake and high prevalence of inflammatory bowel disease in developed countries.ObjectivesTo establish a profile of protein expression in the intestinal epithelium and cellular infiltration in periodontal tissue after chronic consumption of non‐nutritive sweeteners in BALB/c mice.Material and Methods8‐week‐old BALB/c mice were supplemented with non‐nutritive sweeteners (sucralose and steviol glycosides) for 6 weeks. Expression of total and phosphorylated forms of ERK, JNK and p38 in intestinal tissue were evaluated by western‐blot. Jaws were analyzed to determine histopathological changes in oral mucose membranes related to chronic sweetener consumptionResultsDensitometry analysis of protein bands showed a increased expression of ERK, JNK and p38 phosphorylated proteins in mice supplemented with sucralose and steviol glycosides, compared to control groups; however, these differences were not statistically significant. Similarly, histopathological study of the oral cavity showed increased cellularity, vascularity and cell clustering in gingivae in the same groups.ConclusionsThe present study provides evidence that frequent consumption of non‐nutritive sweeteners may generate pro‐inflammatory changes in oral cavity and intestinal mucosae, based on higher expression of phosphorylated ERK, JNK and p38 proteins and increased cellular recruitment at the periodontal level.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of KPC‐2 Backbone Dynamics Upon Avibactam Binding via NMR Spectroscopic Methods
Authors: Shurina Ben , VanPelt Jamie , Ramelot Theresa , Page Richard 
Published: June 2021

Antibiotic resistance is a continual threat in the clinic. One of the resistance modes bacteria employ is the expression of the beta‐lactamases that target most of the clinically relevant antibiotics, such as penicillin. The enzyme hydrolyzes the β‐lactam bond, which renders the antibiotic ineffective. Targeting β‐lactamases with small molecule inhibitors can restore efficacy of current generation antibiotics. Klebsiella pneumoniae carbapenemase (KPC) presents a global threat to public health as it confers multidrug resistance, including resistance to last resort antibiotics. Avibactam is the only FDA approved inhibitor for KPC are on the market, however some variants of KPC are showing increased resistance to avibactam. Futhermore, avibactam can be slowly degraded by KPC‐2 through a de‐sulfonation mechanism. A conserved omega‐loop near the active site of KPC‐2 has been implicated in substrate binding and is of particular interest as a target for future inhibitors. We report the first backbone dynamic characterization of a Class A carbapenemase in the presence and absence of an inhibitor. Backbone dynamic parameters for KPC‐2 obtained through the use of 3D HNCO based T1, T2, and heteronuclear NOE relaxation experiments. Chemical shift perturbations were obtained by comparing avibactam bound and apo states. H/D exchange experiments were also conducted to examine relative rates of exchange in avibactam bound and apo KPC‐2 in order to estimate slow timescale dynamics. The binding of avibactam caused an increase in R1 relaxation rates, suggesting avibactam can increase the dynamic flexibility of KPC‐2. The changes in the R2 relaxation rates suggest that the binding of avibactam might make the Ω‐loop slightly more rigid. These results could be used as a benchmark in measuring interactions with the Ω‐loop, interactions with KPC‐2, or interactions with other β‐lactamases.Support or Funding InformationNIH R35 GM128595This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gut dysbiosis with decreased Akkermansia muciniphila and differential expression of tumor‐associated genes contribute to a tumor‐preventive microenvironment in intestinal epithelial Pten‐deficient mice
Authors: Rhee Sang Hoon , Howe Cody , Kim Su Jin , Mitchell Jonathon , Im Eunok 
Published: June 2021

Phosphatase and tensin homolog (Pten) antagonizes PI3K‐Akt signaling; therefore, Pten impairment causes tumorigenesis. However, the correlation between Pten deficiency and colon cancer has remained elusive due to numerous opposite observations. To study this correlation, we examined whether Pten deficiency in intestinal epithelial cells (IECs) induces tumorigenesis.With mucosal biopsies of human colon cancer and normal colon, Pten mRNA was evaluated by quantitative PCR. Using IEC‐specific Pten knockout mice (PtenΔIEC/ΔIEC), we examined the mitotic activity of IECs; and PtenΔIEC/ΔIEC; Apcmin/+ mice were generated by combining PtenΔIEC/ΔIEC with Apcmin/+ mice. Tumor‐associated gene was evaluated by micro‐array analysis. Fecal microbiome was analyzed through 16S rRNA gene sequencing.We found that Pten mRNA level was reduced in human colon cancer relative to normal tissues. Augmented chromatids, increased Ki‐67 and PCNA expression, and enhanced Akt activation were identified in IECs of PtenΔIEC/ΔIEC mice compared to Pten+/+ littermate. Combining PtenΔIEC/ΔIEC with Apcmin/+ condition caused rapid and aggressive intestinal tumorigenesis. However, PtenΔIEC/ΔIEC mice did not develop any tumors. While maintaining the tumor‐driving potential, these data indicated that IEC‐Pten deficiency alone did not induce tumorigenesis in mice. Furthermore, the expression of tumor‐promoting and tumor‐suppressing genes was decreased and increased, respectively, in the intestine of PtenΔIEC/ΔIEC mice compared to controls. The abundance of Akkermansia muciniphila, capable of inducing chronic intestinal inflammation, was diminished in PtenΔIEC/ΔIEC mice compared to controls.These findings suggested that altered tumor‐associated gene expression and changed gut microbiota shape a tumor‐preventive microenvironment to counteract the tumor‐driving potential, leading to the tumor prevention in PtenΔIEC/ΔIEC mice.Support or Funding InformationThis research was supported by a grant from Oakland University and the National Institutes of Health (DK079015, S.H.R).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fungistatic Effects of 1,4‐dimethylnaphthalene on Pythium ultimum and Aspergillus niger
Authors: Stefanick Veronica , Dobson Kara , Campbell Michael 
Published: June 2021

The compound 1,4‐dimethylnaphthalene (DMN) was originally isolated from dormant potatoes and is currently being used as a commercial sprout inhibitor. In order to determine how the microbiome of potatoes is affected by the compound, two fungal species were grown and quantified under control and DMN conditions: Pythium ultimum and Aspergillus niger. These two fungal species are commonly found on the surface of potatoes and can result, respectively, in Pythium leak and black rot on onions. Fungal cultures were obtained from ATCC (P. ultimum: 58811, A. niger: 16888) and grown on potato dextrose agar (PDA). Mycelial cultures were grown in 9.5 Liter BBL GasPak chambers, exposed to either mineral oil (control) or DMN for two days. The mycelial mats exhibited reduced growth under DMN conditions in comparison to controls. DMN treatment of mature A. niger cultures also prevented sporulation. This indicates that DMN displays fungistatic properties but does not function as a fungicide towards P. ultimum and A. niger. In order to determine how DMN affects the gene expression of these fungi, RNA sequences will be collected and gene expression will be observed between control and DMN treatments.Support or Funding InformationSupported in part by a grant from the 1,4‐Group of Meridian, ID and the Behrend College Undergraduate Research FundThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Understanding Commensal Bacteria Influence Host Heme Metabolism
Authors: DuBois Jennifer Lynn , Coe Genevieve , Celis Arianna I. , Walk Seth T. 
Published: June 2021

Heme serves as a critical source of iron for human pathogens that can access O2 and the abundant heme stores in red blood cells. The lack of a well‐defined cellular heme transporter in the intestinal lumen cells suggests a possible role for microbes in metabolizing the tetrapyrrole on behalf of themselves and the host. A fully anaerobic pathway for heme breakdown (chuA chuW chuS) was recently discovered in E. coli, which could render heme iron bioavailable to the host, or minimize the toxic or inflammatory effects of heme for the microbiome and host, respectively. We tested these hypotheses using a chu‐containing commensal E. coli strain an isogenic DchuW knock out, and mice with defined microbiota. We provide evidence that the microbiome may serve as a missing link in rendering heme iron bioavailable to animal hosts.Support or Funding InformationThis work was supported by the National Institutes of Health, grant R21DK114607.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Comparative Study of Equine Gut Microbiomes Using 16S and 18S rRNA Sequencing
Authors: Jorgenson Jillian K. , Bennett Jennifer A. , Burk Steffanie V. 
Published: June 2021

The gut microbiome of domestic horses (Equus ferus caballus) can be broken down into three primary divisions: (1) bacterial composition, (2) fungal composition, and (3) eukaryotic parasite communities. While each has been the subject of various scientific explorations, in this study we analyze the potential interconnectedness of these three categories. To determine the extent to which each component of the gut microbiome influences the other, we extracted and amplified DNA from equine fecal samples and utilized a combination of 16S and 18S rRNA Illumina Sequencing to illustrate the bacterial and fungal compositions taxonomically with the assistance of QIIME2 software. Meanwhile, fecal egg counts for each sample were completed using the modified Stoll method to estimate the parasitic load for each horse. The identification of relationships between bacterial communities, fungal communities, and parasite load could have clinical applications to future studies of equine health. Additionally, comparisons could be made between domestic horses and exotic hoofstock as a means to improve our understanding of gut microbiome health in lesser studied species.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular mechanism of cereblon‐dependent down regulation of AMP‐activated protein kinase
Authors: Yang Seung‐Joo , Jeon Seung Je , Han Hyemin , Park Chul‐Seung 
Published: June 2021

Cereblon (CRBN), initially identified as a target protein of human mental retardation, is a substrate receptor of cullin‐ring E3 ubiquitin ligase (CRL) and a primary target of thalidomide‐induced teratogenicity. We previously reported that CRBN can negatively regulate the functional activity of AMP‐activated protein kinase (AMPK) by binding to the a subunit of the AMPK complex. We also noticed that the exogenous expression of CRBN reduced the content of g subunit in AMPK complex. However, the molecular mechanism for the CRBN‐dependent reduction of g subunit from AMPK complex was still unclear. Thus, we investigated the molecular mechanism of CRBN‐dependent inhibition of AMPK. We noticed that the amount of g subunit in AMPK complex was increased in Crbn‐knockout (KO) MEFs and the g subunit was degraded more rapidly in wild‐type MEFs than in Crbn‐KO MEFs. After treating MG132, a proteasome inhibitor, the g subunit is accumulated more rapidly in wild‐type MEFs compared to Crbn‐KO MEFs. Moreover, ectopic expression of CRBN promoted the degradation of g subunit in Crbn‐knockout MEFs only in the presence of a subunit. We further found that AMPK g subunit was ubiqutinated by CRBN and ubiquitination persisted in the presence of IMiDs. These results strongly suggest that CRBN negatively regulates the AMPK activity by recruiting AMPK complex to CRL and degrading its g subunit.Support or Funding InformationThis work was supported by GIST Research Institute (GRI) grant funded by the GIST in 2019This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Transcriptional regulation of the yeast TGL3‐encoded triacylglycerol lipase
Authors: Lu Chieh , Ye Bo‐Hong , Su Wen‐Min 
Published: June 2021

The yeast TGL3‐encoded triacylglycerol (TAG) lipase catalyzes the hydrolytic reaction of TAG yielding fatty acid and diacylglycerol (DAG). The reaction products DAG and fatty acid not only can be used as building blocks for the synthesis of phospholipids, but also serve as lipid signaling molecules. In addition, fatty acid can undergo β‐oxidation to generate ATP. During growth resumption, the stored TAG in lipid droplet is rapidly hydrolyzed by Tgl3 to provide precursors for membrane phospholipids and to generate lipid signaling molecules that are essential for normal cell growth and cell cycle progression at the G1/S phase. However, excessed fatty acid and DAG are also detrimental to cell growth as they exhibit detergent‐like properties that can disrupt membrane structure and cause defect in cell integrity. Thus, Tgl3 lipase activity must be tightly controlled and mechanisms of regulation need to be elucidated. In this study, using β‐galactosidase reporter assay, we found a sharp induction of TGL3 expression after cells were transferred into fresh medium from the stationary phase. Maximum expression was observed at first hour where the β‐galactosidase activity was 62% greater than the activity at the stationary phase. Moreover, the TGL3 promoter was shown to interact with the purified Reb1 transcriptional factor in the electrophoretic mobility shift assay. A Reb1 consensus recognition sequence (GGGTAA, −362 to −357) in TGL3 promoter is responsible for its specific binding to the Reb1 protein. The effects of Reb1‐mediated TGL3 regulation on the function of Tgl3 as well as lipid metabolism are currently under investigation using the TGL3 Reb1‐binding site mutations.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparative Analysis of Healthcare Providers and Patient Perspectives on Natural Remedies as Alternative Medicine in Two Communities in Peru
Authors: Simental Brianna N , Loaiza Anaiz 
Published: June 2021

More often than not there is a disconnect between healthcare providers (HCPs) and their patients regarding the use of natural remedies as an alternative medicine. Lack of knowledge and communication between healthcare providers and patients can change the efficacy of the medication and can be a safety hazard for the patient. Examining the relationship between healthcare providers' view on the use of natural remedies is important, especially in developing countries like Peru. Due to unequal access to healthcare people may rely more heavily on alternative medicine. Healthcare providers and patients will have a different perspective on natural remedy use and their effectiveness. This miscommunication could affect patient care and treatment due to uninformed guidance and lack of knowledge of natural remedies. To analyze the perspectives of HCPs and patients about natural remedy use, we designed a cross‐sectional study that was conducted over two summers, among healthcare providers and women in two communities in Peru. Two individual surveys were distributed and used for data collection. Our preliminary results suggest that there is a disconnect between the percentage of patients that use natural remedies reported by healthcare providers and the percentage of women in the community that reported their use of natural remedies. There was also a divide between the perspective of HCP's on whether their patients feel comfortable addressing their health concerns and the perspectives of the patients who felt comfortable addressing their health concerns but did not trust their HCP. Further analysis is necessary to examine other barriers that prevent open communication between healthcare providers and their patients.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Crystallization of SARS Coronavirus 3CL Protease to Identify Inhibitor Targets
Authors: Fernandes Samantha Zinnia , Anson Brandon , Mesecar Andrew 
Published: June 2021

The SARS coronavirus has been identified as the pathogen responsible for the 2003 outbreak of severe acute respiratory syndrome (SARS). This highly contagious virus, which most likely originated in Southern China, has since been found in over 32 countries, affecting about 8,500 people, and killed over 900 patients. Due to SARS ease of transmission through aerosols and the virus's high mortality rate (~10%), its reemergence is recognized as a looming threat to global health.However, neither a vaccine nor effective treatment exists to combat the illness in the event of another outbreak. The virus's 3 Chymotrypsin‐like main protease, is a major target for inhibitor studies, as it is crucial to viral replication through its proteolytic processing of polyprotein 1ab (pp1ab) at 11 different cleavage sites.This project worked to optimize the crystallization conditions for the SARS 3CL protease for use in X‐ray fragment based screening to identify possible inhibitors. The results of this step in the drug discovery study for the SARS‐CoV can be used as a basis for pioneering new scaffolds for inhibitor studies in other coronaviruses.Support or Funding InformationFunding from NSF DBI “REU Site: Molecular and Biochemical Analysis of Proteins” to Purdue University, Purdue College of Biochemistry, and the Purdue College of AgricultureThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.