bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pannexin 1 and Pannexin 3 regulate body fat accumulation in mouse models of dietinduced obesity
Authors: Penuela Silvia , Lee Vanessa R , Barr Kevin , Kelly John J , Johnston Danielle , Brown Cody FC , Robb Kevin P , Sayedyahossein Samar , Huang Kenneth , Gros Robert , Flynn Lauren 
Published: June 2021

Pannexins (Panx) are channel‐forming glycoproteins important in paracrine signaling and cellular development. Panx1 and Panx3 are expressed in adipocytes and adipose stromal cells. We discovered that mice globally lacking Panx1 (KO) have significantly greater total fat mass and reduced lean mass compared to wild type (WT) mice under a normal diet, while Panx3 KO mice have reduced total fat mass and weigh less than WT controls. Despite having higher fat content, Panx1 KO mice on a high fat diet exhibited no differences in weight gain but showed an increase in glucose and insulin levels compared to WT. However, metabolic cage data revealed that these Panx1 KO mice display significantly increased activity levels, higher ambulatory activity, and reduced sleep duration relative to their WT littermates on a high‐fat diet. To uncover the cellular mechanism responsible for the increased fat content in the Panx1 KO, we isolated primary cultures of adipose‐derived stromal cells (ASCs) from WT and KO fat pads. In WT ASCs we observed that Panx1 protein levels increase upon induction into an adipogenic lineage. ASCs isolated from Panx1 KO mice proliferate less but demonstrate enhanced adipogenic differentiation with increased intracellular lipid accumulation, glycerol‐3‐phosphate dehydrogenase (GPDH) enzyme activity, and adipokine secretion, as compared to WT ASCs. This was consistent with the increased adipocyte size (hypertrophy) and decreased adipocyte numbers observed in subcutaneous fat of the Panx1 KO mice compared to WT. Panx3 is also expressed in adipose stromal cells and Panx3 protein levels significantly increase in differentiated adipocytes. However, the effect of the Panx3 KO seems to be the opposite of the Panx1 KO, with decreased weight under a high fat diet and lower fat mass (even under a normal chow diet). We propose that Panx1 and Panx3 may play opposing roles in adipose stromal cells during the early stages of adipogenic proliferation and differentiation, regulating fat accumulation in vivo.Support or Funding InformationFunded by the Canadian Institutes of Health Research (CIHR) grants to SP and LFThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of Proteins Released from the Egg Cell Surface During Fertilization
Authors: Wiseman Emily , Carroll David 
Published: June 2021

Multiple events occur at the interface between the sperm and egg membrane during fertilization, including sperm‐egg binding, species‐specific discrimination, membrane fusion, and egg activation. Each of these events is believed to rely upon discrete cell surface molecules. To date, only a handful of egg membrane proteins have been identified in any species. The process of fertilization as a whole is not well understood on a molecular level, from the initial sperm and egg binding, to the activation of early development. An excellent model organism to address these problems is the Pacific bat star, Patiria miniata, because they produce a large quantity of synchronized oocytes that are large, nearly transparent, and rigid enough to hold their shape despite removal of extracellular layers. Furthermore, we are learning that their genome is similar to humans with less complexity. Recent work suggests that proteases are necessary for successful fertilization, raising the possibility that fragments of egg membrane proteins could be released from the surface during fertilization. This could provide a mechanism for the identification of fertilization‐relevant membrane proteins. Using biotinylation and western blotting, we show that peptides ranging from 50kD to 250kD are being cleaved from the egg surface and released into the sea water within three minutes of introducing sperm. These peptides have been collected by affinity interaction against streptavidin conjugated mag‐sepharose beads and their identities are being determined by tandem mass spectrometry. Since a proteome has not yet been created for P. miniata, a recently developed transcriptome of the mature unfertilized egg (NCBI accession number: PRJNA398668) will be used to identify the peptides revealed by tandem mass spectrometry. Furthermore, this transcriptome is also being examined computationally with the purpose of identifying and grouping transcripts that may be integral to fertilization, as a whole, and with a specific focus on the egg cell surface. Understanding how fertilization takes place at the molecular level in starfish could eventually lead to understanding infertility issues, as well as offer new possible non‐hormonal prophylactics for use in humans.Support or Funding InformationNIH R15This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
GlycoSense™: A Simple Alternative to Existing Methods for Glycosylation Detection and Monitoring
Authors: Woods Robert J. , Saunders Matthew J. , Yang Loretta 
Published: June 2021

Lectenz® Bio has developed an innovative technology called GlycoSense™ that detects binding between glycans and glycan‐specific reagents that are conjugated to spectrally‐unique beads. By combining the individual bead‐based reagents into a multiplex array, a profile of key glycan features can be obtained in less than a minute on a basic cytometer. We illustrate the performance of the method using glycoprotein standards and demonstrate its utility by monitoring changes in glycosylation patterns upon treatment with glycoenzymes. The GlycoSense™ approach addresses the unmet need for a rapid and robust tool to analyze protein glycosylation. In addition to established carbohydrate‐detection reagents such as antibodies and lectins, this approach can employ the engineered proteins (known as Lectenz®), developed by Lectenz® Bio.Support or Funding InformationNIH U01 CA221213, R43 GM123863This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mechanical Stress and Alteration of Cellular Morphology Influences O‐GlcNAc Modification
Authors: Nagy Tamas , Cser Annamária , Kolarovszki Béla , Miseta Attila , Frank Dorottya 
Published: June 2021

Protein O‐linked N‐acetylglucosamine modification (O‐GlcNAc) is the result of a post‐translational modification that involves the reversible transfer of an N‐acetyl‐glucosamine molecule to Ser/Thr OH‐group of intracellular proteins. O‐GlcNAc modification is increasingly recognized as a key regulatory element, capable to modify the functionality and the life‐cycle of hundreds of proteins. O‐GlcNAc's participation in stress adaptation processes was evidenced after several environmental challenges such as heat shock, hypoxia or oxidative stress. However, intracellular O‐GlcNAc modification was not yet studied under mechanical stress, despite the fact that physical stress is a constant challenge for many cells in our body.In our present work, we used human periodontal ligament (PDL) cells isolated from teeth extracted for orthodontic indications. PDL cells were cultivated under standard cell culture conditions and exposed to various amount of compression force for 12 hours.We have analyzed cell survival by flow‐cytometry and protein O‐GlcNAc by Western‐blot. Since cytoskeletal filaments are essential to the structural integrity of the cells both in normal condition and under mechanical stress, we also studied the intracellular distribution of O‐GlcNAc in relation to actin, alpha‐tubulin, vimentin and the cellular shape. We have found that O‐GlcNAc elevated under moderate mechanical load (1.5 – 3 g/cm2) but did not changed significantly under higher load ( ≥ 7 g/cm2). Intracellular distribution of O‐GlcNAc in non‐loaded cell demonstrated an accumulation at several membrane protrusion sites which showed the most overlap with tubulin and no overlap at all with vimentin. Under mechanical stress, the cells retracted these protrusions and O‐GlcNAc distribution became more centralized as well.In conclusion, we found that O‐GlcNAc regulation responds to mechanical challenges and our data suggest that O‐GlcNAc is coupled to cellular morphology.Support or Funding InformationThis work was funded by the Research Fund of the University of Pécs, Faculty of Medicine (grant No.: AOK‐KA 2/2015 (06.30)) and the EU founded Hungarian project GINOP 2.3.2. –15 (grant No.: 2.3.2. –15‐2016‐00050).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Acute Exposure to E‐Cigarette Vapor Causes Changes in Apoptotic Pathways in the Lung
Authors: Heyob Kathryn M , Sugar Sophia S , Johnson Brooke , Mieth Saya , Pavlek Leeann R , Adelstein Jeremy , Schwieterman Neill A , Wold Loren E , Rogers Lynette 
Published: June 2021

BackgroundElectronic cigarette usage is increasing at an alarming rate primarily because of reports implicating their use as safer than traditional cigarettes. However, little is known about the acute effects on lung injury and inflammation. These studies tested the hypothesis that acute exposure to e‐cigarette vapor would result in lung inflammation and injury.MethodsC57Bl/6 mice were exposed to e‐cigarette vapor consisting of 10 mg/mL nicotine in a 50/50 mixture of propylene glycol/vegetable glycerin (PG/VG) or PG/VG alone for 6 hr/day, 5 days/week for either 1 or 2 weeks. Lungs were excised and snap‐frozen for analysis. Lung homogenates were analyzed by western blot for pAKT, pSTAT3, PECAM, BCL‐2, and BAX. RNA was isolated for measurement of MMP9, KC, and VEGF by RT‐PCR.ResultspAKT protein expression was decreased (0.54 vs 1.0 relative density, p=0.02) and pSTAT3 was increased (1.7 vs 1.0 relative density, p=0.02) in the e‐cigarette exposed mice vs PG/VG exposed mice after 1 week of exposure. At 2 weeks, pSTAT3 remained increased (1.9 vs 1.0 relative density, p=0.01) and both BCL‐2 (2.5 vs 1.0 relative density, p=0.02) and BAX (1.7 vs 1.0 relative density, p<0.05) were increased in the exposed mice compared to mice exposed to PG/VG alone. No differences were observed in PECAM, MMP9, KC, or VEGF.ConclusionsAcute exposure to e‐cigarette vapor caused changes in signaling molecules associated with apoptosis. While we did not observe differences in the inflammatory pathways tested, additional inflammatory and apoptotic markers may yield insights into the acute effects of e‐cigarette vapor exposure on the lung.Support or Funding InformationNIH NHLBI R01 HL139348 02This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Involvement of Activated Leukocyte Cell Adhesion Molecule (ALCAM/CD166) in Pulmonary Fibrosis
Authors: Kim Mi Na , Hong Jung Yeon , Kwak Eun Ji , Kim Eun Gyul , Lee Yong Ju , Kim Kyung Won , Sohn Myung Hyun 
Published: June 2021

RationaleIdiopathic pulmonary fibrosis is a chronic, progressive lung disease characterized by fibroproliferative matrix molecule accumulation, collagen deposition and apoptosis, eventually resulting in pulmonary dysfunction and death. Activated leukocyte cell‐adhesion molecule (ALCAM; CD166) is a cell adhesion molecule that has been implicated in adhesive and migratory attribution including leukocyte homing and trafficking and the cancer metastasis. We investigated the involvement of ALCAM on development of bleomycin‐induced pulmonary fibrosis in murine model.MethodsBleomycin‐induced pulmonary fibrosis model was established with wild type and ALCAM knockout mice. Pulmonary fibrosis was also induced in TGF‐ β transgenic mice that conditionally overexpress TGF‐β upon doxycycline administration. Levels of ALCAM were measured in lung tissue and bronchial alveolar lavage fluid (BALF) of wild type mice and TGF‐ β transgenic mice. Histopathologic investigation, collagen deposition were evaluated.ResultThe protein and mRNA levels of ALCAM were decreased in the BALF and lung tissue of bleomycin‐treated wild type mice. Reduced ALCAM level was observed in BALF of doxycycline administered TGF‐β transgenic mice relative to those of control mice. ALCAM knockout mice showed aggravated lung fibrosis response compared with wild type mice upon bleomycin treatment.ConclusionALCAM contributes to bleomycin‐induced pulmonary fibrosis associated with TGF‐β response.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of a Low Fat Diet versus a High Fat Diet on Lymphoid Cell Infiltration in a Mouse Model of Oral Cancer
Authors: Pitstick Lenore , Green Jacalyn M , Goral Joanna , Barakat Ameir , Meyer Alice , Pytynia Matthew , Ciancio Mae , Jham Bruno 
Published: June 2021

IntroductionRisk factors for oral cancer include tobacco use, male sex, and possibly dietary fat. Exposure to 4‐nitroquinoline 1‐oxide (4NQO) in the drinking water causes oral cancer in mice and models the effects of tobacco use. This study was undertaken to investigate the effects of a high saturated fat (HF) or low fat (LF) diet and sex on presence of lymphoid cells in cancerous tongues. We hypothesized that inflammation, characterized by the increased presence of lymphoid cells, would accompany development of tumor lesions and that sex and diet would affect the levels of inflammation.MethodsMale and female C57Bl/6 mice (36 each gender), 5 weeks old, were divided into a low fat (10kcal% fat; LF) or HF (60 kcal% fat) diet. Within each dietary group, mice were randomly assigned to one of 3 water treatment groups for 17 weeks: water alone (control); propylene glycol in water (1.25%; PG‐H2O); or 4NQO in PG‐H2O (50 mg/ml; 4NQO). After 17 weeks, all mice were given water alone for 6 more weeks. Tongues harvested from the euthanized animals were fixed in formalin and processed for histological examination. T cells (CD3+ cells), macrophages (CD68+ cells), and neutrophils (Ly6+ cells) were detected by immunohistochemistry, counted, and normalized to tissue area. Analysis was done using GraphPad Prism 7.0. All animal work was in compliance with the regulations of IACUC at Midwestern University, Downers Grove, IL.ResultsOral cancer developed in all mice exposed to 4NQO and was absent in all controls. All three lymphoid cell types were significantly higher (p<0.05) in the 4NQO mice versus the control mice: CD3+ (11.2 vs 0.9 cells/mm2), CD68+ (3.6 vs 1.5 cells/mm2), and Ly6+ (9.4 vs 0.38 cells/mm2). Among the 4NQO treated mice, there were no significant differences between males and females, or between HF and LF mice for T cells or macrophages. 4NQO treated males had a significantly higher numbers of neutrophils than females (12 vs 6.6 cells/mm2). LF diet increased the levels of neutrophils in both males and females (15 and 7.7 cells/mm2 respectively) when compared to the HF diet in both males and females (9.3 and 5.3 cells/mm2 respectively); however, this trend was not statistically significant. We speculate that the higher carbohydrate content of the LF diet may support increased anaerobic glycolytic metabolism via the Warburg effect.ConclusionDevelopment of 4NQO‐induced oral cancer was accompanied by inflammatory responses as manifested by the presence of T cells, macrophages, and neutrophils. Neutrophils were the only cell type affected by sex and possibly by the content of dietary fat.Support or Funding InformationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Immunological and Bacteriological Assessment of Women with Breast Cancer in Benin, Nigeria
Authors: Okojie Rachel Obhade , Ojo Matilda Adesuwa 
Published: June 2021

Neutrophils and lymphocytes are important cells involved in cellular immune response. Pre‐treatment neutrophil‐lymphocyte ratio (NLR) is a biomarker which can be used to predict disease progression, mortality and morbidity in breast cancer patients. Fungating breast lesions found in advanced stages of breast cancer patients are usually infected with opportunistic mixed bacteria flora and treatment with systemic antibiotics is often geared at alleviating symptoms and optimizing the quality of life of the patients.A total of 60 participants (30 patients and 30 apparently healthy women as controls) were involved in this study. Full blood count analysis was done using Sysmex KX‐21N Automated Hematology Analyzer. Swab samples were also collected from the breast wounds. These were cultured and isolated using appropriate media and standard microbiological techniques. Antibiotic susceptibility test was determined using commonly used antibiotics.It was observed that there is a preponderance of breast cancer within the study area. Majority of the subjects presented at the advanced stages (stage 3 and 4) and they had high values of neutrophil‐lymphocyte ratio. The mean value of the neutrophil‐lymphocyte ratio (NLR) obtained for the women with breast cancer (3.50±2.7 x103/μl) was found to be higher than the ratio of the control group (2.1±0.56 x103/μl). Using 2.3 as the cut‐off marker for NLR, 36.7 % of the subjects had NLR less than 2.3 while 63.3 % were higher or equal to 2.3; which were found to be statistically significant (p < 0.05). It showed that majority (40%) of the cancer subjects presented at stage 4 and out of which 30% had fugitive breast lesions. The predominant bacteria isolated from the lesions were Escherichia coli (35.7 %), Proteus mirabilis (28.6 %), Pseudomonas aeruginosa (21.4 %) and Staphylococcus aureus (14.3 %). All the bacterial isolates showed high susceptibility to pefloxacin and ceftriaxone (78.50 %) respectively and exhibited very low susceptibility to amoxicillin (7 %) and ampiclox (0 %).It is pertinent to increase public awareness of breast cancer, to encourage women to be involved in routine medical examination and to educate women with breast cancer to present early to the hospitals for adequate treatment, as these will help to manage the progression of the disease and also reduce mortality and morbidity. Appropriate use of antibiotics is also very important.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The BTB domain protein‐Cul3 complex is a novel regulator of Collagen expression in Breast Cancer
Authors: kim kyungho , Kim Youn‐Jae 
Published: June 2021

CUL3‐RING ubiquitin ligases are associated with diverse cellular processes through over two hundred BTB‐domain proteins. RHOBTB3, a BTB‐domain protein, harbors capable of assembling Cullin3‐ dependent ubiquitin ligase complexes and recent studies have been suggested to play an essential role in transport vesicle docking at the Golgi complex. Collagen type I alpha 1(Col1A1) reportedly associated with tumor interstitial macromolecular transport and cancer cell dissemination. However, the precise function and mechanism of Col1A1 remain unclear how Col1A1 is regulated in human breast cancer growth and metastasis. To address this question, we analysed RHOBTB3 and Col1A1 gene expression in breast cancer using the public database. We exemplify each three representative studies showing RHOBTB3 and Col1A1 are generally upregulated in breast cancer. To determine the effects of RHOBTB3 and Col1A1 expression on metastasis and involvement of RHOBTB3 and Col1A1, we performed knock‐down of RHOBTB3 in breast cancer cells. The knock‐down of RHOBTB3 significantly decreased expression of COL1A1 mRNA and protein and inhibition of cell growth and invasion in breast cancer. Collectively, these findings demonstrated RHOBTB3 is an important factor for regulation of the COL1A1 synthesis and inhibition of cell growth and invasion in breast cancer cells. Thus, RHOBTB3 is oncogenic and a potential oncotarget for breast cancer therapy.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Overexpression of Elabela Transgene Ameliorates UUO‐Induced Renal Fibrosis through Suppressing TGF‐β in Sparague‐Dawley Rats
Authors: Wu Chunying , Zhu Yeyan , Li Haonan , Wang Lei , Zhu Qing 
Published: June 2021

Elabela (ELA) is a newly discovered endogenous ligand of APJ receptor, which is widely distributed in various tissues and organs including kidney. Accumulated evidences demonstrate that ELA plays a critical role in the regulation of renal physiology and pathophysiology. Our preliminary data showed that ELA expression was significantly inhibited in the kidney of unilateral ureteral obstruction (UUO) rats. However it remains unknown whether ELA has renoprotective effects in UUO model. The present study tested the hypothesis that overexpression of ELA transgenes protect against renal fibrosis in UUO rats. Male Sprague‐ Dawley rats (200–250g body weight) were subjected to UUO surgery and plasmids expressing ELA transgenes or control vectors were transfected into the obstructed kidneys via renal arteries. After 7 days, the rats were scarified and the kidneys were collected for analysis. The ELA mRNA levels were decreased by 70% in UUO rats compared to that of the control rats, which were significantly increased by 2‐fold in the UUO+ELA plasmids‐treated rats, suggesting successful overexpression of ELA in the kidney via transfection of ELA overexpression plasmid. Siris red staining results showed that collagen accumulation was increased in the UUO rats, which was dramatically suppressed in UUO+ELA plasmids‐treated animals. The mRNA levels of Collagen I, Fibronectin and α‐SMA were all elevated in the UUO group and were notably attenuated in the UUO+ELA plasmids‐treated group. Furthermore we examined the expression of TGF‐β, a central mediator of renal fibrosis and the results showed that the mRNA and protein levels of TGF‐β were upregulated in UUO rats but greatly decreased in UUO+ELA plasmids‐treated rats. These results suggest that the impaired activation of ELA in the kidney may represent a novel mechanism for renal fibrosis under pathological condition and that correction of this impairment may serve as a novel therapeutic approach for the treatment of chronic renal fibrotic diseases.Support or Funding InformationThis work was supported by grants from the National Natural Science Foundation of China: No. 81600322 and No. 81770707This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.