bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lysophosphatidic Acid Stimulation of Amphiregulin Shedding Enhanced Paracrine Role of BM CD11+ Cells
Authors: Liu Tianju , De Los Santos Francina Gonzalez , Wu Zhe , Phan Sem H 
Published: November 2023

Amphiregulin (AREG), an epidermal growth factor receptor ligand, plays an important role in tissue repair and fibrosis. Bone marrow (BM) CD11c+ cell‐derived AREG promotes pulmonary fibrosis by activating fibroblast proliferation, migration and collagen expression. Pro‐AREG is a membrane bound protein whose mature soluble form represents the extracellular domain that is cleaved off by ADAM17. However, the precise mechanism of regulation or shedding/release of mature AREG is uncertain. Lysophosphatidic acid (LPA) is a bioactive lipid that mediates diverse cellular responses through interactions with specific G‐protein‐coupled receptors. LPA is implicated in tissue repair and wound healing processes, and elevated in the airspaces of patients with idiopathic pulmonary fibrosis, as well as fibrotic lungs of animal model of fibrosis. LPA is known to be able to induce shedding of TGFβ and AREG from cell surface. The objective of this study was to assess the effects of LPA on shedding of AREG ectodomain from CD11c+ cells, and the consequent effect on fibroblast activation. The results showed that LPA caused rapid shedding of AREG from both mouse BM‐derived dendritic cells (BMDC) and human BM CD11c+ mononuclear cells as revealed by the decrease in cell surface AREG expression analyzed by flow cytometry. AREG shedding was confirmed by the elevated level of released AREG protein by LPA‐stimulated BMDC as measured by ELISA. To investigate the paracrine activity of AREG released from BMDC, mouse lung fibroblasts were cultured in the presence of conditioned media from LPA‐stimulated BMDC cultures. The data showed that the fibroblast proliferation rate was significantly increased by these AREG containing conditioned media. These findings together indicated that induced LPA in fibrotic lung tissue could stimulate the shedding of AREG from CD11c+ BM cells, including dendritic cells, and thus enhanced the paracrine role of CD11c+ cells on fibroblast activation in tissue repair and remodeling. They also suggested that targeting the LPA receptor may further mediate its potential therapeutic efficacy for control of tissue fibrosis by reducing AREG shedding.Support or Funding InformationNIH grants HL052285 and HL112880.
Identifying Key Domains in Endothelial IQGAP1 Critical for Leukocyte Transmigration
Authors: Dalal Prarthana , Sullivan David , Muller William 
Published: November 2023

Inflammation results in the dilation of local blood vessels, the activation of endothelium by cytokines and the recruitment of circulating leukocytes into areas of tissue damage. The majority of leukocytes undergo transendothelial migration (TEM) by using a unique endothelial cell membrane system referred to as the lateral border recycling compartment (LBRC). IQ‐domain GTPase‐activating protein 1 (IQGAP1) was identified as an LBRC‐associated protein and subsequently found to be essential for TEM. IQGAP1 is a scaffolding protein comprised of six major interaction domains. It has been reported to interact productively with over 135 cellular proteins. In endothelial cells IQGAP1 partially localizes to cell borders and knocking down its expression prevents TEM. However, it is not known which of the many proteins it is known to interact with carries out its role in TEM. Since these proteins bind to specific IQGAP1 domains, as a first step toward understanding the mechanism by which IQGAP1 regulates TEM, we explored which of its six protein interactions domains were critical for its function. We knocked down IQGAP1 in endothelial cells and re‐expressed truncated forms of IQGAP1 with specific regions deleted to identify the regions (and thereby the potential binding partners) that are critical for TEM. We show that the actin‐binding domain and the calmodulin‐binding isoleucine‐glutamine (IQ) domain of IQGAP1 are critical for its function during TEM.Human endothelial cells in which >80% of endogenous IQGAP1 was depleted by shRNA did not support TEM; re‐expression of wild‐type IQGAP1 restored TEM to control levels. Re‐expression of IQGAP1 with a GFP‐tagged construct after knockdown also showed IQGAP1 enrichment around transmigrating leukocytes. Re‐expression of IQGAP1 truncation mutants lacking the N‐terminal actin‐binding domain did not localize to endothelial borders and did not support TEM. Re‐expression of truncated IQGAP1 constructs containing the actin‐binding domain and the IQ domain localized to cell borders and supported TEM. Constructs containing the actin‐binding domain but lacking the IQ domain localized to the cell borders but did not restore TEM.After identifying the IQ domain as having a critical role in TEM, we generated specific point mutations in this domain within the full‐length construct. In particular, the isoleucine (I)‐glutamine (Q) motif after which the IQ domain is named has been identified in the literature as having an essential role in IQGAP1 function and we are currently testing whether they are also important in TEM. Using these constructs in future studies will help identify the role of the IQ domain in TEM and its interacting partner(s). Identifying the binding partners of IQGAP1 that are critical for inflammation will provide clues to its mechanism of action and potential therapeutic targets for anti‐inflammatory therapy.Support or Funding InformationFunded by R01 HL046849, R01 HL064774, Alpha Omega Alpha Carolyn Kuckein Student Research Fellowship
Immunoproteomic Identification of Bovine Pericardial Heart Valve Antigens
Authors: Gates Katherine Yvonne , Dalgliesh Ailsa , Griffiths Leigh 
Published: November 2023

American Heart Association estimates the prevalence of valvular heart disease to be 2.5% in the United States. Biological heart valve replacements, such as glutaraldehyde‐fixed bovine pericardium (BP), are considered the current standard of care. The NHLBI xenotransplantation working group identified xenoantigenicity as the major barrier to expanding use of such xenogeneic tissues. Previous research has utilized an immunoproteomic approach based on two‐dimensional western blots to identify a small number of BP xenoantigens, which is time consuming and technically challenging. These limitations have resulted in an ever increasing use of shotgun proteomic approaches due to their superior throughput and reproducibility. However, traditional Liquid Chromatography‐Mass Spectrometry (LC‐MS) fails to separate proteins based on their antigenicity, and therefore it is impossible to determine which identified proteins are antigenic versus not. We hypothesized that an affinity chromatography method utilizing poly‐polyclonal serum would bridge this gap and represent an innovative shotgun proteomic antigen identification approach. Using affinity chromatography rabbit IgG antibodies were isolated from whole serum with Protein A HP SpinTrap (GE Healthcare). IgG capture was confirmed by serum protein electrophoresis and ELISA. Two types of antibody affinity columns were generated, with naive serum (D0) or serum from rabbits immunized with bovine pericardium (D84). Bovine pericardial protein extracts were run through both affinity columns, allowing for the specific capture of antigens, which were later eluted and collected. To confirm specificity of antigen capture, eluants were subjected to western blot, and probed with D84 serum. Affinity enriched antigenic proteins were identified by LC‐MS/MS, with statistical comparisons made between D0 and D84. This approach identified 133 putative antigenic proteins that were enriched on D84 compared to D0 columns. Ten of these antigens corresponded to previously published BP xenoantigens. Of the remaining 123 previously unidentified antigens, 30 have been indicated to be immunogenic in other fields such as cancer and autoimmunity. This method was then applied to three humans who had been implanted with bovine pericardial heart valves. Surprisingly, 300 to 500 proteins were identified from patient columns. Between 57 to 79% of the identified proteins were shared among the three individuals, as well as almost all of the proteins identified in the rabbit model were also present in the human. This work therefore represents the first high throughput method of antigen identification that can now be used to monitor currently implanted patients as well as inform the development of BP‐based biomaterials in the future.Support or Funding InformationResearch supported by grant number R01HL115205 from the National Heart Lung and Blood Institute (NHLBI) at the National Institutes of Health (NIH) and grant number 5T32OD010931‐10 from the NIH.
Effect of Probiotic Pediococcus pentosaceus on intestinal permeability and occludin protein distribution in a murine model of colitis
Authors: Torres‐Aguilar Lorena , Rodriguez‐Fragoso Lourdes , Garcia‐Vazquez Francisco , Reyes‐Esparza Jorge 
Published: November 2023

Ulcerative colitis (UC) is a chronic inflammatory disease of the colon. In individuals with ulcerative colitis, redistribution and downregulation of several tight junction proteins has been observed in active disease as a molecular basis of increased epithelial permeability. A new alternatives for the treatment of UC is the use of probiotics. We have shown recently that Pediococcus pentosaceus treatment at the dose of 1×1010 CFU delayed the onset of some of the symptoms of colitis in a murine model of colitis. The aim of this study is to evaluate the effect of Pediococcus pentosaceus on intestinal permeability and distribution of tight junction occludin. Twenty‐one male BALB/c mice weighing 20–22 g were divided in three groups: Control, Colitis and Colitis+ P. pentosaceus (n = 7 each). Acute colitis was induced by administration of dextran sodium sulfate (DSS) (molecular weight 40 kDa) 3.5% wt/vol in drinking water ad libitum for 6 days. At day 2 during induction of colitis mice were intragastrically gavaged by P.pentosaceus (doses 1×1010 Colony Forming Units). At the seven day of study the intestinal permeability was measured by determination of the fluorescence intensity of FITC‐dextran (molecular weight 4.0 kDa) in serum after 4 hours of oral administration and the occludin distribution were studied by immunohistochemistry. The results showed a decrease of intestinal permeability in the Colitis+P. pentosaceus group with a 39% less of fluorescence intensity in serum compared with Colitis group. In addition, we could see the occluding distribution was exclusively localized at the apical junctional complex both at the surface and in the crypts. In conclusion Pediococcus pentosaceus treatment improve the intestinal barrier function decreasing the intestinal permeability and increasing the occludin distribution epithelial in a murine model of colitis.Support or Funding InformationLorena Torres Aguilar is fellowship holder from CONACYT
PAD4 Deficiency Limits Kidney Dysregulation in a Murine Model of Shock/Sepsis
Authors: Girard Bethany Biron , Chen Yaping , Chung Chun‐Shiang , Reichner Jonathan Biron , Ayala Alfred 
Published: November 2023

Indirect Acute Respiratory Distress Syndrome (iARDS) is caused by a non pulmonary inflammatory process as a result from insults such as non‐pulmonary sepsis, hypotensive shock, etc. Patients with ARDS frequently develop Acute Kidney Injury, which in turn, increases overall mortality. Neutrophils are thought to have a significant role in mediating ARDS, with the development of iARDS being characterized by dysregulation and recruitment of activated neutrophils to the lung. PAD4 is an enzyme responsible for the citrullination of histones and subsequent neutrophil extracellular trap (NET) formation. Components of NETs including neutrophil derived circulating free DNA as well as circulating histones has been implicated in ARDS as well as AKI in ischemia/reperfusion injury models. However, it is unknown if there is any pathological significance of NET formation in AKI as a result from a “2 hit” model of traumatic shock (fixed‐pressure hemorrhage; Hem) followed by septic (CLP) insult (Hem/CLP)(iARDS). Previously, we have demonstrated that PAD4 gene deficiency improves overall survival as well as decreases pro‐inflammatory cytokines in the lung in this model of iARDS. Here we aim to determine if PAD4 deficient mice (PAD4−/− mice vs. WT mice) were also protected from AKI in an iARDS model. After Hem/CLP (the induction of iARDS) WT mice displayed increased vascular leakage in the kidney as well as increased circulating Blood Urea Nitrogen (BUN) levels. In contrast, PAD4 −/− mice were protected against increased vascular leakage within the kidney and displayed significant decreases in BUN levels.ConclusionThese data taken together suggest that PAD4 deficiency protects against kidney dysfunction associated with ARDS, which in turn contributes to the overall improved survival in the Hem/CLP model of iARDS.
A study protocol for the comparative study of biological and metabolism bio‐marker between healthy population and patients with acne vulgaris
Authors: Kim Kyuseok , Ha Injin , Kim Eunok 
Published: November 2023

Recent studies have reported that pro‐inflammatory mediators and cytokines related to innate immune responses and insulin resistance related factors, which induce sebaceous lipogenesis, sebocyte, and keratinocyte proliferation via milk consumption and hyperglycemic diets, are important roles in the pathogenesis of acne vulgaris. However, current level of evidence is poor to sufficiently explain the pathophysiology of acne vulgaris. Therefore, this study was designed to find out the molecular biological and metabolism bio‐marker between healthy population and patients with acne vulgaris. The study will consist of 40 healthy subjects and 60 patients with acne vulgaris. The molecular biological biomarker will be analyzed with the use of several cytokines (IL‐1α, IL‐1β, IL‐12 β, IL‐15, IL‐6 and TNF α) and chemokines (MCP‐1 and IL‐8) examinined by ELISA, real‐time PCR, or Western blot assay, whereas the metabolism biomarker will be assessed with steroidal hormones (DHEA, androstenedione, testosterone, DHT, estrone, and estradiol), saturated / trans‐fats (palmitic acid, elaidic acid, etc.) and untargeted markers related to TCA cycle by using the LC‐MS. We will examine skin sebum, skin surface temperature and skin viscosity elasticity in constant temperature and humidity room at Korean Medicine Clinical Trial Center, Kyung Hee University Korean Medicine Hospital (Seoul, Republic of Korea). In addition, we will collect the information including their diet pattern, menstruation (only female), medication history, smoking, and sleep pattern from subjects through a questionnaire.Support or Funding InformationThis study was supported by a grant from the Traditional Korean Medicine R&D Project, Ministry of Health & Welfare, Republic of Korea. (HI16C0801)
Involvement of RAGE signaling and inflammatory cytokine elaboration following in vitro exposure to electronic cigarette liquid
Authors: Hirschi Kelsey M , Lewis Joshua B , Ostergar Adam S. , Hall Parker D. , Broberg Dallin S. , Arroyo Juan A. , Reynolds Paul R. 
Published: November 2023

The development of the electronic cigarette (e‐cig) has given rise to a new, largely unregulated market within the smoking industry. Proponents suggest they assist with smoking cessation as e‐cigs rely on vapor produced by heating an e‐liquid to deliver flavored nicotine. While generally assumed to be less harmful than traditional tobacco cigarette smoke, knowledge about the toxicity of the e‐liquid consumed via such electronic devices is still scarce. During use of an electronic cigarette, the oral cavity and respiratory system receives direct exposure to the chemical constituents of the e‐liquids. An additional area of concern is the effect of e‐cigs on the placenta due to a potential misconception that e‐cigs are safer to use during gestation than classical tobacco. Traditional cigarette smoke compromises health by increasing inflammation, mediated through receptor for advanced glycation end‐products (RAGE) signaling. Pro‐inflammatory profiles have been suggested following e‐cig exposure; however, the role of RAGE in e‐cig liquid‐induced inflammation has not been resolved. The purpose of this study was to determine the effects of e‐liquid on RAGE expression and levels of several pro‐inflammatory cytokines mediated at least in part by RAGE signaling such as interleukin (IL)‐1b, IL‐6, IL‐8 and tumor necrosis factor (TNF)‐a. Gingival epithelium (Ca9‐22), pulmonary epithelium (A549), and placental trophoblast (BeWo) cells were exposed to 0, 2, or 4% of Cuttwood™ e‐cigarette liquid, a popular national brand, for 24 hours. Cytotoxicity was measured by LDH assays. E‐cig induced RAGE upregulation was confirmed by quantitative RT‐PCR in A549 and Ca9‐22 cells while only a trend in RAGE upregulation was observed in BeWo cells. QPCR revealed increased transcription of IL‐1b, IL‐6, IL‐8 and TNF‐a in each cell type. Finally, a multiplex cytokine array was performed in order to precisely determine the secretion of these and other pro‐inflammatory modulators. In an effort to block RAGE signaling, we have synthesized a family of anionic, partially lipophilic sulfated polysaccharide derivatives known as semi‐synthetic glycosaminoglycan ethers (SAGEs). Co‐treatment of e‐cig liquid and SAGEs resulted in amelioration of inflammatory cytokine secretion. Together, these results provide a new perspective on a possible mechanism whereby e‐cigarette inflammation occurs.Support or Funding InformationThis work was supported by a grant from the Flight Attendant's Medical Research Institute (FAMRI, PRR) and a BYU Mentoring Environment Grant (PRR).
Investigating the Potential Role of North American Animals as Hosts for Zika Virus
Authors: Ragan Izabela , Blizzard Emily , Bowen Richard 
Published: November 2023

The recent emergence of the mosquito‐borne Zika virus (ZIKV) in the Americas has become a global public health concern. While little is known about potential animal reservoirs for ZIKV, nonhuman primates are thought to play a role during enzootic transmission cycles. We describe a series of experimental infections designed to investigate whether species within certain taxonomic groups in North America have the potential to serve as ZIKV amplifying hosts or reservoirs. The objectives of this study were to (1) identify potential animal reservoirs in North America with an emphasis on wildlife and livestock, (2) examine ZIKV pathogenicity in neonatal and adult animals, and (3) evaluate the ability of animal models to recapitulate neonatal birth defects as seen in human patients. Species investigated include armadillos, cottontail rabbits, goats, mink, chickens, pigeons, ground hogs, deer mice, cattle, raccoons, ducks, Syrian Golden hamsters, garter snakes, leopard frogs, house sparrows, and pigs. Animals ranging in age from neonates to adults were inoculated with 105 PFU of one of two ZIKV strains (PRVA BC59 and FSS13025) by subcutaneous and intradermal injection. All animals were monitored daily for clinical signs of disease. Blood samples and body temperatures were collected on days 0–5, 7, 14, 21 and 28 post inoculation. Plaque assays, polymerase chain reaction (PCR), and plaque reduction neutralization tests were performed to detect viral presence and antibody production. Infectious virus was isolated from seven frogs and one armadillo. In addition, neutralizing antibodies were detected in goats, rabbits, ducks, frogs, and pigs. This study indicates that the animals tested to date are unlikely to act as animal reservoirs for ZIKV. However, goats, rabbits, frogs, and pigs could potentially serve as a sentinel species. Future work includes examining the role of amphibians in the transmission dynamics of ZIKV. Understanding the transmission cycle and maintenance of ZIKV in animals will help in developing effective surveillance programs and preventative measures for future outbreaks.Support or Funding InformationThis material is based upon work supported by Colorado State University Animal Models Core and the U.S. Department of Homeland Security under Grant Award Number DHS‐2010‐ST‐061‐AG0001. The views and conclusions contained in this document are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of the U.S. Department of Homeland Security.
Floating microelectrode arrays chronically implanted into the feline peripheral nerve elicit a characteristic tissue response
Authors: Kolarcik Christi L , Puglisi Kelly , Lesniak Andrew , Demetris Anthony J , Fisher Lee E , Gaunt Robert A 
Published: November 2023

Neural interfacing technologies have the potential to significantly improve the quality of life of patients that are severely disabled or suffering from a variety of neurodegenerative conditions. To recapitulate proper physiological function, motor commands and sensory feedback must be considered. The spinal nerve is a unique location at which these complementary systems can be accessed. In this study, the dorsal root ganglion‐ventral root (DRG‐VR) complex was targeted with floating microelectrode arrays (FMAs) designed either to record from the VR or test stimulation paradigms in the DRG. Extensive characterization of the tissue response to these chronically implanted microelectrode arrays was performed to determine the reliability of these peripheral locations for interfacing applications. To accomplish these analyses, hematoxylin and eosin and Nissl/Luxol fast blue staining along with antibody‐based stains [NF200 (axons), S100 (Schwann cells), vimentin (fibroblasts, endothelial cells, astrocytes), MAC387(monocytes/macrophages), GLUT1 (glucose transport proteins)] were employed, regions of interest representing the entire area of the implanted array defined, and pixel‐based image analyses specific for each stain utilized. Implanted roots were compared to the non‐implanted (i.e., contralateral) roots from the same cohort of animals. The results indicate that the inflammatory reaction elicited with array insertion is not significantly greater than what is observed in a control (i.e., non‐implanted) spinal nerve. The absence of a significant immune response to these arrays suggests some level of immune privilege in the spinal nerve and bodes well for long‐term applications. In addition, no significant decreases in neuronal density or myelination were observed in the area associated with the array indicating that both neuronal cell bodies and axonal projections are maintained. Observations related to the status of the blood‐nerve barrier suggest it is preserved and/or reestablished at the chronic time points examined. Our data indicates that targeting the DRG‐VR complex is a viable option and that the associated tissue response in this location is an opportunity to achieve the overall goal of a long‐term, reliable interface with prosthetic limbs.Support or Funding InformationThis work was sponsored by the Defense Advanced Research Projects Agency (DARPA) Microsystems Technology Office under the auspices of Dr. Jack Judy through the Space and Naval Warfare Systems Center, Pacific Grant No. N66001‐11‐C‐4171.