Attenuation of Protein Adsorption on Static and Vibrating Magnetic Nanowires

2005 ◽  
Vol 877 ◽  
Author(s):  
Kristy M. Ainslie ◽  
Gaurav Sharma ◽  
Maureen A. Dyer ◽  
Craig A. Grimes ◽  
Michael V. Pishko
Keyword(s):  
Cell Adhesion ◽  
Host Response ◽  
Nanowire Array ◽  
The Body ◽  
Nanowire Arrays ◽  
Enzyme Linked Immunosorbent Assay ◽  
Magnetic Nanowires ◽  
Nanostructured Surfaces ◽  
Protein Adhesion

AbstractThe research described here investigates the hypothesis that nanoarchitecture contained in a nanowire array is capable of attenuating the adverse host response, biofouling, generated when medical devices, such as sensors, are implanted in the body. This adverse host response generates an avascular fibrous mass transfer barrier between the device and the analyte of interest, disabling the sensor. Numerous studies have indicated that surface chemistry and architecture modulated the host response. These findings lead us to hypothesize that nanostructured surfaces will significantly inhibit the formation of an avascular fibrous capsule. We are investigating whether vibrating magnetostrictive nanowires, formed in nanowire arrays, can prevent protein and cell adhesion. Magnetostrictive nanowires are fabricated by electroplating a ferromagnetic metal alloy into the pores of a nanoporous alumina template. The ferromagnetic nanowires are made to vibrate by altering the magnetic field surrounding the wires. Enzyme-linked immunosorbent assay (ELISA) and other protein assays were used to study protein adhesion on the nanowire arrays. These results display a reduced protein adhesion per surface area of static nanowires. The vibrating nanowires show a further reduction in protein adhesion, compared to static wires. Studies were also preformed to investigate the effects of nanoarchitecture have on cell adhesion. These studies were performed with both static and vibrating nanowires. Preliminary protein adhesion studies have shown that a nanowire arrays modulate protein adhesion in vitro.

Maturation and Terminal Differentiation of Embryonic Erythroid Cells in the Developing Fetal Liver

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1369-1369
Author(s):  
Margaret H. Baron ◽  
Joan Isern ◽  
Stuart T. Fraser ◽  
Zhiyong He
Keyword(s):  
Cell Adhesion ◽  
Transgenic Mouse ◽  
Peripheral Blood ◽  
Fetal Liver ◽  
The Body ◽  
Surface Proteins ◽  
Erythroid Cells ◽  
Transgenic Mouse Line ◽  
Β1 Integrins

Abstract Primitive erythroid cells (EryP) are the earliest differentiated cell type of the mammalian embryo, arising in the yolk sac by the end of gastrulation. They begin to enter the embryonic circulation two days later and continue to mature in a stepwise and synchronous fashion. Around the time the embryonic circulatory system is forming, EryP comprise 25–40% of all cells in the embryo, indicating that huge resources have been set aside specifically for their development. Although, like their adult counterparts, EryP enucleate, they circulate throughout the embryo for several days before the first enucleated forms can be identified in the blood. We have used novel transgenic mouse lines to investigate this seemingly long lag and have identified a previously unappreciated developmental niche for EryP maturation. As they circulate in the peripheral blood (PB), EryP begin to express selected cell adhesion proteins, including α4 and β1 integrins, on their surface and migrate into the fetal liver (FL). They are found in the FL parenchyma as early as E10.5, in contact with macrophages within erythroblastic islands, and continue to accummulate there through E14.5. One day later (E15.5), nearly all EryP have enucleated and re-entered the circulation. Integrins α4, β5 and β1 (but not αV, β2, or β3) are dramatically upregulated on the surface of EryP that have entered the FL, to levels that are much higher than those found on PB EryP at the same stage. The high level cell adhesion molecule expression is reflected in greatly enhanced binding to fetal liver macrophages in vitro. Blocking of VCAM-1, a counter-receptor for α4β1 integrin that is expressed on macrophages, abrogates this adhesion. To examine EryP enucleation at higher resolution, we generated a transgenic mouse line in which the nuclei of EryP are marked by a histone H2B-EGFP fusion protein. Extruded EryP nuclei could be detected in very low numbers in the circulation and in much larger numbers within the FL at E12.5. Enucleated EryP are first detected within the blood at E12.5. These observations, along with the green fluorescence detected in E10.5 FL, may indicate that EryP enucleation begins earlier than previously believed and, therefore, that the resulting reticulocytes may not be released into the circulation immediately. Moreover, the large number of extruded nuclei in FL compared with peripheral blood argues that most enucleation events occur within the FL. The H2B-EGFP reporter allowed us to use flow cytometry (FACS) to isolate and characterize surface protein expression on the extruded nuclei. Adhesion molecules (α4, α5, and β1 integrins) were greatly enriched on the surface of the extruded nuclei, demonstrating that these proteins are selectively partitioned away from the body of the developing reticulocyte. The redistribution of surface proteins to the extruding nucleus may facilitate phagocytosis by macrophages and/or the release of integrin-poor reticulocytes into the circulation. Extruded EryP nuclei can be identified within FL macrophages both in vitro and in vivo. We conclude that EryP home to and enucleate within the fetal liver, a tissue that is just developing as EryP begin to circulate. We are currently using a number of molecular and cellular approaches to determine whether similar mechanisms are involved in the maturation of definitive erythroid cells in the adult bone marrow.

Circulating MMP-7 and VEGF as potential predictive biomarkers for recurrent implantation failures

Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Mustapha Benkhalifa ◽  
Wiem Zidi ◽  
Hatem Bahri ◽  
Sami Mahjoub ◽  
Khaled Boudhraa ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Adhesion Molecules ◽  
Embryo Implantation ◽  
Predictive Biomarkers ◽  
Limiting Factors ◽  
Leukaemia Inhibitory Factor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Implantation Failure ◽  
Roc Curve Analysis

Summary Recurrent implantation failure (RIF) is considered to be one of the major limiting factors of assisted reproductive technology (ART) programme success. The current study focused on the investigation of matrix metalloproteinases (MMPs), tissue inhibitors of MMPs (TIMPs), cytokines and cell adhesion molecules in peripheral blood (PB) and follicular fluid (FF) obtained from 44 women aged between 25 and 39 years old and undergoing intracytoplasmic sperm injection (ICSI). These women were divided into two groups: 22 RIF women with embryo implantation failures after the transfer of at least four fresh or frozen–thawed good quality embryos in a minimum of three ICSI cycles, and 22 ICSI success women (controls) who achieved a clinical pregnancy at their first ICSI attempt. The PB and FF samples were obtained from each patient on the day of oocyte retrieval. MMP-1, -2, -3, -7, -9, TIMP-1, -2, vascular endothelial growth factor (VEGF), leukaemia inhibitory factor (LIF), vascular cell adhesion molecule 1 (VCAM1) and intercellular adhesion molecules 1 (ICAM1) were analyzed using enzyme-linked immunosorbent assay of PB and FF. Our results showed significant decreases in PB MMP-7 and PB VEGF in the RIF group compared with controls [281.11 (33–614) pg/ml vs 119.92 (27–441) pg/ml; P-value = 0.030] and [82.54 (25.94–210.20) pg/ml vs 30.93 (13.62–193.33) pg/ml; P-value = 0.022; respectively]. Receiver operating characteristic (ROC) curve analysis showed informative area under the curve values for PB MMP-7, as well as for PB VEGF, making them able to be proposed as biomarkers of the RIF. Therefore, circulating MMP-7 and VEGF seem to play an interesting role in embryo implantation in in vitro fertilization (IVF)/ICSI cycles and could be proposed as circulating biomarkers of the RIF. These results could be helpful for clinicians and patients to choose the best rescue strategy and treatment to minimize implantation failure in women undergoing IVF/ICSI procedures after the first attempt.

scholarly journals Heparin oligosaccharides bind L- and P-selectin and inhibit acute inflammation

Blood ◽  
1993 ◽  
Vol 82 (11) ◽  
pp. 3253-3258 ◽  
Author(s):  
RM Nelson ◽  
O Cecconi ◽  
WG Roberts ◽  
A Aruffo ◽  
RJ Linhardt ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Acute Inflammation ◽  
Sialyl Lewis X ◽  
Enzyme Linked Immunosorbent Assay ◽  
Carbohydrate Binding ◽  
Cos Cells ◽  
Selectin Ligands ◽  
Heparin Oligosaccharides

Abstract Initial attachment of leukocytes to the vessel wall at sites of inflammation is supported by a family of carbohydrate-binding adhesion molecules called the selectins. Selectin ligands include sialyl-Lewis x (sLex, Neu5Ac alpha 2–3Gal beta 1–4[Fuc alpha 1–3]GlcNAc--) and related structures. We report here that defined heparin oligosaccharides interact with the selectins. Heparin chains containing four or more monosaccharide residues inhibited the function of L- and P-selectin, but not E-selectin, in vitro. In a competition enzyme-linked immunosorbent assay measuring inhibition of solution-phase selectin-Ig fusion proteins (selectin-Ig) binding to immobilized bovine serum albumin-sLex neoglycoprotein, a heparin-derived tetrasaccharide mixture inhibited 50% of L- and P-selectin-Ig binding (IC50) at 200 +/- 40 mumol/L and 850 +/- 110 mumol/L, respectively. A single hexasulfated tetrasaccharide (delta UA2S alpha 1–4GlcNS6S alpha 1–4IdoA2S alpha 1- 4GlcNS6S) was particularly active against L- and P-selectin-Ig (IC50 = 46 +/- 5 mumol/L and 341 +/- 24 mumol/L). By comparison, the tetrasaccharide sLex was not inhibitory at concentrations up to 1 mmol/L. In cell adhesion assays, heparin tetrasaccharides reduced binding of neutrophils to COS cells expressing P-selectin but not to COS cells expressing E-selectin. They also blocked colon cancer cell adhesion to L- and P-selectin but not E-selectin. In a model of acute inflammation, intravenously administered heparin tetrasaccharides diminished influx of neutrophils into the peritoneal cavities of thioglycollate-treated mice. We conclude that heparin oligosaccharides, including non-anticoagulant tetrasaccharides, are effective L- and P- selectin inhibitors in vitro and have anti-inflammatory activity in vivo.

scholarly journals MiR-203-3p inhibits the oxidative stress, inflammatory responses and apoptosis of mice podocytes induced by high glucose through regulating Sema3A expression

2020 ◽  
Vol 15 (1) ◽  
pp. 939-950
Author(s):  
Jingfu Chen ◽  
Qing Xu ◽  
Wei Zhang ◽  
YuLan Zhen ◽  
Fei Cheng ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Inflammatory Cytokines ◽  
High Glucose ◽  
Renal Injury ◽  
Inflammatory Responses ◽  
Cell Model ◽  
The Body ◽  
Enzyme Linked Immunosorbent Assay ◽  
Podocyte Injury

AbstractDiabetic nephropathy (DN) is the most serious long-term microvascular complication of diabetes, which mainly causes podocyte injury. Many studies have shown that microRNAs play a vital role in the development of DN. Studies have shown that miR-203-3p is involved in mesangial cell proliferation and apoptosis of DN mice. Therefore, we speculated that miR-203-3p might be related to the development of DN, but our study does not provide any evidence. In animal experiments, diabetic mice (db/db) were transfected with iR-203-3p overexpression lentiviral vectors (LV-miR-203-3p) and their control (LV-miR-con), with normal mice (db/m) being used as the control. High glucose (HG)-induced podocytes were used to construct a DN cell model in vitro. The expression levels of miR-203-3p, Semaphorin 3A (Sema3A) and inflammatory cytokines were detected by quantitative real-time polymerase chain reaction. Also, serum creatinine and blood urea nitrogen levels were used to evaluate the degree of renal injury in DN mice. Sema3A and apoptosis-related protein levels were assessed by the western blot analysis. Enzyme-linked immunosorbent assay was used to determine the different oxidative stress-related indicators and inflammatory cytokines. Flow cytometry and caspase-3 activity detection were used to analyze the degree of podocyte apoptosis. Our results suggested that the expression of miR-203-3p was lower in DN mice and in HG-induced podocytes. Overexpression of miR-203-3p reduced the body weight, blood glucose and renal injury of DN mice in vivo, as well as relieve the oxidative stress, inflammatory response and apoptosis of HG-induced podocytes in vitro. Functionally, Sema3A was a target of miR-203-3p, and Sema3A overexpression reversed the inhibitory effect of miR-203-3p on HG-induced podocyte injury. Our findings revealed that miR-203-3p alleviated the podocyte injury induced by HG via regulating Sema3A expression, suggesting that miR-203-3p might be a new therapeutic target to improve the progression of DN.

scholarly journals Heparin oligosaccharides bind L- and P-selectin and inhibit acute inflammation

Blood ◽  
1993 ◽  
Vol 82 (11) ◽  
pp. 3253-3258 ◽  
Author(s):  
RM Nelson ◽  
O Cecconi ◽  
WG Roberts ◽  
A Aruffo ◽  
RJ Linhardt ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Acute Inflammation ◽  
Sialyl Lewis X ◽  
Enzyme Linked Immunosorbent Assay ◽  
Carbohydrate Binding ◽  
Cos Cells ◽  
Selectin Ligands ◽  
Heparin Oligosaccharides

Initial attachment of leukocytes to the vessel wall at sites of inflammation is supported by a family of carbohydrate-binding adhesion molecules called the selectins. Selectin ligands include sialyl-Lewis x (sLex, Neu5Ac alpha 2–3Gal beta 1–4[Fuc alpha 1–3]GlcNAc--) and related structures. We report here that defined heparin oligosaccharides interact with the selectins. Heparin chains containing four or more monosaccharide residues inhibited the function of L- and P-selectin, but not E-selectin, in vitro. In a competition enzyme-linked immunosorbent assay measuring inhibition of solution-phase selectin-Ig fusion proteins (selectin-Ig) binding to immobilized bovine serum albumin-sLex neoglycoprotein, a heparin-derived tetrasaccharide mixture inhibited 50% of L- and P-selectin-Ig binding (IC50) at 200 +/- 40 mumol/L and 850 +/- 110 mumol/L, respectively. A single hexasulfated tetrasaccharide (delta UA2S alpha 1–4GlcNS6S alpha 1–4IdoA2S alpha 1- 4GlcNS6S) was particularly active against L- and P-selectin-Ig (IC50 = 46 +/- 5 mumol/L and 341 +/- 24 mumol/L). By comparison, the tetrasaccharide sLex was not inhibitory at concentrations up to 1 mmol/L. In cell adhesion assays, heparin tetrasaccharides reduced binding of neutrophils to COS cells expressing P-selectin but not to COS cells expressing E-selectin. They also blocked colon cancer cell adhesion to L- and P-selectin but not E-selectin. In a model of acute inflammation, intravenously administered heparin tetrasaccharides diminished influx of neutrophils into the peritoneal cavities of thioglycollate-treated mice. We conclude that heparin oligosaccharides, including non-anticoagulant tetrasaccharides, are effective L- and P- selectin inhibitors in vitro and have anti-inflammatory activity in vivo.

New Tools for the Herpes Virologist

1988 ◽  
Vol 4 (4) ◽  
pp. 634-636 ◽  
Author(s):  
Johan Harmenberg ◽  
Barbro Levén ◽  
Jorma Hinkula ◽  
Eva Ohlsson ◽  
Vivi-Anne Sundqvist ◽  
...  
Keyword(s):  
Herpes Virus ◽  
Wide Spectrum ◽  
The Body ◽  
Enzyme Linked Immunosorbent Assay ◽  
Common Belief ◽  
Antiviral Compounds ◽  
Resistance Patterns ◽  
Life Threatening ◽  
Herpès Virus

Herpes viruses are responsible for a wide spectrum of infections ranging from cold sores, genital herpes, and chicken pox to disseminated herpes infections in normal and more commonly in immunocompromised patients. Symptoms range from mildly distressing and uncomfortable to severe and life-threatening. The sophistication of virological methods is increasing. Specific types, classes, and subclasses of antibodies to viruses can now be determined, as well as the reactivity of T-lymphocytes. It is possible to detect herpes virus type-specific antibodies (to HSV-l or HSV-2) in a blood sample using the simple and inexpensive enzyme-linked immunosorbent assay (ELISA). The virus is usually type-determined rapidly and currently susceptibility or resistance to antiviral compounds can be defined in vitro. Such assays of antiviral resistance have been shown to be quick and effective. Effective antiviral therapies have been developed against a number of viral diseases. The concomitant need for isolation and evaluation of resistance patterns of herpes virus against different antiviral compounds appears to be of considerable importance. Pure clinical observations are no longer sufficient to distinguish the types, since HSV-l may be present in the genital region and HSV-2 at upper parts of the body–contrary to common belief.

scholarly journals Single Diameter Modulation Effects on Ni Nanowire Array Magnetization Reversal

Nanomaterials ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3403
Author(s):  
Luis C. C. Arzuza ◽  
Victor Vega ◽  
Victor M. Prida ◽  
Karoline O. Moura ◽  
Kleber R. Pirota ◽  
...  
Keyword(s):  
Aspect Ratio ◽  
Domain Wall ◽  
Long Range ◽  
Magnetization Reversal ◽  
Nanowire Array ◽  
Magnetic Behavior ◽  
Nanowire Arrays ◽  
Magnetic Nanowires ◽  
Range Interaction ◽  
Domain Wall Propagation

Geometrically modulated magnetic nanowires are a simple yet efficient strategy to modify the magnetic domain wall propagation since a simple diameter modulation can achieve its pinning during the nanowire magnetization reversal. However, in dense systems of parallel nanowires, the stray fields arising at the diameter interface can interfere with the domain wall propagation in the neighboring nanowires. Therefore, the magnetic behavior of diameter-modulated nanowire arrays can be quite complex and depending on both short and long-range interaction fields, as well as the nanowire geometric dimensions. We applied the first-order reversal curve (FORC) method to bi-segmented Ni nanowire arrays varying the wide segment (45–65 nm diameter, 2.5–10.0 μm length). The FORC results indicate a magnetic behavior modification depending on its length/diameter aspect ratio. The distributions either exhibit a strong extension along the coercivity axis or a main distribution finishing by a fork feature, whereas the extension greatly reduces in amplitude. With the help of micromagnetic simulations, we propose that a low aspect ratio stabilizes pinned domain walls at the diameter modulation during the magnetization reversal. In this case, long-range axial interaction fields nucleate a domain wall at the nanowire extremities, while short-range ones could induce a nucleation at the diameter interface. However, regardless of the wide segment aspect ratio, the magnetization reversal is governed by the local radial stray fields of the modulation near null magnetization. Our findings demonstrate the capacity of distinguishing between complex magnetic behaviors involving convoluted interaction fields.

In Vitro Cell Adhesion, Proliferation and Differentiation of Adipose Derived Stem Cells on Tulbaghia violacea Loaded Polycaprolactone (PCL) Nanofibers

2019 ◽  
Vol 9 (11) ◽  
pp. 1485-1498 ◽  
Author(s):  
Lerato N. Madike ◽  
M. Pillay ◽  
Ketul C. Popat
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Cell Adhesion ◽  
The Body ◽  
Osteogenic Potential ◽  
Adipose Derived Stem Cells ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Osteocalcin Production

Tissue engineering has been used for decades to restructure, replace and repair damaged tissue in the body. However, there are a number of challenges that have been identified, with the biggest one currently being the development of scaffolds with the ideal properties that can promote cell-scaffold interactions to enhance cell proliferation and differentiation. There is currently very little research on the incorporation of extracts of medicinal plants in scaffold fabrication with the aim of enhancing the surface properties of the scaffold. For this study, Tulbaghia violacea-based PCL scaffolds were fabricated and evaluated for their osteogenic potential on adipose derived stem cells (ADSCs) in osteogenic media. The short-term studies illustrated enhanced cell adhesion and proliferation with low levels of toxicity as well as the formation of elongated cells in the T. violacea-based scaffolds when compared to the control PCL scaffold. The long term studies indicated increased alkaline phosphate activity (ALP) in the T. violacea scaffolds when compared to PCL and overall higher levels of osteocalcin production over a period of 3 weeks. Immunofluorescence imaging of marker proteins also illustrated that the T. violacea incorporated scaffolds supported better osteocalcin production which is a specific extracellular matrix (ECM) marker for cartilaginous tissue. These results support the incorporation of T. violacea plant extracts for the enhancement of nanofiber scaffolds with the potential for tissue engineering applications.

scholarly journals Sigesbeckia glabrescens Makino extract attenuated the collagen-induced arthritis through inhibiting the synovial hyperplasia and inflammation

2020 ◽  
Author(s):  
Qiu Shuo Ma ◽  
Ke-Gang Linghu ◽  
Tian Zhang ◽  
Guan Ding Zhao ◽  
Wei Xiong ◽  
...  
Keyword(s):  
Bone Erosion ◽  
Cartilage Damage ◽  
Synovial Cell ◽  
The Body ◽  
Enzyme Linked Immunosorbent Assay ◽  
Collagen Induced Arthritis ◽  
Synovial Hyperplasia ◽  
Human Synovial Cell ◽  
And Migration

Abstract Background Sigesbeckia glabrescens Makino (SG) has been traditionally used for rheumatism and joint protection. However, the anti-arthritic effects and underling mechanisms of SG have not been demonstrated. In this study, we investigated the anti-arthritic effects and mechanisms of SG extract (SGE) on collagen-induced arthritic rats and interleukin (IL)- 1β-stimulated human synovial SW982 cells. Methods Rats were induced to arthritis by collagen for 15 days and then received SGE treatment for 35 days. The body weight and arthritis severity score of the rats were monitored weekly. At the end of the experiment, the radiographic and histological changes of rats’ hind paw were obtained; the cytokines expression in serum and joint muscles were determined by enzyme-linked immunosorbent assay (ELISA); and the level of regulatory T cells (Tregs) in the spleen was detected using flow cytometry. In addition, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and scratch wound healing assay were used to evaluate the proliferation of SW982 synovial cells. ELISA, western blot and immunofluorescence staining were used to investigate the anti-inflammatory mechanisms of SGE on IL-1β-induced SW982 cells. Results SGE attenuated the collagen-induced hind paw swelling, cartilage damage and bone erosion. SGE inhibited the synovial hyperplasia to the articular cavity in the toe joint and ankle. Moreover, SGE decreased the production of C-reactive protein in serum and the expression of IL-6 and IL-1β in the joint muscle. SGE also recovered the decreased Tregs. Results from the in vitro experiments showed that SGE not only inhibited the proliferation and migration of human synovial cell but also inhibited the IL-1β-induced expression of IL-6 and IL-8. Besides, SGE inhibited the activation of nuclear transcription factor-kappa B and the expression of cyclooxygenase-2. Conclusions SGE attenuated the collagen-induced arthritis through inhibiting the synovial hyperplasia and inflammation.

scholarly journals Increased Levels of Soluble CD14 in Sera of Periodontitis Patients

1999 ◽  
Vol 67 (1) ◽  
pp. 417-420 ◽  
Author(s):  
Joichiro Hayashi ◽  
Tamami Masaka ◽  
Isao Ishikawa
Keyword(s):  
Healthy Subjects ◽  
Chronic Exposure ◽  
Host Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
U937 Cells ◽  
Healthy Controls ◽  
Soluble Cd14 ◽  
Dihydroxyvitamin D ◽  
Membrane Bound

ABSTRACT Soluble CD14 (sCD14) mediates the response to lipopolysaccharide (LPS) in cells lacking membrane-bound CD14. We determined sCD14 concentrations in the sera of 38 periodontitis patients and 25 healthy controls by enzyme-linked immunosorbent assay. The sCD14 levels in the sera of patients with periodontitis were significantly higher than those of healthy subjects and decreased after treatment. Enhanced levels of sCD14 in serum may contribute to the host response to LPS in periodontitis. Furthermore, we showed in vitro that addition of LPS enhanced the release of sCD14 by monoblastic U937 cells treated with 1α,25-dihydroxyvitamin D3. Thus, increased sCD14 levels in periodontitis patients may be due to chronic exposure to LPS.

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