transforming-growth-factor-alpha and Inflammation

Skin wounds datation is one of the most challenging problems in forensic pathology. The vitality of a recent wound cannot be affirmed when no inflammatory cell is visible. There are in the literature numerous studies about wound vitality, looking for markers involved in coagulation or inflammation, using various methods such as enzymology, molecular biology or immunohistochemistry. In this update, we first introduce some methodological principles to respect. Then, we review the main studies available in the literature. We insist on immunohistochemistry, which seems to be the more valuable method, given its easiness to perform and the possibility to analyze the localization of the molecules of interest. Some markers are promising, such as TNFα, IL-6, IL-1β, TGFα or TGFβ1. Before using them in daily practice, these first results need to be confirmed with other studies, driven by independent teams and integrating multiple controls. Most notably, the antibodies have to be tested in numerous post-mortem wounds. Indeed, there is a critical risk of overexpression in post-mortem wounds, and some interesting markers have been secondary invalidated because of post-mortem false positivity (e.g. fibronectin, P-selectin). Finally, optimal sensibility and specificity values would be probably reached by combining several markers, validated with large groups of pre- and post-mortem wounds.

Topics: Biomarkers; Blood Coagulation; Cell Adhesion Molecules; Forensic Pathology; Hemostasis; Humans; Immunohistochemistry; Inflammation; Interleukin-1beta; Interleukin-6; Skin; Transforming Growth Factor alpha; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha; Wounds and Injuries

Injury to the skin initiates a cascade of events including inflammation, new tissue formation, and tissue remodeling, that finally lead to at least partial reconstruction of the original tissue. Historically, animal models of repair have taught us much about how this repair process is orchestrated and, over recent years, the use of genetically modified mice has helped define the roles of many key molecules. Aside from conventional knockout technology, many ingenious approaches have been adopted, allowing researchers to circumvent such problems as embryonic lethality, or to affect gene function in a tissue- or temporal-specific manner. Together, these studies provide us with a growing source of information describing, to date, the in vivo function of nearly 100 proteins in the context of wound repair. This article focuses on the studies in which genetically modified mouse models have helped elucidate the roles that many soluble mediators play during wound repair, encompassing the fibroblast growth factor (FGF) and transforming growth factor-beta (TGF-beta) families and also data on cytokines and chemokines. Finally, we include a table summarizing all of the currently published data in this rapidly growing field. For a regularly updated web archive of studies, we have constructed a Compendium of Published Wound Healing Studies on Genetically Modified Mice which is avaialble at http://icbxs.ethz.ch/members/grose/woundtransgenic/home.html.

Topics: Animals; Cytokines; Fibroblast Growth Factor 7; Fibroblast Growth Factors; Inflammation; Mice; Mice, Knockout; Mice, Transgenic; Signal Transduction; Skin; Time Factors; Transforming Growth Factor alpha; Transforming Growth Factor beta; Wound Healing

Inflammation contributions to the thrombotic response involve both cellular and humoral modulation. Inflammation impacts the initiation, propagation and the inhibitory phases of blood coagulation. Inflammatory mediators like endotoxin and tumor necrosis factor alpha (TNF alpha) elicit the expression of tissue factor on blood cells. Under normal circumstance, negatively charged membrane surfaces are limiting so that, even if some activated coagulation factors are generated, propagation of the coagulant stimulus is minimal. Complement activation, however, or exposure of collagen in combination with thrombin, provides a potent stimulus eliciting the exposure of negatively charged phospholipid membrane surfaces. Natural anticoagulant mechanisms limit the thrombotic response, but these pathways are depressed by inflammatory mediators. The protein C pathway is one of the major targets. Thrombomodulin and the endothelial cell protein C receptor are both required for optimal protein C activation, but both are down regulated by inflammatory mediators. Furthermore, free protein S levels often decrease resulting in impaired anticoagulant function of the activated protein C that is generated. In addition, anti-phospholipid antibodies severely impair the protein C pathway further inhibiting this pathway in inflammatory states associated with auto-immunity. In addition to shifting the hemostatic system in favor of clot formation, inflammation elevates the levels of plasminogen activator inhibitor thereby decreasing fibrinolytic activity. The procoagulant impact of inflammation can also be seen at the cellular level. Inflammatory mediators like interleukin 6 can increase both platelet count and their responsiveness to agonists like thrombin. All of these events tend to shift the hemostatic balance in favor of clot formation.

Topics: Animals; Anticoagulants; Antigens, CD; Autoimmunity; Blood Coagulation; Cell Membrane; Complement Activation; Down-Regulation; Endothelial Protein C Receptor; Fibrinolysis; Glycoproteins; Humans; Inflammation; Interleukin-6; Models, Biological; Models, Molecular; Phospholipids; Protein C; Protein S; Receptors, Cell Surface; Thrombin; Thrombosis; Transforming Growth Factor alpha

Inflammation and genetics are both prominent mechanisms in the pathogenesis of atherosclerosis and arterial thrombosis. Accordingly, a number of population studies have explored the association of ischaemic heart disease with gene polymorphisms of the inflammatory molecules tumour necrosis factors (TNF) alpha and beta, transforming growth factors (TGF) beta1 and 2, interleukin (IL) 1 and its receptor antagonist (IL 1ra), CD14 (the receptor for lipopolysaccharide), P and E selectins, and platelet endothelial cell adhesion molecule (PECAM) 1. Although they are very preliminary and partly conflicting, the data provide some evidence that alterations in the genetics of the inflammatory system may modify the risk of ischaemic heart disease.

Topics: Adult; E-Selectin; Female; Humans; Inflammation; Interleukin-1; Lipopolysaccharide Receptors; Male; Middle Aged; Myocardial Ischemia; P-Selectin; Phenotype; Platelet Endothelial Cell Adhesion Molecule-1; Polymorphism, Genetic; Risk Factors; Transforming Growth Factor alpha; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Topics: Animals; Colony-Stimulating Factors; Fibroblast Growth Factors; Growth Substances; Hematopoiesis; Humans; Inflammation; Interleukins; Macrophages; Nerve Regeneration; Platelet-Derived Growth Factor; Somatomedins; Transforming Growth Factor alpha; Transforming Growth Factor beta; Wound Healing

Prostatic inflammation is the driving force in benign prostatic hyperplasia (BPH). This work investigated the potential modulatory effect of COX-2 inhibition on ADAM-17/EGFR/ERK1/2 axis.. Adult male Wistar rats were used.. Celecoxib (10 and 20 mg/kg; i.p.) was injected i.p. daily for three weeks. Testosterone (TST) (3 mg/kg; s.c.) was used to induce BPH.. Prostatic inflammation and hyperplasia were assessed by organ weight and histopathology. Inflammatory mediators were measured using ELISA technique. Protein analysis was performed using western blotting and immunohistochemistry. Gene expression analysis was performed using qRT-PCR. Statistical analyses included one-way ANOVA and Tukey's multiple comparison test.. Testosterone-treated rats had a marked increase in COX-2, prostate weight, and index. Moreover, TST-induced COX-2 was inferred from cytoskeletal changes and was attributable to the overexpression of PGE2, NF-κB (p65), and IL-6. COX-2-derived PGE2 increased the activity of ADAM-17, TGF-α, and TNF-α. Consequently, EGFR-ERK1/2 pathway was over-activated, disrupting anti-apoptotic Bcl-2, cyclin D1, and pro-apoptotic Bax. Celecoxib reversed these effects.. COX-2 stimulates the ERK1/2 pathway via PGE2-ADAM-17-catalyzed shedding of TGF-α in testosterone-induced BPH. The results indicate a functional correlation between inflammation and hyperplasia in BPH.

Topics: ADAM17 Protein; Animals; Celecoxib; Cyclooxygenase 2; Dinoprostone; ErbB Receptors; Hyperplasia; Inflammation; Male; MAP Kinase Signaling System; Prostatic Hyperplasia; Rats; Rats, Sprague-Dawley; Rats, Wistar; Testosterone; Transforming Growth Factor alpha

The clinical features of SARS-CoV-2 infection range from asymptomatic to severe disease with life-threatening complications. Understanding the persistence of immune responses in asymptomatic individuals merit special attention because of their importance in controlling the spread of the infections. We here studied the antibody and T cell responses, and a wide range of inflammation markers, in 56 SARS-CoV-2 antibody-positive individuals, identified by a population screen after the first wave of SARS-CoV-2 infection. These, mostly asymptomatic individuals, were reanalyzed 7-8 months after their infection together with 115 age-matched seronegative controls. We found that 7-8 months after the infection their antibodies to SARS-CoV-2 Nucleocapsid (N) protein declined whereas we found no decrease in the antibodies to Spike receptor-binding domain (S-RBD) when compared to the findings at seropositivity identification. In contrast to antibodies to N protein, the antibodies to S-RBD correlated with the viral neutralization capacity and with CD4

Topics: Antibodies, Neutralizing; Antibodies, Viral; Asymptomatic Infections; CD4 Lymphocyte Count; CD4-Positive T-Lymphocytes; Coronavirus Nucleocapsid Proteins; COVID-19; Humans; Inflammation; Inflammation Mediators; Interleukin-18; Macrophages; Monocytes; Oncostatin M; Phosphoproteins; Protein Domains; S100A12 Protein; SARS-CoV-2; Spike Glycoprotein, Coronavirus; Transforming Growth Factor alpha

The metalloproteinase ADAM17 contributes to inflammatory and proliferative responses by shedding of cell-surface molecules. By this ADAM17 is implicated in inflammation, regeneration, and permeability regulation of epithelial cells in the colon. ADAM17 maturation and surface expression requires the adapter proteins iRhom1 or iRhom2. Here we report that expression of iRhom2 but not iRhom1 is upregulated in intestinal tissue of mice with acute colitis. Our analysis of public databases indicates elevated iRhom2 expression in mucosal tissue and epithelial cells from patients with inflammatory bowel disease (IBD). Consistently, expression of iRhom2 but not iRhom1 is upregulated in colon or intestinal epithelial cell lines after co-stimulation with tumor necrosis factor (TNF) and interferon gamma (IFNgamma). This upregulation can be reduced by inhibition of Janus kinases or transcription factors NF-kappaB or AP-1. Upregulation of iRhom2 can be mimicked by iRhom2 overexpression and is associated with enhanced maturation and surface expression of ADAM17 which then results in increased cleavage of transforming growth factor (TGF) alpha and junctional adhesion molecule (JAM)-A. Finally, the induction of these responses is suppressed by inhibition of iRhom2 transcription. Thus, inflammatory induction of iRhom2 may contribute to upregulated ADAM17-dependent mediator and adhesion molecule release in IBD. The development of iRhom2-dependent inhibitors may allow selective targeting of inflammatory ADAM17 activities.

Topics: ADAM17 Protein; Animals; Carrier Proteins; Cell Adhesion Molecules; Colon; Computational Biology; Computer Simulation; Cytokines; Epithelial Cells; HT29 Cells; Humans; Inflammation; Inflammatory Bowel Diseases; Interferon-gamma; Intracellular Signaling Peptides and Proteins; Mice; Mice, Inbred C57BL; Receptors, Cell Surface; Surface Properties; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha; Up-Regulation

Fetal swallowing of human amniotic fluid (hAF) containing trophic factors (TFs) promotes gastrointestinal tract (GIT) development. Preterm birth interrupts hAF swallowing, which may increase the risk of necrotizing enterocolitis (NEC). Postnatally, it is difficult to replicate fetal swallowing of hAF due to volume. We aimed to evaluate whether hAF lyophilization is feasible and its effect on hAF-borne TFs.. We collected hAF (n = 16) from uncomplicated pregnancies. hAF was divided into three groups: unprocessed control (C), concentration by microfiltration (F), and by dialysis and lyophilization (L). EGF, HGF, GM-CSF, and TGF-α were measured in each group by multiplex assay. Bioavailability of TFs was measured by proliferation and LPS-induced IL-8 production by intestinal epithelial cells FHs74.. After dialysis/lyophilization, GM-CSF and TGF-α were preserved with partial loss of EGF and HGF. hAF increased cell proliferation and reduced LPS-induced IL-8 production compared to medium alone. Compared to control, dialysis/lyophilization and filtration of hAF increased FHs74 cell proliferation (p < 0.001) and decreased LPS-induced IL-8 production (p < 0.01).. Lyophilization and filtration of hAF is feasible with partial loss of TFs but maintains and even improves bioavailability of TFs measured by proliferation and LPS-induced IL-8 production by FHs74.

Topics: Amniotic Fluid; Cell Proliferation; Cryopreservation; Deglutition; Enterocolitis, Necrotizing; Female; Freeze Drying; Gastrointestinal Tract; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation; Interleukin-8; Pregnancy; Transforming Growth Factor alpha

HIV-associated neurocognitive disorders (HAND) affect over half of HIV-infected individuals, despite antiretroviral therapy (ART). Therapeutically targetable mechanisms underlying HAND remain elusive, partly due to a lack of a representative model. We developed a human-induced pluripotent stem cell (hiPSC)-based model, independently differentiating hiPSCs into neurons, astrocytes, and microglia, and systematically combining to generate a tri-culture with or without HIV infection and ART. Single-cell RNA sequencing analysis on tri-cultures with HIV-infected microglia revealed inflammatory signatures in the microglia and EIF2 signaling in all three cell types. Treatment with the antiretroviral compound efavirenz (EFZ) mostly resolved these signatures. However, EFZ increased RhoGDI and CD40 signaling in the HIV-infected microglia. This activation was associated with a persistent increase in transforming growth factor α production by microglia. This work establishes a tri-culture that recapitulates key features of HIV infection in the CNS and provides a new model to examine the effects of infection, its treatment, and other co-morbid conditions.

Topics: Alkynes; Anti-HIV Agents; Antiretroviral Therapy, Highly Active; Astrocytes; Benzoxazines; CD40 Antigens; Cell Differentiation; Cells, Cultured; Cyclopropanes; Cytokines; Eukaryotic Initiation Factor-2; HIV Infections; Humans; Induced Pluripotent Stem Cells; Inflammation; Microglia; Models, Biological; Neurons; rho-Specific Guanine Nucleotide Dissociation Inhibitors; Signal Transduction; Single-Cell Analysis; Transforming Growth Factor alpha

Diabetic retinopathy (DR) is a frequent complication of diabetes and it can lead to visual impairment and blindness. However, the mechanism of their regulation remains little known. circRNAs can function as crucial competing endogenous RNA, which can sponge corresponding miRNAs and affect mRNA expression in various diseases, including DR. In our current research, we observed that circRNA_0084043 was elevated in high glucose (HG)-incubated ARPE-19 cells. Then, we focused on whether and how circRNA_0084043 participated in retinal vascular dysfunction under conditions diabetes. Apoptosis, inflammation and oxidative stress are hallmark of DR progression. This work was aimed to investigate the signaling mechanisms of circRNA_0084043 in these pathogenesis of DR. We discovered loss of circRNA_0084043 significantly increased cell survival and repressed HG-triggered apoptosis. In addition, knockdown of circRNA_0084043 remarkably reduced oxidative stress as evidenced by the down-regulated malondialdehyde (MDA) content, enhanced activities of Super Oxide Dismutase (SOD) and Glutathione peroxidase (GSH-PX). Addition, silence of circRNA_0084043 effectively restrained HG-stimulated inflammation as proved by repressing inflammatory cytokines Tumor Necrosis Factor α (TNF-α), Interleukin 6 (IL-6) and Cox-2 in ARPE-19 cells. Subsequently, we successfully predicted that miR-140-3p was a downstream target of circRNA_0084043, which could be negatively regulated by circRNA_0084043. Mechanistically, loss of miR-140-3p abrogated the beneficial effects of circRNA_0084043 siRNA on ARPE-19 cells. Transforming Growth Factor alpha (TGFA) can exhibit a role in multiple diseases. Taken these together, these data demonstrated that loss of circRNA_0084043 depressed HG-induced damage via sponging miR-140-3p and regulating TGFA.

Topics: Apoptosis; Base Sequence; Cell Line; Cell Proliferation; Diabetic Retinopathy; Disease Progression; Down-Regulation; Epithelial Cells; Gene Expression Regulation; Glucose; Humans; Inflammation; MicroRNAs; Models, Biological; Oxidative Stress; Retinal Pigment Epithelium; RNA, Circular; Transforming Growth Factor alpha; Up-Regulation

Inflammation induced by Helicobacter pylori infection related to gastric carcinogenesis. In this study, we have investigated the anti-inflammatory effect and its mechanism of kaempferol in the inflammatory response caused by H. pylori infection in vitro. We found that kaempferol reduced the expression of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-8) and production of IL-8 in AGS cells. In addition, kaempferol suppressed translocation of cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA) of H. pylori to AGS cells. It was due to decreased transcription of type IV secretion system (T4SS) components involved in CagA injection and secretion system subunit protein A (SecA) of type V secretion system (T5SS) involved in VacA secretion by kaempferol. In conclusion, kaempferol shows the anti-inflammatory effect by suppressing the translocation of CagA and VacA proteins and leading to the down-regulation of pro-inflammatory cytokines. Abbreviations: CagA: cytotoxin-associated gene A; VacA: vacuolating cytotoxin A; T4SS: type IV secretion systems; SecA: secretion system subunit protein A; T5SS: type V secretion system.

Topics: Anti-Inflammatory Agents; Antigens, Bacterial; Bacterial Proteins; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Inflammation; Inflammation Mediators; Interleukin-1beta; Interleukin-8; Kaempferols; Protein Transport; Transforming Growth Factor alpha

Despite increasing evidence that adults with long-standing atopic dermatitis (AD) have systemic inflammation, little is known about systemic inflammation in recent-onset early pediatric AD.. To analyze blood inflammatory proteins of early pediatric AD.. Using high-throughput proteomics (proximity extension assay), we assessed 257 inflammatory and cardiovascular risk proteins in the blood of 30 children with moderate to severe AD younger than 5 years of age (within 6 months of onset) compared with age-matched pediatric control individuals and adult patients with AD.. In pediatric AD blood, T helper (Th) type 2 (CCL13, CCL22) and Th17 (peptidase inhibitor-3/elafin) markers were increased, together with markers of tissue remodeling (matrix metalloproteinases 3/9/10, urokinase receptor), endothelial activation (E-selectin), T-cell activation (IL2RA), neutrophil activation (myeloperoxidase), lipid metabolism (FABP4), and growth factors (FGF21, transforming growth factor-α). Total numbers of dysregulated proteins were smaller in pediatric AD (n = 22) than in adult AD (n = 61). Clinical severity scores were positively correlated with receptors for interleukins 33 and 36 and inversely correlated with some Th1 markers (interferon gamma, CXCL11).. Different baseline expression levels in healthy pediatric vs adult samples.. Within months of pediatric AD onset, systemic immune activation is present, with Th2/Th17 skewing but otherwise different proteomic patterns from adult AD. Future correlation of proteomic patterns with disease course, comorbidity development, and drug response may yield predictive biomarkers.

Topics: Age Factors; Biomarkers; Case-Control Studies; Chemokines; Child, Preschool; Chronic Disease; Dermatitis, Atopic; E-Selectin; Elafin; Fatty Acid-Binding Proteins; Female; Fibroblast Growth Factors; Humans; Infant; Inflammation; Interleukin-2 Receptor alpha Subunit; Male; Matrix Metalloproteinases; Peroxidase; Proteome; Receptors, Interleukin; RNA, Messenger; Severity of Illness Index; Transforming Growth Factor alpha

Microglia and astrocytes modulate inflammation and neurodegeneration in the central nervous system (CNS)

Topics: Animals; Astrocytes; Cells, Cultured; Central Nervous System; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; ErbB Receptors; Female; Humans; Inflammation; Lipopolysaccharide Receptors; Mice; Mice, Inbred C57BL; Microglia; Multiple Sclerosis; Receptors, Aryl Hydrocarbon; Symbiosis; Transforming Growth Factor alpha; Tryptophan; Vascular Endothelial Growth Factor B; Vascular Endothelial Growth Factor Receptor-1

This study describes specific patterns of elevated inflammatory proteins in clinical subtypes of myasthenia gravis (MG) patients. MG is a chronic, autoimmune neuromuscular disease with antibodies most commonly targeting the acetylcholine receptors (AChRab), which causes fluctuating skeletal muscle fatigue. MG pathophysiology includes a strong component of inflammation, and a large proportion of patients with early onset MG additionally present thymus hyperplasia. Due to the fluctuating nature and heterogeneity of the disease, there is a great need for objective biomarkers as well as novel potential inflammatory targets. We examined the sera of 45 MG patients (40 AChRab seropositive and 5 AChRab seronegative), investigating 92 proteins associated with inflammation. Eleven of the analysed proteins were significantly elevated compared to healthy controls, out of which the three most significant were: matrix metalloproteinase 10 (MMP-10; p = 0.0004), transforming growth factor alpha (TGF-α; p = 0.0017) and extracellular newly identified receptor for advanced glycation end-products binding protein (EN-RAGE) (also known as protein S100-A12; p = 0.0054). Further, levels of MMP-10, C-X-C motif ligand 1 (CXCL1) and brain derived neurotrophic factor (BDNF) differed between early and late onset MG. These novel targets provide valuable additional insight into the systemic inflammatory response in MG.

Topics: Adult; Aged; Aged, 80 and over; Autoantibodies; Biomarkers; Female; Humans; Inflammation; Inflammation Mediators; Male; Matrix Metalloproteinase 10; Middle Aged; Myasthenia Gravis; Phenotype; Receptors, Cholinergic; S100A12 Protein; Transforming Growth Factor alpha

Anti-inflammatory therapy is a logical approach to slowing the inevitable lung function deterioration in cystic fibrosis (CF) patients. This study's aim was to evaluate inflammatory markers and disease progression in paediatric CF patients chronically treated with azithromycin or low-dose prednisolone.. The study included 204 patients with CF and 100 healthy controls; 102 CF patients were treated with basic therapy only (without anti-inflammatory treatment; WAT), and 102 individuals received basic therapy along with azithromycin (n = 59) or low-dose prednisolone (n = 43). The median duration of therapy was 24 months (range 12-82) with azithromycin and 31 months (range 12-180) with prednisolone. A cross-sectional analysis of plasma and sputum biomarkers was performed.. Compared with the healthy controls, the WAT group showed elevated IFN-γ, IL-10 (total), and TGFβ1 concentrations, and decreased TNFα (total) and adrenocorticotropic hormone (ACTH) levels (all p < 0.05). Plasma TNFα (total) concentrations in azithromycin/prednisolone patients were significantly higher than those in WAT patients and similar to those of healthy children. In contrast, IL-10 (total) levels were significantly decreased in azithromycin/prednisolone-treated patients compared with WAT patients. Children from the azithromycin group demonstrated ACTH levels similar to those of healthy controls. Azithromycin-treated patients showed a significantly reduced rate of CF-related liver disease and a significantly increased incidence of glucose metabolism disturbances.. Steady-state anti-inflammatory treatments may have a sustained immunomodulatory action at systemic and local levels in CF patients. Further investigations are needed to assess the effects of supportive azithromycin therapy on the hypothalamic-pituitary-adrenal axis and the incidence of non-pulmonary CF complications.

Topics: Adolescent; Adrenocorticotropic Hormone; Anti-Bacterial Agents; Anti-Inflammatory Agents; Azithromycin; Biomarkers; Child; Cross-Sectional Studies; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Disease Progression; Female; Genotype; Humans; Inflammation; Interferon-gamma; Interleukin-10; Leukocyte Elastase; Male; Prednisolone; Transforming Growth Factor alpha; Transforming Growth Factor beta

Hepatic ischemia/reperfusion (I/R) describes the paradox of additional tissue injury caused by reperfusion. The aim of this survey was to investigate the mRNA expression of genes exerting their inflammatory and regenerative reaction in a porcine model of I/R and extended hepatectomy.. Twelve pigs were used, weighing 30-35 kg in average, which were allocated in two groups: the I/R group with eight pigs and the sham-operated (control) one with four pigs. The I/R group underwent portacaval anastomosis and Pringle maneuver followed by extended hepatectomy. The hepatoduodenal ligament was occluded for 150 min and the liver remnant was reperfused for 24 hours. Blood samples were steadily received throughout the surgical procedure, where hepatic biopsies were taken for pathological evaluation. Animals were sacrificed in 24 hours after the onset of reperfusion.. Between the two groups, statistically significant differences were noticed in serum values of AST, ALT, ALP, and total bilirubin in the early and late phase of reperfusion. The mRNA expression of iNOS, IL-1b, and TGF-a did not increase significantly in the I/R group. Conversely, the mRNA modification of IL-6, STAT-3, and E-selectin demonstrated significantly increased expression in I/R animals.. In the present survey, a new I/R swine model was proposed and specific parameters were analyzed, revealing differences between the study groups.

Topics: Alanine Transaminase; Alkaline Phosphatase; Animals; Aspartate Aminotransferases; Bilirubin; Biopsy; Disease Models, Animal; E-Selectin; Female; Hepatectomy; Inflammation; Interleukin-1beta; Interleukin-6; Ischemia; Liver; Liver Diseases; Liver Regeneration; Nitric Oxide Synthase Type II; Reperfusion Injury; RNA, Messenger; STAT3 Transcription Factor; Swine; Transforming Growth Factor alpha

Injection drug use is a growing major public health concern. Injection drug users (IDUs) have a higher incidence of co-morbidities including HIV, Hepatitis, and other infections. An effective humoral response is critical for optimal homeostasis and protection from infection; however, the impact of injection heroin use on humoral immunity is poorly understood. We hypothesized that IDUs have altered B cell and antibody profiles.. A comprehensive systems biology-based cross-sectional assessment of 130 peripheral blood B cell flow cytometry- and plasma- based features was performed on HIV-/Hepatitis C-, active heroin IDUs who participated in a syringe exchange program (n = 19) and healthy control subjects (n = 19). The IDU group had substantial polydrug use, with 89% reporting cocaine injection within the preceding month. IDUs exhibited a significant, 2-fold increase in total B cells compared to healthy subjects, which was associated with increased activated B cell subsets. Although plasma total IgG titers were similar between groups, IDUs had significantly higher IgG3 and IgG4, suggestive of chronic B cell activation. Total IgM was also increased in IDUs, as well as HIV Envelope-specific IgM, suggestive of increased HIV exposure. IDUs exhibited numerous features suggestive of systemic inflammation, including significantly increased plasma sCD40L, TNF-α, TGF-α, IL-8, and ceramide metabolites. Machine learning multivariate analysis distilled a set of 10 features that classified samples based on group with absolute accuracy.. These results demonstrate broad alterations in the steady-state humoral profile of IDUs that are associated with increased systemic inflammation. Such dysregulation may impact the ability of IDUs to generate optimal responses to vaccination and infection, or lead to increased risk for inflammation-related co-morbidities, and should be considered when developing immune-based interventions for this growing population.

Topics: Adult; B-Lymphocytes; CD40 Ligand; Comorbidity; Cross-Sectional Studies; Female; Hepatitis C; Heroin; HIV Antibodies; HIV Infections; Humans; Immunity, Humoral; Immunoglobulin G; Immunoglobulin M; Inflammation; Interleukin-8; Male; Narcotics; New York; Substance Abuse, Intravenous; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha; Young Adult

Although the cause of Alzheimer's disease (AD) is unknown, glial-induced neuroinflammation is an early symptom. Familial AD is caused by increases in amyloid-beta (Aβ) peptide, particularly soluble oligomeric (oAβ), considered a proximal neurotoxin and neuroinflammatory stimuli. APOE4, a naturally occurring genotype of APOE, is the greatest genetic risk factor for AD; increasing risk up to 12-fold compared to APOE3 and APOE2. oAβ-induced neuroinflammation is greater with APOE4 compared to APOE3 and APOE2. As sinapates and flavonoids have anti-inflammatory properties, a protocol was developed for optimizing polyphenol production in seedlings of Arabidopsis thaliana (A. thaliana). Three mutants (cop1, prn1, xpf3) were identified, and the extracts treated with liver microsomes to mimic physiological metabolism, with HPLC and MS performed on the resulting metabolites for peak identification. These extracts were used to treat primary glial cells isolated from human APOE-targeted-replacement (APOE-TR) and APOE-knock-out (KO) mice, with neuroinflammation induced by lipopolysaccharide (LPS) or oAβ. The dose-response data for TNFα secretion demonstrate the followed the order: APOE-KO > APOE4 > APOE3 > APOE2, with xpf3 the most effective anti-neuroinflammatory across APOE genotypes. Thus, the plant-based approach described herein may be particularly valuable in treating the APOE4-induced neuroinflammatory component of AD risk.

Topics: Alzheimer Disease; Animals; Apolipoproteins E; Arabidopsis; Dose-Response Relationship, Drug; Genotype; In Vitro Techniques; Inflammation; Lipopolysaccharides; Mice; Mice, Knockout; Plant Extracts; Polyphenols; Transforming Growth Factor alpha; Ultraviolet Rays

Chronic kidney disease is characterized by Vitamin D deficiency and activation of the renin-angiotensin-aldosterone system. Increasing data show that vitamin D receptor agonists (VDRAs) exert beneficial effects in renal disease and possess anti-inflammatory properties, but the underlying mechanism remains unknown. Emerging evidence suggests that "a disintegrin and metalloproteinase" (ADAM)/epidermal growth factor receptor (EGFR) signalling axis contributes to renal damage. Aldosterone induces EGFR transactivation regulating several processes including cell proliferation and fibrosis. However, data on tubular epithelial cells is scarce. We have found that, in cultured tubular epithelial cells, aldosterone induced EGFR transactivation via TGF-α/ADAM17. Blockade of the TGF-α/ADAM17/EGFR pathway inhibited aldosterone-induced proinflammatory gene upregulation. Moreover, among the potential downstream mechanisms, we found that TGF-α/ADAM17/EGFR inhibition blocked ERK and STAT-1 activation in response to aldosterone. Next, we investigated the involvement of TGF-α/ADAM17/EGFR axis in VDRA anti-inflammatory effects. Preincubation with the VDRA paricalcitol inhibited aldosterone-induced EGFR transactivation, TGF-α/ADAM-17 gene upregulation, and downstream mechanisms, including proinflammatory factors overexpression. In conclusion, our data suggest that the anti-inflammatory actions of paricalcitol in tubular cells could depend on the inhibition of TGF-α/ADAM17/EGFR pathway in response to aldosterone, showing an important mechanism of VDRAs action.

Topics: ADAM Proteins; ADAM17 Protein; Aldosterone; Cell Line; Cell Proliferation; Epithelial Cells; ErbB Receptors; Ergocalciferols; Gene Expression Regulation; Humans; Inflammation; Kidney Tubules; Receptors, Calcitriol; Renal Insufficiency, Chronic; Renin-Angiotensin System; Signal Transduction; STAT1 Transcription Factor; Transforming Growth Factor alpha; Vitamin D Deficiency

This study aimed to investigate the possible gastroprotective effect of tocotrienol against water-immersion restraint stress (WIRS) induced gastric ulcers in rats by measuring its effect on gastric mucosal nitric oxide (NO), oxidative stress, and inflammatory biomarkers. Twenty-eight male Wistar rats were randomly assigned to four groups of seven rats. The two control groups were administered vitamin-free palm oil (vehicle) and the two treatment groups were given omeprazole (20 mg/kg) or tocotrienol (60 mg/kg) orally. After 28 days, rats from one control group and both treated groups were subjected to WIRS for 3.5 hours once. Malondialdehyde (MDA), NO content, and superoxide dismutase (SOD) activity were assayed in gastric tissue homogenates. Gastric tissue SOD, iNOS, TNF-α and IL1-β expression were measured. WIRS increased the gastric MDA, NO, and pro-inflammatory cytokines levels significantly when compared to the non-stressed control group. Administration of tocotrienol and omeprazole displayed significant protection against gastric ulcers induced by exposure to WIRS by correction of both ulcer score and MDA content. Tissue content of TNF-α and SOD activity were markedly reduced by the treatment with tocotrienol but not omeprazole. Tocotrienol significantly corrected nitrite to near normal levels and attenuated iNOS gene expression, which was upregulated in this ulcer model. In conclusion, oral supplementation with tocotrienol provides a gastroprotective effect in WIRS-induced ulcers. Gastroprotection is mediated through 1) free radical scavenging activity, 2) the increase in gastric mucosal antioxidant enzyme activity, 3) normalisation of gastric mucosal NO through reduction of iNOS expression, and 4) attenuation of inflammatory cytokines. In comparison to omeprazole, it exerts similar effectiveness but has a more diverse mechanism of protection, particularly through its effect on NO, SOD activity, and TNF-α.

Topics: Animals; Antioxidants; Biomarkers; Gastric Mucosa; Inflammation; Interleukin-1beta; Lipid Peroxidation; Male; Malondialdehyde; Nitric Oxide; Omeprazole; Oxidative Stress; Random Allocation; Rats; Rats, Wistar; Stomach; Stomach Ulcer; Superoxide Dismutase; Tocotrienols; Transforming Growth Factor alpha

Despite growing recognition of an etiologic role for inflammation in lung carcinogenesis, few prospective epidemiologic studies have comprehensively investigated the association of circulating inflammation markers with lung cancer.. We conducted a nested case-control study (n = 526 lung cancer patients and n = 592 control subjects) within the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. Control subjects were matched to lung cancer case patients on age, sex, follow-up time (median = 2.9 years), randomization year, and smoking (pack-years and time since quitting). Serum levels of 77 inflammation markers were measured using a Luminex bead-based assay. Conditional logistic regression and weighted Cox models were used to estimate odds ratios (ORs) and cumulative risks, respectively.. Of 68 evaluable markers, 11 were statistically significantly associated with lung cancer risk (P trend across marker categories < .05), including acute-phase proteins (C-reactive protein [CRP], serum amyloid A [SAA]), proinflammatory cytokines (soluble tumor necrosis factor receptor 2 [sTNFRII]), anti-inflammatory cytokines (interleukin 1 receptor antagonist [IL-1RA]), lymphoid differentiation cytokines (interleukin 7 [IL-7]), growth factors (transforming growth factor alpha [TGF-A]), and chemokines (epithelial neutrophil-activating peptide 78 [ENA 78/CXCL5], monokine induced by gamma interferon [MIG/CXCL9], B cell-attracting chemokine 1 [BCA-1/CXCL13], thymus activation regulated chemokine [TARC/CCL17], macrophage-derived chemokine [MDC/CCL22]). Elevated marker levels were associated with increased lung cancer risk, with odds ratios comparing the highest vs the lowest group ranging from 1.47 (IL-7) to 2.27 (CRP). For IL-1RA, elevated levels were associated with decreased lung cancer risk (OR = 0.71; 95% confidence interval = 0.51 to 1.00). Associations did not differ by smoking, lung cancer histology, or latency. A cross-validated inflammation score using four independent markers (CRP, BCA-1/CXCL13, MDC/CCL22, and IL-1RA) provided good separation in 10-year lung cancer cumulative risks among former smokers (quartile [Q] 1 = 1.1% vs Q4 = 3.1%) and current smokers (Q1 = 2.3% vs Q4 = 7.9%) even after adjustment for smoking.. Some circulating inflammation marker levels are associated with prospective lung cancer risk.

Topics: Aged; Biomarkers; Biomarkers, Tumor; C-Reactive Protein; Case-Control Studies; Chemokines; Cytokines; Female; Humans; Inflammation; Logistic Models; Lung Neoplasms; Male; Middle Aged; Odds Ratio; Predictive Value of Tests; Prospective Studies; Risk Assessment; Risk Factors; Serum Amyloid A Protein; Transforming Growth Factor alpha

To investigate the efficacy of combined administration of alpha-tocopherol (AT) and ascorbic acid (AA) in reducing ethanol-induced hepatotoxicity.. Rats were maintained for 90 days and grouped as follows: I-control rats, II-ethanol, III-alpha-tocopherol, IV-ethanol+alpha-tocopherol, V-AA, VI-ethanol+ascorbic acid, VII-alpha-tocopherol+ascorbic acid, VIII-ethanol+alpha-tocopherol+ascorbic acid. At the end of the experimental period, markers of hepatic function, oxidative stress, and the expression of markers of inflammation and fibrosis were assayed.. The markers of hepatic function, lipid peroxidation products, protein carbonyls, and the expression of nuclear factor kappa B, tumor necrosis factor alpha, transforming growth factor beta 1, cytochrome P4502E1, and collagen Type I were elevated after ethanol administration. All these parameters were reduced in the ethanol group administered AT and AA in combination. The activities of antioxidant enzymes which were reduced by ethanol administration were enhanced on combined administration of AT and AA. The reduction in hepatic fibrosis was almost 20% more in AT and AA co-administered group compared with AT and AA alone treated groups.. Combined administration of fat soluble AT and water soluble AA was beneficial against ethanol-induced hepatotoxicity. This may be due to their different subcellular localizations.

Topics: alpha-Tocopherol; Animals; Ascorbic Acid; Biomarkers; Chromatography, High Pressure Liquid; Collagen Type I; Drug Therapy, Combination; Ethanol; Inflammation; Lipid Peroxidation; Liver; Liver Cirrhosis; Male; NF-kappa B; Oxidative Stress; Rats; Rats, Sprague-Dawley; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha

Interleukin (IL)-8 and neutrophil inflammation play a vital role in the pathogenesis of chronic inflammatory airway diseases. A disintegrin and metalloproteinase 17 (ADAM17) cleaves ectodomains of various transmembrane proteins and is an important regulator of almost every cellular event. The aim of this work was to investigate the roles of epidermal growth factor receptor (EGFR) signaling and ADAM17 in IL-8 expression induced by lipopolysaccharide (LPS) in A549 lung epithelial cells. In the present study, we found that transforming growth factor-α (TGF-α)-neutralizing Ab, EGFR-neutralizing Ab, and AG1478 significantly inhibited the phosphorylation of EGFR and IL-8 production induced by LPS. Lentivirus-mediated ADAM-17 RNA interference markedly inhibited ADAM-17 expression, the release of TGF-α, and the phosphorylation of EGFR and IL-8 expression. The results demonstrated that lentivirus-mediated RNA interference targeting ADAM17 suppressed IL-8 expression induced by LPS via inhibiting EGFR signaling, and ADAM17-EGFR signaling cascade mediated IL-8 production induced by LPS in lung epithelial cells.

Topics: ADAM Proteins; ADAM17 Protein; Antibodies, Neutralizing; Cell Line; Epithelial Cells; ErbB Receptors; Gene Transfer Techniques; Genetic Vectors; Humans; Inflammation; Interleukin-8; Lentivirus; Lipopolysaccharides; Lung; Phosphorylation; Quinazolines; Respiratory Mucosa; RNA Interference; RNA, Small Interfering; Signal Transduction; Transforming Growth Factor alpha; Tyrphostins

Certain Bifidobacterium strains have been shown to inhibit inflammatory responses in intestinal epithelial cells. However, the precise mechanisms of these effects, including the chemical nature of the active compounds, remain to be elucidated. Here partial characterization of the anti-inflammatory properties of Bifidobacterium strains isolated from feces of healthy infants is reported. It was found that conditioned media (CM) of all strains studied are capable of attenuating tumor necrosis factor-α (TNF-α) and lipopolysaccharide- (LPS) induced inflammatory responses in the HT-29 cell line. In contrast, neither killed bifidobacterial cells, nor cell-free extracts showed such activities. Further investigations resulted in attribution of this activity to heat-stable, non-lipophilic compound(s) resistant to protease and nuclease treatments and of molecular weight less than 3 kDa. The anti-inflammatory effects were dose- and time-dependent and associated with inhibition of IκB phosphorylation and nuclear factor-κ light chain enhancer of activated B cells (NF-κB)-dependent promoter activation. The combined treatments of cells with CMs and either LPS or TNF-α, but not with CMs alone, resulted in upregulation of transforming growth factor-β1, IκBζ, and p21(CIP) mRNAs. Our data suggest certain species-specificities of the anti-inflammatory properties of bifidobacteria. This observation should prompt additional validation studies using larger set of strains and employing the tools of comparative genomics.

Topics: Apoptosis; Bifidobacterium; Culture Media, Conditioned; Dose-Response Relationship, Immunologic; Escherichia coli; Feces; Gene Expression Regulation, Bacterial; HT29 Cells; Humans; I-kappa B Proteins; Infant; Inflammation; Interleukin-8; Intestines; Lipopolysaccharides; Molecular Weight; NF-kappa B; NF-KappaB Inhibitor alpha; Phosphorylation; Promoter Regions, Genetic; RNA, Messenger; Species Specificity; Time Factors; Transcriptional Activation; Transfection; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha

Sphingosine-1-phosphate (S1P), a multifunctional phospholipid, regulates vascular cell function. Whether S1P influences vascular inflammatory responses, particularly in hypertension, is unclear. We tested the hypothesis that S1P is a proinflammatory mediator signaling through receptor tyrosine kinase transactivation and that responses are amplified in vascular smooth muscle cells from stroke-prone spontaneously hypertensive rats (SHRSPs), a model in which we demonstrated Edg1 (S1P1 receptor) to be a candidate gene for salt-sensitive hypertension. Vascular smooth muscle cell from Wistar-Kyoto rats and SHRSPs were studied. S1P receptor subtypes, S1P1 and S1P2, were similarly expressed in Wistar-Kyoto rats and SHRSPs. S1P induced phosphorylation of epidermal growth factor receptor and platelet-derived growth factor and activation of p38 mitogen-activated protein kinase and c-Jun N-terminal kinase, with amplified effects in SHRSPs versus Wistar-Kyoto rats. Inhibition of epidermal growth factor receptor and platelet-derived growth factor (with AG1478 and AG1296, respectively) abolished S1P-induced phosphorylation of p38 mitogen-activated protein kinase and c-Jun N-terminal kinase in Wistar-Kyoto rats with variable effects in SHRSPs. Vascular smooth muscle cell inflammation was evaluated by expression of adhesion molecules and functional responses assessed by monocyte adhesion. S1P stimulated expression of intercellular adhesion molecule 1 and vascular cell adhesion protein 1 and promoted monocyte adhesion, particularly in SHRSP cells. S1P-mediated inflammation was blunted by AG1478 and AG1296 in SHRSP cells. VPC23019, a S1P1 receptor antagonist, inhibited S1P-induced mitogen-activated protein kinase phosphorylation, intercellular adhesion molecule 1 and vascular cell adhesion protein 1 expression, and monocyte adhesion. Our data indicate that molecular processes underlying vascular inflammation and cell adhesion in SHRSPs involve S1P/S1P1 receptors and phosphorylation of receptor tyrosine kinases. We identify a novel pathway linking S1P/S1P1 receptors to specific proinflammatory signaling pathways through epidermal growth factor receptor and platelet-derived growth factor transactivation, a process that is upregulated in SHRSPs. Such molecular events may contribute to vascular inflammation in hypertension.

Topics: Analysis of Variance; Animals; Blotting, Western; Cell Adhesion; Cells, Cultured; Hypertension; Inflammation; Lysophospholipids; Male; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Phosphorylation; Rats; Rats, Inbred WKY; Rats, Wistar; Receptors, Lysosphingolipid; Receptors, Platelet-Derived Growth Factor; Species Specificity; Sphingosine; Transforming Growth Factor alpha; Up-Regulation

To investigate effects of hepatotropic growth factors on radical production in rat hepatocytes during sepsis.. Rat hepatocytes, isolated by collagenase perfusion, were incubated with a lipopolysaccharide (LPS)-containing cytokine mixture of interleukin-1β, tumor necrosis factor-α and interferon-γ to simulate sepsis and either co-incubated or pre-incubated with hepatotropic growth factors, e.g. hepatocyte growth factor, epidermal growth factor and/or transforming growth factor-α. Cells were analyzed for glutathione levels. Culture supernatants were assayed for production of reactive oxygen intermediates (ROIs) as well as NO(2) (-), NO(3) (-) and S-nitrosothiols. To determine cellular damage, release of aspartate aminotransferase (AST) into the culture medium was analyzed. Activation of nuclear factor (NF)-κB was measured by electrophoretic mobility shift assay.. Rat hepatocytes treated with the LPS-containing cytokine mixture showed a significant increase in ROI and nitrogen oxide intermediate formation. AST leakage was not significantly increased in cells treated with the LPS-containing cytokine mixture, independent of growth-factor co-stimulation. However, pretreatment with growth factors significantly reduced AST leakage and ROI formation while increasing cellular glutathione. Application of growth factors did not result in increased NF-κB activation. Pretreatment with growth factors further increased formation of NO(2) (-), NO(3) (-) and S-nitrosothiols in hepatocytes stimulated with LPS-containing cytokine mixture. Thus, we propose that, together with an increase in glutathione increased NO(2) (-), NO(3) (-) formation might shift their metabolism towards non-toxic products.. Our data suggest that hepatotropic growth factors positively influence sepsis-induced hepatocellular injury by reducing cytotoxic ROI formation via induction of the cellular protective antioxidative systems.

Topics: Animals; Antioxidants; Epidermal Growth Factor; Glutathione; Hepatocyte Growth Factor; Hepatocytes; Inflammation; NF-kappa B; Nitric Oxide; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Transforming Growth Factor alpha; Up-Regulation

Molecular mechanisms underlying modulation of inflammatory responses in primary human keratinocytes by plant polyphenols (PPs), namely the glycosylated phenylpropanoid verbascoside, the stilbenoid resveratrol and its glycoside polydatin, and the flavonoid quercetin and its glycoside rutin were evaluated. As non-lethal stimuli, the prototypic ligand for epidermal growth factor receptor (EGFR) transforming growth factor alpha (TGFalpha), the combination of tumor necrosis factor (TNFalpha) and interferon (IFNgamma) (T/I), UVA+UVB irradiation, and bacterial lipopolysaccharide (LPS) were used. We demonstrated differential modulation of inflammatory responses in keratinocytes at signal transduction, gene transcription, and protein synthesis levels as a function of PP chemical structure, the pro-inflammatory trigger used, and PP interaction with intracellular detoxifying systems. The PPs remarkably inhibited constitutive, LPS- and T/I-induced but not TGFalpha-induced ERK phosphorylation. They also suppressed NFkappaB activation by LPS and T/I. Verbascoside and quercetin invariably impaired EGFR phosphorylation and UV-associated aryl hydrocarbon receptor (AhR)-mediated signaling, while rutin, polydatin and resveratrol did not affect EGFR phosphorylation and further activated AhR machinery in UV-exposed keratinocytes. In general, PPs down-regulated gene expression of pro-inflammatory cytokines/enzymes, except significant up-regulation of IL-8 observed under stimulation with TGFalpha. Both spontaneous and T/I-induced release of IL-8 and IP-10 was suppressed, although 50μM resveratrol and polydatin up-regulated IL-8. At this concentration, resveratrol activated both gene expression and de novo synthesis of IL-8 and AhR-mediated mechanisms were involved. We conclude that PPs differentially modulate the inflammatory response of human keratinocytes through distinct signal transduction pathways, including AhR and EGFR.

Topics: Basic Helix-Loop-Helix Transcription Factors; Dermatitis; Flavonoids; Gene Expression; Humans; Immunoblotting; Inflammation; Interferon-gamma; Keratinocytes; Lipopolysaccharides; NF-kappa B; Phenols; Polyphenols; Receptors, Aryl Hydrocarbon; Reverse Transcriptase Polymerase Chain Reaction; RNA; Signal Transduction; Transforming Growth Factor alpha; Ultraviolet Rays

Monocytes/macrophages and fibroblasts are recruited to the injury site and orchestrate the host response and tissue repair. We have previously shown that polyethylene glycol (PEG)-ylated arginine-glycine-aspartic acid (RGD) sequence grafted onto an extracellular matrix (ECM)-based semi-interpenetrating network (sIPN) enhances monocyte adhesion, and modulates subsequent gene expression and release of inflammatory and matrix remodeling factors. In this study, we investigate the direct influence of fibroblasts on monocyte response to this ECM mimic. Key wound-healing factors in inflammation, matrix remodeling, and regeneration were analyzed to gain insight into the interrelated role of regulation in fibroblast-monocyte interaction. Interleukin-1alpha/-1beta (IL-1alpha/-1beta), interleukin-6 (IL-6), tumor necrosis factor- alpha (TNF-alpha), monocyte inflammatory protein-1alpha/-1beta (MIP-1alpha/-1beta), transforming growth factor-alpha (TGF-alpha), monocyte chemoattractant factor (MCP-1), matrix metalloproteinase-2/-9 (MMP-2/-9), vascular endothelial growth factor (VEGF), granulocyte-macrophage colony-stimulating factor (GM-CSF) were analyzed. Fibroblasts decreased monocyte adhesion onto the RGD-grafted sIPN while increasing monocyte GM-CSF on all surfaces over time except for on RGD and PHSRN-grafted sIPN at 96 h. Monocytes decreased initial fibroblast IL-1alpha and TGF-alpha, but drastically increased fibroblast MMP-2 and GM-CSF. Monocyte IL-1beta, TNF-alpha, MIP-1beta, MCP-1, MMP-9, and GM-CSF expression was increased over time in the presence of all sIPNs, and when the sIPNs were immobilized with ligands, a down-regulation of fibroblast IL-1beta, MIP-1alpha, MIP-1beta compared with unmodified sIPN was observed. When the ligand immobilized was RGD, monocyte TGF-alpha, MIP-1beta, and VEGF expression was increased while monocyte GM-CSF was decreased at selected time points. These results showed a dynamic monocyte response to selected ECM components in the presence of fibroblasts.

Topics: Adult; Cell Adhesion; Chemokine CCL4; Coculture Techniques; Extracellular Matrix; Fibroblasts; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Infant, Newborn; Inflammation; Intercellular Signaling Peptides and Proteins; Interleukin-1alpha; Interleukin-1beta; Ligands; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Monocytes; Oligopeptides; Polyethylene Glycols; Protein Transport; Time Factors; Transforming Growth Factor alpha; Vascular Endothelial Growth Factor A

This symposium was held in Trento, Italy, from June 27 to 29, 2006, and was co-chaired by Robert Weinberg and Enrico Mihich. The interactions between tumor cells and their microenvironment were discussed with particular emphasis on their molecular mechanisms. The roles of transforming growth factor beta signaling, urokinase, and matrix metalloproteinases in matrix remodeling; the effects of matrix-tumor interactions on cell proliferation and migration; the tumor-promoting effects of inflammation and of related host cell and cytokine functions; the signaling mechanisms affecting the biology of the stroma; and the mechanisms governing angiogenesis were discussed.

Topics: Cell Division; Cell Movement; Humans; Inflammation; Matrix Metalloproteinases; Neoplasm Metastasis; Neoplasms; Placenta Growth Factor; Pregnancy Proteins; Research; Transforming Growth Factor alpha

It has been shown that Smad3 exerts both tumor-suppressive and -promoting roles. To evaluate the role of Smad3 in skin carcinogenesis in vivo, we applied a chemical skin carcinogenesis protocol to Smad3 knockout mice (Smad3(-/-) and Smad3(+/-)) and wild-type littermates (Smad3(+/+)). Smad3(-/-) mice exhibited reduced papilloma formation in comparison with Smad3(+/+) mice and did not develop any squamous cell carcinomas. Further analysis revealed that Smad3 knockout mice were resistant to 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced epidermal hyperproliferation. Concurrently, increased apoptosis was observed in TPA-treated Smad3(-/-) skin and papillomas when compared with those of wild-type mice. Expression levels of activator protein-1 family members (c-jun, junB, junD, and c-fos) and transforming growth factor (TGF)-alpha were significantly lower in TPA-treated Smad3(-/-) skin, cultured keratinocytes, and papillomas, as compared with Smad3(+/+) controls. Smad3(-/-) papillomas also exhibited reduced leukocyte infiltration, particularly a reduction of tumor-associated macrophage infiltration, in comparison with Smad3(+/+) papillomas. All of these molecular and cellular alterations also occurred to a lesser extent in Smad3(+/-) mice as compared with Smad3(+/+) mice, suggesting a Smad3 gene dosage effect. Given that TGF-beta1 is a well-documented TPA-responsive gene and also has a potent chemotactic effect on macrophages, our study suggests that Smad3 may be required for TPA-mediated tumor promotion through inducing TGF-beta1-responsive genes, which are required for tumor promotion, and through mediating TGF-beta1-induced macrophage infiltration.

Topics: Animals; Apoptosis; Disease Susceptibility; DNA-Binding Proteins; Epidermis; Female; Inflammation; Keratinocytes; Macrophages; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Skin Neoplasms; Smad3 Protein; Tetradecanoylphorbol Acetate; Trans-Activators; Transcription Factor AP-1; Transforming Growth Factor alpha

During inflammatory skin disorders such as psoriasis, atopic dermatitis, and allergic contact dermatitis, epidermal keratinocytes overexpress large amounts of soluble epidermal growth factor receptor ligands in response to tumor necrosis factor alpha and interferon gamma. These cytokines also promote de novo synthesis of numerous chemokines, including CCL2/MCP-1, CCL5/RANTES, CXCL10/IP-10, and CXCL8/IL-8, in turn responsible for the recruitment of different leukocyte populations. This study demonstrates that stimulation of EGFR down-regulates CCL2, CCL5, and CXCL10, while it increases CXCL8 expression in keratinocytes. Conversely, EGFR signaling blockade produces opposite effects, with increased CCL2, CCL5, and CXCL10, and reduced CXCL8 expression. In a mouse model of contact hypersensitivity, a single topical administration of a selective EGFR kinase blocker before antigen challenge results in a markedly enhanced immune response with increased chemokine expression and heavier inflammatory cell infiltrate. Targeting EGFR on epithelial cells may thus have profound impact on inflammatory and immune responses.

Topics: Adult; Animals; Cells, Cultured; Chemokines; Culture Techniques; Dermatitis, Contact; Disease Models, Animal; ErbB Receptors; Female; Humans; Inflammation; Interferon-gamma; Keratinocytes; Male; Mice; Mice, Inbred BALB C; Signal Transduction; Skin; Transcriptional Activation; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha

While previous studies have indicated that exogenous TGF-alpha stimulates epithelial growth, maintenance, and repair of the gut, roles of endogenous TGF-alpha are less well-defined particularly in the small bowel. The current study examined effects of TGF-alpha knockout on adult small intestinal epithelial cell proliferation, migration, apoptosis, and damage/repair response after methotrexate treatment. Compared to normal mice, TGF-alpha gene knockout did not affect crypt cell production, mitosis position, migration, and apoptosis in non-injured intestine. RT-PCR gene expression analysis revealed presence of four out of six TGF-alpha related EGF family ligands in the normal intestine, suggesting a possible functional redundancy of the EGF family in maintenance of the intestine. Although TGF-alpha gene knockout did not significantly impair the overall mucosal repair in methotrexate-induced acute damage in the small intestine, it resulted in a higher apoptotic response in the early hours following methotrexate challenge, and a delayed and reduced crypt cell proliferation during repair. Consistently, after methotrexate challenge, intestinal TGF-alpha mRNA was found to be markedly upregulated in the early hours and during repair in the wild type, and there were similar profiles in the increased expression of all other ligands (except EGF) between the wild type and knockout intestines. Therefore, despite a possible functional redundancy among the EGF family ligands in the normal small intestine, TGF-alpha may play a role in modulating the early apoptotic events and in enhancing the subsequent reparative proliferative response in the methotrexate-damaged intestine.

Topics: Animals; Antimetabolites, Antineoplastic; Cell Cycle; ErbB Receptors; Inflammation; Intestinal Mucosa; Intestine, Small; Ligands; Methotrexate; Mice; Mice, Knockout; Reference Values; Transforming Growth Factor alpha; Wound Healing

Capsaicin-sensitive nerve fibres protect gastrointestinal mucosa in animal models of mucosal injury by modulation of mucosal blood flow and mucus secretion. The aim of our study was to evaluate the effects of capsaicin-sensitive nerve fibres in rat colonic mucosa on epithelial cell proliferation and transforming growth factor-alpha (TGFalpha) expression, which is important in mucosal defence, protection and repair.. Male Wistar rats received either a capsaicin enema with or without giving antagonists to calcitonin-gene-related-peptide (CGRP) or substance P (SP) i.v. immediately prior to the capsaicin enemas; a capsaicin enema after sensory desensitization as described previously; or a vehicle enema. In all experiments, animals received 50 mg/kg BrdU i.v. and were killed at 2, 4, 8, 12, 24 and 48 h after the various treatments. Colonic mucosal specimens were evaluated microscopically for mucosal damage, changes in the numbers of inflammatory cells and BrdU-immunoreactive epithelial cell nuclei. In the same specimens, TGFalpha-mRNA and -protein expression were evaluated by RT-PCR and Western blot analysis using standardized procedures.. A significant increase in the number of mucosal inflammatory cells and an increase in BrdU-immunoreactive nuclei were detected following mucosal exposure to capsaicin. A 2-fold increase of TGFalpha mRNA and a 10-fold increase of TGFalpha protein expression were obtained 2-12 h after capsaicin enemas. The effects on the invading number of inflammatory cells and on the increase in BrdU immunoreactive epithelial cell nuclei were significantly reduced by both CGRP and SP antagonists and were abolished in rats previously sensory-desensitized.. Capsaicin-sensitive nerve fibres modulate epithelial cell proliferation and TGFalpha expression in colonic mucosa as well as a migration of inflammatory cells into the colonic mucosa. These effects are mediated by the neurotransmitters CGRP and SP.

Topics: Animals; Blotting, Western; Bromodeoxyuridine; Calcitonin Gene-Related Peptide; Capsaicin; Cell Division; Colon; Denervation; Goblet Cells; Immunohistochemistry; Inflammation; Intestinal Mucosa; Male; Nerve Fibers; Peroxidase; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Substance P; Transforming Growth Factor alpha

With the objective of estimating proliferation ability of epidermis of middle ear cholesteatoma, the difference in proliferating cell nuclear antigen (PCNA) staining between the skin of the bone region of the external ear canal (control skin) and cholesteatoma epidermis and the effects on PCNA staining of subepidermal inflammatory cell infiltration of cholesteatoma were immunohistochemically studied using an antibody against PCNA. Transforming growth factor-alpha (TGF-alpha) is known to promote epidermal proliferation based on autocrine mechanism. But it is not clear that cholesteatoma epidermis is actually in the state of hyperproliferation under the effect of this growth factor. To estimate the effect of TGF-alpha on epidermal proliferation ability, the authors compared the location of PCNA and TGF-alpha in the same specimen. Unlike the control skin, not only epidermal basal cell layer and suprabasal cell layer, but also more superior layers were found to have high levels of PCNA staining in the epidermis of cholesteatoma. However, in the same cholesteatoma epidermal tissue, the PCNA staining was varied and the difference was ascribable to subepidermal cell inflammation. It appeared that the proliferation ability was high in regions where subepidermal inflammatory cell infiltration was severe. These differences in microenvironment are inferred to greatly affect proliferation ability of cholesteatoma epidermis.

Topics: Adult; Cell Division; Cholesteatoma, Middle Ear; Coloring Agents; Ear Canal; Epidermis; Fibroblasts; Humans; Immunohistochemistry; In Situ Hybridization; Inflammation; Proliferating Cell Nuclear Antigen; Skin; Temporal Bone; Transforming Growth Factor alpha

Transforming growth factor-alpha (TGF-alpha) exerts several effects on target cells, such as neovascularization promotion and mitogenic signalling. Using immunoelectron microscopy, we show that monocytes and neutrophils, store TGF-alpha in cytoplasmic granules. In monocytes, TGF-alpha did not colocalize with components of peroxidase-positive granules or with albumin of secretory vesicles. Furthermore, no colocalization of TGF-alpha with components of azurophilic or specific granules or secretory vesicles was observed in neutrophils. Activated monocytes and tissue-macrophages contained much less TGF-alpha-positive granules, suggesting TGF-alpha release. Western blot analysis showed a protein of 10 kD in lysates of monocytes. TGF-alpha mRNA was detected in monocytoid cells from the bone marrow by in situ hybridization. This study shows for the first time that monocytes and neutrophils contain TGF-alpha in all stages of maturation and that TGF-alpha in monocytes is stored in a large population of peroxidase-negative granules suggesting a function for these granules. Monocytes and neutrophils are important effector cells in inflammatory reactions. The present finding that these cells contain TGF-alpha might explain complications such as fibrosis and neoplastic transformation, caused by chronic inflammation.

Topics: Bone Marrow; Bone Marrow Cells; Cytoplasmic Granules; Humans; In Situ Hybridization; Inflammation; Microscopy, Immunoelectron; Monocytes; Neutrophils; RNA, Messenger; Transforming Growth Factor alpha

The oviduct provides the environment in which fertilization of the egg and subsequent development of the preimplantation mouse embryo occurs, but little is known about the oviduct's capacity to produce growth factors or cytokines that may influence these preimplantation events. Northern blot analysis and/or immunohistochemistry were employed to examine the expression or cellular distribution, respectively, of the growth factors heparin-binding epidermal-like growth factor (HB-EGF), transforming growth factor (TGF) alpha, epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I), TGF beta 1, TGF beta 2, and TGF beta 3; of estrogen-regulated lactoferrin (LF); and of the cytokines interleukin (IL)-1 alpha and IL-1 beta in the mouse oviduct during the preimplantation period (Days 1-4 [Day 1 = vaginal plug]) and 7 days after ovariectomy. The results demonstrated that, except for EGF, each of the growth factors and the LF genes are expressed in the ampulla and isthmus regions of the oviduct throughout the preimplantation period. Prominent immunostaining in secretory epithelial cells was noted for HB-EGF, TGF alpha, IGF-I, TGF beta 1, and TGF beta 2, and LF. Less intense immunostaining in the serosa and/or smooth muscle was also noted for TGF alpha, IGF-I, and TGF beta 1. In contrast, intense immunostaining in smooth muscle was noted for TGF beta 2, and TGF beta 3 was detected exclusively in smooth muscle cells. The abundance of these mRNAs was relatively constant during the preimplantation period, and ovariectomy did not reduce the levels of these mRNAs. In contrast to these growth factors, the cytokine mRNAs examined (IL-1 alpha and IL-1 beta) were at or below the limits of detection under these experimental conditions, and inflammatory leukocytes (LF-immunopositive neutrophils, IL-1 beta-immunopositive monocytes/macrophages, or peroxidase-positive eosinophils) were not detected in the oviduct, but were abundant in the adjacent uterine stroma on Day 1. These studies show that several growth factors are synthesized by the mouse oviduct and suggest that ovarian steroids do not play a major role in modulating expression of these genes in the oviduct during the preimplantation period. Furthermore, unlike the uterus on Day 1, the oviduct does not exhibit an inflammatory response to mating.

Topics: Animals; Blotting, Northern; Embryonic Development; Eosinophils; Epidermal Growth Factor; Fallopian Tubes; Female; Gene Expression; Growth Substances; Inflammation; Insulin-Like Growth Factor I; Interleukin-1; Lactoferrin; Leukocytes; Mice; Monocytes; Neutrophils; Ovariectomy; Pregnancy; RNA, Messenger; Transforming Growth Factor alpha; Transforming Growth Factor beta