Anti-inflammatory Reactions Research Articles

Anti-inflammatory role of PGD 2 in acute lung inflammation and therapeutic application of its signal enhancement

We investigated the role of prostaglandin D2 (PGD2) signaling in acute lung injury (ALI), focusing on its producer-effector interaction in vivo. Administration of endotoxin increased edema and neutrophil infiltration in the WT mouse lung. Gene disruption of hematopoietic PGD synthase (H-PGDS) aggravated all of the symptoms. Experiments involving bone marrow transplantation between WT and H-PGDS-deficient mice showed that PGD2 derived from alveolar nonhematopoietic lineage cells (i.e., endothelial cells and epithelial cells) promotes vascular barrier function during the early phase (day 1), whereas neutrophil-derived PGD2 attenuates its own infiltration and cytokine expression during the later phase (day 3) of ALI. Treatment with either an agonist to the PGD2 receptor, DP, or a degradation product of PGD2, 15-deoxy-Δ(12,14)-PGJ2, exerted a therapeutic action against ALI. Data obtained from bone marrow transplantation between WT and DP-deficient mice suggest that the DP signal in alveolar endothelial cells is crucial for the anti-inflammatory reactions of PGD2. In vitro, DP agonism directly enhanced endothelial barrier formation, and 15-deoxy-Δ(12,14)-PGJ2 attenuated both neutrophil migration and cytokine expression. These observations indicate that the PGD2 signaling between alveolar endothelial/epithelial cells and infiltrating neutrophils provides anti-inflammatory effects in ALI, and suggest the therapeutic potential of these signaling enhancements.

Critical role of Trib1 in differentiation of tissue-resident M2-like macrophages

Macrophages consist of at least two subgroups, M1 and M2 (refs 1-3). Whereas M1 macrophages are proinflammatory and have a central role in host defence against bacterial and viral infections, M2 macrophages are associated with responses to anti-inflammatory reactions, helminth infection, tissue remodelling, fibrosis and tumour progression. Trib1 is an adaptor protein involved in protein degradation by interacting with COP1 ubiquitin ligase. Genome-wide association studies in humans have implicated TRIB1 in lipid metabolism. Here we show that Trib1 is critical for the differentiation of F4/80(+)MR(+) tissue-resident macrophages--that share characteristics with M2 macrophages (which we term M2-like macrophages)--and eosinophils but not for the differentiation of M1 myeloid cells. Trib1 deficiency results in a severe reduction of M2-like macrophages in various organs, including bone marrow, spleen, lung and adipose tissues. Aberrant expression of C/EBPα in Trib1-deficient bone marrow cells is responsible for the defects in macrophage differentiation. Unexpectedly, mice lacking Trib1 in haematopoietic cells show diminished adipose tissue mass accompanied by evidence of increased lipolysis, even when fed a normal diet. Supplementation of M2-like macrophages rescues the pathophysiology, indicating that a lack of these macrophages is the cause of lipolysis. In response to a high-fat diet, mice lacking Trib1 in haematopoietic cells develop hypertriglyceridaemia and insulin resistance, together with increased proinflammatory cytokine gene induction. Collectively, these results demonstrate that Trib1 is critical for adipose tissue maintenance and suppression of metabolic disorders by controlling the differentiation of tissue-resident M2-like macrophages.

Reduced secretion of the inflammatory cytokine IL-1β by stimulated peritoneal macrophages of radiosensitive Balb/c mice after exposure to 0.5 or 0.7Gy of ionizing radiation

Since the beginning of the 20th century, low dose radiotherapy (LD-RT) has been practiced and established as therapy of inflammatory diseases. Several clinical studies already have proven the anti-inflammatory effect of low doses of ionizing irradiation (LDR). However, further research is inevitable to reveal the underlying immune-biological mechanisms. Focus has been set on the modulation of activated macrophages by LDR, since they participate in both, initiation and resolution of inflammation. Here we examined with an ex vivo peritoneal mouse macrophage model how LDR modulates the secretion of the inflammatory cytokines IL-1β and TNF-α by activated macrophages and whether the basal radiosensitivity of the immune cells has influence on it. Peritoneal macrophages of Balb/c mice responded to exposure of 0.5 or 0.7 Gy of ionizing irradiation (X-ray) with significant decreased release of IL-1β and slightly, but not significantly, reduced release of TNF-α. Macrophages of the less radiosensitive C57BL/6 mice did not show this anti-inflammatory reaction. This was observed in both wild type and human TNF-α transgenic animals with C57BL/6 background. We conclude that only the inflammatory phenotype of more radiosensitive macrophages is reduced by LDR and that ex vivo and in vivo models with primary cells should be applied to examine how the immune system is modulated by LDR.

Host Innate Immune Responses to Microbial Pathogens

Sepsis is among the leading causes of death worldwide and its incidence is increasing. Defined as the host response to infection, sepsis is a clinical syndrome considered to be the expression of a dysregulated immune reaction induced by danger signals that may lead to organ failure and death. Remarkable progresses have been made in our understanding of the molecular basis of host defenses in recent years. The host defense response is initiated by innate immune sensors of danger signals designated under the collective name of pattern-recognition receptors. Members of the family of microbial sensors include the complement system, the Toll-like receptors, the nucleotide-binding oligomerization domainlike receptors, the RIG-I-like helicases and the C-type lectin receptors. Ligand-activated pattern-recognition receptors kick off a cascade of intracellular events resulting in the expression of co-stimulatory molecules and release of effector molecules playing a fundamental role in the initiation of the innate and adaptive immune responses. Fine tuning of proinflammatory and anti-inflammatory reactions is critical for keeping the innate immune response in check. Overwhelming or dysregulated responses induced by infectious stimuli may have dramatic consequences for the host as shown by the profound derangements observed in sepsis. Unfortunately, translational research approaches aimed at the development of therapies targeting newly identified innate immune pathways have not held their promises. Indeed, all recent clinical investigations of adjunctive anti-sepsis treatments had little, if any, impact on morbidity and all-cause mortality of sepsis. Dissecting the mechanisms underlying the transition from infection to sepsis is essential for solving the sepsis enigma. Important components of the puzzle have already been identified, but the hunt must go on in the laboratory and at the bedside.

Tempol reduces injury area in rat model of spinal cord contusion injury through suppression of iNOS and COX-2 expression

The present study focused on the biologic effects of tempol on anti-inflammatory and nitric oxide generation in contusion spinal cord injury (SCI). The animal model of SCI was induced by dropping a 10-g rod (2.0 mm in diameter) at a height of 25 mm. Tempol was injected intraperitoneally a dose of 100 mg/kg at 15 min before SCI. Controls was injected with saline. The contused spinal segments were removed according to time courses, and the expression level of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was analyzed along with the size of irreversibly damaged region. After SCI, the relative amounts of COX-2 and iNOS mRNA were peaked at 8 h after post-injury, and then decreased up to 7 days post-injury, and normal level at 14 days. Expression of COX-2 protein was peaked at 8 h post-injury. With the tempol pre-treatment, the immunoreactivity of COX-2 and nitrotyrosine in paraffin-embedded tissue slices was profoundly decreased. The irreversibly damaged area of the spinal cord was peaked at 3 days after SCI. With tempol pre-treatment, the irreversibly damaged area shows a statistically significant decrease at 3 days after SCI. These evidences indicate that tempol pre-treatment reduces irreversibly damaged area on the contusion SCI in rat. The mechanisms of biologic reactions of tempol might be related to the decreased expressions of COX-2 and iNOS in spinal cord cells, neurons and glia. It is expected that the tempol effect on the SCI is not only antioxidant activity but also anti-inflammatory reaction.

Labeling Acetyl- and Butyrylcholinesterase Using Semiconductor Nanocrystals for Biological Applications

Colloidal semiconductor nanocrystals (SC-NC) present unique and well-controlled physical and chemical properties that make them most suitable, as fluorescent agents, for high resolution imaging and direct follow-up in biological milieus. SC-NC present intense fluorescence that is tunable by size, composition, and shape. In addition, they have large molar extinction coefficients and thermal and photochemical stability, thus avoiding the relatively fast photobleaching characteristic of traditional fluorescent dyes. Nevertheless, constructing SC-NC–labeled, biologically active macromolecules and especially enzymes is challenging. Ensuring the enzyme’s stability, sustaining its biological functions, and achieving the desired application all require specific design of the nanoparticles (NPs) properties, their surface coating, and the labeling process. Here, we report an in-depth investigation of the experimental steps involved in the development of a flexible SC-NC labeling toolkit for recombinant human cholinesterases (ChEs), including acetylcholinesterase-R (AChE-R), and the homologous enzyme butyrylcholinesterase (BChE). AChE and BChE, the major acetylcholine (ACh) hydrolyzing enzymes, regulate ACh-mediated neurotransmission, signal transduction, and anti-inflammatory reactions. Impaired ChE functioning is causally involved in many pathologies, including Alzheimer’s and Parkinson’s diseases, cardiovascular disorders, myasthenia gravis, and Sjogren’s syndrome. In addition, exposure to anti-ChE drugs, insecticides, and poisonous nerve agents present prevalent health and security issues. Therefore, recombinant ChEs have been developed both as therapeutic agents and for biomedical research purposes. The labeling toolkit presented here enables optimization of highly emitting SC-NC- ChE conjugates with hydrolytic activity and capacity to bind anti-ChEs, suitable for use for biomedical applications and allows sensitive long-term follow-up of the locations and interactions of these important enzymes in biological systems.

Visual outcome after macular grid laser and intravitreal triamcinolone acetonide in diabetic macular edema

Introduction: Macular edema affects approximately 29% of diabetic patients and constitutes the primary cause of visual impairment in this population. The combination therapy of grid laser and intravitreal triamcinolone acetonide (IVTA), the reaction mechanism of each therapy to macular edema is different; hence, not only greater effects on the reduction of macular edema can be anticipated, but also the stimulation of inflammatory reaction induced by grid laser could be prevented by anti-inflammatory reaction of steroids as investigated in several studies. Purpose: The purpose of this study was to assess the best corrected visual acuity after IVTA and macular grid laser in combination in cases of diabetic macular edema. Materials and Methods: A total of 50 patients of diabetic macular edema were enrolled in our prospective, randomized clinical trial study, grid laser, and IVTA was given to each patient in combination. The pre-intervention and post-intervention best corrected visual acuity was studied. Results: We observed 35 (70%) patients improved by 1 or more line on Snellen's chart, 12 (24%) remain unchanged and 3 (6%) deteriorated from baseline. The overall mean improvement in visual acuity was 1.08 with SD of ± 0.16. Conclusion: The results of present study showed a favorable influence of grid laser and intravitreal triamcinolone acetonide in diabetic macular edema. It improves and stabilizes the visual acuity in patients with the diabetic macular edema.

MicroRNA-132 Modulates Cholinergic Signaling and Inflammation in Human Inflammatory Bowel Disease

A feedback loop, termed the anti-inflammatory reflex, exists between the immune system and the nervous system. Inflammation triggers an afferent vagal response that is transmitted to the hypothalamus, where it stimulates the efferent vagal nerve to release the neurotransmitter acetylcholine. Acetylcholine activates its nicotinic receptor on macrophages through which it inhibits production of pro-inflammatory cytokines. Multi-leveled evidence suggests a role for the anti-inflammatory reflex in the pathogenesis of inflammatory bowel disease (IBD). Electrical stimulation of the vagus nerve attenuated DSS-induced colitis in rats. Similarly, in vagotomized mice, DSS-induced colitis was more severe compared to control mice and sympathectomy improved experimental colitis. Although, the anti-inflammatory reflex has not been studied in human IBD, enteric nervous system abnormalities in IBD patients, may support its potential role in the pathogenesis of this disease. MicroRNA-132 (miR-132) targets acetylcholinesterase (AChE) and potentiates the cholinergic blockade of inflammatory reactions in cultured cells and experimental mice, but the implications of this interaction to human inflammatory disease remained unexplored. This study aimed to test whether vagal tone (assessed through AChE activity) correlates with inflammation and whether miR-132 is causally involved in anti-inflammatory reactions of patients with IBD and modulates vagal tone and consequently inflammation in IBD patients compared to healthy controls (HC). We prospectively collected clinical data and measured inflammation readouts and the cholinergic status (total capacity for hydrolyzing acetylcholine in one’s circulation), as well as AChE activity in two independent cohorts of IBD patients and HC (Cohort 1: n = 77 IBD, n = 74 HC and cohort 2: n = 33 IBD, n = 16 HC participants). We quantified miR-132 levels systemically in peripheral blood and locally in intestinal tissue biopsies removed at colonoscopy from inflamed and apparently quiescent tissues of tested volunteers. MiR-132 levels are higher in inflamed compared to apparently quiescent intestinal biopsies from IBD patients, while systemic levels were unchanged among groups. Correspondingly, the cholinergic status as well as AChE activity was significantly lower in IBD patients suffering from moderate-severe disease as compared to HC or IBD patients presenting low disease severity. IBD patients (n = 33) presented lower AChE activity compared to HC (n = 16) (289± 128 AU vs 391± 102AU, P = 0.001), and a negative correlation between AChE activity and C-reactive protein (CRP) levels (r = -0.47, P = 0.01). Corroborating these observations in an additional cohort of participants, CRP and AChE activity were negatively correlated in patients with moderate-severe disease (n = 16; r = -0.6, P = 0.04) and positively correlated in HC (n = 74, r = 0.24, P = 0.046). We demonstrated a strong negative correlation between AChE activity and hs-CRP that is disease severity dependent in IBD patients and maybe locally regulated by colonic miR-132, suggesting a new insight to inflammation regulation in IBD. This may signify dual protective role for cholinergic status on inflammation where in acute inflammation cholinergic status enhances the inflammatory process to overcome triggering events such as infections, but in chronic inflammatory states the cholinergic status promotes amelioration of the inflammatory process to restore homeostasis. This may implicate a novel homeostatic mechanism involving a miR-132-mediated change of the cholinergic status in IBD.

Renal Cell Apoptosis and New Treatment Options in Sepsis-Induced Acute Kidney Injury

Sepsis is a common and important cause of mortality in critically ill patients. Acute kidney injury (AKI) is one of the most important factors determining morbidity and mortality in the prognosis of sepsis. Recent studies have indicated that the pathogenetic mechanism in septic AKI is totally different from that in non-septic AKI. Our understanding of sepsis-associated AKI pathophysiology is shifting from renal vasoconstriction, ischemia, and acute tubular necrosis to heterogeneous vasodilation, hyperemia, and acute tubular apoptosis. Especially, apoptosis is gradually gaining importance in the understanding of the development of renal injury. The frequency of renal tubular apoptosis on biopsies of septic patients has been pointed out in recently published studies. Apoptosis can be triggered by ischemia, exogen toxins, or endogen mediators. It has been shown in some animal models that hyperglycemia, which is common in critically ill patients, causes apoptosis in renal tubular cells. New treatment options have emerged in the light of recent findings. Ghrelin that inhibits pro-inflammatory cytokines, caspase inhibitors that block the apoptotic pathway, and suppression of anti-inflammatory reactions are under study. Among the existing methods of treatment, usage of arginine, which is a vasopressor agent, ventilation with a low tidal volume, and hemofiltration methods cleaning toxic mediators from the circulation should be considered in the first place. Hyperglycemia treatment is of major importance, since, besides its anti-inflammatory effect, it has a protective role on the kidney. Regarding pathogenesis, rates of morbidity and mortality are aimed to be reduced through the new agents of therapy that have been studied on.

Differential recognition by tick-resistant cattle of the recombinantly expressed Rhipicephalus microplus serine protease inhibitor-3 (RMS-3)

Rhipicephalus microplus is an important bovine ectoparasite, widely distributed in tropical and subtropical regions of the world causing large economic losses to the cattle industry. Its success as an ectoparasite is associated with its capacity to disarm the antihemostatic and anti-inflammatory reactions of the host. Serpins are protease inhibitors with an important role in the modulation of host–parasite interactions. The cDNA that encodes for a R. microplus serpin was isolated by RACE and subsequently cloned into the pPICZαA vector. Sequence analysis of the cDNA and predicted amino acid showed that this cDNA has a conserved serpin domain. B- and T-cell epitopes were predicted using bioinformatics tools. The recombinant R. microplus serpin (rRMS-3) was secreted into the culture media of Pichia pastoris after methanol induction at 0.2mgl−1. qRT-PCR expression analysis of tissues and life cycle stages demonstrated that RMS-3 was mainly expressed in the salivary glands of female adult ticks. Immunological recognition of the rRMS-3 and predicted B-cell epitopes was tested using tick-resistant and susceptible cattle sera. Only sera from tick-resistant bovines recognized the B-cell epitope AHYNPPPPIEFT (Seq7). The recombinant RMS-3 was expressed in P. pastoris, and ELISA screening also showed higher recognition by tick-resistant bovine sera. The results obtained suggest that RMS-3 is highly and specifically secreted into the bite site of R. microplus feeding on tick-resistant bovines. Capillary feeding of semi-engorged ticks with anti-AHYNPPPPIEFT sheep sera led to an 81.16% reduction in the reproduction capacity of R. microplus. Therefore, it is possible to conclude that R. microplus serpin (RMS-3) has an important role in the host–parasite interaction to overcome the immune responses in resistant cattle.

Regulatory Cells in Allergen-Specific Immunotherapy

Allergen-specific immunotherapy (SIT) is currently the best available curative treatment in allergies and has been used for the treatment of patients for the past 100 years. The formation of a Th2 cell predominant inflammation in addition to production of allergen-specific IgE, the attraction of proinflammatory cells and the degranulation of effector cells, such as mast cells, are essential mechanisms in allergy development. Tregs aim to diminish these effects by IL-10- and TGF-β-mediated anti-inflammatory reactions and therefore are one of the main targets in SIT. The induction of allergen tolerance is the key to successful SIT. With a special focus on Tregs, this review aims to clarify what is currently known about allergy development and the mode of action in allergen-SIT, which helps to develop further therapeutic strategies in the fight against allergic diseases.

The cell-penetrating peptide domain from human heparin-binding epidermal growth factor-like growth factor (HB-EGF) has anti-inflammatory activity in vitro and in vivo

A heparin-binding peptide (HBP) sequence from human heparin-binding epidermal growth factor-like growth factor (HB-EGF) was identified and was shown to exhibit cell penetration activity. This cell penetration induced an anti-inflammatory reaction in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. HBP penetrated the cell membrane during the 10min treatment and reduced the LPS-induced production of nitric oxide (NO), inducible nitric oxide synthase (iNOS), and cytokines (TNF-α and IL-6) in a concentration-dependent manner. Additionally, HBP inhibited the LPS-induced upregulation of cytokines, including TNF-α and IL-6, and decreased the interstitial infiltration of polymorphonuclear leukocytes in a lung inflammation model. HBP inhibited NF-κB-dependent inflammatory responses by directly blocking the phosphorylation and degradation of IκBα and by subsequently inhibiting the nuclear translocation of the p65 subunit of NF-κB. Taken together, this novel HBP may be potentially useful candidate for anti-inflammatory treatments and can be combined with other drugs of interest to transport attached molecules into cells.

The changes in subgroup of helper T lymphocyte and calcitonin and their correlation in patients with severe hospital acquired pneumonia

To observe the changes in subgroups of helper T cells and serum calcitonin (PCT) levels in patients with hospital acquired pneumonia (HAP) and their correlation. Eighty-nine patients with diagnosis of HAP (severe in 59 patients, mild in 30 cases) were included, with 20 healthy adults as control. Percentage of CD4(+), Th1 and Th2 cells, and Th1/Th2 ratio were determined, and sandwich immune luminescence was used to detect the level of serum PCT. The percentage of Th1 cells [(8.40 ± 3.01)%] was significantly lower in patients with severe HAP compared with that of mild pneumonia group [(13.90 ± 2.37)%, P < 0.05] and healthy controls [(17.40 ± 4.20)%, P < 0.01]. Percentage of Th2 cells was obviously higher in patients with severe HAP [(17.30 ± 5.74)%] than mild pneumonia group [(7.70 ± 2.35)%, P < 0.05] and healthy controls [(7.90 ± 1.92)%, P < 0.01]. Th1/Th2 ratio was also obviously lower in severe pneumonia group (0.57 ± 0.15) than that of mild pneumonia group (2.80 ± 0.46, P < 0.01) and healthy controls (3.11 ± 0.87, P < 0.01). Compared with healthy controls, Th1 cells in mild pneumonia patients were reduced significantly (P < 0.05), but there was no significant difference in Th2 cells and Th1/Th2 ratio (both P > 0.05). There was no difference in CD4(+) among severe, mild pneumonia and healthy controls [(30.20 ± 10.83)%, (34.70 ± 13.57)%, (28.80 ± 9.61)%, respectively, all P >0.05]. The level of PCT (μg/L) was significantly elevated in mild and severe pneumonia patients compared with that of healthy controls (1.73 ± 0.88, 3.51 ± 2.66 vs. 0.30 ± 0.10, both P < 0.01), and the level of PCT in severe pneumonia was significantly higher than that of mild pneumonia (P < 0.05). Regression analysis of Th1/Th2 and PCT revealed a significant negative correlation, with regression equation Y = -0.937x (F=236.23,P = 0.0000). Patients with severe HAP had obvious imbalance of Th1/Th2. The suppression of cellular immune function, reduction in Th2 cells and exacerbation of anti-inflammatory reaction intensify the infection leading to multiple organ dysfunction syndrome. There is obvious negative correlation between Th1/Th2 and PCT. PCT could be used as an indicator of immune response in reflecting cellular and humoral immunity of the patient.

Signaling by Myeloid C-Type Lectin Receptors in Immunity and Homeostasis

Myeloid cells are key drivers of physiological responses to pathogen invasion or tissue damage. Members of the C-type lectin receptor (CLR) family stand out among the specialized receptors utilized by myeloid cells to orchestrate these responses. CLR ligands include carbohydrate, protein, and lipid components of both pathogens and self, which variably trigger endocytic, phagocytic, proinflammatory, or anti-inflammatory reactions. These varied outcomes rely on a versatile system for CLR signaling that includes tyrosine-based motifs that recruit kinases, phosphatases, or endocytic adaptors as well as nontyrosine-based signals that modulate the activation of other pathways or couple to the uptake machinery. Here, we review the signaling properties of myeloid CLRs and how they impact the role of myeloid cells in innate and adaptive immunity.

The Clinical Course of Acute Pancreatitis and the Inflammatory Mediators That Drive It

Acute pancreatitis (AP) is a common emergency condition. In the majority of cases, it presents in a mild and self-limited form. However, about 20% of patients develop severe disease with local pancreatic complications (including necrosis, abscess, or pseudocysts), systemic organ dysfunction, or both. A modern classification of AP severity has recently been proposed based on the factors that are causally associated with severity of AP. These factors are both local (peripancreatic necrosis) and systemic (organ failure). In AP, inflammation is initiated by intracellular activation of pancreatic proenzymes and/or nuclear factor-κB. Activated leukocytes infiltrate into and around the pancreas and play a central role in determining AP severity. Inflammatory reaction is first local, but may amplify leading to systemic overwhelming production of inflammatory mediators and early organ failure. Concomitantly, anti-inflammatory cytokines and specific cytokine inhibitors are produced. This anti-inflammatory reaction may overcompensate and inhibit the immune response, rendering the host at risk for systemic infection. Currently, there is no specific treatment for AP. However, there are several early supportive treatments and interventions which are beneficial. Also, increasing the understanding of the pathogenesis of systemic inflammation and the development of organ dysfunction may provide us with future treatment modalities.

Roles of PI3K/AKT/GSK3/mTOR Pathway in Cell Signaling of Mental Illnesses.

Several pharmacological agents acting on monoamine neurotransmission are used for the management of mental illnesses. Regulation of PI3K/AKT and GSK3 pathways may constitute an important signaling center in the subcellular integration of the synaptic neurotransmission. The pathways also modulate neuronal cell proliferation, migration, and plasticity. There are evidences to suggest that inflammation of neuron contributes to the pathology of depression. Inflammatory activation of neuron contributes to the loss of glial elements, which are consistent with pathological findings characterizing the depression. A mechanism of anti-inflammatory reactions from antidepressant medications has been found to be associated with an enhancement of heme oxygenase-1 expression. This induction in brain is also important in neuroprotection and neuroplasticity. As enzymes involved in cell survival and neuroplasticity are relevant to neurotrophic factor dysregulation, the PI3K/AKT/GSK3 may provide an important signaling for the neuroprotection in depression. In this paper, we summarize advances on the involvement of the PI3K/AKT/GSK3 pathways in cell signaling of neuronal cells in mental illnesses.

Effects of Bifidobacterium breve on inflammatory gene expression in neonatal and weaning rat intestine

To examine the immune-modulatory effects of probiotics during early infancy, Bifidobacterium breve M-16V (B. breve) was administered to rat pups during the newborn or weaning period, and the expression of inflammatory genes was investigated using a cDNA microarray and real-time PCR. After B. breve administration, significant increases in the numbers of Bifidobacterium in both the cecum and colon were confirmed during the newborn period. The numbers of upregulated and downregulated genes were greater during the weaning period than in the newborn period and were greatest in the colon, with fewer genes altered in the small intestine and the fewest in the spleen. The expression of inflammation-related genes, including lipoprotein lipase (Lpl), glutathione peroxidase 2 (Gpx2), and lipopolysaccharide-binding protein (Lbp), was significantly reduced in the colon during the newborn period. In weaning rat pups, the expression of CD3d, a cell surface receptor-linked signaling molecule, was significantly enhanced in the colon; however, the expression of co-stimulatory molecules was not enhanced. Our findings support a possible role for B. breve in mediating anti-inflammatory and antiallergic reactions by modulating the expression of inflammatory molecules during the newborn period and by regulating the expression of co-stimulatory molecules during the weaning period. Gene expression in the intestine was investigated after feeding 5 × 10(8) cfu of B. breve every day to the F344/Du rat from days 1 to 14 (newborn group) and from days 21 to 34 (weaning group). mRNA was extracted from intestine, and the expression of inflammatory gene was analyzed by microarray and real-time PCR.

Damage associated molecular patterns within xenogeneic biologic scaffolds and their effects on host remodeling

The immune response is an important determinant of the downstream remodeling of xenogeneic biologic scaffolds in vivo. Pro-inflammatory responses have been correlated with encapsulation and a foreign body reaction, while anti-inflammatory reactions are associated with constructive remodeling. However, the bioactive and bioinductive molecules within the extracellular matrix (ECM) that induce this polarization are unclear, although it is likely that cellular remnants such as damage associated molecular patterns (DAMPs) retained within the scaffold may play a role. The present study investigated the immunomodulatory effects of common ECM scaffolds. Results showed that tissue source, decellularization method and chemical crosslinking modifications affect the presence of the well characterized DAMP - HMGB1. In addition, these factors were correlated with differences in cell proliferation, death, secretion of the chemokines CCL2 and CCL4, and up regulation of the pro-inflammatory signaling receptor toll-like receptor 4 (TLR4). Inhibition of HMGB1 with glycyrrhizin increased the pro-inflammatory response, increasing cell death and up regulating chemokine and TLR4 mRNA expression. The present study suggests the importance of HMGB1 and other DAMPS as bioinductive molecules within the ECM scaffold. Identification and evaluation of other ECM bioactive molecules will be an area of future interest for new biomaterial development.

Increase of high molecular weight adiponectin in hypertensive pregnancy was correlated with brain-type natriuretic peptide stimulation on adipocyte

BackgroundHigh-molecular weight (HMW)-adiponectin is an active multimer for insulin sensitivity and anti-inflammatory reactions. We compared the ratio of serum total and HMW-adiponectin with brain-type natriuretic peptide (BNP) and other adipocytokines in normal pregnancy and pregnancy-induced hypertension (PIH). Effect of BNP on the secretion of adiponectin from cultured adipocytes was also examined. MethodsThe three study groups consisted of 44 non-pregnant women, 40 normal (healthy) pregnant women over 28weeks gestation and 29 patients with severe PIH. Adiponectin (protease-pretreated for HMW), BNP-N-terminal, leptin, and monocyte chemoattractant protein (MCP)-1 were measured with ELISA. Pre-adipocytes were differentiated to matured adipocytes and cultured with recombinant-BNP addition. ResultsHMW-to-total adiponectin ratio (HMW-ratio) was lower in normal pregnancy than in non-pregnant, and significantly higher in PIH than normal pregnancies. BNP-N-terminal showed positive correlation with HMW-adiponectin and HMW-ratio. Leptin and MCP-1 showed positive correlation with HMW-adiponectin, but not with HMW-ratio. Adiponectin in the supernatant of adipocyte cultures and intracellular cyclic-GMP was increased in dose-dependent manner in response to BNP. ConclusionThe observed increase in the HMW-adponectin ratio in subjects with PIH may reflect a functional increase of adiponectin in the pathophysiology of PIH. Additionally, this increase seemed to be related to BNP via stimulation of adipocytes.

Inhibitory Effect of a Glycoprotein Isolated from Golden Oyster Mushroom (Pleurotus citrinopileatus) on the Lipopolysaccharide-Induced Inflammatory Reaction in RAW 264.7 Macrophage

Mushrooms have become an important source of natural antitumor, antiviral, antibacterial, immunomodulatory, and anti-inflammatory agents. Golden oyster mushroom, Pleurotus citrinopileatus , is a common mushroom in oriental countries for human consumption. The present study investigated the anti-inflammatory reaction of the bioactive nonlectin glycoprotein (PCP-3A) isolated from the fresh fruiting body of this mushroom. Western blot analysis on LPS-induced iNOS, COX-2, and NF-κB expressions in RAW 264.7 cells as affected by PCP3-A was performed to elucidate the mechanism of NO and PGE2 reduction. The results showed that PCP-3A failed to affect RAW 264.7 viability at a concentration up to 6.25 μg/mL, but inhibited LPS (1 μg/mL)-induced expression, and that PCP-3A inhibited the production of NO and PGE2 in LPS-activated macrophages via the down-regulation of certain pro-inflammatory mediators, including iNOS and NF-κB.