Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P43026 (lipopolysaccharide)
 62,215 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Ehrlichia canis (E. canis) is a lipopolysaccharide (LPS)-deficient obligatory intracellular bacterium that causes canine monocytic ehrlichiosis, a chronic febrile disease accompanied with hematological abnormality. This study analyzed temporal expression levels of IL-1beta, IL-2, IL-6, IL-8, IFN-gamma, and TNF-alpha mRNA by peripheral blood leukocytes from dogs experimentally infected with a new virulent strain of E. canis by using real-time RT-PCR. Relative levels of IL-1beta and IL-8 transcripts normalized by the beta-actin transcript levels, were significantly upregulated, whereas those of TNF-alpha and IFN-gamma transcripts were only weakly upregulated in all three infected dogs, starting from 2 days up to 52 days post inoculation. The expressions of IL-2 and IL-6 genes were extremely low compared with the positive control (ConA-stimulated canine peripheral blood leukocytes). This study showed that E. canis can induce chronic expression of a subset of proinflammatory cytokine genes: balance, timing, and duration of these cytokine generations may contribute to the progression of canine ehrlichiosis.
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PMID:Cytokine gene expression by peripheral blood leukocytes in dogs experimentally infected with a new virulent strain of Ehrlichia canis. 1711 59

Proinflammatory phenotype activation in macrophages (MPhis) after sepsis orchestrates an inflammatory response leading to multiple organ dysfunction. Trehalose preserves cell viability during exposure to a range of environmental stresses. We investigated whether trehalose may inhibit endotoxin-induced activation of the inflammatory phenotype in MPhis. Rat peritoneal MPhis were stimulated with 50 microg/mL of Salmonella enteritidis lipopolysaccharide (LPS). Stimulated MPhis were coincubated with trehalose (25, 50, and 100 mmol), sucrose (100 mmol), or RPMI alone. Macrophages cultures were used for Western blot analysis of extracellular-regulated kinase, c-jun-N terminal kinase, and inducible nitric oxide synthase; interleukin (IL) 1beta, IL-6, and tumor necrosis factor alpha (TNF-alpha) gene expression by real-time reverse transcriptase-polymerase chain reaction, and supernatants for measuring the release of inflammatory cytokines and nitrite content. In vitro trehalose significantly blunted LPS-induced extracellular-regulated kinase (LPS = 21 +/- 6 integrated intensity; LPS + trehalose 100 mmol = 2 +/- 0.3 integrated intensity), c-jun-N terminal kinase (LPS = 15 +/- 5 integrated intensity; LPS + trehalose 100 mmol = 3.5 +/- 0.9 integrated intensity), and inducible nitric oxide synthase activation (LPS = 12 +/- 3 integrated intensity; LPS + trehalose 100 mmol = 1 +/- 0.09 integrated intensity), blunted IL-1beta (LPS = 5 +/- 1.9 n-folds/beta-actin; LPS + trehalose 100 mmol = 1.5 +/- 0.8 n-folds/beta-actin), IL-6 (LPS = 4 +/- 1.5 n-folds/beta-actin; LPS + trehalose 100 mmol = 1.4 +/- 0.5 n-folds/beta-actin), and TNF-alpha (LPS = 4.2 +/- 1.6 n-folds/beta-actin; LPS + trehalose 100 mmol = 1.1 +/- 0.7 n-folds/beta-actin) gene expression, and markedly reduced the release of inflammatory cytokines and nitrite content. Furthermore, in vivo trehalose prevented mortality in rats challenged with a lethal dose (20 mg/kg; LD90) of LPS (80% survival rate and 70% survival rate 24 and 72 h after LPS injection, respectively) and reduced serum TNF-alpha. Sucrose did not modified inflammatory phenotype in vitro nor in vivo protected against endotoxin-induced mortality. Our study suggests that trehalose inhibits proinflammatory phenotype activation in MPhis and prevents endotoxin-induced mortality.
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PMID:The disaccharide trehalose inhibits proinflammatory phenotype activation in macrophages and prevents mortality in experimental septic shock. 1717 86

In the present study, we compared the levels of intravascular tissue factor (TF) present in cord versus adult whole blood (WB) prior and after lipopolysaccharide (LPS) stimulation. High levels of intravascular TF might help to explain the clinically observed efficient clotting of cord blood despite low levels of procoagulatory factors. Quantitative reverse transcription-polymerase chain reaction revealed same (basal) TF mRNA expression levels in both native cord and adult WB, and approximately same increase in TF mRNA expression owing to LPS incubation in both cord and adult WB (normalized to the housekeeping gene beta-actin). Flow-cytometric (fluorescence activated cell sorting) analysis revealed significantly higher surface TF antigen exposure on the neonatal monocyte membrane in native WB samples, and approximately same ability of neonatal and adult monocytes to express TF upon LPS-stimulation. Thrombelastography revealed significantly shorter clotting times of native cord versus adult WB (527+/-41 vs. 592+/-23 s, P<0.05). Moreover, shortening of clotting times owing to LPS-stimulation was significantly more pronounced in cord versus adult WB (29.65+/-3.35% vs. 12.03+/-6.23%, P<0.05). Because both quantitative reverse transcription-polymerase chain reaction and fluorescence activated cell sorting analysis revealed same capability of both neonatal and adult monocytes to express TF upon LPS-stimulation, this efficient shortening effect in cord WB might be explained by the constitutively high number of monocytes present in neonates. We suggest that the high levels of intravascular TF present in neonates (prior and after LPS-stimulation) might help to explain the clinically observed efficient clotting of cord blood despite low levels of procoagulatory factors.
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PMID:High availability of intravascular tissue factor in neonates. 1748

The tissue expressions of nine immune related genes in apparently healthy Pacific white shrimp Litopenaeus vannamei were analyzed by conventional RT-PCR, quantitative real time PCR (qPCR) and in situ hybridisation. The nine genes were beta-glucan binding protein-high density lipoprotein (BGBP-HDL), lipopolysaccharide-beta-glucan binding protein (LGBP), haemocyanin, prophenoloxidase (proPO), transglutaminase (TGase), crustins, penaeidins (PEN), cytosolic manganese superoxide dismutase (cMnSOD), and lysozyme. Transcripts of all nine genes were detected in all tissues with differential expression levels when examined by RT-PCR and qPCR. BGBP-HDL, LGBP and haemocyanin were mainly expressed in the hepatopancreas and their expressions levels were about 1/10-1/3 those of beta-actin. Their expressions in other tissues were relatively limited. ProPO, TGase, crustins, PEN-3, and lysozyme showed the highest levels of expression in haemocytes and the lowest in hepatopancreas. Their expression levels in the haemocytes were 3 (PEN-3) to 10(-2) (proPO) times those of beta-actin. In contrast to the other genes, cMnSOD showed higher expression levels in haemolymph related organ, stomach and muscle; and lower expression levels in haemocyte, migut, neural ganglion and hepatopancreas. When examined by in situ hybridisation, hepatopancreatic F cells were found to be the major cell type that produced transcripts of BGBP-HDL, LGBP and haemocyanin. On the other hand, circulatory haemocytes and haemocytes infiltrated in various tissues contributed to the expressions of proPO, TGase, crustins, PEN-3 and lysozyme. Both hepatopancreatic F cell and haemocyte generated cMnSOD transcripts. Using in situ hybridisation, the present study is the first to show the tissue distributions of BGBP-HDL, LGBP, haemocyanin, TGase, crustins and cMnSOD in healthy white shrimp. The present results provide a baseline data of physiological expressions for the genes that are important in immune activation and modulation in Pacific white shrimp and a guideline of tissue or organ sampling for effective gene expression analyses for future immunological studies.
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PMID:Tissue expressions of nine genes important to immune defence of the Pacific white shrimp Litopenaeus vannamei. 1796 9

Real-time PCR has become a powerful tool for the detection of inflammatory parameters, including cytokines. Reference or housekeeping genes are used for the normalization of real-time RT-PCR results. In order to obtain reliable results, the stability of these housekeeping genes needs to be determined. In this study the stability of five genes, including beta-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine phophoribosyl-transferase (HPRT), ubiquitin (UB) and glucose-6-phosphate dehydrogenase (G6PDH), was determined in a lipopolysaccharide inflammation model in chickens. beta-Actin appeared to be the most stable single gene in our model. Because the use of a single gene for normalization can lead to relatively large errors, the use of the geometric mean of multiple reference genes or normalization factor is preferred. The most stable combination for gene expression analysis in this lipopolysaccharide inflammation model in chickens is G6PDH and UB, since their correlation coefficients were 0.953 and 0.969, respectively (BestKeeper) and an M value of 0.34 and a low V(2/3) value of 0.155 (geNorm) were obtained. The use of HPRT and GAPDH should be avoided. The stable housekeeping genes, G6PDH and UB together, can be used to normalize the expression of pro-inflammatory cytokines in a lipopolysaccharide inflammation model in chickens.
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PMID:Identification and validation of housekeeping genes as internal control for gene expression in an intravenous LPS inflammation model in chickens. 1827 35

To accurately measure the relative expression of a target gene, mRNA expression data is routinely normalized with reference to an internal control gene. We examined the transcriptional stability of four internal control genes, beta-actin, glyceraldehyde-3 phosphate dehydrogenase (GAPDH), elongation factor1-alpha (EF1-alpha), and 18S ribosomal RNA (18S rRNA) while measuring the mRNA expression of a gene encoding a pattern recognition protein, lipopolysaccharide and glucan binding protein (LGBP) gene, in healthy and white spot syndrome virus (WSSV) infected shrimp (Penaeus stylirostris) before and after (4, 8, 16 and 32 h) challenge using real-time RT-PCR. Here, we describe a method to rank the internal control genes based on a linear regression analysis. This analysis enables us to analyze the multivariate data set, e.g. time course study samples with control and treatment groups. Using the linear regression analysis and the WSSV-challenged time course samples, GAPDH was found to be the most stable internal control gene followed by the genes EF1-alpha, 18S rRNA and beta-actin. Using the program geNorm, GAPDH was also found to be the most stable gene followed by the genes EF1-alpha, beta-actin and 18S rRNA. Using the program NormFinder, the ranking of the internal control genes were in the order of EF1-alpha>GAPDH>18S rRNA>beta-actin. The ability to compare the healthy and WSSV infected samples in parallel by the regression analysis makes this method a very useful approach while identifying the optimal reference gene for gene expression analysis.
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PMID:Validation of reference genes for quantitative measurement of immune gene expression in shrimp. 1929 25

A single-tube real-time reverse transcription-polymerase chain reaction (RT-PCR) method has been developed which makes it possible to conduct the entire procedure, from nucleic acid extraction to product detection, in a single PCR tube. In this study, we developed the method using an oligodeoxythymidine-immobilized PCR tube, which enables simple and rapid mRNA extraction and quantification of target genes in blood and other tissues. The beta-actin gene was analyzed from lysates of blood and various tissues using this method. The data showed a good correlation between the plotted threshold cycle values and log(10) of blood and tissue amounts without a reduction in PCR efficiency. Gene expression of interleukin-1beta in blood from lipopolysaccharide (LPS)-stimulated rats and of beta(3)-adrenoceptors in adipose tissue from SHRSP.Z-Lepr (fa)/IzmDmcr (obese SHRSP) rats was also analyzed using the single-tube method, as well as a general real-time RT-PCR method, using RNA purified with a silica membrane column. In both methods, the copy number ratio of interleukin-1beta to beta-actin in LPS-stimulated rats was higher than in control rats, and the ratio of beta(3)-adrenoceptors to beta-actin in obese SHRSP rats was lower than in lean littermates. These results indicate that the single-tube method can provide results equivalent to those from general real-time RTPCR methods in gene expression analysis.
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PMID:Gene expression assay in blood and various tissues using a single-tube real-time reverse transcription-polymerase chain reaction method using an oligodeoxythymidine-immobilized polymerase chain reaction tube. 1964 36

The signal regulatory protein-beta1 (SIRPbeta1) is a DAP12-associated transmembrane receptor expressed in a subset of hematopoietic cells. Recently, it was shown that peritoneal macrophages express SIRPbeta1, which positively regulated phagocytosis. Here, we found that SIRPbeta1 was up-regulated and acted as a phagocytic receptor on microglia in amyloid precursor protein J20 (APP/J20) transgenic mice and in Alzheimer's disease (AD) patients. Interferon (IFN)-gamma and IFN-beta stimulated gene transcription of SIRPbeta1 in cultured microglia. Activation of SIRPbeta1 on cultured microglia by cross-linking antibodies induced reorganization of the cytoskeleton protein beta-actin and suppressed lipopolysaccharide-induced gene transcription of tumor necrosis factor-alpha and nitric oxide synthase-2. Furthermore, activation of SIRPbeta1 increased phagocytosis of microsphere beads, neural debris, and fibrillary amyloid-beta (Abeta). Phagocytosis of neural cell debris and Abeta was impaired after lentiviral knockdown of SIRPbeta1 in primary microglial cells. Thus, SIRPbeta1 is a novel IFN-induced microglial receptor that supports clearance of neural debris and Abeta aggregates by stimulating phagocytosis.
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PMID:Signal regulatory protein-beta1: a microglial modulator of phagocytosis in Alzheimer's disease. 1989 26

p38 MAP kinase mediates a signal pathway that is involved in many physiological and pathological processes such as inflammation, cellular stress, apoptosis, cell cycle and growth, ischemia/re-perfusion, and myocardium hypertrophy. To determine the molecular and regulative mechanism of p38 signal pathway, we used in vitro binding methods to screen the proteins that interact with p38. Here we report two proteins from mouse macrophage RAW264.7 strain treated with lipopolysaccharide (LPS) or ultraviolet radiation (UV), binding directly to p38. One of them is beta-actin identified by peptide mass spectrum and ProFound program. Actin can inhibit the autophosphorylation of p38 and the phosphorylation of ATF by p38. It suggests that the binding of actin to p38 in vitro may represent a negative feedback to the kinase activity of p38, which leads to the regulation of p38 pathway and cellular function.
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PMID:The binding of actin to p38 MAPK and inhibiting its kinase activity in vitro. 2021 65

beta-Actin has been frequently used as an internal control (gene) or as a housekeeping gene to normalize the expression of the target gene(s) or mRNA levels between different samples. However, the beta-actin expression has been shown to be influenced by the sample type and experimental conditions. If beta-actin could be used as a reference gene for the half-smooth tongue sole Cynoglossus semilaevis remains ill-defined. Here we evaluate the tissue-specific beta-actin gene expression pattern in C. semilaevis when challenged with antigenic agents namely, lipopolysaccharide (LPS) or Vibrio anguillarum employing absolute quantitative real-time PCR. The real-time PCR was performed based on the standard curve generated from recombinant plasmids. No significant differences in beta-actin expression were found between treated and untreated tissue samples. We thus conclude that beta-actin could be used as a reliable internal reference gene for real-time PCR based quantitation of gene expression studies in various tissue samples of C. semilaevis challenged with LPS or pathogenic bacteria.
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PMID:beta-Actin is a useful internal control for tissue-specific gene expression studies using quantitative real-time PCR in the half-smooth tongue sole Cynoglossus semilaevis challenged with LPS or Vibrio anguillarum. 2022 7


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