CLINICAL SIGNIFICANCE OF CALPROTECTIN IN RHEUMATIC DISEASES

Calprotectin (CP) belongs to the S100 leukocyte protein family, consists of two calcium-binding protein molecules, S100A8 and S100A9 (MRP8/14, calgranulin A/B). MRP8/14 is the main intracellular protein of neutrophil granulocytes and monocytes, which exerts a proinflammatory effect on phagocytes, endothelial cells in vitro and favors the development of the inflammatory process in vivo; it is an important mediator of many regulatory functions, such as chemotaxis; activation of neutrophil degranulation and phagocytosis; inhibition of immunoglobulin synthesis, cell proliferation and differentiation. CP is currently considered as a potential acute-phase marker in many inflammatory and autoimmune diseases. In rheumatoid arthritis (RA), CP maintains chronic inflammation, by activating the endothelium and increasing the migration of neutrophils into the inflamed synovial fluid. A number of studies have demonstrated the role of CP in monitoring RA activity, detecting subclinical inflammation, and predicting disease exacerbations. CP is of great importance in monitoring the efficiency of RA therapy with disease-modifying antirheumatic drugs and biological agents. The value of CP in spondyloarthritis (SpA) is ambiguous: on the one hand, this marker is highly expressed in the synovial tissue of patients with SpA and correlates with the level of acute-phase reactants; on the other hand, its correlation with disease activity indices is very contradictory and requires further clarification. By and large, CP is a promising laboratory marker of great clinical significance.

Download Full-text

Extract of the plant Cotinus coggygria Scop. attenuates pyrogallol-induced hepatic oxidative stress in Wistar rats

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y11-043 ◽

2011 ◽

Vol 89 (6) ◽

pp. 401-411 ◽

Cited By ~ 8

Author(s):

Sanja Matić ◽

Snežana Stanić ◽

Desanka Bogojević ◽

Melita Vidaković ◽

Nevena Grdović ◽

...

Keyword(s):

Oxidative Stress ◽

Acute Phase ◽

Wistar Rats ◽

Thiobarbituric Acid ◽

Hepatoprotective Activity ◽

Acute Phase Reactants ◽

Markers Of Oxidative Stress ◽

Cotinus Coggygria

To examine the protective potential of the Cotinus coggygria Scop. methanol extract, Wistar rats were treated with the hepatotoxic compound pyrogallol, which possesses a potent ability to generate free radicals and induce oxidative stress. The ability of the extract to counteract the oxidative stress was examined in rats that were injected with the extract intraperitoneally (500 mg·(kg body weight)–1) either 2 or 12 h before the pyrogallol treatment. The extract possesses a reducing activity in vitro and an ability to chelate the ferrous ion both in vivo and in vitro. Application of the extract prior to pyrogallol treatment led to a decrease in the levels of thiobarbituric acid-reactive substances, aspartate aminotransferase, and alanine aminotransferase, increased activities of antioxidant enzymes and attenuation of DNA damage, as well as increased Akt activity and inhibition of NF-κB protein expression. Treatment with the extract 12 h prior to pyrogallol administration was more effective in suppressing pyrogallol-induced oxidative damage than the 2 h pretreatment. Extract administration promoted an increase in acute phase reactants haptoglobin and α2-macroglobulin that was short of a full-fledged acute phase response. Administration of the extract considerably improved the markers of oxidative stress, thus revealing a potential hepatoprotective activity. Our results suggest that Akt activation, NF-κB inhibition, and induction of the acute phase play important roles in mediating hepatic protection by the extract. The greater effectiveness of the 12 h pretreatment with extract points to the important role that preconditioning assumes in improving resistance to subsequent exposure to oxidative stress.

Download Full-text

Regulation of Spi 2.1 and 2.2 gene expression after turpentine inflammation: discordant responses to IL-6

AJP Cell Physiology ◽

10.1152/ajpcell.1999.276.6.c1374 ◽

1999 ◽

Vol 276 (6) ◽

pp. C1374-C1382 ◽

Cited By ~ 29

Author(s):

Susan A. Berry ◽

Pearl L. Bergad ◽

Allison M. Stolz ◽

Howard C. Towle ◽

Sarah Jane Schwarzenberg

Keyword(s):

Gene Expression ◽

Acute Phase ◽

Gene Promoter ◽

Rat Hepatocytes ◽

Acute Phase Reactants ◽

Primary Rat Hepatocytes ◽

Fold Induction ◽

Vitro Model

The rat serine protease inhibitor (Spi) 2 gene family includes both positive (Spi 2.2) and negative (Spi 2.1) acute phase reactants, facilitating modeling of regulation of hepatic acute phase response (APR). To examine the role of signal transducer and activation of transcription (STAT) proteins in the divergent regulation of these model genes after induction of APR, we evaluated the proximal promoters of the genes, focusing on STAT binding sites contained in these promoter elements. Induction of APR by turpentine injection includes activation of a STAT3 complex that can bind to a γ-activated sequence (GAS) in the Spi 2.2 gene promoter, although the Spi 2.2 GAS site can bind STAT1 or STAT5 as well. To create an in vitro model of APR, primary hepatocytes were treated with combinations of cytokines and hormones to mimic the hormonal milieu of the whole animal after APR induction. Incubation of primary rat hepatocytes with interleukin (IL)-6, a critical APR cytokine, leads to activation of STAT3 and a 28-fold induction of a chloramphenicol acetyltransferase reporter construct containing the −319 to +85 region of the Spi 2.2 promoter. This suggests the turpentine-induced increase of Spi 2.2 is mediated primarily by IL-6. In contrast, although turpentine treatment reduces Spi 2.1 mRNA in vivo and IL-6 does not increase Spi 2.1 mRNA in primary rat hepatocytes, treatment of hepatocytes with IL-6 results in a 5.4-fold induction of Spi 2.1 promoter activity mediated through the paired GAS elements in this promoter. Differential regulation of Spi 2.1 and 2.2 genes is due in part to differences in the promoters of these genes at the GAS sites. IL-6 alone fails to reproduce the pattern of rat Spi 2 gene expression that results from turpentine-induced inflammation.

Download Full-text

Exosomal S100A4 derived from highly metastatic hepatocellular carcinoma cells promotes metastasis by activating STAT3

Signal Transduction and Targeted Therapy ◽

10.1038/s41392-021-00579-3 ◽

2021 ◽

Vol 6 (1) ◽

Author(s):

Haoting Sun ◽

Chaoqun Wang ◽

Beiyuan Hu ◽

Xiaomei Gao ◽

Tiantian Zou ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cancer Progression ◽

Calcium Binding ◽

Tumor Metastasis ◽

Metastatic Potential ◽

Functional Roles ◽

Stat3 Phosphorylation ◽

Hcc Cells

AbstractIntercellular cross-talk plays important roles in cancer progression and metastasis. Yet how these cancer cells interact with each other is still largely unknown. Exosomes released by tumor cells have been proved to be effective cell-to-cell signal mediators. We explored the functional roles of exosomes in metastasis and the potential prognostic values for hepatocellular carcinoma (HCC). Exosomes were extracted from HCC cells of different metastatic potentials. The metastatic effects of exosomes derived from highly metastatic HCC cells (HMH) were evaluated both in vitro and in vivo. Exosomal proteins were identified with iTRAQ mass spectrum and verified in cell lines, xenograft tumor samples, and functional analyses. Exosomes released by HMH significantly enhanced the in vitro invasion and in vivo metastasis of low metastatic HCC cells (LMH). S100 calcium-binding protein A4 (S100A4) was identified as a functional factor in exosomes derived from HMH. S100A4rich exosomes significantly promoted tumor metastasis both in vitro and in vivo compared with S100A4low exosomes or controls. Moreover, exosomal S100A4 could induce expression of osteopontin (OPN), along with other tumor metastasis/stemness-related genes. Exosomal S100A4 activated OPN transcription via STAT3 phosphorylation. HCC patients with high exosomal S100A4 in plasma also had a poorer prognosis. In conclusion, exosomes from HMH could promote the metastatic potential of LMH, and exosomal S100A4 is a key enhancer for HCC metastasis, activating STAT3 phosphorylation and up-regulating OPN expression. This suggested exosomal S100A4 to be a novel prognostic marker and therapeutic target for HCC metastasis.

Download Full-text

Cloning of mouse ptx3, a new member of the pentraxin gene family expressed at extrahepatic sites

Blood ◽

10.1182/blood.v87.5.1862.1862 ◽

1996 ◽

Vol 87 (5) ◽

pp. 1862-1872 ◽

Cited By ~ 147

Author(s):

M Introna ◽

VV Alles ◽

M Castellano ◽

G Picardi ◽

L De Gioia ◽

...

Keyword(s):

Gene Family ◽

Tertiary Structure ◽

Helical Structure ◽

Distinct Pattern ◽

Mononuclear Phagocytes ◽

Acute Phase Reactants ◽

Inflammatory Reactions ◽

Reactive Protein

Abstract Pentraxins, which include C reactive protein (CRP) and serum amyloid P component (SAP), are prototypic acute phase reactants that serve as indicators of inflammatory reactions. Here we report genomic and cDNA cloning of mouse ptx3 (mptx3), a member of the pentraxin gene family and characterize its extrahepatic expression in vitro and in vivo. mptx3 is organized into three exons on chromosome 3: the first (43 aa) and second exon (175 aa) code for the signal peptide and for a protein portion with no high similarity to known sequences the third (203 aa) for a domain related to classical pentraxins, which contains the “pentraxin family signature.” Analysis of the N terminal portion predicts a predominantly alpha helical structure, while the pentraxin domain of ptx3 is accommodated comfortably in the tertiary structure fold of SAP. Normal and transformed fibroblasts, undifferentiated and differentiated myoblasts, normal endothelial cells, and mononuclear phagocytes express mptx3 mRNA and release the protein in vitro on exposure to interleukin-1beta (IL-1beta) and tumor necrosis factor (TNF)alpha. mptx3 was induced by bacterial lipopolysaccharide in vivo in a variety of organs and, most strongly, in the vascular endothelium of skeletal muscle and heart. Thus, mptx3 shows a distinct pattern of in vivo expression indicative of a significant role in cardiovascular and inflammatory pathology.

Download Full-text

Neuroprotective Effect of Optimized Yinxieling Formula in 6-OHDA-Induced Chronic Model of Parkinson’s Disease through the Inflammation Pathway

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/2529641 ◽

2019 ◽

Vol 2019 ◽

pp. 1-11 ◽

Cited By ~ 1

Author(s):

Renrong Wei ◽

Cuiping Rong ◽

Qingfeng Xie ◽

Shouhai Wu ◽

Yuchao Feng ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Calcium Binding ◽

Dopaminergic Neurons ◽

Neuroprotective Effect ◽

Interleukin 1 ◽

Mrna Levels ◽

Cox 2

Parkinson’s disease (PD) is characterized by progressive degeneration of dopaminergic neurons in the substantia nigra (SN)-striatum circuit, which is associated with glial activation and consequent chronic neuroinflammation. Optimized Yinxieling Formula (OYF) is a Chinese medicine that exerts therapeutical effect and antiinflammation property on psoriasis. Our previous study has proven that pretreatment with OYF could regulate glia-mediated inflammation in an acute mouse model of PD induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Given that PD is a chronic degeneration disorder, this study applied another PD animal model induced by striatal injection of 6-hydroxydopamine (6-OHDA) to mimic the progressive damage of the SN-striatum dopamine system in rats. The OYF was administrated in the manner of pretreatment plus treatment. The effects of the OYF on motor behaviors were assessed with the apomorphine-induced rotation test and adjusting steps test. To confirm the effect of OYF on dopaminergic neurons and glia activation in this model, we analyzed the expression of tyrosine hydroxylase (TH) and glia markers, ionized calcium-binding adapter molecule 1 (Iba-1), and glial fibrillary acidic protein (GFAP) in the SN region of the rat PD model. Inflammation-associated factors, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), were further evaluated in this model and in interferon-γ- (INF-γ-) induced murine macrophages RAW264.7 cells. The results from the in vivo study showed that OYF reversed the motor behavioral dysfunction in 6-OHDA-induced PD rats, upregulated the TH expression, decreased the immunoreactivity of Iba-1 and GFAP, and downregulated the mRNA levels of TNF-α and COX-2. The OYF also trended to decrease the mRNA levels of IL-1β and iNOS in vivo. The results from the in vitro study showed that OYF significantly decreased the mRNA levels of TNF-α, IL-1β, IL-6, iNOS, and COX-2. Therefore, this study suggests that OYF exerts antiinflammatory effects, which might be related to the protection of dopaminergic neurons in 6-OHDA-induced chronic neurotoxicity.

Download Full-text

Effect of microbial isolates on microbial yield estimation in RUSITEC system

BSAP Occasional Publication ◽

10.1017/s0263967x0003295x ◽

1998 ◽

Vol 22 ◽

pp. 306-308

Author(s):

M. D. Carro ◽

E. L. Miller

Keyword(s):

Protein Synthesis ◽

Liquid Phase ◽

Solid Phase ◽

Liquid Fraction ◽

Microbial Protein ◽

Microbial Protein Synthesis ◽

Continuous Culture System ◽

The One

The estimation of rumen microbial protein synthesis is one of the main points in the nitrogen (N)-rationing systems for ruminants, as microbial protein provides proportionately 0.4 to 0.9 of amino acids entering the small intestine in ruminants receiving conventional diets (Russell et al., 1992). Methods of estimating microbial protein synthesis rely on marker techniques in which a particular microbial constituent is related to the microbial N content. Marker : N values have generally been established in mixed bacteria isolated from the liquid fraction of rumen digesta and it has been assumed that the same relationship holds in the total population leaving the rumen (Merry and McAllan, 1983). However, several studies have demonstrated differences in composition between solid-associated (SAB) and fluid-associated bacteria in vivo (Legay-Carmier and Bauchart, 1989) and in vitro (Molina Alcaide et al, 1996), as well in marker : N values (Pérez et al., 1996). This problem could be more pronounced in the in vitro semi-continuous culture system RUSITEC, in which there are three well defined components (a free liquid phase, a liquid phase associated with the solid phase and a solid phase), each one having associated microbial populations.The objective of this experiment was to investigate the effect of using different bacterial isolates (BI) on the estimation of microbial production of four different diets in RUSITEC (Czerkawski and Breckenridge, 1977), using (15NH4)2 SO4 as microbial marker, and to assess what effects any differences would have on the comparison of microbial protein synthesis between diets.This study was conducted in conjunction with an in vitro experiment described by Carro and Miller (1997). Two 14-day incubation trials were carried out with the rumen simulation technique RUSITEC (Czerkawski and Breckenridge, 1977). The general incubation procedure was the one described by Czerkawski and Breckenridge (1977) and more details about the procedures of this experiment are given elsewhere (Carro and Miller, 1997).

Download Full-text

Microtubule Actin Cross-Linking Factor (Macf)

The Journal of Cell Biology ◽

10.1083/jcb.147.6.1275 ◽

1999 ◽

Vol 147 (6) ◽

pp. 1275-1286 ◽

Cited By ~ 150

Author(s):

Conrad L. Leung ◽

Dongming Sun ◽

Min Zheng ◽

David R. Knowles ◽

Ronald K.H. Liem

Keyword(s):

Calcium Binding ◽

Coiled Coil ◽

Specific Protein ◽

Binding Domain ◽

Full Length ◽

Actin Binding ◽

Cross Linking ◽

Actin Binding Domain

We cloned and characterized a full-length cDNA of mouse actin cross-linking family 7 (mACF7) by sequential rapid amplification of cDNA ends–PCR. The completed mACF7 cDNA is 17 kb and codes for a 608-kD protein. The closest relative of mACF7 is the Drosophila protein Kakapo, which shares similar architecture with mACF7. mACF7 contains a putative actin-binding domain and a plakin-like domain that are highly homologous to dystonin (BPAG1-n) at its NH2 terminus. However, unlike dystonin, mACF7 does not contain a coiled–coil rod domain; instead, the rod domain of mACF7 is made up of 23 dystrophin-like spectrin repeats. At its COOH terminus, mACF7 contains two putative EF-hand calcium-binding motifs and a segment homologous to the growth arrest–specific protein, Gas2. In this paper, we demonstrate that the NH2-terminal actin-binding domain of mACF7 is functional both in vivo and in vitro. More importantly, we found that the COOH-terminal domain of mACF7 interacts with and stabilizes microtubules. In transfected cells full-length mACF7 can associate not only with actin but also with microtubules. Hence, we suggest a modified name: MACF (microtubule actin cross-linking factor). The properties of MACF are consistent with the observation that mutations in kakapo cause disorganization of microtubules in epidermal muscle attachment cells and some sensory neurons.

Download Full-text

Hierarchy of evidence in interpretation of clinical significance of drug interactions

Medicinski pregled ◽

10.2298/mpns1202045n ◽

2012 ◽

Vol 65 (1-2) ◽

pp. 45-49

Author(s):

Bozana Nikolic ◽

Miroslav Savic

Keyword(s):

Drug Metabolism ◽

Drug Interactions ◽

Clinical Significance ◽

Health Professionals ◽

Molecular Entity ◽

Population Pharmacokinetic ◽

Pharmacokinetic Studies ◽

Potential Interactions

Introduction. Since drug interactions may result in serious adverse effects or failure of therapy, it is of huge importance that health professionals base their decisions about drug prescription, dispensing and administration on reliable research evidence, taking into account the hierarchy of data sources for evaluation. Clinical Significance of Potential Interactions - Information Sources. The sources of data regarding drug interactions are numerous, beginning with various drug reference books. However, they are far from uniformity in the way of choosing and presenting putative clinically relevant interactions. Clinical Significance of Potential Interactions - Interpretation of Information. The difficulties in interpretation of drug interactions are illustrated through the analysis of a published example involving assessment made by two different groups of health professionals. Systematic Evaluation of Drug-Drug Interaction. The potential for interactions is mainly investigated before marketing a drug. Generally, the in vitro, followed by in vivo studies are to be performed. The major metabolic pathways involved in the metabolism of a new molecular entity, as well as the potential of induction of human enzymes involved in drug metabolism are to be examined. In the field of interaction research it is possible to make use of the population pharmacokinetic studies as well as of the pharmacodynamic assessment, and also the postregistration monitoring of the reported adverse reactions and other literature data. Conclusion. In vitro and in vivo drug metabolism and transport studies should be conducted to elucidate the mechanisms and potential for drug-drug interactions. The assessment of their clinical significance should be based on well-defined and validated exposure-response data.

Download Full-text

Study of yolk-sac endoderm organogenesis in the chick using a specific enzyme (cysteine lyase) as a marker of cell differentiation

Development ◽

10.1242/dev.29.1.159 ◽

1973 ◽

Vol 29 (1) ◽

pp. 159-174

Author(s):

Nelly Bennett

Keyword(s):

Cell Differentiation ◽

Blood Cells ◽

Primitive Streak ◽

Yolk Sac ◽

Specific Enzyme ◽

Cell Proliferation And Differentiation ◽

Lyase Activity ◽

Morphological Specialization

The detection of a specific enzyme (cysteine lyase) of the yolk-sac endoderm by a very sensitive method is employed to characterize cell differentiation during the early stages of endoderm organogenesis in the chick. The first cells to contain active cysteine lyase are found in the germ wall at the primitive streak stage. In vivo observations establish a relation between the morphological specialization and organization of endodermal cells, their loss of mitotic activity and the increase in cysteine lyase activity. They suggest an influence of the mesoderm on endoderm differentiation. In vitro experiments confirm the existence in the yolk-sac endoderm of an incompatibility between cell proliferation and differentiation, as well as the action of the mesoderm on both the structural organization of the endoblast and the appearance of cysteine lyase; this last action seems to be due mainly to blood cells; chicken and rabbit blood cells are equally active. The problems of the origin of the endoderm and of the interactions occurring during the organogenesis of the yolk-sac endoderm are discussed.

Download Full-text

Mechanics of the cellular microenvironment as perceived by cells in vivo

10.1101/2021.01.04.425259 ◽

2021 ◽

Author(s):

Alessandro Mongera ◽

Marie Pochitaloff ◽

Hannah J. Gustafson ◽

Georgina A. Stooke-Vaughan ◽

Payam Rowghanian ◽

...

Keyword(s):

Stress Relaxation ◽

Mechanical Parameters ◽

Cellular Microenvironment ◽

Stem Cell Maintenance ◽

Quantitative Studies ◽

Cell Proliferation And Differentiation ◽

3D Microenvironment ◽

Proliferation And Differentiation

Tissue morphogenesis and repair, as well as organ homeostasis, require cells to constantly monitor their 3D microenvironment and adapt their behaviors in response to local biochemical and mechanical cues1-6. In vitro studies have shown that substrate stiffness and stress relaxation are important mechanical parameters in the control of cell proliferation and differentiation, stem cell maintenance, cell migration 7-11, as well as tumor progression and metastasis12,13. Yet, the mechanical parameters of the microenvironment that cells perceive in vivo, within 3D tissues, remain unknown. In complex materials with strain- and time-dependent material properties, the perceived mechanical parameters depend both on the strain and timescales at which the material is mechanically probed14. Here, we quantify in vivo and in situ the mechanics of the cellular microenvironment that cells probe during vertebrate presomitic mesoderm (PSM) specification. By analyzing the magnitude and dynamics of endogenous, cell-generated strains, we show that individual cells preferentially probe the stiffness associated with deformations of the supracellular, foam-like tissue architecture. We reveal how stress relaxation leads to a perceived microenvironment stiffness that decreases over time, with cells probing the softest regime. While stress relaxation timescales are spatially uniform in the tissue, most mechanical parameters, including those probed by cells, vary along the anteroposterior axis, as mesodermal progenitors commit to different lineages. Understanding the mechanical parameters that cells probe in their native 3D environment is important for quantitative studies of mechanosensation in vivo2-4,6,15 and can help design scaffolds for tissue engineering applications16-18.

Download Full-text