Degenerative alterations in articular cartilage are involved in the pathogenesis of osteoarthritis. The present study aimed to evaluate the role of <em>complement</em> <em>component</em> <em>5a</em> (C<em>5a</em>) in osteoarthritic alterations in the articular cartilage and synovialis via a joint immobilization (IM) rat model. Rats were assigned to three groups: Control, IM and IM+anti‑C<em>5a</em> antibody (IM+anti‑C<em>5a</em>) groups. A terminal deoxynucleotidyl transferase dUTP nick end labeling assay and hematoxylin and eosin staining were used to evaluate the morphological alterations in the articular cartilage and synovialis. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis, immunohistochemical analysis and western blotting were used to evaluate C<em>5a</em> expression in the articular cartilage and synovialis. An ELISA was used to evaluate C<em>5a</em>‑induced alterations in interleukin (IL)‑1β, IL‑17A and tumor necrosis factor (TNF)‑α levels in the serum and joint fluid. The results demonstrated that knee joint immobilization induced destruction of knee joint synovial fluid and cartilage in the IM and IM+anti‑C<em>5a</em> antibody groups. Immobilization significantly increased the expression levels of C<em>5a</em> in serum and joint fluid in the IM group. Immunohistochemistry, western blotting and RT‑qPCR analysis illustrated markedly increased expression of C<em>5a</em> in the IM group. Immobilization markedly increased the IL‑1β, IL‑17A and TNF‑α expression levels in the serum and joint fluid in the IM group. Anti‑C<em>5a</em> was able to decrease immobilization‑induced alterations in morphology and cytokines compared with the IM group. The expression of C<em>5a</em> was increased in synoviocytes and joint cartilage in the IM model. Pro‑inflammatory cytokines, including TNF‑α and IL‑1β were released in the activated synoviocytes via the induction of C<em>5a</em>, suggesting that C<em>5a</em> serves an important role in joint inflammatory processes.

EP67 is a <em>complement</em> <em>component</em> <em>5a</em> (C<em>5a</em>)-derived peptide agonist of the C<em>5a</em> receptor (CD88) that selectively activates DCs over neutrophils. Systemic administration of EP67 covalently attached to peptides, proteins, or attenuated pathogens generates TH1-biased immunogen-specific humoral and cellular immune responses with little inflammation. Furthermore, intranasal administration of EP67 alone increases the proportion of activated APCs in the airways. As such, we hypothesized that EP67 can act as a mucosal adjuvant. Intranasal immunization with an EP67-conjugated CTL peptide vaccine against protective MCMV epitopes M84 and pp89 increased protection of naïve female BALB/c mice against primary respiratory infection with salivary gland-derived MCMV and generated higher proportions of epitope responsive and long-lived memory precursor effector cells (MPEC) in the lungs and spleen compared to an inactive, scrambled EP67-conjugated CTL peptide vaccine and vehicle alone. Thus, EP67 may be an effective adjuvant for mucosal vaccines and warrants further study.

Patients with a relapse of idiopathic nephrotic syndrome have significantly increased levels of serum <em>complement</em> <em>component</em> <em>5a</em> (C<em>5a</em>), and proteinuria has been noted in mice treated with C<em>5a</em> via changes in permeability of kidney endothelial cells (KECs) in established animal models. However, the apoptosis of KECs treated with high concentrations of C<em>5a</em> has also been observed. As mitochondrial damage is known to be important in cell apoptosis, the aim of this study was to examine the association between C<em>5a</em>-induced mouse KEC apoptosis and mitochondrial damage. Mouse KECs were isolated and treated with different concentrations of C<em>5a</em>. Cell viability assays showed that a high-concentration mouse recombinant protein C<em>5a</em> (rmC<em>5a</em>) treatment reduced mouse KEC growth. Cell cycle phase analysis, including apoptosis (sub-G1 phase) showed an increased percentage of the subG1 phase with a high-concentration rmC<em>5a</em> treatment. Cytochrome c and caspase 3/9 activities were significantly induced in the mouse KECs after a high-dose rmC<em>5a</em> (50 ng/mL) treatment, and this was rescued by pretreatment with the C<em>5a</em> receptor (C<em>5a</em>R) inhibitor (W-54011) and N-acetylcysteine (NAC). Reactive oxygen species (ROS) formation was detected in C<em>5a</em>-treated mouse KECs; however, W-54011 or NAC pretreatment inhibited high-dose rmC<em>5a</em>-induced ROS formation and also reduced cytochrome c release, apoptotic cell formation, and apoptotic DNA fragmentation. These factors determined the apoptosis of mouse KECs treated with high-dose C<em>5a</em> through C<em>5a</em>R and subsequently led to apoptosis via ROS regeneration and cytochrome c release. The results showed that high concentrations of C<em>5a</em> induced mouse KEC apoptosis via a C<em>5a</em>R/ROS/mitochondria-dependent pathway. These findings may shed light on the potential mechanism of glomerular sclerosis, a process in idiopathic nephrotic syndrome causing renal function impairment.

Extremely potent <em>complement</em> activating (anti-<em>complement</em>ary) polysaccharides from roots of Lithospermum euchromum were fractionated in a novel way and also assayed for mitogenic and enhancing activity of immune complex binding to macrophages in vitro. Anti-<em>complement</em>ary activity and enhancing activity of immune complex binding were observed in the different polysaccharide fractions, but no mitogenic activity was found. The acidic polysaccharide fraction (LR-2) which had the most potent anti-<em>complement</em>ary activity was highly heterogeneous, and on further fractionation gave active polysaccharides. Extremely potent anti-<em>complement</em>ary polysaccharides, LR-2IId-1a, LR-2IId-1b, LR-2IId-3a and LR-2IId-<em>5a</em> were characterized chemically. These consisted mainly of mannose, galactose, glucose and glucuronic acid, in addition to rhamnose, fucose and arabinose as minor <em>components</em>. Methylation analysis showed that the four polysaccharides contained mainly (1-->3)-linked galactose, (1-->3)-linked fucose and (1-->4)-linked glucose. (1-->3)-Linked mannose was also detected in LR-2IId-1a, 1b and <em>5a</em>, and (1-->4)-linked glucuronic acid in LR-2IId-1a and 1b as the major glycosidic linkages.

Allergen-specific immunotherapy is the only treatment that can alter the natural course of allergic disease. We performed long-term sublingual immunotherapy (SLIT) for patients with seasonal allergic rhinitis caused by Japanese cedar pollen (SAR-JCP), screened molecules as candidate biomarkers, and investigated serum IL-17A and complement components 3a (C3a) and C5a in order to evaluate whether these molecules show changes correlated to symptom scores. In this study, we found that the long-term SLIT reduced the serum levels of IL-17A and C3a and C5a. The levels of C3a in the patients significantly decreased from year 1 compared with those at the baseline, and their levels of IL-17A significantly decreased from year 2 compared with those at baseline. The levels of IL-17A, C3a, and C5a at year 4 of SLIT were significantly lower than not only those at baseline, but also those at year 1. A significant positive correlation was found between the symptom medication scores and the levels of IL-17A at year 4. The symptom medication scores in the group in which IL-17A levels decreased at year 4 were significantly lower than those in the group without such a decrease. The serum level of IL-17A might prove useful as a biological parameter to ascertain the effectiveness of SLIT for patients with SAR-JCP. It is necessary to produce new therapeutics for non-responders in whom serum IL-17A levels are still higher against long-term SLIT.

The role of the complement cascade in the pathophysiology of cerebral arteriovenous malformation (AVM) is largely undefined. Complement subcomponents, C3a and C5a, are potent anaphylatoxins and key mediators of immuno-inflammatory response. Complement activation may contribute to the pro-inflammatory state observed in AVM. Thus, we sought to determine the systemic levels of C3a and C5a and their response to treatments in patients with AVM. Blood samples of 18 patients undergoing treatment for unruptured AVM, and from 30 healthy control participants, were obtained at four times: (i) pre-treatment, (ii) 24-hours post-embolization, (iii) 24-hours post-resection, and at 1-month follow-up. Plasma concentrations of C3a and C5a were measured using enzyme-linked immunosorbent assay. The pre-treatment mean plasma C3a level was significantly higher in patients with AVM (1817±168 ng/mL) compared to controls (1126±151 ng/mL). The mean C3a level decreased 24-hours after embolization (1482±170 ng/mL) and remained at statistically similar levels 24-hours after resection (1511±149 ng/mL) and at 1-month follow-up (1535±133 ng/mL). Mean C3a levels at the three time points were higher than control levels.The baseline mean plasma C5a level was significantly elevated in patients with AVM (13.1±2.2 ng/mL) compared to controls (3.9±1.5 ng/mL).Mean C5a level decreasedpost-embolization (8.2±2.3 ng/mL) and remained at similar levels post-resection (8.5±3.0 ng/mL) and at 1-month follow-up (7.7±2.9 ng/mL). Mean C5a levels at the three time points were significantly higher than the control levels. We conclude that systemic C3a and C5a levels in patients with AVM are elevated at baseline, decrease significantly after embolization, and remain at the new baseline levels after surgery and 1-month follow-up.

The purpose of this study was to reveal possible consequences of long-bone fracture on cardiac tissue and to analyze the role of systemically elevated danger associated molecular patterns, <em>complement</em> anaphylatoxins and cytokines. Blood samples of mice, pigs, and humans after a fracture were analyzed by ELISAs for <em>complement</em> <em>component</em> <em>5a</em> (C<em>5a</em>), tumor necrosis factor (TNF) and extracellular histones. In vivo results were completed by in vitro experiments with human cardiomyocytes treated with TNF and extracellular histones. The influence of histones and human plasma after fracture on isolated human polymorphonuclear leukocytes (PMNs) was investigated. An elevation of TNF, C<em>5a</em> and extracellular histones after long bone fracture was measured. Moreover, the appearance of systemic troponin I levels was observed and structural changes in connexin 43 and desmin were detected. Further, the presence of TNF lead to elevation of reactive oxygen species, troponin I release and histone appearance in supernatant of human cardiomyocytes. Incubation of human PMNs with histones and plasma of patients after fracture lead to formation of neutrophil extracellular traps. Present results suggest that structural alterations in the heart might be consequences of the <em>complement</em> activation, the release of extracellular histones and the systemic TNF elevation in the context of a long bone fracture.

The phenomena of allosterism continues to advance the field of drug discovery, by illuminating gainful insights for many key processes, related to the structure-function relationships in proteins and enzymes, including the transmembrane G-protein coupled receptors (GPCRs), both in normal as well as in the disease states. However, allosterism is completely unexplored in the native protein ligands, especially when a small covalent change significantly modulates the pharmacology of the protein ligands toward the signaling axes of the GPCRs. One such example is the human C5a ((h)C5a), the potent cationic anaphylatoxin that engages C5aR and C5L2 to elicit numerous immunological and non-immunological responses in humans. From the recently available structure-function data, it is clear that unlike the mouse C5a ((m)C5a), the (h)C5a displays conformational heterogeneity. However, the molecular basis of such conformational heterogeneity, otherwise allosterism in (h)C5a and its precise contribution toward the overall C5aR signaling is not known. This study attempts to decipher the functional role of allosterism in (h)C5a, by exploring the inherent conformational dynamics in (m)C5a, (h)C5a and in its point mutants, including the proteolytic mutant des-Arg(74)-(h)C5a. Prima facie, the comparative molecular dynamics study, over total 500 ns, identifies Arg(74)-Tyr(23) and Arg(37)-Phe(51) "cation-π" pairs as the molecular "allosteric switches" on (h)C5a that potentially functions as a damper of C5aR signaling.

Tumor necrosis factor-α (TNF-α), caspase-8, and <em>complement</em> <em>component</em> <em>5a</em> receptor (C<em>5a</em>R) are known to play a crucial role in the myocardial ischemia/reperfusion (I/R) injury in cardiac transplantation. We hypothesized that the intracoronary infusion of TNF-α, caspase-8, and C<em>5a</em>R small interfering RNAs (siRNA) would protect cardiac allograft function and improve graft survival from I/R injury-induced organ failure. I/R injury of cardiac allograft was induced by syngeneic rat cardiac transplantation, in which the transplanted hearts were infused with saline or different amounts of siRNA cocktail solution targeting TNF-α, caspase-8, and C<em>5a</em>R via coronary arteries, and subsequently subjected to 18 h of preservation at 4 °C in histidine-tryptophan-ketoglutarate (HTK) solution. The effects of siRNA cocktail solution on prolonged cold I/R injury were determined by assessing graft survival, histopathological changes, myeloperoxidase (MPO) activity, and malondialdehyde (MDA) concentration. The perfused siRNA cocktail solution successfully knocked down the expression of TNF-α, caspase-8, and C<em>5a</em>R in vitro and in vivo. Approximately 91.7% of control hearts that underwent 18 h of cold ischemia ceased their function after transplantation; however, 87.5% of cardiac allografts from the highest-dose siRNA cocktail solution-pretreated hearts survived >14 days and exhibited minimal histological changes, with minimal cellular infiltration, interstitial edema, and inflammation and maximal reduced MPO activity and MDA concentration in the cardiac allograft. We demonstrated the feasibility and efficiency of infusion of TNF-α, caspase-8, and C<em>5a</em>R siRNA via the intracoronary route as a promising strategy for gene silencing against I/R injury in cardiac transplantation.

Keywords: Cardiac transplantation; Graft survival; Intracoronary infusion; Oxidative stress damage; Small interfering RNAs.

Psoriasis is one of the most common chronic inflammatory skin diseases, affecting ~2% of the population. The lack of characterization of the pathogenesis of psoriasis has hindered efficient clinical treatment of the disease. In our study, we observed that expression of <em>complement</em> <em>component</em> <em>5a</em> receptor 1(C<em>5a</em>R1) was significantly increased in skin lesions of both imiquimod (IMQ) and IL23-induced psoriatic mice and patients with psoriasis. C<em>5a</em>R1 deficiency or treatment with C<em>5a</em> receptor 1 antagonist (C<em>5a</em>R1a) in mice significantly attenuated psoriasis-like skin lesions and expression of inflammatory cytokines and chemokines. Moreover, C<em>5a</em>R1 deficiency significantly decreased IMQ-induced infiltration of plasmacytoid dendritic cells (pDCs), monocytes and neutrophils in psoriatic skin lesions and functions of pDCs, evidenced by the remarkable reduction in the IMQ-induced production of interferon-α (IFN-α) and tumor necrosis factor α (TNF-α), and FMS-like tyrosine kinase 3 ligand (FLT3L)-dependent pDCs differentiation. Accordingly, <i>in vitro</i> treatment with recombinant C<em>5a</em> accelerated pDCs migration and the differentiation of bone marrow cells into pDCs. Furthermore, biopsies of psoriatic patients showed a dramatic increase of C<em>5a</em>R1⁺ pDCs infiltration in psoriatic skin lesions, compared to healthy subjects. Our results provide direct evidence that C<em>5a</em>/C<em>5a</em>R1 signaling plays a critical role in the pathogenesis of psoriasis. Inhibition of C<em>5a</em>/C<em>5a</em>R1 pathway is expected to be beneficial in the treatment of patients with psoriasis.

Pancreatitis is the inflammation of the pancreas. However, little is known about the genes associated with pancreatitis severity. Our microarray analysis of pancreatic tissues from mild and severe acute pancreatitis mice models identified angiopoietin-like 4 (ANGPTL4) as one of the most significantly upregulated genes. Clinically, ANGPTL4 expression was also increased in the serum and pancreatic tissues of pancreatitis patients. The deficiency in ANGPTL4 in mice, either by gene deletion or neutralizing antibody, mitigated pancreatitis-associated pathological outcomes. Conversely, exogenous ANGPTL4 exacerbated pancreatic injury with elevated cytokine levels and apoptotic cell death. High ANGPTL4 enhanced macrophage activation and infiltration into the pancreas, which increased <em>complement</em> <em>component</em> <em>5a</em> (C<em>5a</em>) level through PI3K/AKT signaling. The activation of the C<em>5a</em> receptor led to hypercytokinemia that accelerated acinar cell damage and furthered pancreatitis. Indeed, C<em>5a</em> neutralizing antibody decreased inflammatory response in LPS-activated macrophages and alleviated pancreatitis severity. In agreement, there was a significant positive correlation between C<em>5a</em> and ANGPTL4 levels in pancreatitis patients. Taken together, our study suggests that targeting ANGPTL4 is a potential strategy for the treatment of pancreatitis.

<strong class="sub-title"> Keywords: </strong> ANGPTL4; C<em>5a</em>; acute pancreatitis; macrophage.

<b>Background:</b> Preterm birth is the most frequent cause of neonatal death, but its aetiology remains unclear. It has been suggested that the imbalance of immunological mechanisms responsible for maintaining pregnancy is contributing to preterm birth pathogenesis. We aimed to investigate global gene expression and the levels of several <em>complement</em> system <em>components</em> in umbilical cord blood samples from preterm neonates and compare them to term newborns. We sought to examine how differentially expressed genes could affect various immune-related pathways that are believed to be crucial factors in preterm birth. <b>Material and methods:</b> We enrolled 27 preterm infants (<37 weeks GA) and 52 term infants (>37 weeks GA), from which umbilical cord blood samples were collected. From these samples, peripheral blood mononuclear cells were isolated and subsequent RNA isolation was performed. We used Affymetrix Human Gene 2.1 ST Array Strip for microarray experiment and DAVID resources for bioinformatics analysis of the obtained data. Concentrations of C2, C3a, C5/C<em>5a</em>, C9, FactorD, Properdin were measured in umbilical cord blood plasma samples using multiplex fluorescent bead-based immunoassays using Luminex technology. <b>Results:</b> The levels of C3a and C5/<em>5a</em> were significantly elevated in preterm neonates compared to term babies, whereas C9 concentration was evidently increased in term infants. The expression of 250 genes was upregulated at least 2-fold and 3781 genes were downregulated at least 2-fold in preterm neonates in comparison with term infants. Functional annotation analysis revealed that in preterm infants in comparison to term babies there was a significant downregulation of genes encoding several Toll-like receptors, interleukins and genes involved in major signalling pathways (e.g. NF-κB, MAPK, TNF, Notch, JAK) and vital cellular processes (e.g. intracellular signal transduction, protein ubiquitination, protein transport, RNA splicing, DNA-templated transcription). <b>Conclusions:</b> Preterm birth results in immediate and long-term complications. Our results indicate that infants born prematurely show significant differences in <em>complement</em> <em>components</em> concentration and a downregulation of over 3,000 genes, involved mainly in various immune-related pathways, including innate immune response, phagocytosis and TLR function, when compared to full-term babies. Further studies on larger cohorts are needed to elucidate the role of immunity in prematurity.

Keywords: complement system proteins; gene expression; immunity; premature birth.

This paper reports the synthesis and the bioassay of 4-methoxy- and 4-hydroxyspiro[benzofuran-2(3H)-cyclohexane] partial analogues (5) of the <em>complement</em> inhibitory sesquiterpene fungal metabolite 6,7-diformyl-3',4',4a',5',6',7',8',8a'-octahydro-4,6',7'-trihydroxy-2',5',5',8a'-tetramethylspiro[1'(2'H)-naphthalene-2(3H)-benzofuran] (1a, K-76) and its silver oxide oxidized product (1b, K-76COOH). The described target compounds represent spirobenzofuran B/C/D-ring analogues lacking the A-ring <em>component</em> of the prototype structure. The target compounds were evaluated by the inhibition of total hemolytic <em>complement</em> activity in human serum. It was observed that the structurally simplified analogue 4-methoxyspiro[benzofuran-2(3H)-cyclohexane]-6-carboxylic acid (<em>5a</em>) exhibited an IC(50) = 0.53 mM similar to the IC(50) = 0.57 mM that was observed for the natural product derivative 1b. Exhibiting an IC(50) = 0.16 mM, the three-ringed partial structure 6-carboxy-7-formyl-4-methoxyspiro[benzofuran-2(3H)-cyclohexane] (5k)was found to be the most potent target compound. Like the natural product, 5k appears to inhibit primarily at the C5 activation step and inhibits both the classical and alternative human <em>complement</em> pathways. Several other analogues inhibited <em>complement</em> activation in vitro at concentrations similar to those required for inhibition by the natural product 1b.

The <em>complement</em>-activated product, <em>complement</em> <em>component</em> <em>5a</em> (C<em>5a</em>), is a potent inflammatory peptide with a broad spectrum of functions. In vivo and in vitro studies have demonstrated that C<em>5a</em> serves an important role in inflammation; however, the role of C<em>5a</em> in the pathogenesis of inflammatory bowel disease (IBD) is not known. The purpose of the current study was to investigate the role of C<em>5a</em> in IBD using an experimental mouse model of colitis. Colitis was induced in mice using 2,4,6-trinitrobenzene sulfonic acid (TNBS), and C<em>5a</em> aptamers were subsequently administered via intraperitoneal injection. Clinical symptoms of the disease, histopathological analysis of the colon and the level of inflammatory <em>components</em> were examined. The symptoms of colitis, including changes in behavior, weight loss, colon damage and an increase in inflammatory cytokines, were attenuated following the treatment of mice with TNBS-induced colitis with C<em>5a</em> aptamers. The aptamer-treated mice exhibited a marked attenuation of colitis when compared with untreated mice, as demonstrated by the phenotypic observations, histological examinations and inflammatory cytokine levels. Colitis is characterized by an imbalance between pro-inflammatory and anti-inflammatory mediators. The results of the current study suggest that C<em>5a</em> may serve a critical role in inflammation in IBD.

A quantitative transmigration system, permitting the harvest of transmigrated cells for further analysis, was used to study carp head kidney (HK) granulocyte migration in vitro. Pooled carp serum and leukotriene B4 (LTB-4), but not recombinant human C-X-C chemokine ligand 8 (rhCXCL8), recombinant human <em>complement</em> <em>component</em> <em>5a</em> (rhC<em>5a</em>) or N-formyl-methionyl-leucyl-phenylalanine (FMLP) induced strong migration (up to 70%) of carp HK granulocytes. The transmigrated cells were viable >>or=96%) and uniform (purity>>or=97%). After serum- as well as LTB-4-induced transmigration granulocytes produced the same amounts of reactive oxygen species (ROS) as non-migrated cells in HK cell suspension. Their morphology, staining characteristics and flow cytometric scatter characteristics, plus their ability to produce ROS characterised the transmigrated granulocytes as neutrophils. The quantitative transmigration system described here could also serve as an excellent tool for the selective attraction and isolation of highly purified carp neutrophils from HK cell suspensions.

UNASSIGNED

Neuropathic pain and inflammatory pain are two common types of pathological pain in human health problems. To date, normal painkillers are only partially effective in treating such pain, leading to a tremendous demand to develop new chemical entities to combat pain and inflammation. A promising pharmacological treatment is to control signal transduction via the inflammatory mediator-coupled receptor protein C5aR by finding antagonists to inhibit C5aR activation. Here, we report the first computational study on the identification of non-peptide natural compound inhibitors for C5aR by homology modeling and virtual screening. Our study revealed a novel natural compound inhibitor Acteoside with better docking scores than all four existing non-peptidic natural compounds. The MM-GBSA binding free energy calculations confirmed that Acteoside has a decrease of ~39 kcal/mol in the free energy of binding compared to the strongest binding reference compound. Main contributions to the higher affinity of Acteoside to C5aR are the exceptionally strong lipophilic interaction, enhanced electrostatics and hydrogen bond interactions. Detailed analysis on the physiochemical properties of Acteoside suggests further directions in lead optimization. Taken together, our study proposes that Acteoside is a potential lead molecule targeting the C5aR allosteric site and provides helpful information for further experimental studies.

UNASSIGNED

High strength oxide ceramic materials like alumina and zirconia are frequently used for artificial joints because of their biocompatibility and high wear resistance. Their suitability as materials for implants and biomedical devices with direct blood contact, such as cardiovascular implants or <em>components</em> for blood pumps and dialyzers, has not been confirmed to date. The objective of this study was to investigate whether oxide ceramics show sufficient hemocompatibility. Dense specimens were made out of alumina, zirconia, titanium oxide, and aluminum titanate. Polyvinylchloride and silicone were additionally tested as reference materials. Interactions of human blood with the surfaces were studied by investigating partial thromboplastin time (PTT), thrombin antithrombin III complex (TAT), free plasma hemoglobin concentration, complete blood count, <em>complement</em> factor <em>5a</em>, and protein adsorption. The results from the PTT and TAT tests clearly indicated higher blood activation by the ceramic materials when compared to the two polymer materials. However, alumina and zirconia showed lower C<em>5a</em> concentrations and less protein adsorption than the reference materials. Our results revealed that oxide ceramic materials alone cannot be used for implants in direct blood contact without modification of the ceramic surface, for example, by made-to-measure inert nanocoatings.

Fallopia sachalinensis, regarded as an invasive plant in Europe and designated for disposal, is traditionally used in Japan and China as herbal medicine. Attempted for valorization of the leaves, this paper reports on two protein-free polysaccharide fractions, a neutral (FS-<em>5A</em>) and an acidic (FS-5B) one, obtained via alkali extraction and consecutive purification. Both fractions were characterized by chemical, molecular, structural and bioactive properties. FTIR and 1D/2D NMR analyses revealed that FS-<em>5A</em> consisted of a fucogalactoxyloglucan, whereas, glucuronoxylan was the major hemicellulose in FS-5B accompanied with low proportions of fucosylated xyloglucan and pectic RG-I. Both hemicellulose fractions exhibited significant immunostimulating activity in the <em>complement</em>-fixation test and the latter had noticeable DPPH radical-scavenging ability. The results completed information about neutral and acidic bioactive polysaccharide <em>components</em> present in the leaves of F. sachalinensis.

Hydrophobic uremic toxins accumulate in patients with chronic kidney disease, contributing to a highly increased cardiovascular risk. The clearance of these uremic toxins using current hemodialysis techniques is limited due to their hydrophobicity and their high binding affinity to plasma proteins. Adsorber techniques may be an appropriate alternative to increase hydrophobic uremic toxin removal. We developed an extracorporeal, whole-blood bifunctional adsorber particle consisting of a porous, activated charcoal core with a hydrophilic polyvinylpyrrolidone surface coating. The adsorption capacity was quantified using analytical chromatography after perfusion of the particles with an albumin solution or blood, each containing mixtures of hydrophobic uremic toxins. A time-dependent increase in hydrophobic uremic toxin adsorption was depicted and all toxins showed a high binding affinity to the adsorber particles. Further, the particle showed a sufficient hemocompatibility without significant effects on <em>complement</em> <em>component</em> <em>5a</em>, thrombin-antithrombin III complex, or thrombocyte concentration in blood in vitro, although leukocyte counts were slightly reduced. In conclusion, the bifunctional adsorber particle with cross-linked polyvinylpyrrolidone coating showed a high adsorption capacity without adverse effects on hemocompatibility in vitro. Thus, it may be an interesting candidate for further in vivo studies with the aim to increase the efficiency of conventional dialysis techniques.

The aim of the study was to evaluate the significance of metalloproteinase 3 (MMP-3), chemokine CXC ligand 13 (CXCL-13) and <em>complement</em> <em>component</em> <em>5a</em> (C<em>5a</em>) in different stages of ANCA associated vasculitis (AAV). 89 adults were included into the study. 28 patients with active AAV (Birmingham Vasculitis Activity Score, BVAS > 3) formed the Active Group. 24 individuals who were in remission after 6 months of induction therapy formed the Short R Group, while 34 patients with longitudinal remission formed the Long R Group. 28 patients without autoimmune diseases similar in terms of age, gender and stage of kidney disease formed the Control Group. Receiver operating characteristic curve analysis (ROC) was used to evaluate MMP-3, CXCL-13 and C<em>5a</em> as markers of the different phases of vasculitis. In ROC analysis, MMP-3, CXCL-13 and C<em>5a</em> presented a good ability in distinguishing active vasculitis (Active Group) from the Control Group (AUC > 0.8), whereas only CXCL-13 displayed potential ability in distinguishing active vasculitis (Active Group) from long term remission (Long R Group, AUC = 0.683). MMP-3 significantly and positively correlated with serum creatinine concentration (r = 0.51, p = 0.011; r = 0.44, p = 0.009; r = -0.66, p < 0.001) and negatively with eGFR (r = -0.5, p = 0.012; r = -0.35, p = 0.039; r = -0.63, p < 0.001) in the Short R, Long R and Control Groups. MMP-3, CXCL-13, C<em>5a</em> can be potential markers in differentiating an active phase of vasculitis from other pathologies. However they can be treated as <em>complement</em>ary to the well-known markers. CXCL-13 seems to be a potential marker in distinguishing active vasculitis from long term remission. MMP-3 level can be related to kidney function expressed by eGFR, therefore its elevation should be interpreted with caution in patients with kidney failure.

Lodging is one of the main factors in reducing the yield and grain quality of winter and spring wheat varieties. The resistance of wheat cultivars to lodging largely depends on environmental factors, biological and morphological features of the stem and root systems. Selection of the varieties for resistance to lodging is relevant in many countries of the world and has a number of achievements. Plant height is one of the most important morphological characters associated with lodging resistance. Breeding of the varieties carrying the dwarfing genes (Rht) is the main direction to reduce the risk of lodging. The Rht-B1b, Rht-D1b, Rht8 and Rht11 genes are widely used throughout the world due to their significant influence on agronomically valuable traits, including lodging. It turned out to be important to study the anatomical and morphological features and chemical composition of stem tissues, which <em>complement</em> the assessment of resistance to lodging and allow the varietal material to be more fully characterized. The thickness of stem internodes and their anatomical structure play an important role in the stem strength. The diameter of the stem, its thickness and weight, a large number of vascular bundles and a wide ring of mechanical tissues correlate with resistance to lodging. The content of lignin, silicon and cellulose are important structural <em>components</em> and provide the stem strength of wheat plants. Molecular genetic analysis and mapping of genes and quantitative trait loci are of great importance in identifying the genetic basis of the relationship between the anatomical and morphophysiological characters of the stem and root system and lodging. Genetic factors reflecting correlations between the lodging and the thickness of the stem wall, the number of vascular bundles and other characters were mapped to chromosomes 1A, 1B, 2A, 2D, 3A, 4B, 4D, <em>5A</em>, 5D, 6D and 7D. It has been found that loci with high phenotypic effects on lodging tolerance are colocalized with loci responsible for plant height, stem diameter and stem strength. To increase resistance to lodging, it is necessary to develop a set of agrotechnical methods that reduce the influence of soil and climatic factors and create wheat varieties tolerant to lodging.

Полегание является одной из основных проблем снижения урожайности и качества зерна озимой и яровой пшеницы. Устойчивость этой культуры к полеганию в значительной степени зависит от факторов внешней среды, биологических и морфологических особенностей стебля и корневой системы. Селекция сортов на устойчивость к полеганию актуальна во многих странах мира, и в данном направлении получен ряд достижений. Высота растений – важный морфологический признак, связанный с устойчивостью к полеганию. Основным направлением для снижения риска возникновения полегания стало выведение сортов, несущих гены короткостебельности (Rht). Гены Rht-B1b, Rht-D1b, Rht8, Rht11 получили широкое распространение во всем мире среди сортов мягкой пшеницы благодаря значительному влиянию на хозяйственно ценные признаки, включая полегание. Немаловажным оказалось изучение анатомо-морфологических особенностей и химического состава тканей стебля, которые дополняют оценку устойчивости к полеганию и позволяют более полно характеризовать изучаемый сортовой материал. Особенно большую роль в прочности стебля многие исследователи отводят толщине стенок междоузлий и их анатомическому строению. Диаметр соломины, ее толстостенность и вес, большое количество сосудистых пучков и широкое кольцо механических тканей коррелируют с устойчивостью к полеганию. Важными структурными компонентами, обеспечивающими прочность стебля у пшеницы, являются содержание лигнина, кремния и целлюлозы. Большое значение в выявлении генетической основы взаимоотношений между анатомическими и морфофизиологическими признаками стебля и корневой системы и полеганием имеют молекулярно-генетический анализ и картирование генов и локусов количественных признаков. Генетические факторы, отражающие корреляции между полеганием и толщиной стенки стебля, числом проводящих пучков и другими параметрами, были картированы в хромосомах 1А, 1B, 2A, 2D, 3A, 4B, 4D, <em>5A</em>, 5D, 6D и 7D. Установлено, что локусы с высоким фенотипическим эффектом в отношении толерантности к полеганию колокализуются с локусами, ответственными за высоту растения, диаметр и прочность стебля. Для повышения устойчивости к полеганию необходимы разработка комплекса агротехнических методов, снижающих влияние почвенноклиматических факторов, и создание толерантных к полеганию сортов.

Keywords: Rht genes; anatomical and morphological characters; lignin; lodging; stem; wheat.

<strong class="sub-title"> Background: </strong> <em>Complement</em> <em>component</em> <em>5a</em> (C<em>5a</em>) is a highly potent anaphylatoxin with a variety of pro-inflammatory effects. C<em>5a</em> contributes to progression of atherosclerosis and inhibition of the receptor (C<em>5a</em>R) might offer a therapeutic strategy in this regard. Single nucleotide polymorphisms (SNPs) of the C5 gene may modify protein expression levels and therefore function of C<em>5a</em> and C<em>5a</em>R. This study aimed to examine associations between clinically relevant C<em>5a</em> SNPs and the prognosis of patients with symptomatic coronary artery disease (CAD). Furthermore, we sought to investigate the influence of C5 SNPs on C<em>5a</em>R platelet surface expression and circulating C<em>5a</em> levels.

<strong class="sub-title"> Methods: </strong> C5 variants (rs25681, rs17611, rs17216529, rs12237774, rs41258306, and rs10985126) were analyzed in a consecutive cohort of 833 patients suffering from symptomatic coronary artery disease (CAD). Circulating C<em>5a</em> levels were determined in 116 patients whereas C<em>5a</em>R platelet surface expression was measured in 473 CAD patients. Endpoints included all-cause mortality, myocardial infarction (MI), and ischemic stroke (IS). Homozygous carriers (HC) of the minor allele (rs10985126) showed significantly higher all-cause mortality than major allele carriers. While we could not find significant associations between rs10985126 allele frequency and C<em>5a</em>R platelet surfazl ce expression, significantly elevated levels of circulating C<em>5a</em> were found in HC of the minor allele of the respective genotype. rs17216529 allele frequency correlated with the composite combined endpoint and bleeding events. However, since the number of HC of the minor allele of this genotype was low, we cannot draw a robust conclusion about the observed associations.

Conclusion: In this study, we provide evidence for the prognostic relevance of rs10985126 in CAD patients. C5 rs10985126 may serve as a prognostic biomarker for risk stratification in high-risk CAD patients and consequently promote tailored therapies.

Keywords: SNPs; complement C5; coronary artery disease; prognostic factors.

Avacopan (TAVNEOS™) is a <em>complement</em> <em>5a</em> receptor (C<em>5a</em>R) antagonist developed by ChemoCentryx for the treatment of autoimmune diseases including anti-neutrophil cytoplasmic autoantibody (ANCA)-associated vasculitis. The therapeutic effects of avacopan are attributed to the inhibition of C<em>5a</em>R activity on neutrophils, however, the exact mechanism of therapeutic efficacy in patients with ANCA-associated vasculitis has not been established. In September 2021, avacopan received its first approval in Japan for the treatment of microscopic polyangiitis (MPA) and granulomatosis with polyangiitis (GPA), the two most common forms of ANCA-associated vasculitis, where it is being commercialized by Kissei Pharmaceutical through a partnership with Vifor Pharma. In October 2021, avacopan was approved in the USA as an adjunctive treatment in adults for severe active ANCA-associated vasculitis (specifically MPA and GPA) in combination with standard therapy including glucocorticoids (avacopan does not eliminate glucocorticoid use). Avacopan has received a positive opinion in the EU, and is also undergoing regulatory review in Switzerland and Canada. Avacopan is being investigated for the treatment of <em>complement</em> <em>component</em> 3 glomerulopathy, hidradenitis suppurativa, lupus nephritis and IgA nephropathy. This article summarizes the milestones in the development of avacopan leading to these first approvals in Japan and the USA.

The <em>complement</em> system (CS) plays a pivotal role in Coronavirus disease 2019 (COVID-19) pathophysiology. The objective of this study was to provide a comparative, prospective data analysis of CS <em>components</em> in an all-comers cohort and COVID-19 patients. Patients with suspected COVID-19 infection admitted to the Emergency department were grouped for definite diagnosis of COVID-19 and no COVID-19 accordingly. Clinical presentation, routine laboratory and von Willebrand factor (vWF) antigen as well as CS <em>components</em> 3, 4 and activated 5 (C<em>5a</em>) were assessed. Also, total <em>complement</em> activity via the classical pathway (CH50) was determined. Levels of calprotectin in serum were measured using an automated quantitative lateral flow assay. We included 80 patients in this prospective trial. Of those 19 (23.7%) were tested positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Patients with COVID-19 had higher levels of CS <em>components</em> <em>5a</em> and 4 (54.79 [24.14-88.79] ng/ml vs. 35 [23.15-46.1] ng/ml; p = 0.0433 and 0.3772 [± 0.1056] g/L vs. 0.286 [0.2375-0.3748] g/L; p = 0.0168). COVID-19 patients had significantly higher levels of vWF antigen when compared to the control group (288.3 [± 80.26] % vs. 212 [151-320] %; p = 0.0469). There was a significant correlation between CS C3 and <em>5a</em> with vWF antigen (r_s = 0.5957 [p = 0.0131] and r_s = 0.5015 [p = 0.042]) in COVID-19 patients. There was no difference in calprotectin plasma levels (4.786 [± 2.397] µg/ml vs. 4.233 [± 2.142] µg/ml; p = 0.4175) between both groups. This prospective data from a single centre all-comers cohort accentuates altered levels of CS <em>components</em> as a distinct feature of COVID-19 disease. Deregulation of CS <em>component</em> 3 and C<em>5a</em> are associated with increased vWF antigen possibly linking vascular damage to alternative CS activation in COVID-19.

Keywords: COVID-19; Calprotectin; Coagulopathy; Complement components; Von Willebrand factor.