Factor B Research Articles

Alternative complement pathway is activated in the brains of scrapie-infected rodents.

Activation of complement system in central nervous system (CNS) of the patients suffering from prion diseases or animal models infected with prion agents experimentally is reported repeatedly, but which pathways are involved in the complement system during prion infection is not well documented. Here, we evaluated the level of complement factor B (CFB), which is the key factor that triggers alterative pathway (AP) of complement in the brain tissues of scrapie-infected mice with various methodologies. We found that the levels of mRNA and protein of CFB significantly increased in the brain tissues of scrapie-infected mice. Morphologically, the increased CFB-specific signal overlapped with the elevated C3 signal in brain sections of scrapie-infected mice, meanwhile overlapped with damaged neurons and activated microglia, but not with the proliferative astrocytes. Additionally, the level of complement factor P (CFP), the key positive regulator of AP, also increased remarkably in the brain tissues of infected mice. The transcriptional levels of CD55 and CD46, two negative regulators of AP, decreased without significance in brain tissues of scrapie-infected mice at the terminal stage. However, the mRNA and protein levels of CFH, another negative regulator of AP, increased. Through the dynamic analyses of the expressions of CFB, CFP, and CFH in brain sections of 139A-infected mice, which were collected at different time-points during incubation period, illustrated time-dependent increase levels of each factor during the incubation period of scrapie infection. Taken together, our data here demonstrate that the AP of complement cascade is activated in the CNS microenvironment during prion infection.

Normal range of complement components during pregnancy: A prospective study.

ProblemThe complement system plays a key role in normal placentation, and delicate regulation of complement system activation is critical for successful pregnancy. Therefore, establishing a normal range of complement components during pregnancy is important for clinical evaluation and research.MethodsWe performed a prospective study to investigate the normal range of complement components in circulation during different stages of pregnancy. Plasma concentrations of complement factor B (CFB), C1q, complement factor H (CFH), C3, C3c, and C4 were measured using an immunoturbidimetric assay; mannan‐binding lectin (MBL), C3a, C5a, and soluble C5b‐9 (sC5b‐9) levels at different time points of pregnancy were determined by enzyme‐linked immunosorbent assay (ELISA).ResultsA total of 733 plasma samples were collected from 362 women with a normal pregnancy and 65 samples from non‐pregnant women. In the first trimester of pregnancy, the levels of CFB, CFH, MBL, C3c, C4, and C3a were 414.5 ± 85.9 mg/L (95% CI for mean: 402.4‐426.6 mg/L), 381.0 ± 89.0 mg/L (95% CI for mean: 368.5‐393.6 mg/L), 4274.5 ± 2752 ng/mL (95% CI for mean: 3881.1‐4656.4 ng/mL), 1346.9 ± 419.8 mg/L (95% CI for mean: 1287.7‐1406.0 mg/L), 357.4 ± 101.8 mg/L (95% CI for mean: 343.0‐371.7 mg/L), and 182.5 ± 150.0 ng/mL (95% CI for mean: 186.9‐229.1 ng/mL), respectively. The levels of C3 and C4 increased gradually throughout pregnancy. The levels of C1q, C5a, and sC5b‐9 in the first and second trimesters were nearly the same as those in non‐pregnant women.ConclusionThe results of this study show that pregnancy itself may influence the plasma levels of complement system components.

Cerebrospinal fluid complement system biomarkers in demyelinating disease

Background: Multiple sclerosis (MS) can be difficult to differentiate from other demyelinating diseases, notably neuromyelitis optica spectrum disorder (NMOSD). We previously showed that NMOSD is distinguished from MS by plasma complement biomarkers. Objective: Here, we measure cerebrospinal fluid (CSF) complement proteins in MS, NMOSD and clinically isolated syndrome (CIS), a neurological episode that may presage MS, to test whether these distinguish NMOSD from MS and CIS. Materials and methods: CSF (53 MS, 17 CIS, 11 NMOSD, 35 controls) was obtained; complement proteins (C4, C3, C5, C9, C1, C1q, Factor B (FB)), regulators (Factor I (FI), Factor H (FH), FH-Related Proteins 1, 2 and 5 (FHR125), C1 Inhibitor (C1INH), Properdin) and activation products (terminal complement complex (TCC), iC3b) were quantified by ELISA and results expressed relative to CSF total protein (μg/mg). Results: Compared to control CSF, (1) levels of C4, C1INH and Properdin were elevated in MS; (2) TCC, iC3b, FI and FHR125 were increased in CIS; and (3) all complement biomarkers except TCC, FHR125, Properdin and C5 were higher in NMOSD CSF. A statistical model comprising six analytes (C3, C9, FB, C1q, FI, Properdin) plus age/gender optimally differentiated MS from NMOSD.

Poor allograft outcome in Indian patients with post-transplant C3 glomerulopathy.

BackgroundComplement 3 glomerulopathy (C3G) results from dysfunction of the alternative complement pathway (ACP). No data are available on post-transplant C3G in South Asia.MethodsIn this study, renal allograft biopsies of C3G patients performed from 2012 to 2017 were analysed for ACP functional assay (APFA), serum complement levels, complement factor H (CFH), complement factor B (CFB) and autoantibodies to CFH and CFB. Limited genetic screening for CFH/CFHR5 genes was carried out. All study patients were also followed up.ResultsA total of 21 cases of C3G were included, of which 11 had native C3G disease (that is, recurrent C3G). Of these 11 recurrent cases, 7 presented with allograft dysfunction and 4 with proteinuria and renal dysfunction. Early post-transplant recurrence (<1 month) was noted in six patients, whereas recurrence in five patients occurred within 8–17 months of transplant. Biopsies showed mild focal mesangial expansion with or without endocapillary proliferation and thrombotic microangiopathy. Rejection was also noted in six patients. APFA/C3 levels were low in all cases. Serum CFH levels were low [dense deposit disease (DDD), 44%; C3 glomerulonephritis (C3GN), 25%], whereas CFB levels were normal. Autoantibodies to CFH, CFB and C3 nephritic factor were present in 11, 0 and 44% of DDD cases, respectively, and in 17, 17 and 33% of C3GN cases, respectively. Genetic analysis revealed only non-pathogenic CFH gene variants (93%). No novel mutation was found. At follow-up (140 months), stable graft was noted in 28% of cases, progressive renal failure in 19%, graft loss in 34%, and 19% of patients died.ConclusionPost-transplant C3G can present with graft dysfunction and/or proteinuria. Subtle histological findings demand careful interpretation of immunofluorescence results. Autoantibodies to complement pathway regulatory proteins are common, and no novel mutation has been found from limited genetic workup. Clinical outcome is poor.

Increased Complement Factor B and Bb Levels Are Associated with Mortality in Patients with Severe Aortic Stenosis

Inflammation is involved in initiation and progression of aortic stenosis (AS). However, the role of the complement system, a crucial component of innate immunity in AS, is unclear. We hypothesized that circulating levels of complement factor B (FB), an important component of the alternative pathway, are upregulated and could predict outcome in patients with severe symptomatic AS. Therefore, plasma levels of FB, Bb, and terminal complement complex were analyzed in three cohorts of patients with severe symptomatic AS and mild-to-moderate or severe asymptomatic AS (population 1, n = 123; population 2, n = 436; population 3, n = 61) and in healthy controls by enzyme immunoassays. Compared with controls, symptomatic AS patients had significantly elevated levels of FB (2.9- and 2.8-fold increase in population 1 and 2, respectively). FB levels in symptomatic and asymptomatic AS patients were comparable (population 2 and 3), and in asymptomatic patients FB correlated inversely with valve area. FB levels in population 1 and 2 correlated with terminal complement complex levels and measures of systemic inflammation (i.e., CRP), cardiac function (i.e., NT-proBNP), and cardiac necrosis (i.e., Troponin T). High FB levels were significantly associated with mortality also after adjusting for clinical and biochemical covariates (hazard ratio 1.37; p = 0.028, population 2). Plasma levels of the Bb fragment showed a similar pattern in relation to mortality. We concluded that elevated levels of FB and Bb are associated with adverse outcome in patients with symptomatic AS. Increased levels of FB in asymptomatic patients suggest the involvement of FB from the early phase of the disease.

The effect of complement factor B gene variation on age-related macular degeneration in Iranian patients

PurposeTo determine the possible association of rs4151667 (L9H) complement factor B (CFB) gene with age-related macular degeneration (AMD). The L9H is one of the functional variations of the CFB. CFB gene encodes the most important protein of the complement system. MethodsTwo hundred sixty-six patients with AMD and 194 unrelated age/sex-matched controls were genotyped for CFB gene (rs4151667) using the polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) method. All research subjects were selected from three regions of Iran (Tehran, Tabriz, and Gonabad). ResultsThe results showed a significant difference between the frequency of non-TT genotype in total patients and controls [odds ratio (OR) = 0.424, P = 0.038]. The analysis for each studied region showed that in patients originating from the Gonabad population, the frequency of TT and non-TT genotypes between patients and the control group were significantly different (OR = 2.894, P = 0.046 for TT genotype and OR = 0.346, P = 0.026 for non-TT genotype). In patients originating from Tabriz population, TT and non-TT genotypes and A allele revealed considerably different frequencies between the patient and control groups (OR = 3.043, P = 0.017; OR = 0.329, P = 0.013 and OR = 0.347, P = 0.048, respectively). Analysis of patients from Tehran also showed that there was a significant difference in the frequency of TT genotype between patients and controls (OR = 2.168, P = 0.04). ConclusionsThe results of the current study indicated a possible protective role for non-TT genotype in L9H variation CFB gene against AMD in a sample of the Iranian population. The region segregation results showed that TT genotype might be a risk factor for susceptibility to AMD.

Magnetized Carbon Nanotube Based Lateral Flow Immunoassay for Visual Detection of Complement Factor B.

The authors describe a magnetized carbon nanotube (MCNT) based lateral flow immunoassay (LFI) for visual detection of complement factor B (CFB) in blood. MCNT was prepared by decorating magnetic Fe3O4 nanoparticles on multi-walled CNT surface and used as a colored tag for LFI. Monoclonal antibody (mAb, Ab1) of CFB was covalently immobilized on the MCNT surface via diimide-activated conjugation between the carboxyl groups on the MCNT surface and amino groups of antibodies. Polyclonal antibody of CFB (Ab2) and the secondary antibody were used to prepare the lateral flow test strips. The assay involved: (1) the capture of CFB in blood with the mAb-functionalized MCNT; (2) magnetic separation of the formed CFB-mAb-MCNT and excess of mAb-MCNT from the blood with an external magnet; (3) lateral flow test to capture the CFB-mAb-MCNT complex on the test zone and the excess of mAb-MCNT on the control zone; (4) Recording the intensities of the produced the characteristic brown bands with a portable strip reader and quantitating the concentration of CFB. The proof-of-concept was demonstrated by testing CFB in the buffer, and the detection limit was 5 ng mL−1 under the optimized analytical parameters. CFB in 1 μL of human blood was detected successfully in 30 min with this LFI and the results had a high correlation with commercial ELISA kit. Thence, the MCNT-based LFI offers a rapid and low-cost tool for detecting CFB in human blood directly.

Isobaric tag for relative and absolute quantitation based quantitative proteomics reveals unique urinary protein profiles in patients with preeclampsia.

Preeclampsia (PE) is one of the most significant pregnancy‐related hypertensive disorders. Currently, there are no useful markers to predict the onset of the condition in pregnant women. To provide further insights into the pathogenesis of PE and identify biomarkers of the condition, we used isobaric tags for relative and absolute quantitation (iTRAQ) proteomics coupled with 2‐D LC‐MS/MS, to analyze urinary protein profiles from 7 PE patients and 7 normotensive pregnant women. A total of 294 proteins were abnormally expressed in PE patients. Of these, 233 were significantly down‐regulated and 61 proteins were significantly up‐regulated. Bioinformatics analysis using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) database, found that the most differentially expressed proteins (DEPs) were involved in coagulation and complement pathways, the renin‐angiotensin system and cell adhesion molecules (CAMs) pathways. We further validated three of the DEPs, including serotransferrin (TF) and complement factor B (CFB) by immunoblottingand serum paraoxonase/arylesterase 1 (PON1) by ELISA using 14 pairs of urine samples from PE patients and normal pregnant women. Taken together, our results provide the basis for further understanding the pathogenesis of PE and identifying predictive biomarkers.

Prognostic potential of the preoperative plasma complement factor B in resected pancreatic cancer: A pilot study.

For patients with pancreatic cancer, a preoperative assessment of prognosis is crucial to predict cancer recurrence and to prepare a postoperative adjuvant strategy and appropriate patient-counsel. We evaluated the prognostic predictive power of complement factor B (CFB) by comparing it to that of other known tumor markers in resected pancreatic cancer patients. From 2012 to 2013 period, we retrospectively reviewed the plasma CFB levels of 35 pancreatic cancer patients. The patients were divided into two groups according to serologic CFB values. Disease-free survival (DFS) and overall survival (OS) rates were analyzed. Based on the cut-off values of plasma CFB, 15 patients were placed in the low CFB group and the other 20 patients were placed in the high CFB group. There was a significant difference in DFS between the two groups (Low CFB vs. High CFB: 36.9 months vs. 13.9 months, p: 0.007). In the OS analysis, there was also a significant difference in the survival rates of the two groups (Low CFB vs. High CFB: 49.7 months vs. 29.0 months, p: 0.048). Preoperative plasma CFB can be used to predict the prognosis of resectable pancreatic cancers; it outperforms both CA 19-9 and CEA.

Mass spectrometry-based analysis of cerebrospinal fluid from arthritis patients\u2014immune-related candidate proteins affected by TNF blocking treatment

BackgroundSigns of inflammation in cerebrospinal fluid (CSF) of rheumatoid arthritis patients correlate positively with fatigue, a central nervous system (CNS)-related symptom that can be partially suppressed by TNF blockade. This suggests a possible role for CNS inflammation in arthritis that may be affected by TNF blockade. We therefore investigated the effects of TNF blockade on the arthritis CSF proteome and how candidate proteins related to clinical measures of disease activity and inflammation.MethodsMass spectrometry-based quantitative proteomic analysis was performed on CSF from seven polyarthritis patients before and during infliximab treatment. Treatment-associated proteins were identified using univariate (Wilcoxon signed rank test) and multivariate (partial least squares discriminant analysis (PLS-DA)) strategies. Relations between selected candidate proteins and clinical measures were investigated using the Spearman correlations. Additionally, selected proteins were cross-referenced to other studies investigating human CSF in a thorough literature search to ensure feasibility of our results.ResultsUnivariate analysis of arthritis CSF proteome revealed a decrease of 35 proteins, predominantly involved in inflammatory processes, following TNF blockade. Seven candidate proteins, Contactin-1 (CNTN1), fibrinogen gamma chain (FGG), hemopexin (HPX), cell adhesion molecule-3 (CADM3), alpha-1B-glycoprotein (A1BG), complement factor B (CFB), and beta-2-microglobulin (B2M), were selected for further studies based on identification by both univariate and multivariate analyses and reported detection in human CSF and known associations to arthritis. Decreased levels of FGG and CFB in CSF after treatment showed strong correlations with both erythrocyte sedimentation rate and disability scores, while CNTN1 and CADM3 were associated with pain.ConclusionSeveral immune-related proteins in the CSF of arthritis patients decreased during TNF blockade, including FGG and CFB that both correlated strongly with systemic inflammation. Our findings stress that also intrathecal inflammatory pathways are related to arthritis symptoms and may be affected by TNF blockade.

Mutations resulting in the formation of hyperactive complement convertases support cytocidal effect of anti-CD20 immunotherapeutics

Anti-CD20 monoclonal antibodies (mAbs) rituximab and ofatumumab are potent activators of the classical complement pathway, and have been approved for the treatment of B-cell malignancies. However, complement exhaustion and overexpression of complement inhibitors by cancer cells diminish their therapeutic potential. The strategies of targeting membrane complement inhibitors by function-blocking antibodies and the supplementation with fresh frozen plasma have been proposed to overcome tumour cell resistance. We present a novel approach, which utilizes gain-of-function variants of complement factor B (FB), a component of alternative C3/C5 convertases, which augment mAb-activated reactions through a positive feedback mechanism called an amplification loop. If complement concentration is limited, an addition of quadruple gain-of-function FB mutant p.D279G p.F286L p.K323E p.Y363A (or selected single mutants) results in significantly increased complement-mediated lysis of ofatumumab-resistant tumour cells, as well as the complete lysis of moderately sensitive cells. Importantly, this effect cannot be achieved by further increasing ofatumumab concentration. Potentiation of cytotoxic effect towards moderately sensitive cells was less apparent at physiological serum concentration. However, an addition of hyperactive FB could compensate the loss of cytotoxic potential of serum collected from the NHL and CLL patients after infusion of rituximab. Residual levels of rituximab in such sera, in combination with added FB, were able to efficiently lyse tumour cells. We suggest that the administration of gain-of-function variants of FB can restore cytotoxic potential of complement-exhausted serum and maximize the therapeutic effect of circulating anti-CD20 mAbs.

A new complement factor B mutation associated with crescentic C3 glomerulopathy; a case report

Background: C3 glomerulopathy is a recently described entity classified as complementassociated glomerular disease. Case Presentation: We report a case of a 48-year-old man referred to the nephrology department for nephrotic syndrome with rapidly progressive kidney failure, acquired partial lipodystrophy and drusen in Bruch’s membrane of the retina. Blood tests showed low C3 and no evidence for autoimmune diseases, monoclonal gammopathy or infection. The renal biopsy revealed a proliferative endocapillary and crescentic glomerulonephritis with glomerular deposits exclusively of C3 and significant interstitial fibrosis. The electronic microscopy was consistent with dense deposit disease. The complement analysis revealed a pathogenic mutation of the complement factor B (CFB) gene not previously described in literature. Conclusions: The authors report a new mutation of CFB, in a dense deposit disease patient; this finding brings a new insight to the pathogenic pathway of C3 glomerulopathy and possibly to other complement dysregulation associated glomerular diseases. More clinical trials are needed to clarify both the pathogenicity and the optimal treatment for these entities.

Diferencias fisicoquímicas de suelo suplementado con pilas eléctricas

Existen prácticas de campesinos en el departamento del Caquetá y testimonios de personas en la ciudad de Florencia que involucran el uso de pilas eléctricas como base del abono orgánico para plantas. Frente a la hipótesis que las pilas eléctricas pueden contribuir al desarrollo saludable de una planta, se planteó como objetivo del presente estudio, evaluar la incidencia de las pilas eléctricas en las propiedades físicas y químicas de una muestra de suelo destinado a la siembra de fríjol. El experimento consistió en un diseño completamente al azar (DCA) con arreglo factorial 1 x 4, donde el factor A corresponde fríjol y el factor B a la variedad de pilas eléctricas (genéricas y alcalinas) con tres repeticiones. En cada tratamiento se determinó: pH, acidez intercambiable (A.I.), carbono orgánico (C.O), cenizas, color y conductividad. El estudio permitió evidenciar que la adición de pilas eléctricas al suelo favorece la absorción de la energía solar, haciendo que la temperatura media del suelo tienda a ser más elevada lo que tendería a promover una germinación rápida en las semillas, aspecto positivo en la etapa germinativa de estas. No obstante, puede incidir en la baja liberación de nitrógeno al suelo, en sus fases posteriores al crecimiento.

Clinical application evaluation of serum C1q and other complement factors in the diagnosis and prediction of preeclampsia

Objective To observe the levels of serum complement C1q, C3, C4 and factor B in different phases during normal pregnancy; To evaluate the diagnostic value and the predictive value of serum complement C1q, C3, C4 and factor B in preeclampsia (PE). Methods Three groups of subjectes were enrolled from January 2017 to March 2018 in Department of Obstetrics and Gynecology, Peking University Third Hospital. (1) 30 pregnant women in each group at 8-14 weeks, 20-26 weeks and 28-36 weeks were retrospectively selected, and the serum levels of complement C1q, C3, C4 and B factors were measured and compared. (2)Selecting 17 cases of early-onset mild PE, 47 cases of early-onset severe PE, 24 cases of late-onset mild PE, 27 cases of late-onset severe PE, and 30 normal pregnant cases of the same gestational stage as early-onset / late-onset controls, through ANOVA analysis and comparison between two groups, this study evaluated the diagnostic value of serum complement C1q, C3, C4 and factor B in PE. (3)To evaluate the predictive effect in PE, it analyzed serum C1q and factor B levels of pregnant women at 20-26 gestation weeks through prospective nested case-control study of 214 cases. Results The levels of serum C1q remained stable in the whole pregnancy. The levels of C3 and factor B increased at the early stage of pregnancy and remained stable after the middle stage. C4 increased early in pregnancy and then remained stable. Compared with the control group, the levels of serum C1q in all four types of PE patients were significantly decreased (median: 169 mg/L, 161 mg/L, 165 mg/L, 163 mg/L; early-onset, late-onset control group: 187 mg/L, 194 mg/L; U=130.500, 426.500, 159.500, 130.500, all P 0.05). 33 (15.4%) cases developed PE out of 214 pregnant women with PE risk factors. Compared to those who didn′t develop PE, it showed no statistical difference of serum C1q, C3, C4, and factor B levels at 20-26 gestational weeks of the women who subsequently developed PE (C1q: 175 mg/L vs. 184 mg/L; C3: 1 523 mg/L vs. 1 467 mg/L; C4: 230 mg/L vs. 229 mg/L; FB: 344 mg/L vs. 320 mg/L; U=2 090.000, 1 575.000, 2 058.500, 1 362.000, all P>0.05). Compared to those of the healthy pregnant controls, it showed no statistical difference of serum C1q, C3 and C4 levels of 20-26 gestational weeks of the women who subsequently developed PE (C1q: 175 mg/L vs. 190 mg/L; C3: 1 523 mg/L vs. 1 428 mg/L; C4: 230 mg/L vs. 227 mg/L; U=353.000, 395.000, 493.500, all P>0.05), while it showed statistical difference (344 mg/L vs. 306 mg/L; U= 233.500, P=0.007) for factor B. Conclusions Serum C1q level of PE patients significantly decreased, which can be used as potential indicators of PE diagnosis, but serum C1q, C3, C4 level of 20-26 gestational weeks cannot predict risk of PE. Factor B cannot serve as serum index of PE diagnosis, but its serum levels at 20-26 gestational weeks werer higher than those of normal pregnant controls, factor B may be a potential predictor, but need further verification. (Chin J Lab Med, 2018, 41: 934-942) Key words: Preeclampsia; Complement; C1q; C3; C4; factor B

An analysis of plasma reveals proteins in the acute phase response pathway to be candidate diagnostic biomarkers for depression

Globally, depression is one of the most serious debilitating psychiatric mental disorders. In this study, we validated the expression levels of fibrinogen alpha (FGA), fibrinogen beta (FGB), fibrinogen gamma (FGG), Complement factor B (CFB) and serpin family D member 1(SERPIND1) in the acute phase response signaling pathway in plasma samples using enzyme-linked immunosorbent assay (ELISA).Then illuminate the roles of FGA, FGB, FGG, CFB, SERPIND1 in depression using microarray data.Gene expression dataset GSE98793 was downloaded from the Gene Expression Omnibus database. There were 128 whole blood samples included 64 patients with major depressed patients and 64 healthy controls. Differentially expressed genes (DEGs) were identified, and then protein-protein interaction (PPI) network was constructed to screen crucial genes associated with FGA, FGB, FGG, CFB and SERPIND1. Moreover, gene ontology (GO) biological processes analyses was performed.The ELISA data showed that the expression levels of FGA, FGB, FGG, CFB and SERPIND1 were up-regulated in depressed patients. The enriched GO terms were predominantly associated with the biological processes including more genes were inflammation related. The PPI network was found these five genes interacted with 11 genes. FGA, FGB, FGG, CFB and SERPIND1 may be important in the pathogenesis of depression.

Продуктивність сортів пшениці озимої залежно від фону живлення в умовах Південного Степу України

У статті наведені результати досліджень, проведених в 2011-2016 рр. на чорноземі південному в умовах Південного Степу України, з вивчення ефективності оброблення посівів пшениці озимої сучасними рістрегулюючими препаратами по фону внесення мінеральних добрив. Вивчали вплив сортових особливостей пшениці озимої та варіантів живлення на формування показників структури врожаю та рівень урожайності зерна культури. Визначено, що за внесення під передпосівну культивацію пшениці озимої мінерального добрива в дозі N 30 P 30 (фон) та застосування позакореневого підживлення посівів на початку відновлення весняної вегетації та початку виходу рослин у трубку комплексними органо-мінеральними добривами Органік Д2 та Ескор – біо створюються сприятливі умови для формування оптимальних показників структури врожаю та відповідно найвищого рівня врожайності зерна досліджуваних сортів. Так, у середньому за роки досліджень, рослинами пшениці озимої сорту Кольчуга у даних варіантах удобрення сформовано 4,42-4,48 т / га зерна, а сорту Заможність 4,96-4,99 т / га, що перевищило показники контролю на 52,9-55,0 та 62,6-63,6 % відповідно. Із досліджуваних сортів пшениці озимої за комплексом показників кращим визначено сорт Заможність.

Effects of different seed treatments, provenance and size on germination and early establishment of Olea europaea

Knowledge and understanding of the factors affecting germination and growth of tree species is of paramount importance for enhancing the efforts towards afforestation. Hence, the prime objective of this study is to evaluate the effects of seed treatment methods, provenance and size on germination and early nursery performance of Olea europaea. The experiment was arranged in a factorial combination and laid out in randomized complete block design with three replications. Factor A was seed treatment methods (soaking in three levels of hot water, soaking in four levels of concentrated H2SO4, heating with fire, de-coating and control). Factor B was two seed provenances and factor C was two seed size classes. Interaction of seed size and provenance had no significant effect (p > 0.05) on all parameters. Different seed soaking method showed highly significant difference (p < 0.01) in all parameters. Seed de-coating had maximum value for all parameters but seeds treated with fire had lowest value. Hence, the germination percentages under nursery condition were 85.8 and 2.5% for de-coated and fire treated seeds, respectively. It was concluded that seeds from both provenances and seed size class germinated and were established equally; de-coating improved germination and early nursery performance; whereas, fire affected germination and early nursery performance negatively. Seed de-coating has to be done widely for the improvement of germination and early establishment of Olea europaea and further research is needed on the use of fire for breaking dormancy. Key words: Dormancy, germination, Olea europaea, provenance, seed treatment, seed.

Importance of the Complement Alternative Pathway in Serum Chemotactic Activity During Sepsis.

Serum chemotactic activity is important in regulating neutrophil migration. The ability of neutrophils to migrate to infectious site is crucial for host effective pathogen control, but unregulated neutrophil activation can also cause tissue damage. During bacterial sepsis, the complement alternative pathway (AP) is massively activated in blood and tissues and reportedly contributes to sepsis pathogenesis. Complement factor B (FB) is an essential component of the AP activation. However, the impact of FB/AP activation on blood chemotactic activity during bacterial infection is unclear. In this study, we found that sera of septic mice following cecal ligation and puncture (CLP) had much higher chemotactic activities on neutrophils than those of sham animals. Compared with wild-type (WT) mice, FB mice had significantly attenuated serum chemotactic activity, under both nonseptic and septic conditions. Moreover, sera with the activated AP by zymosan and cobra venom factor (CVF) in vitro induced a significant increase in neutrophil migration compared with sera without the AP activation. Complement activation generates complement cleavage fragment such as Ba, C3a, and C5a. To delineate the contribution of these downstream effectors, we incubated AP-active sera (AP activated by zymosan/CVF) or sera from sham and septic mice with anti-C5a or mAb1379 (anti-Ba) neutralizing antibody. We found that anti-C5a, but not mAb1379, markedly attenuated the neutrophil chemotactic effect of the AP-activated sera and that of septic sera. Taking together, these data suggest that the complement AP activation during bacterial sepsis plays a pivotal role in promoting blood chemotactic activity through a C5a-dependent mechanism.

Auto-antibodies against factor B in post infectious

Auto-antibodies against factor B in post infectious

RA10.02: A NOVEL ROLE FOR THE COMPLEMENT CASCADE IN CHEMORADIATION THERAPY RESISTANT OESOPHAGEAL ADENOCARCINOMA

Abstract Background Resistance to chemoradiation therapy (CRT) in oesophageal adenocarcinoma (OAC) is an important clinical challenge. Elucidating underlying mechanisms of treatment resistance is crucial to improving survival rates. Methods OAC tumour biopsies were collected prior to neoadjuvant CRT. Tumour conditioned media was generated by culturing human explants for 24 h. Response to treatment was determined pathologically using the Mandard Grading System. C3, CFB and MCP-1 mRNA expression was assessed by qPCR. Secreted C3, C3a and MCP-1 were measured by ELISA. An isogenic model of radioresistant OAC was established by chronically irradiating OE33 cells with clinically-relevant doses of 2 Gy X-ray radiation. C5aR and C3aR expression was determined by flow cytometry. Results We demonstrate, for the first time, that the central factor of the complement cascade, complement component 3 (C3), is expressed in OAC tumours and is increased in pre-treatment OAC biopsies from patients (n = 13) who have a subsequent poor response to neoadjuvant CRT (P &lt; 0.05). In addition, C3 is secreted from ex vivo human OAC tumour explants (n = 13) and correlates with levels of the monocyte chemoattractant MCP-1 (P &lt; 0.008) and complement factor B (CFB) a component of the alternative pathway of complement activation (P &lt; 0.0001). In vitro, radioresistant OE33 R cells had increased mRNA levels of C3 (P &lt; 0.01), CFB (P &lt; 0.05) and MCP-1 (P &lt; 0.05) compared to radiosensitive OE33 P cells. OEE3 R cells expressed increased levels of secreted C3 and the anaphylatoxin C3a, compared to radiosensitive OE33P cells (both P &lt; 0.001). Furthermore, we demonstrate for the first time that the anaphylatoxin receptor C5aR is expressed by OAC cells, suggesting that OAC cells can respond to complement anaphylatoxins. Conclusion This study highlights, for the first time, a novel role for the complement cascade in the resistance of OAC to CRT and highlights C3 as a novel predictive marker of response to CRT in OAC. Disclosure All authors have declared no conflicts of interest.