Hemolytic Complement Research Articles

The Possible Effects of Galectin-3 on Mechanisms of Renal and Hepatocellular Injury Induced by Intravascular Hemolysis.

Intravascular hemolysis is a central feature of congenital and acquired hemolytic anemias, complement disorders, infectious diseases, and toxemias. Massive and/or chronic hemolysis is followed by the induction of inflammation, very often with severe damage of organs, which enhances the morbidity and mortality of hemolytic diseases. Galectin-3 (Gal-3) is a β-galactoside-binding lectin that modulates the functions of many immune cells, thus affecting inflammatory processes. Gal-3 is also one of the main regulators of fibrosis. The role of Gal-3 in the development of different kidney and liver diseases and the potential of therapeutic Gal-3 inhibition have been demonstrated. Therefore, the objective of this review is to discuss the possible effects of Gal-3 on the process of kidney and liver damage induced by intravascular hemolysis, as well as to shed light on the potential therapeutic targeting of Gal-3 in intravascular hemolysis.

Detection of genetic mutations underlying early-onset systemic lupus erythematosus.

We aimed to investigate the presence of monogenic causes of systemic lupus erythematosus (SLE) in our early-onset SLE patients. Fifteen pediatric SLE cases who had early disease onset (≤6 years) were enrolled in this study. All patients fulfilled the Systemic Lupus International Collaborating Clinics (SLICC) criteria. Genomic DNA was used for whole exome sequencing (WES). Pathogenic variants were confirmed by Sanger sequencing. The median age at diagnosis of 15 early-onset SLE patients included in the study was 4 (2-6) years (F/M = 12/3). Significant gene mutations were detected in five of these patients (33.3%). Patients 1 and 2 with homozygous DNASE1L3 mutations [c.320+4_320+7del and G188A (c.563G>C) variants] had skin involvement and oral ulcers. One of them (patient 1) had arthritis and nephritis, and another (patient 2) had nonscarring alopecia and thrombocytopenia. They are currently clinically inactive but have positive serological findings. Patient 3 with homozygous pathogenic ACP5 mutation [G109R (c.325G>A) variant] had arthritis, nephritis, short stature, and skeletal dysplasia. Patient 4 with a heterozygote novel IFIH1 mutation [L809F (c.2425C>T) variant] had skin findings and leukopenia. Patient 5 with novel C1S variant [homozygous C147W (c.441C>G) variant] had marked skin findings, oral ulcers, nonscarring alopecia, pancytopenia, and low total hemolytic complement CH50 level. All patients have responded to the treatments and have low Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) scores, on therapy. Genetic causes should be investigated in early-onset SLE, for better management and genetic counseling. On the other hand, multicenter studies may help to further define genotype-phenotype associations.

Dietary administration impact of olive pulp on growth performance, metabolic profile, immune status, and antioxidant potential of gilthead seabream (Sparus aurata L.)

The use of natural immunostimulants in aquaculture is intended to foster overall health and bolster resilience against diseases in farmed fish populations. It constitutes a crucial strategy that can contribute to securing the sustainability of the aquaculture industry, and it is an area that warrants ongoing exploration and development. Therefore, the aim of the present work was to evaluate the effect of dietary administration of olive pulp on growth rate, metabolic profile, serum antioxidant potential, and humoral and cellular innate immune parameters of gilthead seabream (Sparus aurata L.). For this, fish were fed control diet or olive pulp enriched diets (50, 100, and 200 mg kg feed−1) for 4 weeks. Our results demonstrated that the highest inclusion level improved the growth rates and the biological antioxidant potential in the serum of fish. However, after 4 weeks of feeding, most of the assayed metabolic parameters (Ca2+, TP, ALB, K+, and Na+) were increased in the serum of fish fed with a diet containing the lowest level of olive pulp (50 mg kg feed−1). Regarding the innate immune parameters, the IgM levels decreased in the serum of fish fed 50′s diet after 2 and 4 weeks of trial. However, the serum of fish fed with diets containing 100 and 200 showed an increase in hemolytic complement activity after 2 weeks whilst this increase was only sustained in the 200′s group after 4 weeks. After 2 weeks of feeding, the serum of the fish showed an increase in peroxidase activity due to the highest olive inclusion. Concerning cellular innate parameters, peroxidase activity, respiratory burst, and phagocytic ability were increased in head-kidney leucocytes of fish fed 100′s diet at 2 weeks compared to values from control fish. These results suggest that the administration of olive pulp-enriched diets can benefit fish growth, antioxidant, and immune status of gilthead seabream.

The time course of complement blockage by ravulizumab in neuromyelitis optica spectrum disorder

AbstractThe classical 50% hemolytic complement test is commonly used to monitor complement function. However, its use in monitoring the effect of ravulizumab on complement assays is unknown. A 47‐year‐old man with anti‐aquaporin 4 antibody‐positive neuromyelitis optica spectrum disorder who was treated with oral prednisolone and azathioprine experienced several episodes of optic neuritis or myelitis. After his latest relapse, ravulizumab was administered to prevent relapse. Serum classical 50% hemolytic complement activity decreased below detectable levels (<10 U/mL) within 1 h of ravulizumab injection. Eighteen weeks later, the patient showed no adverse events or relapse, suggesting that serum classical 50% hemolytic complement activity can monitor ravulizumab effect. In Japan, where 3.5% of the population shows a reduced response to anti‐C5 therapy, serum classical 50% hemolytic complement activity proves useful in confirming ravulizumab efficacy.

Effects of dietary sodium acetate on growth, intestinal microbiota composition, and ammonia tolerance of juvenile yellow catfish Pelteobagrus fulvidraco

Short-chain fatty acids (SCFAs), such as acetates, play a role in modifying metabolic disorders and regulating the homeostasis of the organism. In this study, the impact of dietary sodium acetate (SA) on growth, intestinal microbiota composition, and ammonia tolerance of yellow catfish. During a 56-day study, four groups of yellow catfish weighing an average of 2.72 ± 0.11 g were provided with diets containing varying proportions of SA. The diets included 0%, 0.1%, 0.2%, and 0.4% SA. Each batch consisted of three replicates, with each replicate containing 40 fish. The results indicated that dietary supplemented with 0.2% SA had significantly higher weight gain and specific growth rates than those in 0% and 0.1% groups. The feed efficiency ratio in 0% groups was significantly the highest. Dietary SA can significantly increase the activities of intestinal lipase, amylase, and pepsin, and the contents of liver total protein, immunoglobulin M, and 50% hemolytic complement, while decreasing the contents of liver triglyceride, glucose, high-density lipoprotein cholesterol, aspartate aminotransferase, alanine aminotransferase, tumor necrosis factor-α, interleukin-1, and interleukin-8, and the activities of liver superoxide dismutase, and catalase. In addition, dietary SA significantly increased the relative abundance of beneficial bacteria (Achromobacter) and production of butyrate, meanwhile decreasing the harmful bacterial (Rhodobacter) abundance. After the challenge, cumulative mortality in fish fed 0% SA diets was higher than in fish fed 0.1–0.5% SA diets after 96 h ammonia stress. Correlation analysis showed that the SA induced the enhancement of the metabolic efficiency of carbohydrates, amino acids, and lipids were also associated with the increased relative abundance of Achromobacter. In general, SA can function as a beneficial additive for enhancing growth, intestinal health, and ammonia tolerance in yellow catfish.

Safety, tolerability, and activity of the active C1s antibody riliprubart in cold agglutinin disease: a phase 1b study

AbstractCold agglutinin disease is a rare autoimmune hemolytic anemia characterized by complement pathway-mediated hemolysis. Riliprubart (SAR445088, BIVV020), a second-generation classical complement inhibitor, is a humanized monoclonal antibody that selectively inhibits only the activated form of C1s. This Phase 1b study evaluated the safety, tolerability, and effect on hemolysis of riliprubart in adult patients with cold agglutinin disease. On day 1, 12 patients received a single IV dose of either 30 mg/kg (n = 6) or 15 mg/kg (n = 6) of riliprubart and were subsequently followed for 15 weeks. Riliprubart was generally well tolerated; there were no treatment-emergent serious adverse events, or treatment-emergent adverse events leading to death or permanent study discontinuation. There were no reports of serious infections, encapsulated bacterial infections including meningococcal infections, hypersensitivity, or thromboembolic events. Rapid improvements in hemoglobin (day 5) and bilirubin (day 1) were observed in both treatment cohorts. Mean hemoglobin levels were maintained at >11.0 g/dL from day 29 and mean levels of bilirubin were normalized by day 29; both responses were maintained throughout the study. Improvements in clinical markers closely correlated with a sustained reduction in the 50% hemolytic complement (CH50) throughout the study. Mean C4 levels, an in vivo marker of treatment activity, increased 1 week after treatment with either dose of riliprubart and were sustained throughout the study. In conclusion, a single IV dose of riliprubart was well tolerated, and led to rapid classical complement inhibition, control of hemolysis, and improvement in anemia, all of which were sustained over 15 weeks. This trial was registered at www.ClinicalTrials.gov as #NCT04269551.

Evaluation of Complement Level, Serum Electrophoretic Panel, and Anti-glomerular Basement Membrane in Coronavirus Disease 2019: Focus on Likelihood of Hypersensitivity Type III

<strong data-sider-select-id="e0ba6517-9e2c-4620-8827-36ac66373c0f">Background: As an inflammatory process, the emergence of autoimmunity in COVID-19 is likely through molecular mimicry. In autoimmune hypersensitivity-III, immune complex (IC) formation leads to complement activation, inflammation amplification, and tissue damage. Having accumulated IC, lesions in organs such as the lung with tiny vessels are formed, and the complement is depleted. As a systemic disease, COVID-19 has similar therapies and laboratory findings with hypersensitivity-III. <strong data-sider-select-id="09e60b77-93f6-4c2f-b1e8-fa341b0790ee">Objectives: This study was designed to evaluate complement level, serum electrophoretic pattern, and anti-glomerular basement membrane (GBM) in COVID-19 patients. <strong data-sider-select-id="55fd5e75-5740-416a-8241-47d724fa5d6c">Materials and Methods: Blood was collected, and complement component 3 (C3), C4, 50% hemolytic complement (CH50), anti-GBM level, as well as serum electrophoresis were evaluated. <strong data-sider-select-id="5792ee88-f1ad-4772-83e6-41da9ca0927f">Results: C3/C4 levels were normal in 70% and increased in 30% of the subjects. Albumin was normal in 76.67% and decreased in 23.33% of the patients. Furthermore, alpha-1 and alpha-2 were normal in 33.33% and 53.33% of the patients, but their levels increased in 66.67% and 46.67% of patients, respectively. Moreover, gamma-band was normal in 70% and increased in 30% of the participants. Additionally, all of the subjects had normal beta-band, CH50, and anti-GBM. <strong data-sider-select-id="5b52d788-9f72-4772-b070-52de86d41206">Conclusion: The possibility of autoimmune hypersensitivity-III in the lung and kidneys is not understood. Furthermore, alpha band may be a candidate red flag showing changes from moderate to severe forms of the disease.

A comprehensive model for assessing and classifying patients with thrombotic microangiopathy: the TMA-INSIGHT score

BackgroundThrombotic Microangiopathy (TMA) is a syndrome characterized by the presence of anemia, thrombocytopenia and organ damage and has multiple etiologies. The primary aim is to develop an algorithm to classify TMA (TMA-INSIGHT score).MethodsThis was a single-center retrospective cohort study including hospitalized patients with TMA at a single center. We included all consecutive patients diagnosed with TMA between 2012 and 2021. TMA was defined based on the presence of anemia (hemoglobin level < 10 g/dL) and thrombocytopenia (platelet count < 150,000/µL), signs of hemolysis, and organ damage. We classified patients in eight categories: infections; Malignant Hypertension; Transplant; Malignancy; Pregnancy; Thrombotic Thrombocytopenic Purpura (TTP); Shiga toxin-mediated hemolytic uremic syndrome (STEC-SHU) and Complement Mediated TMA (aHUS). We fitted a model to classify patients using clinical characteristics, biochemical exams, and mean arterial pressure at presentation.ResultsWe retrospectively retrieved TMA phenotypes using automatic strategies in electronic health records in almost 10 years (n = 2407). Secondary TMA was found in 97.5% of the patients. Primary TMA was found in 2.47% of the patients (TTP and aHUS). The best model was LightGBM with accuracy of 0.979, and multiclass ROC-AUC of 0.966. The predictions had higher accuracy in most TMA classes, although the confidence was lower in aHUS and STEC-HUS cases.ConclusionSecondary conditions were the most common etiologies of TMA. We retrieved comorbidities, associated conditions, and mean arterial pressure to fit a model to predict TMA and define TMA phenotypic characteristics. This is the first multiclass model to predict TMA including primary and secondary conditions.

Sodium butyrate improves the growth performance, intestinal immunity, and ammonia tolerance of yellow catfish Pelteobagrus fulvidraco by increasing the relative abundance of Candidatus_Arthromitus

The study aimed to analyze the effect of dietary sodium butyrate (SB) on intestinal digestive enzyme activity, liver physiological status, immune parameter, intestinal microbiota structure and composition, and ammonia tolerance in yellow catfish. Over the course of an 8-week experiment, four separate groups including 480 healthy male yellow catfish (1.52 ± 0.66 g) were fed diets containing varying concentrations of SB: 0, 0.05, 0.1, and 0.2%. Following this, an acute ammonia stress experiment that lasted for 96-h was conducted. The study showed that dietary supplementation of SB can improve growth performance and feed utilization (weight gain rate increased from 163.66 to 226.1%; specific growth rate increased from 1.73 to 2.11%/d; feed conversion rate decreased from 2.02 to 1.34), and increase intestinal digestive enzyme activity (lipase increased from 22.66 to 41.46 U/mg prot; amylase increased from 0.38 to 0.46 U/mg prot; pepsin increased from 10.13 to 12.83 U/mg prot). Plasma total protein (from 510.79 to 523.02 μg/mL), 50% hemolytic complement (from 35.46 to 40.46 U/mL), and immunoglobulin M (from 0.45 to 0.48 g/L) increased as dietary SB levels. On the contrary, plasma glucose (from 6.2 to 3.39 mmol/mL), high-density lipoprotein cholesterol (from 0.38 to 0.25 mmol/mL), aspartate aminotransferase (from 55.66 to 37.36 U/L), alanine aminotransferase (from 358.82 to 283.25 U/L), superoxide dismutase (from 163.84 to 147.13 U/mL), catalase (from 152.89 to 135.07 U/mL), tumor necrosis factor-α (from 67.58 to 62.13 pg/mL), interleukin-1 (from 40.07 to 30.96 pg/mL), and interleukin-8 (from 57.16 to 52.16 pg/mL) decreased with dietary SB levels. In addition, dietary supplementation of 0.2% SB significantly increased the relative abundance of Candidatus_Arthromitus in intestine by approximately 22-fold. To sum up, the utilization of SB has potential to enhance fish growth performance, elevate its physiological status, and improve its ability to tolerate acute ammonia toxicity. This is achievable by increasing the prevalence of Candidatus_Arthromitus in the intestine.

Biomarker Analyses in Patients with Paroxysmal Nocturnal Hemoglobinuria (PNH) Treated with Crovalimab and Eculizumab: Results from the Phase III Randomized COMMODORE 2 Trial

Introduction PNH is a hematopoietic stem cell disorder characterized by hemolytic anemia, hemoglobinuria, and increased risk of thromboembolic events, and is associated with bone marrow failure. Crovalimab (crova) is a novel complement C5 inhibitor engineered with recycling technology, with a long half-life and high solubility that enables low-volume subcutaneous (SC) administration every 4 weeks (q4w). The global Phase III COMMODORE 2 study (NCT04434092) in C5 inhibitor-naive patients (pts) with PNH demonstrated non-inferiority of crova vs eculizumab (ecu) for hemolysis control, transfusion avoidance, breakthrough hemolysis, and hemoglobin stabilization, and a safety profile that was consistent between crova and ecu (Röth EHA 2023; #S181). Here we report results of the exploratory biomarker analyses in COMMODORE 2. Study Design and Methods In COMMODORE 2, C5 inhibitor-naive pts were randomized 2:1 to receive a weight-based tiered crova regimen or ecu. Regimens included loading doses, followed by maintenance dosing (crova: SC injection q4w; ecu: intravenous infusion every 2 weeks). Co-primary efficacy endpoints were proportion of pts with hemolysis control (central lactate dehydrogenase [LDH] ≤1.5×upper limit of normal [ULN]) from Week (W)5 through W25 and proportion of pts with transfusion avoidance from baseline through W25. Exploratory biomarker analyses included terminal complement activity per 50% hemolytic complement (CH50) liposome immunoassay, free hemoglobin, additional exploration of LDH levels, and flow cytometry of the PNH clone size/C3d levels. Additional descriptive analyses of hemoglobin data compared pts with and without transfusions. All biomarker data were assessed from baseline to W25. Results Data forCH50 and LDH were available for all 204 randomized pts (crova: 135; ecu: 69) and data for free hemoglobin, PNH clone size and C3d were available for 126 pts (crova: 85; ecu: 41). At baseline, mean (standard deviation [SD]) LDH levels were 7.6×ULN (3.4) in crova pts and 7.8×ULN (3.5) in ecu pts. 77% of crova pts and 74% of ecu pts had ≥1 packed red blood cell transfusion ≤12 months before screening. 39% of crova pts and 38% of ecu pts had a history of aplastic anemia; 4% and 9% had myelodysplastic syndrome, respectively. Inhibition of complement activity (per CH50) to below the lower limit of quantification was reached at W2 (first post-dose pharmacodynamic sample) in 93% and 46% of pts in the crova and ecu arms, respectively, and in 90% and 73% at W25 (post-dose) (Figure). With regards to intravascular hemolysis markers, the proportion of pts with LDH ≤1.5×ULN was 81% at W5 and 78% at W25 in crova pts, and 84% at W5 and 77% at W25 in ecu pts. Free hemoglobin, another intravascular hemolysis marker, showed a similar initial decrease from baseline in both arms; however, crova pts consistently showed numerically lower levels than ecu pts from W13 to W25. Mean (SD) increase in hemoglobin level from baseline to W25 was 1.2 g/dL (2.0) in the crova arm (baseline: 8.7 g/dL [1.4]) and 0.8 g/dL (1.9) in the ecu arm (baseline: 8.9 g/dL [1.4]). In both arms, a greater increase in mean hemoglobin level from baseline was observed in pts who achieved transfusion avoidance vs those requiring ≥1 transfusion up to W25 (crova: 1.7 g/dL [SD, 1.8; n=88] vs 0.2 g/dL [SD, 1.9; n=40]; ecu: 1.3 g/dL [SD, 1.6; n=47] vs −0.4 g/dL [SD, 2.1; n=20]). Both arms showed a similar increase from baseline to W25 in the mean (SD) percentage of C3d-positive PNH erythrocytes (crova: 0.7 [1.5] at baseline to 20.8 [19.1] at W25; ecu: 0.6 [0.9] to 22.3 [15.0]). Conclusions Treatment with crova or ecu reduced terminal complement activity effectively, with a higher proportion of crova than ecu pts having undetectable terminal complement activity. However, this did not translate into significant differences in other biomarkers (LDH and free hemoglobin) or in the clinical efficacy between crova and ecu arms. Hemoglobin levels increased in both crova and ecu arms. This improvement was more pronounced in pts not receiving a transfusion from baseline to W25; additional analysis is needed to elucidate the impact of bone marrow failure status. Similar increases in C3d level in each arm were expected, as this is a C5 inhibitor class phenomenon. Overall, biomarker data from COMMODORE 2 support the non-inferior efficacy of crova vs ecu in patients with PNH. Further analyses correlating additional biomarkers with clinical outcomes is warranted.

Relevance of complement immunity with brain fog in patients with long COVID

IntroductionThis study aimed to elucidate the prevalence and clinical characteristics of patients with long COVID (coronavirus disease 2019), especially focusing on 50% hemolytic complement activity (CH50). MethodsThis retrospective observational study focused on patients who visited Okayama University Hospital (Japan) for the treatment of long COVID between February 2021 and March 2023. CH50 levels were measured using liposome immunometric assay (Autokit CH50 Assay, FUJIFILM Wako Pure Chemical Corporation, Japan); high CH50 was defined as ≥59 U/mL. Univariate analyses assessed differences in the clinical background, long COVID symptoms, inflammatory markers, and clinical scores of patients with normal and high CH50. Logistic regression model investigated the association between high CH50 levels and these factors. ResultsOf 659 patients who visited our hospital, 478 patients were included. Of these, 284 (59.4%) patients had high CH50 levels. Poor concentration was significantly more frequent in the high CH50 group (7.2% vs. 13.7%), whereas no differences were observed in other subjective symptoms (fatigue, headache, insomnia, dyspnea, tiredness, and brain fog). Multivariate analysis was performed on factors that could be associated with poor concentration, suggesting a significant relationship to high CH50 levels (adjusted odds ratio [aOR], 2.70; 95% confidence interval [CI], 1.33–5.49). Also, high CH50 was significantly associated with brain fog (aOR, 1.66; 95% CI, 1.04–2.66). ConclusionsHigh CH50 levels were frequently reported in individuals with long COVID, indicating a relationship with brain fog. Future in-depth research should examine the pathological role and causal link between complement immunity and the development of long COVID.

Dietary inulin benefits on growth, digestive ability and intestinal microbiota in yellow catfish exposed to ammonia

Inulin can be selectively utilized by the host intestinal microbiota to promote growth and stress resistance. Although inulin is rarely reported in aquatic animal, it has been widely used in feed production. This study evaluated the effects of dietary inulin on growth, digestive ability, and intestinal microbiota of yellow catfish exposed to ammonia. The results showed that ammonia stress resulted in growth retardation (weight gain and specific growth rate decreased), which may be related to reduced feed utilization (feed intake and feed conversion ratio increased). Dietary inulin supplementation can improve intestinal digestive enzyme activity (lipase and pepsin), blood health and immune response (total protein, 50 % hemolytic complement, immunoglobulin M increased; total cholesterol, glucose, aspartate aminotransferase, and alanine aminotransferase decreased), and liver antioxidant enzyme activity (superoxide dismutase and catalase), but reduce inflammatory response (interleukin-1 and tumor necrosis factor-α) of yellow catfish under ammonia stress. In addition, inulin can increase Turicibacter abundance, and reduced Achromobacter abundance in intestinal microbiota, which was positive correlation with improved growth of yellow catfish exposed to ammonia. This study suggests that inulin can be used as a potential feed additive to help yellow catfish cope with ammonia stress.

#3141 AN ATYPICAL EXPERIENCE FOR THE WEST OF SCOTLAND

Abstract Background and Aims Haemolytic uraemic syndrome (HUS) is a disease which affects the kidneys presenting with acute kidney injury (AKI), microangiopathic haemolytic anaemia (MAHA) and thrombocytopenia. In 90% of cases it is precipitated by shiga-toxin producing E coli [1]. The remaining 10% of cases have been termed atypical haemolytic uraemic syndrome (aHUS) and complement system overactivation is the underlying mechanism [2]. This can take the form of autoantibody production or complement gene mutations [2]. Eculizumab has been proven to significantly ameliorate disease progression [3]. In this series, we sought to explore the West of Scotland's experience with this ultra-rare population evaluating outcomes and access to therapies. Method This is a retrospective case series. We extracted data from the west of Scotland renal electronic patient records database Strathclyde Electronic Renal Patient Record (“SERPR”) provided by VitalDataClient. We ran a query to identify patients in whom TMA and/or MAHA and/or HUS was inputted as a diagnosis. 363 patients were identified. Filtering for aHUS in the adult population, 22 patients were identified spanning 19 years. Results The incidence rate is 0.43/million of the population. 14 patients had either genetic mutations (n=13) or acquired antibodies. Genetic mutations: C3, n=5; Complement Factor H (CFH) n=5; Complement Factor I (CFI)+CD46 n=1; CFI alone n=1, variants of uncertain significance n=1. One patient with CFH mutation was positive for the nephritic factor; another with CFH mutation had detectable Factor H autoantibodies. The only acquired patient without a genetic mutation had Factor H autoantibodies. The mean levels of biochemical markers at presentation are: Haemoglobin (g/L) 72; Creatinine, (μmol/L) 603; Platelet count, (×109/L) 89; Lactate dehydrogenase (IU/L) 1853; and Total Bilirubin, (µmol/L) 30. Eculizumab is an available treatment for aHUS utilised in coordination with The National Renal Complement Therapeutics Centre. The outcomes of patients with and without genetic defects and/or antibodies, with the number in parenthesis denoting those who received Eculizumab, are listed below, stratified by CKD staging. Those with identifiable genetic/acquired defects: Recovered 2(2); CKD3 3(3); CKD4 1(0); CKD5 1(0); Renal replacement therapy dependent 7(6)*. Total 14(11). Those with no identifiable defect: Recovered 2(2); CKD3 2(0); CKD4 0; CKD5 0; Renal replacement therapy dependent 4(3)**. Total 8(5). *4 of the 6 have successfully functioning transplants. **Of the 3 patients who received Eculizumab, 2 had a brief trial. The third received it after transplantation but the transplant failed. Conclusion It is clear from the above data that long term renal outcomes are poor for these patients with the majority being left with residual impairment to some degree. Furthermore, in those for whom transplantation is being considering, it is imperative that Eculizumab is introduced, if already withdrawn, to cover transplantation and is maintained to prevent recurrence in the new organ. Even in those without RRT requirements, use of the C5 inhibitor is necessary to stabilise and allow for recovery of at least some native function. Therefore, clinicians in encountering MAHA and TMA in their practice should always consider aHUS and liaise with the Newcastle centre promptly to ensure diagnostics are sent and treatment can be commenced, otherwise the outcomes can be catastrophic.

Peripheral Complement Factor-Based Biomarkers for Patients with First-Episode Schizophrenia.

Schizophrenia (SCZ) is a severe, protracted neurological disorder that causes disruptive conduct in millions of individuals globally. Discovery of potential biomarkers in clinical settings would lead to the development of efficient diagnostic techniques and an awareness of the disease's pathogenesis and prognosis. The aim of the present study was to discover and identify serum complement factor-based biomarkers in discriminating patients with first-episode SCZ from healthy controls. Eighty-nine patients with first-episode SCZ and 89 healthy controls were included in this study. Psychiatric symptom severity of patients with SCZ was measured with the Brief Psychiatric Rating Scale-18 Item Version (BPRS) and the Scales for the Assessment of Negative/Positive Symptoms (SANS/SAPS). A total of 5 complement factors including complement component 1 (C1), C2, C3, C4, and 50% hemolytic complement (CH50) were measured using commercially available enzyme-linked immunosorbent assay (ELISA) kits. The levels of serum complement factors in the SCZ and control groups were compared, and the receiver operating characteristic (ROC) curve method was used to assess the diagnostic values of various complement factors for separating SCZ patients from healthy controls. Pearson's correlation test was used to assess the relationships between serum complement factor concentrations and the psychiatric symptom severity. Therewasanincreaseinserum levels of C1, C2, C3, C4, andCH50amongpatientswith SCZ. Moreover, based on ROC curve analysis, the AUC value of a combined panel of C1, C2, C3, C4, and CH50 was 0.857 when used to discriminate patients with SCZ from healthy controls. Furthermore, serum C2, C3, and CH50 levels were positively correlated to the scores of SANS, SAPS, and BPRS in patients with SCZ, respectively. These results suggested that circulating complement factors including C1, C2, C3, C4, and CH50 may have potential in discovering biomarkers for diagnosing first-episode SCZ.

Dietary N-carbamylglutamate supplementation improves ammonia tolerance of juvenile yellow catfish Pelteobagrus fulvidraco.

Introduction: Ammonia has been of concern for its high toxicity to animals. N-carbamylglutamate (NCG) can reduce blood ammonia levels in mammals, but studies on ammonia tolerance in fish are insufficient. Methods: Juvenile yellow catfish were fed two levels of NCG (0.00% and 0.05%) for 84days under three ammonia levels (0.00, 0.08, and 0.16mg/L NH3). Results and Discussion: The results showed that survival rate (SUR), final body weight (FBW), weight gain (WG), and serum total protein (TP), triglycerides (TG), glucose (Glu), ornithine (Orn), citrulline (Cit) contents, and liver superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), arginase (ARG), ornithine transcarbamylase (OTC) activities decreased with the increase of ammonia levels, on the contrary, feed conversion ratio (FCR), hepatosomatic index (HSI), and serum ammonia, urea, alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutamine (Gln), arginine (Arg) contents, and liver malondialdehyde (MDA), tumor necrosis factor (TNF), interleukin (IL) 1, IL 8 contents, and mRNA expressions of cu/zn sod, cat, gpx, gr, tnf ɑ, il 1, and il 8 were significantly increased. Dietary 0.05% NCG supplementation had higher SUR, FBW, WG, feed intake (FI), whole-body protein, and serum TP, total cholesterol (TC), Glu, citrulline (Cit) contents, and liver SOD, GPx, argininosuccinate synthetase (ASS), argininosuccinate lyase (ASL), inducible nitric oxide synthase (iNOS) activities compared to 0.00% NCG group, but had lower serum ammonia, urea, ALT, AST, Gln, Arg contents, and liver MDA, TNF, IL 1, IL 8 contents, and neuronal nitric oxide synthase activity. At the end of bacterial challenge, cumulative mortality (CM) increased with ammonia levels increased, but serum antibody titer (AT), lysozyme (LYZ) activity, 50% hemolytic complement, immunoglobulin (Ig) contents, respiratory burst (RB), phagocytic indices decreased with ammonia levels increased. CM in 0.05% NCG group was lower than that in 0.00% NCG group, but serum AT, LYZ activity, Ig content, RB in 0.05% NCG group were significantly higher. The correlation analysis found that iNOS was positively correlated with ASS activity. This study indicates that dietary NCG supplementation can improve the ammonia tolerance of yellow catfish, and ASS may also be the target of NCG to activate the urea cycle.

Dietary powder and molecular imprinted polymer nanoencapsulated sodium propionate to enhance growth performance, digestive enzymes activity, antioxidant defense, and mucosal immune response in African cichlid (Labidochromis lividus) fingerlings

Abstract This study was conducted to examine the effects of powder sodium propionate (P-SP) and SP- loaded molecular imprinted polymer (MIP) nanoparticles (MIP-SP NPs) on the growth, skin mucosal immune parameters, and digestive and liver enzymes activities of African cichlid (Labidochromis lividus) fingerlings. Fish with an average weight of 500±2 mg were stocked into 12 experimental units and fed on experimental diets prepared by supplementing the basal diet (control) with MIP NPs, P-SP (5 g SP kg−1 of dry diet), and MIP-SP NPs for 8 weeks. The findings demonstrated that growth indices improved in the MIP-SP NPs followed by the P-SP dietary group compared to the control groups (P&lt;0.05). The activity of digestive enzymes of lipase, trypsin, protease, and alkaline phosphatase was higher in the fish fed SP-supplemented diets than in the controls (P&lt;0.05). The protease and lipase activities in the MIP-SP NPs dietary group increased by 29.33% and 48.81% compared to the control, respectively (P&lt;0.05). In addition, the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels of liver tissue decreased in the SP dietary groups, while the catalase (CAT ), superoxide dismutase (SOD), and alkaline phosphatase (ALP) levels increased compared to the control groups (P&lt;0.05). The highest SOD and ALP levels were observed in the fish fed on the MIP-SP NPs-supplemented diet (P&lt;0.05). Furthermore, the skin mucosal immune indices, including alternative hemolytic complement activity (ACH50), lysozyme, and total immunoglobulin (Ig) levels increased in the MIP-SP NPs and P-SP dietary groups compared to the controls (P&lt;0.05). The findings indicated that sodium propionate encapsulated in molecularly imprinted polymer nanoparticles could enhance the efficiency of dietary SP in African cichlid fish.

Comparison of dietary arginine or/and inulin supplementation on growth, digestive ability and ammonia tolerance of juvenile yellow catfish Pelteobagrus fulvidraco

Most fish have limited use of arginine, and increasing arginine availability by adding other nutrients is an effective method. Inulin is thought to have potential synergistic effects with arginine. Four experiment groups were carried out to test the hypothesis that inulin can increase arginine availability of yellow catfish, including control, arginine, inulin and arginine mixed with inulin groups. The results showed that dietary arginine supplementation improved growth (weight gain and whole-body protein increased; IGF 1, MyoD, MyoG and Myf5 genes up-regulated), digestive ability (intestine protease and lipase increased), blood health (total protein and high-density lipoprotein cholesterol increased) and ammonia tolerance (cumulative mortality reduced; serum lysozyme, 50% hemolytic complement, respiratory burst and phagocytic indices increased) of yellow catfish, and reduced intestine inflammation (tumor necrosis factor and interleukin 1 decreased). Meanwhile, the addition of inulin to arginine-rich diets was further enhanced growth, digestive ability, intestine internal environment (butyric acid and total volatile fatty acids increased), blood health and ammonia tolerance (argininosuccinate synthetase, argininosuccinate lyase, arginase and ornithine transcarboxylase increased) of yellow catfish, and further relieved intestine inflammation. This study indicates that inulin can promote the utilization of dietary arginine; the potential synergistic effects between arginine and inulin were found in growth, digestive ability and ammonia tolerance of yellow catfish.

ВЛИЯНИЕ НИЗКОИНТЕНСИВНОГО ЛАЗЕРНОГО ИЗЛУЧЕНИЯ НА ИММУНОЛОГИЧЕСКИЕ ПОКАЗАТЕЛИ У ПАЦИЕНТОВ С ФЛЕГМОНАМИ ЧЕЛЮСТНО-ЛИЦЕВОЙ ОБЛАСТИ И ШЕИ

Background. The problem of treatment of phlegmon of the maxillofacial region and neck remains one of the most urgent problems of maxillofacial surgery. Aim. To determine the degree of influence of local Low Level Laser Therapy (LLLT) on the quantitative and functional characteristics of immunocytes and humoral factors in peripheral blood and lavage fluid in phlegmon of the maxillofacial region and neck. Material and methods. We have examined and treated 60 patients with phlegmon of the maxillofacial region and neck. Of these, 30 people made up group 1 – traditional treatment and 30 people were included into group 2 (traditional treatment supplemented with local LLLT). Also, 12 practically healthy individuals were examined to determine the regional norm of the selected indicators (control group). The total number of leukocytes and their subpopulations were determined in the blood and lavage fluid, immunophenotyping of lymphocytes was performed using monoclonal antibodies (CD3+, CD4+, CD8+, CD16+, CD19+, CD25+, CD95+), determination of the phagocytic index (PhI) and phagocytic number (PhN), the study of hemolytic complement activity (CH50), the presence of circulating immune complexes (CIC) and the level of immunoglobulins (Ig) of classes M, A, G. Results. Weekly local exposure of LLLT contributed to a decrease in leukocytes and neutrophils in the blood with an increase in their functional activity (PhN), an increase in the number of lymphocytes, levels of IgM, IgG, while reducing CH50 in blood serum. In the lavage fluid under the influence of LLLT, a decrease in leukocytes, neutrophils, activation of phagocytosis, an increase in lymphocytes due to the number of CD3+ and CD4+ T-lymphocytes, a decrease in CD8+ of cytostatic T-lymphocytes, CD25+ activated T- and B-lymphocytes, CD95+T- and B-lymphocytes, decrease in CH50, IgM level and increase in IgG were revealed. Conclusion. Comprehensive treatment of patients with phlegmon of the maxillofacial region supplemented with LLLT has a modulating effect on cellular and humoral immunity factors by enhancing the local immune response.

Autoantibodies against complement factor B in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disorder affecting the joints. Many patients carry anti-citrullinated protein autoantibodies (ACPA). Overactivation of the complement system seems to be part of the pathogenesis of RA, and autoantibodies against the pathway initiators C1q and MBL, and the regulator of the complement alternative pathway, factor H (FH), were previously reported. Our aim was to analyze the presence and role of autoantibodies against complement proteins in a Hungarian RA cohort. To this end, serum samples of 97 ACPA-positive RA patients and 117 healthy controls were analyzed for autoantibodies against FH, factor B (FB), C3b, C3-convertase (C3bBbP), C1q, MBL and factor I. In this cohort, we did not detect any patient with FH autoantibodies but detected C1q autoantibodies in four patients, MBL autoantibodies in two patients and FB autoantibodies in five patients. Since the latter autoantibodies were previously reported in patients with kidney diseases but not in RA, we set out to further characterize such FB autoantibodies. The isotypes of the analyzed autoantibodies were IgG2, IgG3, IgGκ, IgGλ and their binding site was localized in the Bb part of FB. We detected in vivo formed FB-autoanti-FB complexes by Western blot. The effect of the autoantibodies on the formation, activity and FH-mediated decay of the C3 convertase in solid phase convertase assays was determined. In order to investigate the effect of the autoantibodies on complement functions, hemolysis assays and fluid phase complement activation assays were performed. The autoantibodies partially inhibited the complement-mediated hemolysis of rabbit red blood cells, inhibited the activity of the solid phase C3-convertase and C3 and C5b-9 deposition on complement activating surfaces. In summary, in ACPA-positive RA patients we identified FB autoantibodies. The characterized FB autoantibodies did not enhance complement activation, rather, they had inhibitory effect on complement. These results support the involvement of the complement system in the pathomechanism of RA and raise the possibility that protective autoantibodies may be generated in some patients against the alternative pathway C3 convertase. However, further analyses are needed to assess the exact role of such autoantibodies.

Performance of classification and diagnostic criteria for IgG4-related disease and comparison of patients with and without IgG4-related disease

IgG4-related disease (IgG4-RD) was recently described in Japan. It is characterised by extensive organ involvement with tissue fibrosis. We assessed the performance of the 2019 American College of Rheumatology and European League Against Rheumatism (ACR/EULAR) classification criteria and the 2020 revised comprehensive diagnostic (RCD) criteria as well as differences between patients with and without IgG4-RD. In this retrospective, single-centre study of 50 patients admitted with suspected IgG4-RD, we evaluated the sensitivity and specificity of both criteria. We also compared clinical characteristics and laboratory data of patients with IgG4-RD (n = 42) and patients without IgG4-RD (n = 8). The ACR/EULAR classification criteria had 88.1% sensitivity and 87.5% specificity for IgG4-RD diagnosis. The RCD criteria had 100% sensitivity and 50% specificity. Patients with IgG4-RD had significantly more affected organs (p = 0.002). Patients with a single affected organ and IgG4-RD had significantly higher serum IgG4/IgG ratios (p = 0.027), lower serum C-reactive protein levels (p = 0.020), and lower total haemolytic complement activity (p = 0.044) than those without IgG4-RD. The ACR/EULAR classification criteria have high specificity and the RCD criteria have high sensitivity for diagnosing IgG4-RD. The number of affected organs is important for diagnosing IgG4-RD.