Complement 3 Research Articles

Enhancement of growth, antioxidative status, nonspecific immunity, and disease resistance in gibel carp (Carassius auratus) in response to dietary Flos populi extract

This study investigated the effects of dietary Flos populi extract (FPE) on the growth, antioxidation capability, innate immune response, and disease resistance in gibel carp. A total of 480 fish were fed with five different diets containing 0, 0.5, 1.0, 1.5, or 2.0 g kg−1 FPE (designated as control, D0.5, D1.0, D1.5, or D2.0 groups) for 45 days. The fish were challenged with A. hydrophila after the feeding trial. Compared with the control, the feed efficiency (FE), weight gain (WG), final body weight (FBW), and specific growth rate (SGR) were significantly improved in groups D1.0 and D1.5. Dietary FPE significantly increased serum superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities, as well as glutathione (GSH) content. The contents of protein carbonyl (PCC) and malondialdehyde (MDA) in serum decreased significantly. Additionally, FPE supplementation in diets resulted in significant improvement in serum lysozyme (LZM) and myeloperoxidase (MPO) activities, as well as immunoglobulin M (IgM) and complement 3 (C3) concentrations. The hepatic antioxidant enzymes (CAT and SOD) activities increased, whereas content of MDA decreased in fish treated with dietary FPE than those of control both pre- and post-challenged. After 12 h-challenge, an obvious downregulation of hepatic Kelch-like-ECH-associated protein 1 (Keap1), splenic tumor necrosis factor-α (TNF-α), interleukin (IL)-8, IL-1β, and toll-like receptor 2 (TLR2) mRNA levels was observed in fish treated with dietary FPE, whereas hepatic Nrf2 transcription level was upregulated compared to the control. Furthermore, compared to group D0.5, higher relative percent survival (RPS) was observed in gibel carp fed dietary 1.0–2.0 g/kg FPE. Our results reveal that FPE supplemented diet has a stimulatory effect on antioxidant capacity and nonspecific immune response, along with improved growth performance and enhanced resistance against A. hydrophila infection in juvenile gibel carp.

Healing through faith: meeting a chaplain coupled with biblical readings could produce lymphocyte changes that correlate with brain activity (HEALING study)

Introduction: Faith and systems of beliefs are known to impact not only the emotional, but also the immunological state of believers in ways that we are just starting to understand. Moreover, clinical implications of previous studies are limited. The aim of the “HEALING” (Hospital-based Ecumenical and Linguistic Immuno-NeuroloGic) Study was to examine immunological and neurological changes in hospitalized patients after meeting a chaplain coupled with biblical readings. Methods: Hospitalized patients were pre-screened to find those who were the most in need of an intervention. A passage from the Bible was read to them during a meeting with the chaplain at the bedside (n= 20) or in the chapel (n= 18). No meeting occurred in the randomized control group (n=19). Blood samples were taken 30 minutes prior, and 60 minutes after the meeting to measure white blood cells (WBC), interferon gamma (IFN-γ), immunoglobulin M (IgM), IgA, IgG, and complement 3 (C3). A subgroup of the visited patients was subjected to functional magnetic resonance imaging (fMRI), where they were played an audiotape of readings of the same passage from the Bible (n=21). Results: Lymphocyte counts increased more often after the more successful visits, but the immunological changes were not significant. Conversely, a significant (pfwe=0.003) correlation was revealed between changes in lymphocytes and activation of the angular gyrus (left BA39) during fMRI, a brain area involved in word recognition. Conclusions: Although limited by the sample size and cohort study design, the findings suggest the depth of psycho-immunological changes could depend on the degree to which the chaplains’ main message is understood.

The prevalence of hereditary angioedema in a Chinese cohort with decreased complement 4 levels

ObjectiveHereditary angioedema (HAE) is a rare, life-threatening autosomal dominant disorder. We aimed to investigate the prevalence of HAE in a Chinese population with a decreased Complement 4 (C4) level. MethodsAll the patients present in Tongji Hospital with C4 below lower normal range were included from January 2019 to June 2020. The individual data were extracted from the database and categorized by diagnosis. Patients suspected of HAE were further evaluated by C1 inhibitor level and function test to confirm the HAE diagnosis. ResultsA total of 8226 patients were enrolled in our study, among whom 18 had symptoms similar to HAE and received C1 inhibitor level and function tests. Two (1 male and 1 female) of the 18 patients were identified as HAE patients. This means the prevalence of HAE was 2.43/10 000 among the C4-decreased population and 10.1/10 000 in the C4-decreased population with etiology undetermined. The 2 HAE patients had experienced skin and oropharynx edema attack and received tracheotomy. The female patient had a family history. Laboratory tests showed significant decrease of C4 and C1 inhibitor levels in the 2 patients, both of whom were diagnosed as type 1 HAE. ConclusionThe prevalence of HAE is low in C4-decreased patients. In a large cohort, C4 level can serve as a practical indicator to screen the HAE patients, but further testing of C1 inhibitor activity and levels is needed to confirm the diagnosis of HAE.

Effects of dietary pyridoxine on the skin immunity, tight junctions, antioxidants and apoptosis of grass carp (Ctenopharyngodon idella) infected withAeromonas hydrophila

In this study, fish were fed six diets containing different levels of pyridoxine (PN) for 10 weeks, and then infected with Aeromonas hydrophila for 2 weeks. The results showed that dietary supplementation with PN had a powerful effect on the inhibition of skin haemorrhage and lesion morbidity, and the enhancement of immunity after fish were infected with A. hydrophila. For example, PN increased the levels of immunoglobulin M (IgM), acid phosphatase (ACP), complement 3 (C3), lysozyme (LZ), C4, antimicrobial peptides, target of rapamycin (TOR), anti-inflammatory cytokines, ribosomal protein S6 kinases 1 (S6K1) and inhibitor of κBα (IκBα), while decreasing the levels of proinflammatory cytokines [except interleukin-12 p40 (IL-12p40)], eIF4E-binding proteins (4E-BP), IκB kinase β (IKKβ) and nuclear factor κB p65 (NF-κBp65) in grass carp skin. Additionally, dietary supplementation with PN increased the levels of tight junction proteins (not claudin-15 and claudin-12), antioxidant enzymes [except manganese superoxide dismutase (MnSOD) activity and gene expression, glutathione peroxidase 4a (GPx4a) and GPx4b gene expression], NF-E2-related Factor 2 (Nrf2), myeloid cell leukaemia-1 (Mcl-1) and inhibitor of apoptosis proteins (IAP), while decreasing the levels of malondialdehyde (MDA), reactive oxygen species (ROS), protein carbonyl (PC), Kelch-like ECH-associating protein 1a (Keap1a) myosin light chain kinase (MLCK), cysteinyl aspartic acid protease-9 (caspase-9), caspase-8, caspase-7, caspase-3, Bcl-2-associated X protein (Bax), p38 mitogen-activated protein kinase (p38MAPK) and apoptotic protease-activating factor 1 (Apaf-1) in grass carp skin. In summary, dietary PN can enhance the immunity of fish skin via elevated immune substances, an eased inflammatory response and an improved barrier integrity.

Determination of the Risk Factors Contributing to the Development of Neuropsychiatric Lupus in a Systemic Lupus Erythematosus Cohort.

BackgroundSystemic lupus erythematosus (SLE) is a chronic autoimmune disease with a complex, varied clinical presentation that is both more common and has poor outcomes in women of color. SLE outcomes also seem to be influenced by socioeconomic factors. Neuropsychiatric lupus (NPL) is a common manifestation of SLE that is difficult to diagnose and treat and has poor clinical outcomes. There is no clear relationship between NPL and SLE-related autoantibodies, and this contributes to the difficulty in diagnosing NPL. As a result, NPL is a significant contributor to morbidity and mortality in patients with SLE.ObjectiveThe purpose of this study was to examine the relationship between serological and socioeconomic factors in the development of NPL in our patient cohort and determine the risk factors for the development of NPL.MethodsThis was an SLE single-center, retrospective chart review study that was performed at a university-based tertiary referral center. Patients aged 18 and older who meet the American College of Rheumatology (ACR) 1997 criteria and were seen between June 1st, 2015, and June 1st, 2019, were included in this study. Overall, 629 patients with SLE were identified, and 263 patients were included. Demographic and serological data were collected. Supplemental socioeconomic information for each zip code in Southwest Virginia was obtained from the United States Government Census website. Continuous variables were analyzed using the T-test or Mann-Whitney U test. Categorical variables were analyzed using chi-square tests or Fisher’s exact tests. Statistical analysis was performed using SAS9.4, and p-value < 0.05 was considered statistically significant.ResultsWe reviewed a number of risk factors including age, sex, race, and median household income (MHI), noting no statistical relationship between these factors and the diagnosis of NPL. We did find that the presence of antiphospholipid antibodies (aPL) was significantly associated with a diagnosis of NPL and that complement 4 (C4) levels trended toward statistical significance.ConclusionIn our cohort of patients, there was no relationship between age, sex, race, and median household income, and the diagnosis of NPL. There was a statistically significant relationship between aPL and the diagnosis of NPL. Other SLE-related antibodies showed no statistical relationship with the diagnosis of NPL. Although not statistically significant, there was a trend toward significance between complement 4 (C4) levels and the diagnosis of NPL.

The indirect method in the establishment of reference intervals for complement 3 and complement 4: A retrospective study

ObjectiveThe establishment of reference intervals (RIs) for complement 3 (C3) and complement 4 (C4) is rare, especially by indirect methods. Therefore, this study aims to establish regional RIs for C3 and C4 by an indirect method, using relevant statistical methods. MethodsTotal of 12,313 data points for C3 and 12,125 data points for C4 were obtained from the First Hospital of Jilin University's database in China and standardised using the Tukey and Box–Cox statistical methods. The coefficients of the skewness-median-coefficient of variation curves (LMS) were used to determine the critical value for age, and a subsequent z test used to compare the differences. A non-parametric method was used to establish the RIs. ResultsThe C3 and C4 concentrations showed no significant differences by sex, and a weak correlation with age. No significant difference was found after calculating the z value for the age points on the LMS curves. The RIs for C3 and C4 were 0.83–1.58 g/L and 0.15–0.40 g/L, respectively. The RIs all passed verification. ConclusionSuitable RIs for C3 and C4 were established for the local population, and will benefit clinical diagnosis. The feasibility and practicability of the indirect method were demonstrated.

Carboxymethyl chitosan nanoparticles loaded with bioactive protein CiCXCL20a effectively prevent bacterial disease in grass carp (Ctenopharyngodon idella)

Oral antimicrobial proteins have been widely studied for the prevention of bacterial diseases in aquaculture. However, the shortcomings of antimicrobial proteins that are easily degraded in vivo limits the application. To address these challenges, we expressed the antimicrobial chemokine Ctenopharyngodon idella CXCL20a (CiCXCL20a) by Pichia pastoris and loaded it by carboxymethyl chitosan (CMCS). We used dynamic light scattering (DLS) and transmission electron microscopy (TEM) for characterization and confirmation, obtained stable and slow-release nanoparticle CMCS-20a. In vitro experiments, CMCS-20a can effectively kill bacteria by destroying the bacterial membrane and binding DNA, and reduce the influence of enzyme, salt ion and serum on antibacterial activity of CiCXCL20a. Furthermore, CMCS-20a was almost nontoxic to piscine cells and bound to bacteria when eukaryotic cells coexist with bacteria. More importantly, oral CMCS-20a (mixed with feed) can increase the activities of total superoxide dismutase (SOD), lysozyme (LZM), complement 3 (C3) in sera. After Aeromonas hydrophila infection, oral CMCS-20a enhances grass carp survival rate. To uncover the protective mechanism, we investigated the inflammatory response, bacterial loads and tissue injury. The results indicated that oral CMCS-20a can efficiently regulate inflammation responses by swiftly upregulating and recovering mRNA expressions of inflammation-related genes (IL-1β, TNF-α, IL-2, IFN-γ2) in head kidney tissue, significantly reduce bacterial burdens in intestine, head kidney, hepatopancreas, spleen, blood tissues, and obviously alleviate tissue lesions in spleen and intestine tissues. In conclusion, nanoparticle CMCS-20a has a good biosafety and can effectively prevent bacterial disease in grass carp.

Predicting Severe Renal and Gastrointestinal Involvement in Childhood Immunoglobulin A Vasculitis with Routine Laboratory Parameters

Background: Immunoglobulin A vasculitis (IgAV) is the most common vasculitis in children. Although childhood IgAV is generally considered as a self-limited disease, progressive course and poor prognosis could occur in some cases which mostly result from severe renal involvement and gastrointestinal (GI) involvement. Methods: We performed a retrospective study of pediatric patients diagnosed as IgAV in our institution from 2016 to 2019. Patients were divided into groups based on the occurrence and severity of GI and renal involvement. Analysis of variance (ANOVA) and Kruskal-Wallis test were used to compare results of laboratory parameters among groups and prediction models were built by using logistic regression analysis. Results: A total of 286 patients were enrolled. GI involvement occurred in 148 (51.7%) patients, 30 (20.3%) of which were severe cases. Renal involvement developed in 120 (42.0%) patients, 22 (18.3%) of which were severe cases. Compared with patients with only cutaneous manifestations, white blood cell (WBC) count, neutrophil-to-lymphocyte ratio (NLR), and D-dimer levels were higher in those with GI involvement, and D-dimer level was found to be positively associated with severity. Increased NLR and lower complement 3 (C3) were found in patients with renal involvement, but only C3 was relevant in distinguishing moderate and severe cases. The prediction model for severe renal involvement was: Logit (P) = 6.820 + 0.270 (age) + 0.508 (NLR) − 16.130 (C3), with an AUC of 0.914. The prediction model for severe GI involvement was: Logit (P) = −5.459 + 0.005 (WBC) + 1.355 (D-dimer) – 0.020 (NLR), with an AUC of 0.849. Conclusion: Our data suggest C3 to be an exclusive predictor for severe renal involvement and D-dimer level to be positively associated with the severity of GI involvement. Prediction models consisting of the above parameters were built for obtaining prognostic information in the early phase of IgAV.

Serum interleukin-18 and its relationship with subclinical atherosclerosis in systemic lupus erythematosus

BACKGROUNDInterleukin-18 (IL-18) is a pro-inflammatory and pro-atherogenic factor, and its blood level has shown a direct correlation with atherosclerosis. We aimed to evaluate the serum IL-18 level in patients with systemic lupus erythematosus (SLE) and its relationship with the intima-media thickness (IMT) of the carotid artery in these patients, as an indicator of atherosclerosis.METHODSIn this cross-sectional study, 60 patients as the patient group and 30 healthy volunteers as the control group [matched sex, age, and body mass index (BMI)] were selected, and their disease status and general data were gathered using the Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K) form. A blood sample was also obtained from all participants to determine the serum level of IL-18 and other metrics, including high-sensitivity C-reactive protein (hs-CRP), cholesterol, triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), anti-double stranded deoxyribonucleic acid (anti-dsDNA), complement 3 (C3), and C4. The IMT of the carotid artery was calculated in both groups. We also evaluated the clinical cardiovascular manifestations.RESULTSThe serum IL-18 levels in patients were significantly higher than in the control group (P ˂ 0.005). It had no significant correlation with disease activity (P = 0.10). The patients with SLE with high IL-18 serum levels (> 280 pg/ml) had higher SLEDAI-2K (P = 0.02) than the patients with a low level (< 280), where 280 was the median of the IL-18 levels. The serum IL-18 level had no significant correlation with the carotid artery IMT.CONCLUSIONA high level of IL-18 reflects the disease activity, but it was not significantly correlated with subclinical atherosclerosis, denoted by the carotid artery IMT.

Host‐associated Bacillus siamensis and Lactococcus petauri improved growth performance, innate immunity, antioxidant activity and ammonia tolerance in juvenile Japanese seabass ( Lateolabrax japonicus )

Five possible probiotic strains (Bacillus siamensis LF4, Micrococcus yunnanensis LD5, Micrococcus aloeverae LC3, Mitsuaria chitosanitabida LD3 and Lactococcus petaurid LF3) were previously isolated from the intestine of Japanese seabass (Lateolabrax japonicus). A feeding trial was performed to evaluate the effects of dietary administration of the five potential probiotic bacteria on growth performance, digestive enzyme activities, immune response, antioxidant capability and ammonia tolerance in juvenile Japanese seabass (L. japonicus). The fish were fed for six weeks with control diet (C, without probiotics) or diets supplemented with 1 × 108 CFU/g of B. siamensis LF4 (T1), Lc. petaurid LF3 (T2), M. chitosanitabida LD3 (T3), M. aloeverae LC3 (T4) and M. yunnanensis LD5 (T5), respectively. Above 93.33% survival rate was in the control and treatments, indicating that the five potential probiotics are safe for Japanese seabass. Supplementation of probiotics (except M. aloeverae LC3) also significantly improved weight gain rate and specific growth rate of Japanese seabass, and B. siamensis LF4 significant enhanced the feed intake of fish (p < .05). Significantly increased trypsin activity was detected in fish fed B. siamensis LF4 (T1) and Lc. petaurid LF3 (T2)-containing diets, while B. siamensis LF4 significantly boosted amylase activity (p < .05). Overall, B. siamensis LF4 and Lc. petaurid LF3-supplemented groups (T1 and T2) were noticed with improved antioxidant and innate immunity by evaluating relative indictors, that is total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), lysozyme, complement 3 (C3), glutamic–oxalacetic transaminase (GOT) and glutamic–pyruvic transaminase (GPT) levels. The cumulative mortality of Japanese seabass, experimentally exposed to ammonia (80 mg/L) for 96 h, was significantly lower in fish fed diets T1 and T2 as compared to control and other experimental diets. In summary, host-associated B. siamensis LF4 and Lc. petauri LF3 could be applied as probiotics for improving growth performance, innate immunity, antioxidant activity and ammonia tolerance in juvenile Japanese seabass (L. japonicus).

Lung mesenchymal stromal cells influenced by Th2 cytokines mobilize neutrophils and facilitate metastasis by producing complement C3

Pre-metastatic niche formation is critical for the colonization of disseminated cancer cells in distant organs. Here we find that lung mesenchymal stromal cells (LMSCs) at pre-metastatic stage possess potent metastasis-promoting activity. RNA-seq reveals an upregulation of complement 3 (C3) in those LMSCs. C3 is found to promote neutrophil recruitment and the formation of neutrophil extracellular traps (NETs), which facilitate cancer cell metastasis to the lungs. C3 expression in LMSCs is induced and sustained by Th2 cytokines in a STAT6-dependent manner. LMSCs-driven lung metastasis is abolished in Th1-skewing Stat6-deficient mice. Blockade of IL-4 by antibody also attenuates LMSCs-driven cancer metastasis to the lungs. Consistently, metastasis is greatly enhanced in Th2-skewing T-bet-deficient mice or in nude mice adoptively transferred with T-bet-deficient T cells. Increased C3 levels are also detected in breast cancer patients. Our results suggest that targeting the Th2-STAT6-C3-NETs cascade may reduce breast cancer metastasis to the lungs.

Dietary α-lipoic acid can alleviate the bioaccumulation, oxidative stress, cell apoptosis, and inflammation induced by lead (Pb) in Channa argus

This study aims to evaluate the effects of dietary α-lipoic acid (α-LA) on bioaccumulation, oxidative stress, apoptosis, and inflammation in Channa argus after 28 d of lead (Pb) exposure. A total of 300 fish were divided into five groups: the first group was the control group and the other four groups were exposed to waterborne Pb (800 ppb) and fed α-LA diets supplemented with 0, 300, 600, and 900 mg/kg. The results demonstrated that dietary α-LA effectively reduced the Pb accumulation in the liver, kidney, gill, intestine, and muscle of C. argus after exposure to Pb. Meanwhile, dietary α-LA reversed alterations in the biochemical parameters (Alanine aminotransferase (ALT), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), blood urea nitrogen (BUN), cortisol (COR), and creatinine (CRE)) and immunity parameters (myeloperoxidase (MPO), complement 3 (C3), lysozyme (LYS), complement 4 (C4), C-reactive protein (CRP), and immunoglobulin M (IgM)) in the serum of fish caused by Pb. Pb-induced reduction of antioxidant enzyme activities (Catalase (CAT), glutathione reductase (GR), superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-Px)) was inhibited by dietary α-LA. And malondialdehyde (MDA) and protein carbonyl (PC) content exhibited an opposite trend. Meanwhile, dietary supplemented with α-LA was found to relieve Pb-induced oxidative stress by downregulating Keap1 mRNA expression levels and upregulating the expression levels of CAT, nuclear factor erythroid 2-related factor 2 (Nrf2), GSH-Px, and Cu/Zn SOD. Furthermore, α-LA supplementation reversed Pb-induced upregulation of pro-inflammatory genes (interleukin (IL)-6, IL-1β, tumor necrosis factor α (TNF-α), and nuclear factor kappa B (NF-κB)), Pro-apoptotic genes (Bcl-2-associated X (Bax), caspase (Cas)-3, and tumor protein p53 (p53)) and Hsp70, and downregulation of anti-inflammatory genes (IL-10, inhibitor of κBα (IκBα), and transforming growth factor β (TGF-β)) and anti-apoptosis gene (B-cell lymphoma-2 (Bcl-2)). Overall, dietary α-LA supplementation could enhance the innate immunity and antioxidant capacity of fish, attenuating the Pb accumulation, and cell apoptosis after being exposed to Pb. Furthermore, dietary α-LA could relieve Pb-induced inflammatory response and oxidative stress of fish via regulating NF-κB and Nrf2 signaling, respectively.

The role of C1 inhibitor and complement as acute phase reactants: are we missing the diagnosis of hereditary angioedema?

BackgroundC1 inhibitor (C1-INH) and complement 4 (C4) have historically been referred to as positive acute phase reactants, however this has never been evaluated in hereditary angioedema (HAE) patients. Low function of C1-INH and low levels of C4 are important in the diagnosis of HAE type 1 and 2. If C1-INH and/or C4 are significant acute phase reactants, their levels may be falsely “normal” in patients with HAE when measured during times of infection or inflammation resulting in missed or delayed diagnosis.Case presentationWe present a case series of four HAE patients who had C4, C1-INH, c-reactive protein (CRP) and ferritin measured at baseline and again during a self-reported upper respiratory tract infection (URTI) or flu-like illness. We did not identify any HAE patients who had a significant change in their C1-INH functional level in the context of a mild infection. However, the C4 level did increase into the normal range on three occasions (2 patients, with 1 patient having elevation during two separate illnesses).ConclusionsC1 inhibitor may not be a clinically significant acute phase protein and appears to still be a reliable diagnostic marker of hereditary angioedema, even in times of modest acute inflammation, unlike complement C4 which can be elevated in this setting.

TWIST1 transcriptionally upregulates complement 3 in glomerular mesangial cells from spontaneously hypertensive rats.

We have shown that complement 3 (C3) is upregulated in cardiovascular and renal organs, which induces the synthetic phenotype and exaggerates the growth of mesenchymal cells from spontaneously hypertensive rats (SHRs). However, the mechanisms of the upregulation of C3 have remained unclear. In the present study, we investigated the role of TWIST1, a transcription factor that regulates mesodermal embryogenesis, in the upregulation of C3 in glomerular mesangial cells (GMCs) from SHRs and Wistar-Kyoto (WKY) rats. Immunocytochemical staining and western blot analysis showed that the expression of TWIST1 in GMCs from SHRs was higher than that in GMCs from WKY rats in vivo and in vitro. Real-time PCR analysis showed increases in the expression of Twist1 mRNA with attenuated expression of miR-151-3p in GMCs from SHRs compared to that in cells from WKY rats. Chromatin immunoprecipitation assays showed increases in TWIST1 binding to the C3 promoter in GMCs from SHRs compared to that in cells from WKY rats. Transfection of Twist1 cDNA by a lentiviral vector increased the expression of C3 mRNA in GMCs from WKY rats. TWIST1 siRNA significantly decreased the mRNA expression of C3 and osteopontin in GMCs from SHRs. These results indicate that the increases in TWIST1 expression, attenuation of miR-151-3p, and strong binding of TWIST1 upregulate C3 gene expression in GMCs from SHRs. The enhanced TWIST1-C3 system induces the synthetic phenotype of mesenchymal tissue that may be associated with cardiovascular and renal remodeling in hypertension.

Lead impaired immune function and tissue integrity in yellow catfish (Peltobargus fulvidraco) by mediating oxidative stress, inflammatory response and apoptosis

Lead (Pb) widely exists in the water environment and has severe toxic effects on aquatic organisms. The yellow catfish (Pelteobagrus fulvidraco) is one of the most important commercial species in China, and moreover, its natural populations are declining with the degradation of environmental water quality. However, little is known about the toxic effects of Pb on its immune organs. This study was performed to determine waterborne Pb exposure on bioaccumulation, histomorphology, antioxidant status, apoptotic and immune response in the head kidney and spleen of yellow catfish. Experimental fish were randomly allocated into twelve tanks (3 tanks per group), and the Pb concentrations of the four groups were 0, 5, 50, and 500 μg/L, respectively. The results reflected that the Pb bioaccumulation of the head kidney and spleen increased with increasing Pb exposure dose and time. Severe histological alterations in the head kidney and spleen were observed at concentration 500 ug/L. With increasing Pb exposure concentrations, the plasma activity of superoxide dismutase (SOD) and catalase (CAT) significantly increased after exposure 7 days and 14 days, and the levels significantly decreased after exposure 28 days. The change trend of glutathione (GSH) levels was opposite to that of SOD and CAT at corresponding exposure time. The plasma malondialdehyde (MDA) levels together with the activities of plasma alkaline phosphatase (AKP) and acid phosphatase (ACP) increased significantly with the increasing Pb concentrations. In contrast, the levels of lysozyme (LYZ), complement 3 (C3) and immunoglobulin M (IgM) decreased significantly with increasing Pb concentrations. Moreover, Pb exposure induced transcriptional upregulation of heat shock protein 70 (hsp70), metallothionein (mt), sod, cat, interleukin-10 (il-10), transforming growth factor-β (tgf-β), and tumor necrosis factor-α (tnf-α), bcl-2-associated X protein (bax), and cysteinyl aspartate specific proteinase -9 (caspase-9), genes in the head kidney and spleen tissues, while downregulating the levels of the lyz, c3, igm and B-cell lymphoma-2 (bcl-2) genes. Our data provide evidence that Pb impaired immune function and tissue integrity in yellow catfish through oxidative stress, inflammatory and apoptosis, and the results can serve as reference data to better protect water environments from Pb eco-toxicants.

Effect of jujube (Ziziphus jujuba) extract on intestinal flora, digestion, and immunity of rainbow trout (Oncorhynchus mykiss)

Jujube (Ziziphus jujuba), as an edible-medicinal fruit, is an important Chinese herbal medicine, which has several beneficial effects on host such as immunomodulation, antioxidant, and gastrointestinal protection. In the study, the beneficial effects of dietary jujube extract supplementation on digestion, nonspecific immunity and intestinal microbial diversity of rainbow trout (Oncorhynchus mykiss) were investigated. The fish were fed for 56 days with diets containing 0 (control), 0.25%, 0.50% and 1.00% jujube extract, and then gastrointestinal digestive enzyme activities, nonspecific immune genes expressions and intestinal microbial diversity were measured. Intestinal protease and α-amylase activities in 0.25% and 0.50% groups, and lactase activity in 0.50% group were significantly improved. The expression of Cu/Zn-superoxide dismutase (SOD) gene was significantly increased in 0.50% and 1.00% groups. Catalase (CAT) and glutathione S-transferase A (GSTA) were significantly increased in 0.50% and 0.25% groups, respectively, while glutathione peroxidase 1 (GPX1) in 0.50% group, complement 3 (C3) in 1.00% group and transforming growth factor-β (TGF-β) in jujube extract groups were all significantly down-regulated. In the gut microflora, there were significant differences in the gut abundances of three phyla (Bacteroidetes, Tenerictes, and Actinobacteria) and eight genera (Dinghuibacter, Lachnospiraceae_ NK4A136_group, Aeromonas, Pseudarthrobacter, Acetatifactor, uncultured_ bacterium mycobacteriumand Rikenellaceae_RC9) among the four experiment groups. Moreover, the abundances of Aeromonas in 0.25% and 0.50% groups were significantly decreased compared with the control group, while Acetatifactor in 0.25% group and Dinghuibacter in 1.00% group were significantly increased. The clusters of orthologous groups of proteins (COG) functional prediction analysis indicated that these differences of intestinal microflora were mainly in metabolism, cellular processes and signaling, and information storage and processing. These results showed that jujube extract can improve the digestive and immune ability, and induce a positive effect on the intestinal flora of rainbow trout.

Application of Sigma metrics in the quality control strategies of immunology and protein analytes

BackgroundSix Sigma (6σ) is an efficient laboratory management method. We aimed to analyze the performance of immunology and protein analytes in terms of Six Sigma.MethodsAssays were evaluated for these 10 immunology and protein analytes: Immunoglobulin G (IgG), Immunoglobulin A (IgA), Immunoglobulin M (IgM), Complement 3 (C3), Complement 4 (C4), Prealbumin (PA), Rheumatoid factor (RF), Anti streptolysin O (ASO), C‐reactive protein (CRP), and Cystatin C (Cys C). The Sigma values were evaluated based on bias, four different allowable total error (TEa) and coefficient of variation (CV) at QC materials levels 1 and 2 in 2020. Sigma Method Decision Charts were established. Improvement measures of analytes with poor performance were recommended according to the quality goal index (QGI), and appropriate quality control rules were given according to the Sigma values.ResultsWhile using the TEaNCCL, 90% analytes had a world‐class performance with σ>6, Cys C showed marginal performance with σ<4. While using minimum, desirable, and optimal biological variation of TEa, only three (IgG, IgM, and CRP), one (CRP), and one (CRP) analytes reached 6σ level, respectively. Based on σNCCL that is calculated from TEaNCCL, Sigma Method Decision Charts were constructed. For Cys C, five multi‐rules (13s/22s/R4s/41s/6X, N = 6, R = 1, Batch length: 45) were adopted for QC management. The remaining analytes required only one QC rule (13s, N = 2, R = 1, Batch length: 1000). Cys C need to improve precision (QGI = 0.12).ConclusionsThe laboratories should choose appropriate TEa goals and make judicious use of Sigma metrics as a quality improvement tool.

Synergistic Effects Between Dietary Zinc Form Supplementation and Dietary Protein Levels on Performance, Intestinal Functional Topography, Hemato-biochemical Indices, Immune, Oxidative Response, and Associated Gene Expression of Nile Tilapia Oreochromis niloticus.

The present study investigates the effect of different dietary protein levels suboptimum level (25%) and optimum level (35%), different Zn forms bulk zinc oxide (BZnO) or nanoparticles zinc oxide (NZnO), and their interaction on performance, intestinal topography, hematology, serum biochemical, antioxidant-immune responses, and related gene expression of Nile tilapia. Six experimental diets were formulated to contain approximately 25% and 35% crude protein and supplemented with Zn forms with 0 (normal level in ingredients), 60mgkg-1 BZnO and 60mgkg-1 nanoparticles of NZnO. Nile tilapia, Oreochromis niloticus, fingerlings (7.53 ± 0. 06g) were fed on one of tested diets in triplicates with 5% of total biomass three times a day for 84days. Results showed that, fish fed diet containing 35% crude protein and supplemented with NZnO form recorded the highest final body weight (FBW), weight gain (WG), and specific growth rate (SGR). However, no significant (P > 0.05) differences were recorded in FBW, WG, SGR, feed conversion ratio (FCR), and protein efficiency ratio (PER) between fish fed diet containing 35% crude protein without Zn supplementation and fish fed diet containing 25% crude protein supplemented with NZnO form. Either fish fed diet containing 25% or 35% crude protein and supplemented with NZnO exhibited the highest values of villi height/width. The highest absorption surface area (ASA) was obtained in fish fed diet containing 25% or 35% crude protein and supplemented with BZnO. Hemoglobin (Hb), hematocrit (Hct), and red blood cell count (RBCs) highest values were obtained for fish fed diet containing protein level 35% supplemented with NZnO. Fish fed diet containing protein level 35% and supplemented with NZnO had the lowest value of alanine amino transferase (ALT) and aspartate amino transferase (AST). The highest globulin value was recorded for fish provided with diet containing 35% crude protein and supplemented with BZnO followed by those fed diet containing 35% crude protein and supplemented with NZnO. Fish fed diet containing protein level 25% with NZnO supplementation recorded the highest super oxide dismutase (SOD), catalase (CAT), glutathione reductase (GSH), and glutathione peroxidase (GPX), with decreasing malondialdehyde (MAD) values. The highest values of immunoglobulin g (IgG), immunoglobulin M (IgM), complement 4 (C4), and complement 3 (C3) were obtained for diet containing 35% crude protein and supplemented with NZnO form. Growth hormone gene (GH) was upregulated in fish fed 25% dietary protein without Zn supplementation, while it was downregulated in fish fed 25% dietary protein and supplemented with NZnO. Transcription of insulin-like growth factor-1 (IGF-I) gene recorded the highest value for fish fed 35% crude protein and supplemented with BZnO. This is although the diet of 35% crude protein + NZnO induced significant (IGF-I) gene expression compared with 25% crude protein with or without BZnO. Therefore, nano zinc is useful as a feed supplement for Nile tilapia (Oreochromis niloticus).

Genes and Pseudogenes: Complexity of the RCCX Locus and Disease.

Copy Number Variations (CNVs) account for a large proportion of human genome and are a primary contributor to human phenotypic variation, in addition to being the molecular basis of a wide spectrum of disease. Multiallelic CNVs represent a considerable fraction of large CNVs and are strictly related to segmental duplications according to their prevalent duplicate alleles. RCCX CNV is a complex, multiallelic and tandem CNV located in the major histocompatibility complex (MHC) class III region. RCCX structure is typically defined by the copy number of a DNA segment containing a series of genes – the serine/threonine kinase 19 (STK19), the complement 4 (C4), the steroid 21-hydroxylase (CYP21), and the tenascin-X (TNX) – lie close to each other. In the Caucasian population, the most common RCCX haplotype (69%) consists of two segments containing the genes STK19-C4A-CYP21A1P-TNXA-STK19B-C4B-CYP21A2-TNXB, with a telomere-to-centromere orientation. Nonallelic homologous recombination (NAHR) plays a key role into the RCCX genetic diversity: unequal crossover facilitates large structural rearrangements and copy number changes, whereas gene conversion mediates relatively short sequence transfers. The results of these events increased the RCCX genetic diversity and are responsible of specific human diseases. This review provides an overview on RCCX complexity pointing out the molecular bases of Congenital Adrenal Hyperplasia (CAH) due to CYP21A2 deficiency, CAH-X Syndrome and disorders related to CNV of complement component C4.

Pharmacotherapy for Behçet's Disease and the Risk of Malignancy.

Background: Behçet’s disease (BD) is associated with an increased risk of cancer. Few reports have been published on the relationship between drug exposure and the risk of cancer in patients with BD. Herein, we explored the relationship between pharmacologic interventions for BD and the risk of cancer. Methods: we carried out a retrospective nested case-control study in a cohort of BD patients from attending our institution. Among 1,148 patients, 22 cancer patients were individually 1:2 matched to 44 cancer-free controls. The following biochemical indicators were evaluated: routine blood analysis, liver and kidney function tests, inflammatory indexes, blood gas analysis, blood electrolyte and previous pharmacologic interventions to manage BD including systemic glucocorticoids, methotrexate, cyclosporine-A, azathioprine, cyclophosphamide (CYC), and thalidomide, which are considered the primary medicines used for the management of BD. Results: Among the 22 BD patients with cancers, myelodysplastic syndrome (MDS) (22.72%) was the most common type. Furthermore, CYC administration was significantly higher in BD patients with cancer compared with the cancer-free matched control group. Further, we observed that complement 4 (C4) (odds ratio [OR] = 0.0001, 95% confidence interval [CI]: 0.001–0.065) and hemoglobin (Hb) (OR = 0.891, 95% CI: 0.795–0.998) levels were independent protective factors for predicting cancer risk in BD patients on multivariate analyses. Conclusion: Our study revealed that CYC was associated with a high risk of cancer in BD patients. Furthermore, C4 and Hb are independent protective factors for oncogenesis in BD patients. These findings may provide references and suggestions for clinicians to select appropriate treatments and for the early recognition of high-risk patients to reduce cancer incidence in BD patients.