Component 1 Research Articles

Association of complement components with risk of colorectal cancer: A systematic review and meta-analysis.

Complement components could contribute to the tumor microenvironment and the systemic immune response. Nevertheless, their role in colorectal cancer (CRC) remains a contentious subject. To elucidate the relationship between complement components and CRC risk and clinical characteristics. Searches were conducted in PubMed, the Cochrane Library, and the China National Knowledge Infrastructure database until June 1, 2023. We included cohort studies encompassing participants aged ≥ 18 years, investigating the association between complement components and CRC. The studies were of moderate quality or above, as determined by the Agency for Healthcare Research and Quality. The meta-analysis employed fixed-effects or random-effects models based on the I² test, utilizing risk ratio (RR) and their corresponding 95% confidence interval (CI) for outcomes. Sensitivity and subgroup analyses were performed to validate the robustness of the collective estimates and identify the source of heterogeneity. Data from 15 studies, comprising 1631 participants that met the inclusion criteria, were included in the meta-analysis. Our findings indicated that protein levels of cluster of differentiation 46 (CD46) (RR = 3.66, 95%CI: 1.75-7.64, P < 0.001), CD59 (RR = 2.86, 95%CI: 1.36-6.01, P = 0.005), and component 1 (C1) (RR = 5.88, 95%CI: 1.75-19.73, P = 0.004) and serum levels of C3 (standardized mean difference = 1.82, 95%CI: 0.06-3.58, P = 0.040) were significantly elevated in patients with CRC compared to healthy controls. Strong expression of CD55 or CD59 was associated with a higher incidence of lymph node metastasis, whereas strong CD46 expression correlated with a higher incidence of tumor differentiation compared to low CD46 expression (P < 0.05 for all). Although specific pooled results demonstrated notable heterogeneity, subgroup analyses pointed to regional differences as the primary source of inconsistency among the studies. Our analysis underscores that increased levels of specific complement components are associated with a heightened risk of CRC, emphasizing the potential significance of monitoring elevated complement component levels.

Analysing N-nitrosamine occurrence and sources in karst reservoirs, Southwest China.

N-nitrosamines in reservoir water have drawn significant attention because of their carcinogenic properties. Karst reservoirs containingdissolved organic matter(DOM) are important drinking water sources and are susceptible to contamination because of the fast flow of various contaminants. However, it remains unclear whether N-nitrosamines and their precursor, DOM, spread in karst reservoirs. Therefore, this study quantitatively investigated the occurrence and sources of N-nitrosamines based on DOM properties in three typical karst reservoirs and their corresponding tap water. The results showed that N-nitrosamines were widely spread, with detection frequencies > 85%. Similar dominant compounds, including N-nitrosodimethylamine, N-nitrosomethylethylamine, N-nitrosopyrrolidine, and N-nitrosodibutylamine, were observed in reservoirs and tap water, with average concentrations of 4.7-8.9 and 2.8-6.7ng/L, respectively. The average carcinogenic risks caused by these N-nitrosamines were higher than the risk level of 10-6. Three-dimensional fluorescence excitation-emission matrix modeling revealed that DOM was composed of humus-like component 1 (C1) and protein-like component 2 (C2). Fluorescence indicators showed that DOM in reservoir water was mainly affected by exogenous pollution and algal growth, whereas in tap water, DOM was mainly affected by microbial growth with strong autopoietic properties. In the reservoir water, N-nitrosodiethylamine and N-nitrosopiperidine were significantly correlated with C2 and biological indicators, indicating their endogenously generated sources. Based on the principal component analysis and multiple linear regression methods, five sources of N-nitrosamines were identified: agricultural pollution, microbial sources, humus sources, degradation processes, and other factors, accounting for 46.8%, 36.1%, 7.82%, 8.26%, and 0.96%, respectively. For tap water, two sources, biological reaction processes, and water distribution systems, were identified, accounting for 75.7% and 24.3%, respectively. Overall, this study presents quantitative information on N-nitrosamines' sources based on DOM properties in typical karst reservoirs and tap water, providing a basis for the safety of drinking water for consumers.

Water quality parameters from artesian wells located in Lagarto-SE

The objective of this work was to analyse the water quality of eight artesian wells in the municipality of Lagarto/ SE to determine if they comply with the standards of potability of the Ministry of Health Decree No. 888/2021 and the classification of Resolution CONAMA 357/2005. The methodology followed was the evaluation of the physicochemical parameters of the water samples. The results demonstrate some non-conformities in the physico-chemical analyses. Through methods and statistical tools, ANOVA (analysis of variance) and PCA (principal component analysis), it was possible to explain the variation of the data collected in this work, where the main component 1 (CP1) explained 54,41% of the variance of the data, (CP2) explained 20,43% and, at last (CP3) explained 15,17%, totaling 90,01% of the explanation of the variation of the data collected.

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.

Abstract 154: Crossroads Of Immunity And Coagulation: Protease Activity Of Complement Component C1s Inhibits Protein C Activation In Covid-19

Introduction: There is an abundance of crosstalk between innate immunity and coagulation in thrombo-inflammatory diseases such as COVID-19. Several potential procoagulant effects of complement activation have been described, but whether complement proteases affect antithrombotic pathways in is not known. Aims: We tested the hypothesis that complement component 1 (C1) impairs the protein C antithrombotic pathway. Methods: Plasma levels of the C1 complex were measured in plasma collected from healthy individuals and COVID-19 patients hospitalized early in the COVID-19 pandemic (NCT04360824). Activation of protein C by recombinant thrombomodulin was measured in a two-stage assay. In the first stage, protein C, thrombin, and thrombomodulin were incubated to generate activated protein C. The reaction mix was incubated with either healthy or COVID-19 plasma in the presence or absence of C1 inhibitor (which binds to C1 to inhibit its activity) or sutimlimab (a specific C1s inhibitor). During the second stage, protein C activation was measured using spectrophotometry with the chromogenic substrate S-2366. Results: In the inflammatory condition COVID-19, plasma levels of the C1 complex were increased compared to healthy controls ( p = 0.004). Generation of activated protein C by recombinant thrombomodulin was decreased in the presence of COVID-19 plasma ( p = 0.001), whereas the addition of healthy plasma had no significant effect ( p = 0.469). Addition of C1 inhibitor or sutimlimab normalized protein C activation in the presence of COVID-19 plasma ( p = 0.022 and 0.039, respectively). Addition of recombinant C1s protease, but not C1r protease or C1q, impaired the generation of activated protein C in a dose dependent fashion. Conclusions: We have uncovered a novel role of the protease C1s in regulating the generation of anticoagulant activated protein C. These findings suggest that the classical pathway of complement activation may impair anticoagulant responses in COVID-19.

Complement component C1q initiates extrinsic coagulation via the receptor for the globular head of C1q in adventitial fibroblasts and vascular smooth muscle cells.

Vascular diseases are highly associated with inflammation and thrombosis. Elucidating links between these two processes may provide a clearer understanding of these diseases, allowing for the design of more effective treatments. The activation of complement component 1 (C1) is a crucial contributor to innate immunity and is associated with significant concentrations of circulating C1q. Many pathological pathways initiate when C1q interacts with gC1qR. This interaction plays a major role in inflammation observed during atherosclerosis and the initiation of intrinsic coagulation. However, the effects of C1 and the role of C1q/gC1qR on extrinsic coagulation, which is the more physiologically relevant coagulation arm, has not been studied. We hypothesized that C1q binding to gC1qR enhances the expression of tissue factor (TF) in adventitial fibroblastsand vascular smooth muscle cells, the primary TF bearing cells in the body. Using an enzyme-linked immunosorbent assay approach, TF expression and the role of gC1qR was observed. Cells were conditioned for 1 hwith C1q or a gC1qR blocker and C1q, to assess the role of gC1qR. Additionally, cell growth characteristics were monitored to assess changes in viability and metabolic activity. Our results indicate that the expression of TF increased significantly after incubation with C1q as compared with unconditioned cells. Cells conditioned with gC1qR blockers and C1q exhibited no change in TF expression when compared with cells conditioned with the blocking antibodies alone. Our results show no significant differences in metabolic activity or cell viability under these conditions. This indicates that gC1qR association with C1q induces TF expression and may initiate extrinsic coagulation. Overall, this data illustrates a role for C1q in the activation of extrinsic coagulation and that gC1qR activity may link inflammation and thrombosis.

High exogenous humus inhibits greenhouse gas emissions from steppe lakes

Although freshwater lakes are considered to be an important source of greenhouse gas (GHG) emissions, the potential driving mechanisms of such emissions are not well understood, especially in steppe lakes. In this study, the GHG emission characteristics in Hulun Lake Basin, including Hulun Lake, Beier Lake, Wulannuoer Lake, and their surrounding watersheds were investigated. The average methane (CH4) and nitrous oxide (N2O) emission fluxes released from rivers were 67.84 ± 20.53 and 0.11 ± 0.04 μg m−2·min−1, which were larger than those of lakes, with values of 28.60 ± 13.02 and 0.06 ± 0.02 μg m−2·min−1, respectively. Conversely, the average carbon dioxide (CO2) emission flux from lakes (1816.58 ± 498.98 μg m−2·min−1) was higher than that of rivers of (1795.41 ± 670.49 μg m−2·min−1). The water in Hulun Lake Basin was rich in organic matter and had a high chemical oxygen demand (COD). Three-dimensional fluorescence combined with a parallel factor analysis (3D-EEM-PARAFAC) demonstrated that the organic matter was composed of four humus types (from Component 1 (C1) to Component 4 (C4)), of which, C1 and C4 were terrestrial humus. The fluorescence index (FI) and humification index (HIX) indicated that the organic matter in the water was mainly imported from exogenous humus. The GHG emission fluxes were negatively correlated with these four components, indicating that GHG emissions were mainly affected by the organic matter source and components, and humus was the most important factor that inhibited GHG emissions in steppe lakes. However, the GHG emission flux was relatively high in some areas of the lake, especially in areas with high nutrient levels or where algal blooms occurred, as evidenced by the significantly positive correlations with total nitrogen (TN), total phosphorous (TP), and chlorophyll-a (chl-a) (p < 0.01). The algae-derived organic matter simulated the decomposition of refractory humus, thus, promoting GHG emissions. These findings are crucial for accurately evaluating the GHG emission fluxes, understanding the carbon cycle, and proposing future management strategies for steppe lakes.

C1q/gC1qR‐Mediated Activation of Extrinsic Coagulation

Vascular diseases can be characterized by and are highly associated with inflammation and thrombosis. Understanding the relationship between these two processes would allow us to treat vascular diseases more effectively. Activation of complement component 1 (C1) is an important contributor to innate immunity and drives the lysis of pathogens. Upon C1 activation, significant concentrations of circulating C1q are observed. C1q has an affinity for gC1qR, a cell surface receptor molecule that is commonly found on cell types associated with cardiovascular disease development, including vascular smooth muscle cells and adventitial fibroblasts. When C1q interacts with gC1qR, many pathological pathways initiate including the inflammation observed during atherosclerosis and the initiation of intrinsic coagulation. The intrinsic cascade plays a minor role in physiological and pathological thrombosis as compared with the extrinsic cascade; however, the effects of C1 and the role of C1q/gC1qR on extrinsic coagulation have not been studied. We hypothesized that C1q association with gC1qR would increase the amount of sub‐endothelial tissue factor expressed by both aortic adventitial fibroblasts (HAAFs) and coronary artery smooth muscle cells (HCASMCs). We investigated these cells as they are the primary TF bearing cells. Further, we hypothesized that the newly expressed TF would bind Factor VII (FVII), which is a crucial step in the initiation of the extrinsic coagulation cascade. The objective of this study was to elucidate the role of C1q/gC1qR‐mediated TF expression and its ability to bind FVII. Using a solid‐phase ELISA, TF expression was observed on both HAAFs and HCASMCs. Cells were conditioned for one hour with either platelet poor plasma (PPP), purified human C1q, LPS, or a combination of anti‐gC1qR antibodies (60.11 or 74.5.2 clones) and purified human C1q, PPP, or LPS. A capture ELISA approach was used to evaluate the binding of FVII to TF following exposure after the same conditions. Further, a capture ELISA was used to assess released TF. Finally, a live/dead assay to assess cell viability and cell density, as well as an MTT assay to assess cell metabolic activity were performed to monitor cell culture conditions. Our results indicate that HAAF and HCASMC expression of TF increased significantly by approximately 15% after incubation with C1q as compared with unconditioned cells. Cells conditioned with gC1qR blocking antibodies prior to conditioning with C1q exhibited no change in TF expression when compared to unconditioned cells. Additionally, a 20‐30% increase in bound FVII was observed in both cell types when conditioned with C1q as compared to unconditioned cells. Our results show no significant differences in metabolic activity or cell viability under these conditions; however, cell density decreased by approximately 25% when conditioned with C1q. Overall, this data illustrates a role for C1q in activation of extrinsic coagulation. Furthermore, our data suggests that gC1qR acts a mediator for both inflammation and thrombosis. This work serves as a meaningful step in identifying a convergence of inflammatory and thrombotic pathways that can be therapeutically targeted to mitigate vascular diseases. Future work will determine if the expressed TF and the binding of FVII can support downstream extrinsic coagulation activity. Thanks to NIH for supporting this research.

Multi-spectroscopic investigation of the molecular weight distribution and copper binding ability of dissolved organic matter in Dongping Lake, China

The properties and metal-binding abilities of dissolved organic matter (DOM) rely on its molecular weight (MW) structure. In this study, the spatial differences of DOM in compositions, MW structures, and binding mechanisms with copper (Cu2+) in Dongping Lake were investigated by applying excitation-emission matrix combining parallel factor analysis (EEM-PARAFAC), synchronous fluorescence (SF) spectra, two-dimensional correlation spectra (2D-COS), and Fourier transform infrared (FTIR) spectra. The EDOM for the entrance of the Dawen River and PDOM for the macrophyte-dominated region were divided from DOM of Dongping Lake based on hierarchical clustering analysis (HCA) and principal component analysis (PCA) and were size-fractioned into MW < 500 kDa and <100 kDa fractions. According to EEM-PARAFAC, Dongping Lake was dominated by tryptophan-like substances with MW < 500 kDa. The concentration of PDOM was higher than that of EDOM (p < 0.05). 2D-COS showed that protein-like components preceded humic-like components binding to Cu2+ regardless of sample type (215 nm > 285 nm > 310–360 nm). The Cu2+ binding capacity of DOM exhibited specific differences in space, components, and molecular weights. The humic-like component 1 (C1) and tryptophan-like component 4 (C4) of PDOM showed stronger binding abilities than those of EDOM. Endogenous tryptophan-like component 4 (C4) had a higher binding affinity for Cu2+ than humic-like components (logKa: C4 > C1 > C2) in PDOM irrespective of MW. Humic-like components with MW < 500 kDa displayed higher binding potentials for Cu2+. FTIR spectra showed that the main participants of DOM-Cu complexation included aromatic hydrocarbons, aliphatic groups, amide Ⅰ bands, and carboxyl functional groups. This study provides spatial-scale insights into the molecular weight structure of DOM in influencing the behavior, fate, and bioavailability of heavy metals in lakes.

Characterization of colored dissolved organic matter along the western continental shelf of India during the seasonal hypoxia

In recent years the number of hypoxic zones in the world is rising, which has been attributed to the increase in nutrients and organic matter due to anthropogenic activities. Here we present results on the variations in absorption and fluorescent characteristics of colored dissolved organic matter (CDOM) along the western continental shelf of India (WCSI) during the southwest monsoon (SWM). WCSI hosts the world's largest, natural, seasonal hypoxic zone covering 180,000 km2. Water samples were collected along 4 transects off Kochi, Mangalore, Karwar, and Goa during SWM (June–July 2018) for CDOM and other hydrographic parameters. Also, the Goa transect was sampled during late SWM (September–November 2016 and September 2018) when the bottom waters experienced low oxygen conditions. The onset of upwelling was observed in the hydrographical parameters with low temperature and dissolved oxygen (DO) in the sub-surface waters along the Kochi transect. DO decreased with depth and was hypoxic to suboxic in the bottom waters at most of the stations. The Highest CDOM absorption was observed at the nearshore stations of the Goa transect during July 2018. The waters along the Goa transect were highly stratified during the late SWM (September 2018) with low saline waters at the surface (due to heavy rainfall), with the development of hypoxia in the bottom waters. High CDOM absorption (ag412 m-1) was observed in the bottom waters, which coincided with hypoxic/suboxic conditions. Spectral slope S250-600 also showed a significant difference between the oxygen rich and low oxygenated waters. Three fluorescent DOM components were identified from PARAFAC analysis. Component 1 (C1) and 2 (C2) were humic-like with emissions in the longer wavelength, while component 3 (C3) was protein-like with excitation and emission in the UV region. CDOM absorption (ag412) and humic-like fluorescent component (C1) showed a positive correlation with apparent oxygen utilization (AOU), indicating the role of microbial respiration for CDOM production. Laboratory experiments also shed light on the CDOM production in low oxygenated waters when sufficient organic matter is available. This study gives insight into understanding dissolved organic matter's cycling under varying oxygen conditions prevailing in these waters.

Complement Component C1q Initiates Extrinsic Coagulation via gC1qR

Vascular diseases can be characterized by and are highly associated with inflammation and thrombosis. Understanding the relationship between these two processes would allow us to treat vascular diseases more effectively. Activation of complement component 1 (C1) is an important contributor to innate immunity and drives the lysis of pathogens. Upon C1 activation, significant concentrations of circulating C1q are observed. When C1q interacts with gC1qR, many pathological pathways initiate and C1q/gC1qR plays a role in inflammation observed during atherosclerosis and the initiation of intrinsic coagulation. While this link between inflammation and thrombosis is important, the intrinsic cascade plays a minor role in physiological and pathological thrombosis. The effects of C1 and the role of C1q/gC1qR on extrinsic coagulation, which is more physiologically relevant, has not been studied. Extrinsic coagulation initiates upon vessel damage, exposing tissue factor (TF) to the blood. Adventitial fibroblasts and vascular smooth muscle cells are the primary TF bearing cells. We hypothesized that C1q binding to gC1qR enhances the expression of sub-endothelial TF in these cells. The objective of this study was to elucidate the role of gC1qR-mediated TF expression. Using a solid-phase ELISA, TF expression was observed on aortic adventitial fibroblasts (HAAFs) and coronary artery smooth muscle cells (HCASMCs). Cells were conditioned for one hour with either platelet poor plasma (PPP), purified human C1q, LPS, or a combination of anti-gC1qR (60.11 or 74.5.2 clones) and purified human C1q, PPP, or LPS. Further, a capture ELISA was used to assess released TF under these same conditions. Finally, a live/dead assay to assess cell viability and cell density, as well as an MTT assay to assess cell metabolic activity were performed to monitor cell culture conditions. Our results indicate that HAAF and HCASMC expression of TF increased significantly by approximately 35% after incubation with C1q as compared with unconditioned cells. Cells conditioned with gC1qR blocking antibodies exhibited a marked decrease in TF expression (by approximately 10-20%) when compared to cells conditioned with C1q alone. Our results show no significant differences in metabolic activity or cell viability under these conditions; however, cell density decreased by approximately 25% when conditioned with C1q. Overall, this data illustrates a role for C1q in activation of extrinsic coagulation. Furthermore, our data suggests that gC1qR acts a mediator for both inflammation and thrombosis. This work serves as a meaningful step in identifying common inflammatory and thrombotic pathways that can be therapeutically targeted to mitigate vascular diseases. Future work will determine if the expressed TF can support extrinsic coagulation activity.

A solar photo-thermochemical hybrid system using peroxydisulfate for organic matters removal and improving ultrafiltration membrane performance in surface water treatment

Solar energy is considered one of the most promising energy sources for the degradation of pollutants in the water treatments. An innovative solar photo-thermochemical system involving peroxydisulfate (PDS) as an oxidant and xenon lamp as a solar irradiation light source was applied with hopes to degrade organic matters and alleviate the ultrafiltration (UF) membrane fouling when treating the real surface water. Moreover, heat-activated PDS pretreatment was used as a comparison to explore the respective proportions of solar light and heating effects, finding that solar light effect dominated the activation of PDS to degrade natural organic matters (NOMs) when the reaction temperature was below 50 °C and they both contributed to the production of free radicals at the temperature of >50 °C. The results indicated that solar-activated PDS pretreatment significantly alleviated membrane fouling caused by Songhua River water with the highest transmembrane pressure (TMP) reduction of approximately 69.6% at 70 °C. Organic substances (characterized by DOC, UV254 and the maximum florescent intensity) and micropollutant (atrazine) in the feed water were better degraded in the presence of solar light. Both total fouling index (TFI) and hydraulic irreversible fouling index (HIFI) were moderate correlated with the UV254 and DOC, whereas remarkably correlated with the Fmax of component 2 (C2) and component 3 (C3). In addition, no significant correlation was observed between fouling indexes (TFI and HIFI) and the Fmax of component 1 (C1). The membrane irreversible fouling was attributed to the accumulation of cake layers mainly composed of protein-like substances on the membrane surface. Solar-activated PDS pretreatment would efficiently degrade the protein-like substances and terrestrially derived humic-like matters to control UF membrane fouling. The findings are beneficial to develop new strategies for membrane fouling alleviation based on the solar irradiation and PDS oxidation.

The Inhibition of Complement System in Formal and Emerging Indications: Results from Parallel One-Stage Pairwise and Network Meta-Analyses of Clinical Trials and Real-Life Data Studies

This manuscript presents quantitative findings on the actual effectiveness of terminal complement component 5 (C5) inhibitors and complement component 1 (C1) esterase inhibitors through their formal and common “off-label” (compassionate) indications. The results emanated from pairwise and network meta-analyses to present evidence until September 2019. Clinical trials (CT) and real-life non-randomized studies of the effects of interventions (NRSI) are consistent on the benefits of C5 inhibitors and of the absence of effects of C1 esterase inhibitors (n = 7484): Mathematically, eculizumab (surface under the cumulative ranking area (SUCRA) >0.6) and ravulizumab (SUCRA ≥ 0.7) were similar in terms of their protective effect on hemolysis in paroxysmal nocturnal hemoglobinuria (PNH), thrombotic microangiopathy (TMA) in atypical hemolytic uremic syndrome (aHUS), and acute kidney injury (AKI) in aHUS, in comparison to pre-/off-treatment state and/or placebo (SUCRA < 0.01), and eculizumab was efficacious on thrombotic events in PNH (odds ratio (OR)/95% confidence interval (95% CI) in CT and real-life NRSI, 0.07/0.03 to 0.19, 0.24/0.17 to 0.33) and chronic kidney disease (CKD) occurrence/progression in PNH (0.31/0.10 to 0.97, 0.66/0.44 to 0.98). In addition, meta-analysis on clinical trials shows that eculizumab mitigates a refractory generalized myasthenia gravis (rgMG) crisis (0.29/0.13 to 0.61) and prevents new acute antibody-mediated rejection (AMR) episodes in kidney transplant recipients (0.25/0.13 to 0.49). The update of findings from this meta-analysis will be useful to promote a better use of complement inhibitors, and to achieve personalization of treatments with this class of drugs.

Effect of swine biogas slurry application on soil dissolved organic matter (DOM) content and fluorescence characteristics

The application of biogas slurry as an organic fertilizer is a promising method for utilizing breeding manure wastewater. At present, the impact of biogas slurry on the properties of organic matter in soil is not clear. In this study, a pot experiment in which chemical fertilizers were replaced with biogas slurry from a swine farm was performed. The fluorescence spectra combined with parallel factor (PARAFAC) analysis and principal component analysis (PCA) were used to explore the influence of biogas slurry on the protein and humic substance contents in the dissolved organic matter (DOM) in soil. The results showed that there were two proteins (component 3 (C3) and component 4 (C4)) and two humic substances ( component 1 (C1) and component 2 (C2)) in the DOM of the experimental soil. The application of swine biogas slurry can significantly increase the content of DOM in soil, but the increase was weakened with extended time. Compared with the CKA, the biogas slurry significantly increased the C1, C2, C3 and C4 contents in the initial stage by 116.17%, 76.41%, 578.71% and 278.13%, respectively. Within 28 days of planting corn, proteins with simple molecular structure in the DOM in the soil began to be transformed into humic substances with high molecular weight and more complex molecular structures. On the 60th day, the contents of C1 and C2 in the DOM of the treated treatments soil increased by 13.72%–34.40% and 5.05%–17.78% respectively, and tyrosine content decreased by 90.11%–94.41%. This study provides a new perspective on the effects of biogas slurry application on soil properties and sustainable utilization of soil.

In vitro evaluation of complement deposition and opsonophagocytic killing of Rhodococcus equi mediated by poly-N-acetyl glucosamine hyperimmune plasma compared to commercial plasma products.

BackgroundThe bacterium Rhodococcus equi can cause severe pneumonia in foals. The absence of a licensed vaccine and limited effectiveness of commercial R. equi hyperimmune plasma (RE‐HIP) create a great need for improved prevention of this disease.HypothesisPlasma hyperimmune to the capsular polysaccharide poly‐N‐acetyl glucosamine (PNAG) would be significantly more effective than RE‐HIP at mediating complement deposition and opsonophagocytic killing (OPK) of R. equi.AnimalsVenipuncture was performed on 9 Quarter Horses.MethodsThe ability of the following plasma sources to mediate complement component 1 (C1) deposition onto either PNAG or R. equi was determined by ELISA: (1) PNAG hyperimmune plasma (PNAG‐HIP), (2) RE‐HIP, and (3) standard non‐hyperimmune commercial plasma (SP). For OPK, each plasma type was combined with R. equi, equine complement, and neutrophils isolated from horses (n = 9); after 4 hours, the number of R. equi in each well was determined by quantitative culture. Data were analyzed using linear mixed‐effects regression with significance set at P < .05.ResultsThe PNAG‐HIP and RE‐HIP were able to deposit significantly (P < .05) more complement onto their respective targets than the other plasmas. The mean proportional survival of R. equi opsonized with PNAG‐HIP was significantly (P < .05) less (14.7%) than that for SP (51.1%) or RE‐HIP (42.2%).Conclusions and Clinical ImportancePlasma hyperimmune to PNAG is superior to RE‐HIP for opsonizing and killing R. equi in vitro. Comparison of these 2 plasmas in field trials is warranted because of the reported incomplete effectiveness of RE‐HIP.

Initiation of Extrinsic Coagulation via Complement

Thrombosis and inflammation are prevalent during and characteristic of vascular diseases. Understanding the interplay of these processes would likely allow us to treat vascular diseases more effectively. Activation of complement component 1 (C1) is an important contributor to innate immunity and ultimately drives the lysis of pathogens. Upon C1 activation, significant concentrations of circulating C1q are observed. C1q interacting with gC1qR, one of its cell‐surface receptors, initiates many pathological pathways. There is considerable evidence that C1q/gC1qR plays a role in inflammatory processes observed during atherosclerosis and in initiating intrinsic coagulation. While this link between inflammation and thrombosis is important, the intrinsic cascade plays a minor role in physiological and pathological thrombosis. The effects of C1 and the role of C1q on extrinsic coagulation, which is more physiologically relevant, has not been studied. Extrinsic coagulation initiates upon vessel damage, which exposes blood to sub‐endothelial constituents, especially tissue factor. Adventitial fibroblasts and vascular smooth muscle cells are the primary tissue factor bearing cells. We hypothesized that C1q binding to gC1qR initiates this pathway through the enhanced expression of sub‐endothelial tissue factor (TF) in these cells. The objective of this study was to elucidate the role of C1q‐mediated TF expression. Using a solid‐phase ELISA, TF expression was observed on aortic adventitial fibroblasts (HAAFs) and aortic smooth muscle cells (HASMCs) under various conditions. Cells were conditioned for one hour with either platelet poor plasma (PPP), purified human C1q, LPS, anti‐gC1qR (60.11), or a combination of 60.11 and purified human C1q. After the exposure conditions, cells were incubated for one hour with one of the following primary antibodies: anti‐human gC1qR (74.5.2), anti‐human TF, or anti‐human CD54 (ICAM‐1). Following primary antibody incubation, cells were incubated with an appropriate alkaline phosphatase conjugated secondary antibody for one hour. PNPP was added to the cells and allowed to develop for 30 minutes. Absorbance was documented at 405 nm. Our results suggest that HAAF and HASMC expression of TF increased significantly by approximately 35% after incubation with C1q as compared with unconditioned cells. We also observed increased ICAM expression, 20% above baseline, in HASMCs. Cells conditioned with gC1qR blocking antibodies exhibited a marked decrease in TF and ICAM expression (by approximately 10%) when compared to cells conditioned with C1q alone. Overall, this data illustrates a role for C1q in activation of extrinsic coagulation. Furthermore, our data suggests that gC1qR acts a mediator for both inflammatory and thrombotic reactions that may occur during vascular diseases. This work illustrates a step in identifying common inflammatory and thrombotic pathways that can be therapeutically targeted to mitigate vascular diseases and may provide new information regarding off‐target effects. Future work will determine if the expressed TF can support extrinsic coagulation activity and new therapeutics can be designed to mitigate vascular disease processes.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

PNAG-specific equine IgG1 mediates significantly greater opsonization and killing of Prescottella equi (formerly Rhodococcus equi) than does IgG4/7

Prescottella equi (formerly Rhodococcus equi) is a facultative intracellular bacterial pathogen that causes severe pneumonia in foals 1–6 months of age, whereas adult horses are highly resistant to infection. We have shown that vaccinating pregnant mares against the conserved surface polysaccharide capsule, β-1 → 6-linked poly-N-acetyl glucosamine (PNAG), elicits opsonic killing antibody that transfers via colostrum to foals and protects them against experimental infection with virulent.R. equi. We hypothesized that equine IgG1 might be more important than IgG4/7 for mediating protection against R. equi infection in foals. To test this hypothesis, we compared complement component 1 (C1) deposition and polymorphonuclear cell-mediated opsonophagocytic killing (OPK) mediated by IgG1 or IgG4/7 enriched from either PNAG hyperimmune plasma (HIP) or standard plasma. Subclasses IgG1 and IgG4/7 from PNAG HIP and standard plasma were precipitated onto a diethylaminoethyl ion exchange column, then further enriched using a protein G Sepharose column. We determined C1 deposition by enzyme-linked immunosorbent assay (ELISA) and estimated OPK by quantitative microbiologic culture. Anti-PNAG IgG1 deposited significantly (P < 0.05) more C1 onto PNAG than did IgG4/7 from PNAG HIP or subclasses IgG1 and IgG4/7 from standard plasma. In addition, IgG1 from PNAG HIP mediated significantly (P < 0.05) greater OPK than IgG4/7 from PNAG HIP or IgG1 and IgG4/7 from standard plasma. Our findings indicate that anti-PNAG IgG1 is a correlate of protection against R. equi in foals, which has important implications for understanding the immunopathogenesis of R. equi pneumonia, and as a tool for assessing vaccine efficacy and effectiveness when challenge is not feasible.

Using fluorescence-parallel factor analysis for assessing disinfection by-product formation and natural organic matter removal efficiency in secondary treated synthetic drinking waters

Parallel factor (PARAFAC) analysis of fluorescence excitation-emission matrices (EEMs) was used to investigate the organic matter and DBP formation characteristics of untreated, primary treated (enhanced coagulation; EC) and secondary treated synthetic waters prepared using a Suwannee River natural organic matter (SR-NOM) isolate. The organic matter was characterised by four different fluorescence components; two humic acid-like (C1 and C2) and two protein-like (C3 and C4). Secondary treatment methods tested, following EC treatment, were; powdered activated carbon (PAC), granular activated carbon (GAC), 0.1% silver-impregnated activated carbon (SIAC), and MIEX® resin. Secondary treatments were more effective at removing natural organic matter (NOM) and fluorescent DBP-precursor components than EC alone. The formation of a suite of 17 DBPs including chlorinated, brominated and iodinated trihalomethanes (THMs), dihaloacetonitriles (DHANs), chloropropanones (CPs), chloral hydrate (CH) and trichloronitromethane (TCNM) was determined after chlorinating water sampled before and after each treatment step. Regression analysis was used to investigate the relationship between peak component fluorescence intensity (FMAX), DBP concentration and speciation, and more commonly used aggregate parameters such as DOC, UV254 and SUVA254. PARAFAC component 1 (C1) was in general a better predictor of DBP formation than other aggregate parameters, and was well correlated (R ≥ 0.80) with all detected DBPs except dibromochloromethane (DBCM) and dibromoacetonitrile (DBAN). These results indicate that the fluorescence-PARAFAC approach could provide a robust analytical tool for predicting DBP formation, and for evaluating the removal of NOM fractions relevant to DBP formation during water treatment.

Avaliação sazonal e espacial da qualidade das águas superficiais da bacia hidrográfica do rio Longá, Piauí, Brasil

Este artigo teve como objetivo avaliar a qualidade das águas superficiais da bacia hidrográfica do rio Longá no Estado do Piauí, Brasil. Sete pontos foram monitorados, com base na proximidade de estações meteorológicas e acessibilidade, para obter melhores caracterizações espaciais e de ocupação da bacia hidrográfica. As amostras de água foram coletadas mensalmente (01/2015-12/2015). Foram medidas as variáveis: temperatura, pH, turbidez, condutividade elétrica, sólidos totais, fósforo total, nitrato, oxigênio dissolvido, demanda bioquímica de oxigênio (DBO) e Escherichia coli (E. coli). Os resultados foram avaliados individualmente e comparados aos padrões brasileiros para águas doces de classe 2, conforme Resolução nº 357/2005 do Conselho Nacional do Meio Ambiente (CONAMA). Os dados foram avaliados utilizando-se análises multivariadas (agrupamento e análise de componentes principais - ACP). As variáveis E. coli, pH, turbidez, nitrato, DBO e condutividade elétrica variaram entre os dois períodos analisados ​​(seco e chuvoso). As variáveis DBO, E. coli estiveram em desacordo com os padrões do CONAMA nos pontos urbanos P2, P3 e P5, devido à poluição difusa observada no período chuvoso. O agrupamento mostrou uma tendência espacial, apresentando dois grupos distintos (rural e urbano). A ACP identificou quatro componentes principais que explicaram uma variação de 58,64% nos dados. O Componente 1 (CP1) refletiu uma contribuição de poluentes relacionados à poluição difusa de áreas agrícolas e urbanas. O Componente 2 (CP2) foi fortemente associado à poluição, especialmente por esgoto doméstico não tratado. As outras ACPs também refletiram pressões de atividades antropogênicas nos corpos d’água.

Encapsulation of the reductase component of p-hydroxyphenylacetate hydroxylase in poly(lactide-co-glycolide) nanoparticles by three different emulsification techniques.

p-Hydroxyphenylacetate 3-hydroxylase component 1 (C1) is a useful enzyme for generating reduced flavin and NAD+ intermediates. In this study, poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) were used to encapsulate the C1 (PLGA-C1 NPs). Enzymatic activity, stability, and reusability of PLGA-C1 NPs prepared using three different methods [oil in water (o/w), water in oil in water (w/o/w), and solid in oil in water (s/o/w)] were compared. The s/o/w provided the optimal conditions for encapsulation of C1(PLGA-C1,s NPs), giving the highest enzyme activity, stability, and reusability. The s/o/w method improves enzyme activity ∼11 and 9-fold compared to w/o/w (PLGA-C1,w NPs) and o/w (PLGA-C1,o NPs). In addition, s/o/w prepared PLGA-C1,s NPs could be reused 14 times with nearly 50% activity remaining, a much higher reusability compared to PLGA-C1,o NPs and PLGA-C1,w NPs. These nanovesicles were successfully utilised to generate reduced flavin mononucleotide (FMN) and supply this cofactor to a hydroxylase enzyme that has application for synthesising anti-inflammatory compounds. Therefore, this recycling biocatalyst prepared using the s/o/w method is effective and has the potential for use in combination with other enzymes that require reduced FMN. Application of PLGA-C1,s NPs may be possible in additional biocatalytic processes for chemical or biochemical production.