Cytochrome Content Research Articles

ASSOCIATIONS BETWEEN BLOOD CONCENTRATIONS OF CYTOTOXIC CD8+ CELLS AND LYMPHOCYTE APOPTOSIS IN HEALTHY HUMANS

Introduction: Cytotoxic T-lymphocytes play an important role in a specific immune response via a lytic effect in relation to abnormal cells. The number of these cells greatly increases sin pathological states. In addition, an increased cytotoxic activity is a characteristic of the immune response of people living in the Far NorthTherefore, it is important to understand what function cytotoxic T-lymphocytes predominantly perform in healthy people in the North - cytolytic or suppressor? Aim: To assess associations between CD8+ concentration and immune response and apoptotic deaths of the lymphocytes in healthy individuals. Methods: Ninety-three healthy adult residents of the Russian North comprised the sample. Apoptotic lymphocyte death was studied by flow cytometry. FITC-annexine-V and propidium iodide labelled cells were detected. Concentrations of cytokines and apoptosis mediators were assessed by a solid-phase enzyme immunoassay. Neutrophiles, monocytes and phagocytic activity of neutrophils were studied in blood smears stained by Romanowsky's - Giemsa. The level of phenotypic activity of lymphocytes was assessed by double peroxidase labeling using monoclonal antibodies. The data were presented using means, standard deviations, medians, the 1st and the 3rd quartiles. All study participants were divided into two groups: with normal- (0.2-0,4 х109 kl/l) and elevated (more 0,6 х109 kl/l) blood cytotoxic lymphocyte levels. The groups were similar in terms of by age- and gender distribution. Continuous variables were analyzed using Mann-Whitney tests. Results: In individuals with an increased level of cytotoxic CD8+ lymphocytes in peripheral venous blood had greater concentrations of leukocytes (7.4 ± 0.49 х109 cells/l vs. 5.5 ± 0.23 х109, p = 0.003), lymphocytes (2.8 ± 0.17 х109 cells/l vs.1.8 ± 0.07 х109 cells/t p = 0.005), and mature neutrophils (4.1 ± 0.19 х109 cells/l vs. 3.4 ± 0.49 х109 cells/l, p = 0.013). No associations between the level of apoptosis of lymphocytes (AnV+/PI-) and concentrations of sFasL, TRAIL, TNFa, and cytochrome C were observed. Conclusions: Lymphocytes CD8+ in healthy residents of the Russian North perform mainly cytotoxic function, which is not related to apoptotic cellular death.

Periplaneta americana extract ameliorates lipopolysaccharide-induced liver injury by improving mitochondrial dysfunction via the AMPK/PGC-1α signaling pathway.

Periplaneta americana (PA) extract acts clinically as a therapeutic treatment in various diseases; it enhances liver function in mouse models and mitigates the pathological condition of liver fibrosis. The present study aimed to investigate the role and potential mechanisms underlying the action of the PA extract, xinmailong (XML), in lipopolysaccharide (LPS)-induced liver injury. Following the treatment of AML12 cells with LPS, the content of cytochrome c in the cytoplasm and mitochondria, and the level of ATP synthesis were detected using corresponding kits. The relative mRNA expression levels of nuclear respiratory factor 1 and transcription factor A, mitochondrial were investigated using reverse transcription-quantitative (RT-q)PCR analysis. The MTT assay was performed to detect the viability of AML12 cells following treatment with XML, in the absence or presence of LPS. Western blot analysis was performed to determine the expression levels of proteins in the AMP-activated protein kinase (AMPK)/proliferator-activated receptor γ coactivator-1α (PGC-1α) pathway. Following treatment with compound C, an inhibitor of AMPK, the expression levels of inflammatory cytokines were determined using ELISA and RT-qPCR analysis. The levels of oxidative stress-related markers were detected using corresponding kits following treatment with compound C. In addition, TUNEL staining was performed to detect the apoptosis of AML12 cells, and western blot analysis was performed to investigate the expression levels of apoptosis-related proteins. Mitochondrial dysfunction was induced by LPS in AML12 cells. LPS stimulation significantly downregulated the expression of proteins in the AMPK/PGC-1α pathway, which was reversed following treatment with XML. In addition, inflammation, oxidative stress and mitochondrial dysfunction induced by LPS were alleviated by XML in AML12 cells. However, the addition of compound C and XML to LPS-induced AML12 cells resulted in the aggravation of cell injury. Collectively, the results of the present study indicated that XML suppressed mitochondrial dysfunction induced by LPS by activating AMPK/PGC-1α signaling. Thus, the results of the present study may contribute to further understanding of the underlying mechanism via which XML alleviates liver injury.

Hypericin Enhances Paclitaxel-Induced B16-F10 Cell Apoptosis by Activating a Cytochrome c Release–Dependent Pathway

The enhanced inhibitory effect of paclitaxel (PTX) combined with hypericin (HY) on B16-F10 cells may be realized through the ROS-related cytochrome c release pathway. The apoptotic characteristics of the B16-F10 cells, such as DNA fragmentation, chromatin condensation, and apoptotic body formation, were all enhanced in the combined treatment group. Further investigation showed that the combination of paclitaxel and HY could increase the level of mitochondrial damage and the concentration of cytochrome c, causing the expression of caspase-3 and the cleavage of PARP.1. Compared with paclitaxel or HY alone, the level of reactive oxygen species (ROS) increased significantly, while glutathione reductase (GR) activity and intracellular glutathione (GSH) levels decreased significantly in the combination group.

Functional Characterization of 21 Rare Allelic CYP1A2 Variants Identified in a Population of 4773 Japanese Individuals by Assessing Phenacetin O-Deethylation.

Cytochrome P450 1A2 (CYP1A2), which accounts for approximately 13% of the total hepatic cytochrome content, catalyzes the metabolic reactions of approximately 9% of frequently used drugs, including theophylline and olanzapine. Substantial inter-individual differences in enzymatic activity have been observed among patients, which could be caused by genetic polymorphisms. Therefore, we functionally characterized 21 novel CYP1A2 variants identified in 4773 Japanese individuals by determining the kinetic parameters of phenacetin O-deethylation. Our results showed that most of the evaluated variants exhibited decreased or no enzymatic activity, which may be attributed to potential structural alterations. Notably, the Leu98Gln, Gly233Arg, Ser380del Gly454Asp, and Arg457Trp variants did not exhibit quantifiable enzymatic activity. Additionally, three-dimensional (3D) docking analyses were performed to further understand the underlying mechanisms behind variant pharmacokinetics. Our data further suggest that despite mutations occurring on the protein surface, accumulating interactions could result in the impairment of protein function through the destabilization of binding regions and changes in protein folding. Therefore, our findings provide additional information regarding rare CYP1A2 genetic variants and how their underlying effects could clarify discrepancies noted in previous phenotypical studies. This would allow the improvement of personalized therapeutics and highlight the importance of identifying and characterizing rare variants.

Optimization of Multiparameters for Increased Yields of Cytochrome B5 in Bioreactors.

The production of recombinant proteins is gaining increasing importance as the market requests high quality proteins for several applications. However, several process parameters affect both the growth of cells and product yields. This study uses high throughput systems and statistical methods to assess the influence of fermentation conditions in lab-scale bioreactors. Using this methodology, it was possible to find the best conditions to produce cytochrome b5 with recombinant cells of Escherichia coli. Using partial least squares, the height-to-diameter ratio of the bioreactor, aeration rate, and PID controller parameters were found to contribute significantly to the final biomass and cytochrome concentrations. Hence, we could use this information to fine-tune the process parameters, which increased cytochrome production and yield several-fold. Using aeration of 1 vvm, a bioreactor with a height-to-ratio of 2.4 and tuned PID parameters, a production of 72.72 mg/L of cytochrome b5 in the culture media, and a maximum of product to biomass yield of 24.97 mg/g could be achieved.

Мітохондріальні цитохроми й ензими метаболізму гему в печінці щурів за умов різної забезпеченості раціону сахарозою та протеїном

The content of mitochondrial cytochromes and the activity of key enzymes of heme metabolism in the liver of rats under conditions of different dietary supply of protein and sucrose were investigated. The quantitative determination of mitochondrial cytochrome was performed by differential spectrophotometry, δ-aminolevulinate synthase activity was determined spectrophotometrically taking into account the molar extinction coefficient of 0.023x10(3) M(-1)sm(-1). Hemoxygenase activity was determined using the amount of formed bilirubin. It was found that under conditions of consumption of high-sucrose diet a significant decrease in the content of all mitochondrial cytochromes is noted: the content of cytochromes aa3, b and c1 decreases within 1.2-1.7 times, and content of cytochrome c decreases in two times. In the case of excessive consumption of sucrose on the background of alimentary protein deprivation the content of cytochromes b and c1 in the liver of rats does not differ statistically from similar indicators of the group of animals kept on a high-sucrose diet. At the same time, the content of cytochromes aa3 and c is significantly reduced. According to the activity of δ-aminolevulinate synthase under conditions of consumption of a high-sucrose diet, the studied enzymatic activity decreases by about 1.5 times with a simultaneous increase in the activity of heme oxygenase. Thus, there is a marked decrease in heme synthesis against the background of increased catabolism, which explains the decrease in the content of cytochromes in the mitochondria of the liver of rats under conditions of excess sucrose in the diet. The maximum increase in the activity of heme oxygenase (almost threefold) is observed in animals that were kept on a high-sugar diet deficient in protein content. Thus, dietary protein deficiency is a critical factor affecting the heme metabolism in the mitochondria of liver cells. The established changes in the content of mitochondrial cytochromes and the activities of key enzymes of heme metabolism in the liver could be considered as prerequisites for deepening its energy imbalance in conditions of different supply of sucrose and protein in diet.

Combined effects of carbon nanotubes and cadmium on the photosynthetic capacity and antioxidant response of wheat seedlings.

A detailed study of nanomaterials has revealed their broad application prospects. However, the presence of carbon nanotubes (CNTs) in the environment has been increasing and has aroused concerns regarding their toxicity to crops when combined with heavy metals. In the present study, the effects of Cd on the photosynthetic capacity and antioxidant activity of wheat seedlings in the presence of single-walled CNTs (SW) and multi-walled CNTs (MW) were investigated. Our results indicated that SW (5-40 mg L-1) and MW (10-40 mg L-1) significantly increased the oxidative stress response of wheat seedlings to Cd. Compared with Cd alone, CNTs combined with Cd decreased net photosynthetic rate, stomatal conductance, transpiration rate, primary maximum photochemical efficiency of photosystem II, actual quantum yield, photosynthetic electron transport rate, root canal protein, and ribulose-1,5-bisphosphate carboxylase/oxygenase content. Moreover, combined treatments increased the content of superoxide anion, superoxide dismutase, guaiacol peroxidase, cytochrome, and malondialdehyde in wheat seedlings. Moreover, membrane lipid peroxidation was aggravated, causing serious damage to the wheat membrane system. In addition, the toxicity of the SW treatment and the combined treatment with SW and Cd was higher than that of the MW treatment.

The Impact of the Humate Nature Feed Additives on the Antioxidative Status of Erythrocytes, Liver, and Muscle in Chickens, Hens, and Gerbils

Our study's goal was to determine the oxidation states of formed elements of the blood, and mitochondria of liver and muscles of chicken and gerbils could help establish and compare the humic substance's impact. The experiment was carried out on hens of the Lohmann Braun, chicken-broilers of the Cobb 500 cross, and 6-months-old gerbils, which were kept under standard conditions. Experimental hens, chicken-broilers, and gerbils were divided into 2 groups: 1 – intact animals (control), 2 – animals, which added to the drinking water a 1% solution of the fodder biologically active additive of humic nature (Humilid or Hydrohumate) in the optimal amount for 21 days. Results showed that Humilid and Hydrohumate tested in our experiment activated the protective systems in cells against toxic radicals. The species-specific effects of the components of studied humates were established: the activation of the glutathione blood protection system of hens; catalase activation in mitochondria of white muscles of chicken-broilers; activation of catalase and increase of concentration of cytochrome C in liver mitochondria of gerbils. Our results indicate the elucidation of the mechanisms of humic.

Mitochondrial biochemical and histopathological defects induced by the herbicide pendimethalin in tilapia fish (Oreochromis niloticus)

The mitochondrial defects were evaluated after administering tilapia fish, Oreochromis niloticus to sublethal doses (1.02 and 5.10 mg kg−1) of the herbicide pendimethalin (PD). All treatments exhibited a decrease in the cytochrome contents of gills, liver, and brain samples after 12, 24, and 48 h compared with the untreated individuals. However, malondialdehyde (MDA) levels were significantly increased in gills and liver samples. Also, the histopathological profiles showed significant swelling in mitochondria and intracellular spaces in cytoplasm of gills samples. The mitochondrial defects in the treated fish showed a slight decline in cytoplasm/mitochondria ratio (0.92-fold) compared to the control. In hepato-sections of treated fish, destructed mitochondria with less dense matrix as well as some vacuolated mitochondria with matrix disoriented cristae were noted. Similar patterns were observed in brain sections, where destructed axons and a significant decline in cytoplasm/mitochondria ratio (0.52-fold) were found. Therefore, the use of mitochondrial defects and histopathological alterations might represent good markers to assess the impact of herbicides on aquatic organisms. Moreover, the disorganization of cell components is considered an important sign of organ dysfunction.

Novel sulfonamide porphyrin TBPoS-2OH used in photodynamic therapy for malignant melanoma

The application of photodynamic therapy (PDT) for the treatment of skin diseases has been receiving much attention. Here, we examined the anti-tumor effect of a novel porphyrin-based photosensitizer TBPoS-2OH in the malignant melanoma A375 and B16 cells. TBPoS-2OH has obvious cell photo-cytotoxicity, but it has low cell dark-cytotoxicity. Further research showed that TBPoS-2OH is enriched in lysosomes after being taken up by cells. Subsequently, the apoptotic rates were significantly increased in TBPoS-2OH-treated A375 and B16 cells. The specific mechanism may be that after receiving light stimulation, TBPoS-2OH could effectively increase the level of intracellular reactive oxygen species (ROS), thereby activating mitochondrial apoptosis pathway-related proteins in A375 and B16 cells. We found an increase in the content of cytochrome C in the cytoplasm, and the levels of related proteins, such as cleaved caspase-3, cleaved caspase-9, and cleaved PARP1, were significantly increased in TBPoS-2OH-treated cells. These results indicated that the new compound TBPoS-2OH could be developed and become an alternative drug for the treatment of melanoma. Some reference ideas for the development of new photosensitizers are also provided.

The Interplay among Subunit Composition, Cardiolipin Content, and Aggregation State of Bovine Heart Cytochrome c Oxidase.

Mitochondrial cytochrome c oxidase (CcO) is a multisubunit integral membrane complex consisting of 13 dissimilar subunits, as well as three to four tightly bound molecules of cardiolipin (CL). The monomeric unit of CcO is able to form a dimer and participate in the formation of supercomplexes in the inner mitochondrial membrane. The structural and functional integrity of the enzyme is crucially dependent on the full subunit complement and the presence of unperturbed bound CL. A direct consequence of subunit loss, CL removal, or its oxidative modification is the destabilization of the quaternary structure, loss of the activity, and the inability to dimerize. Thus, the intimate interplay between individual components of the complex is imperative for regulation of the CcO aggregation state. While it appears that the aggregation state of CcO might affect its conformational stability, the functional role of the aggregation remains unclear as both monomeric and dimeric forms of CcO seem to be fully active. Here, we review the current status of our knowledge with regard to the role of dimerization in the function and stability of CcO and factors, such as subunit composition, amphiphilic environment represented by phospholipids/detergents, and posttranslational modifications that play a role in the regulation of the CcO aggregation state.

Metabolic rewiring in drug resistant cells exhibit higher OXPHOS and fatty acids as preferred major source to cellular energetics

Alteration in metabolic repertoire is associated with resistance phenotype. Although a common phenotype, not much efforts have been undertaken to design effective strategies to target the metabolic drift in cancerous cells with drug resistant properties. Here, we identified that drug resistant AML cell line HL-60/MX2 did not follow classical Warburg effect, instead these cells exhibited drastically low levels of aerobic glycolysis. Biochemical analysis confirmed reduced glucose consumption and lactic acid production by resistant population with no differences in glutamine consumption. Raman spectroscopy revealed increased lipid and cytochrome content in resistant cells which were also visualized as lipid droplets by Raman mapping, electron microscopy and lipid specific staining. Gene set enrichment analysis data from sensitive and resistant cell lines revealed significant enrichment of lipid metabolic pathways in HL-60/MX2 cells. Further, HL-60/MX2 possessed higher mitochondrial activity and increased OXPHOS suggesting the role of fatty acid metabolism as energy source which was confirmed by increased rate of fatty acid oxidation. Accordingly, OXPHOS inhibitor increased sensitivity of resistant cells to chemotherapeutic drug and fatty acid oxidation inhibitor Etomoxir reduced colony formation ability of resistant cells demonstrating the requirement of fatty acid metabolism and dependency on OXPHOS by resistant leukemic cells for survival and tumorigenicity.

The Effect of Aerobic Training and Tribulus terrestris Extract on Some Oxidative Stress Indices in the Heart Tissue of Male Wistar Rats Poisoned with Hydrogen Peroxide

Background: Oxidative stress plays a key role in cardiovascular diseases. Hydrogen peroxide poisoning also increases oxidative stress. In contrast, aerobic exercise and the use of Tribulus terrestris (Tt) extract can individually play an antioxidant defense balance. Objectives: The aim of this study was to investigate the simultaneous effect of aerobic training (AT), and alcoholic extract of Tt on some oxidative stress indices in the heart tissue of hydrogen peroxide poisoned male Wistar rats. Methods: Forty-two male Wistar rats were randomly divided into seven groups, including (1) healthy-control, (2) H2O2, (3) H2O2 + AT, (4) H2O2 + Tt 1 (5 mg/kg), (5) H2O2 + Tt 2 (10 mg/kg), (6) H2O2 + AT + Tt 1, and (7) H2O2 + AT + Tt 2. The AT protocol consisted of eight weeks (5 sessions per week) of treadmill running at 20 m/min for 60 minutes per session. Tissue samples were collected to measure oxidative stress indices. Two-way analysis of variance was used for statistical analysis. Results: A significant decrease in tissue concentration of cytochrome C (ng/mL), MDA (Pmol/mL), PAB index (HK), while an increase in tissue concentration of ATP (µM) was observed in the H2O2 + AT + Tt 2 group compared to other groups (P ≤ 0.05). Conclusions: Combining AT and Tt can be a good strategy to reduce the complications of hydrogen peroxide poisoning. Because the observed changes were far from complete improvement compared to the healthy control group, it is likely that longer treatment periods and higher drug doses should be used. Future studies should use Tribulus terrestris (Tt) supplementation in addition to Tribulus terrestris (Tt) intake prior to training sessions.

The effects of two iso-volume endurance training protocols on mitochondrial dysfunction in type 2 diabetic male mice.

Type 2diabetes(T2D) is one of the more common diseases in the world and has been widely spread. One of the suggested mechanisms in development of T2D, is mitochondrial dysfunction. The purpose of this study is to compare the effects of two endurance training protocols with low and moderate intensity on biogenesis and mitochondrial function, in Diabetic mice induced by high fat diet and Streptozotocin(STZ). 40 five week old mice divided to four groups including: health control (HC, n = 7), diabetic control (DC, n = 7), low endurance training (DLT, n = 7) and moderate endurance training (DMT, n = 7). DMT group ran at 5m/min for an hour, 3 days a week on a treadmill, and DLT group ran at 3m/min for an hour, 5 days a week on a treadmill for 8 weeks. The cytosolic content of PGC1α, Tfam and mitochondrial content of citrate synthase(Cs) and cytochrome c oxidase(Cox) in DC was significantly reduced compared to HC(P˂0.05). All of the parameters except for Cs in both DLT and DMT were increased compared to DC (P˂0.05), but there was no difference between them and the HC (P˃0.05). There was no difference in Cs enzyme between the DC and the DLT(P˃0.05), but it was significantly increased in the DMT(P˂0.05). There was a significantly difference between Cs enzyme in HC and DLT(P˂0.05), but there wasn't any significant difference between HC and DMT(P˃0.05). The results showed that in same volume condition, both endurance training protocols improved the proteins involved in biogenesis and mitochondrial function in T2D mice and there was no significant difference between them.

Syringic acid mitigates myocardial ischemia reperfusion injury by activating the PI3K/Akt/GSK-3β signaling pathway

Syringic acid is an abundant phenolic acid compound that possesses anti-oxidant, anti-microbial, anti-inflammatory, and anti-endotoxic properties. However, the research of pretreatment with syringic acid against myocardial ischemia reperfusion is still limited. Thus, our research revealed the protective effect of syringic acid in the rat model with myocardial ischemia reperfusion injury. Histological analysis was performed by hematoxylin and eosin (H&E). The myocardial systolic function was detected by echocardiographic. Myocardial infarct size was measured by Evans blue and 2,3,5-triphenyltetrazolium chloride (TTC) double staining. The apoptosis index was recorded by Terminal deoxynucleotidyl transferase dUTP nick end labeling staining (TUNEL). The contents of creatine kinase MB (CK-MB) and lactate dehydrogenase (LDH) in the serum were determined by a commercial kit. The expression of the PI3K/Akt/GSK-3β signaling pathway-related molecules and apoptosis-associated indicators was detected by western blotting or real-time PCR. We found that pretreatment with syringic acid obviously increased the myocardial systolic function (LVEF and LVFS) and decreased the infarct size, the apoptosis index as well as the serum level of CK-MB and LDH. Meanwhile, syringic acid also remarkably augmented the contents of p-PI3K, p-Akt, p-GSK-3β, Bcl-2 and mitochondria cytochrome c. However, the expression of caspase-3, -9 and Bax significantly reduced. Interestingly, co-treatment with PI3K inhibitor of LY294002 counteracted those effects induced by syringic acid. In conclusion, pretreatment with syringic acid can mitigate myocardial ischemia reperfusion injury by inhibiting mitochondria-induced apoptosis which is regulated by the PI3K/Akt/GSK-3β signaling pathway.

Woodchip-sulfur packed biological permeable reactive barrier for mixotrophic vanadium (V) detoxification in groundwater

Groundwater vanadium (V) (V(V)) contamination is ubiquitous in vanadium mining/smelting region and development of novel strategy for its remediation is of particular significance. Herein woodchip-sulfur packed biological permeable reactive barrier (bio-PRB) is established towards successful V(V) bio-detoxification. V(V) removal was accelerated under such mixotrophic condition, compared with heterotrophic and autotrophic V(V) reductions. The performance of bio-PRB was relatively steady with V(V) removal efficiency of 68.5%–98.2% under fluctuant geochemical and hydrodynamic conditions. Microbial community analysis indicated that heterotrophic Geobacter was the main reducer to convert V(V) to insoluble V(IV), by consumption of organic source attributed to woodchip hydrolysis and sulfur anabolism of autotrophs (e.g., Sulfuricurvum and Thiobacillus). V(V) reduction and elemental sulfur oxidation were regulated by genes as omcA, omcB and mtrC and soxB, respectively. The elevated contents of cytochrome c and nicotinamide adenine dinucleotide implied that improved electron transfer facilitated V(V) reduction. This study provides a cost-effective, robust and sustainable route for V(V)-polluted aquifer remediation.

Inhibitory effect of Lycium barbarum polysaccharide on sperm damage during cryopreservation.

With the development of cryopreservation technology, marked progress has been made regarding sperm cryopreservation. However, as conventional cryopreservation agents are not effective at freezing weak sperm, improved cryopreservation agents are in demand. In the present study, the addition of Lycium barbarum polysaccharides to glycerol-egg-yolk-citrate sperm cryopreservation agent was determined to improve sperm forward speed, reduce the DNA fragmentation index and improve the mitochondrial membrane potential. Furthermore, during the freezing and thawing of sperm, the improved cryopreservative increased the content of Bcl-2 while reducing the content of Bax, cytochrome C and caspase-3. These results indicated that polysaccharides added as a protective agent preserved the normal function of sperm mitochondria. Transmission electron microscopy also confirmed the protective effect of the polysaccharides on the structure of mitochondria. It was also indicated that improved cryopreservative lowered the levels of reactive oxygen species (ROS) during the freeze-thawing process. Therefore, it is hypothesized that improved cryopreservative agents may be beneficial for maintaining the structure and function of the mitochondria of weak sperm when cryopreserved, which may be facilitated via reducing the production of ROS in the freezing-thawing process, thus avoiding activation of the apoptotic pathway in sperm mitochondria and protecting mitochondrial structure and sperm function.

Deciphering correlation between chromaticity and activity of anammox sludge

The red color is the most striking character of anaerobic ammonium-oxidizing bacteria (AnAOB) which has been used to estimate the anammox activity roughly. However, the quantitative relationship between the color and activity of anammox sludge still remains unknown. In this study, the chromaticity, activity and their correlation were systematically investigated at different steady-state nitrogen loading rates. The chromaticity of anammox sludge was digitalized by the CIE L*a*b* color space. The results revealed that the average chroma value was found to be significantly correlated with specific anammox activity (r = 0.940, p < 0.01) and the cluster centers of chromaticity coordinates (a*, b*) of anammox sludge were established to define the typical working states of anammox system. The visible spectra of anammox sludge were proved to originate from the cytochrome c. The correlation between chroma and heme c concentration of anammox sludge was consistent with the fully-reduced cytochrome c and the chroma was determined by both content and redox ratio of cytochrome c. The chromaticity of anammox sludge was able to be linked with the anammox activity via reduced cytochrome c content. The gene abundance of cytochrome c synthetase linked the chromaticity with AnAOB quantity via total cytochrome c content, while the enzyme activity of octaheme hydrazine dehydrogenase linked the chromaticity with AnAOB activity via reduced cytochrome c ratio. Moreover, the redundancy analysis proved that heme c, as the key component of cytochrome c, was the most important explanatory variable accounting for the maximum 69.6% of the total variation of the anammox community, which correlated positively with the relative abundance of dominant AnAOB (Candidatus Kuenenia). This work aimed at demonstrating the chromaticity of anammox sludge could be developed as an alternative intuitive anammox activity indicator which will promote the monitoring and optimization of anammox process.

Urinary Cytochrome C and Caspase-3 as Novel Biomarker of Renal Function Impairment in Unilateral Ureteropelvic Junction Obstruction Model of Wistar Rats.

IntroductionProlonged obstruction in UPJO would lead to kidney destruction. It is important to find a non-invasive biomarker for early detection of renal impairment before definitive treatment for UPJO. In this study, we aim to evaluate the role of urinary cytochrome c and caspase-3 as a novel biomarker to predict renal function impairment in a UPJO model in Wistar rats.MethodsTwenty-five male Wistar rats were separated into 3 groups. Group I consists of 5 rats without any treatment or model, group II consists of 5 rats (sham group), and group III consists of 15 rats with the unilateral partial ureteral obstruction model. After 4, 15, and 21 days of observation, the urine was collected and rats were sacrificed to collect the glomerular count in the kidney. Measurement of cytochrome c and caspase-3 was done using the ELISA method, while the glomerular count was done using a light microscope. Data were analyzed using factorial repeated measures ANOVA test and correlation test with Pearson and processed using SPSS version 20.0.ResultsThe UPJO group has a significant increase in cytochrome c concentration and caspase-3 concentration compared to the control group and sham group (p<0.05). There was a significant decrease in the normal glomerulus count of the UPJO group (p<0.05). There was a significant relationship between the decrease in glomerulus count with the concentration of cytochrome c and caspase-3 in the UPJO group.ConclusionThere was a significant relationship between the decrease in glomerulus count with the increase in the concentration of cytochrome c and caspase-3 in the UPJO group.

Apoptosis in cerebrospinal fluid as outcome predictors in severe traumatic brain injury: An observational study.

Traumatic brain injury (TBI), due to its high mortality and morbidity, is an important research topic. Apoptosis plays a pathogenic role in a series of neurological disorders, from neurodegenerative diseases to acute neurological lesions.In this study, we analyzed the association between apoptosis and the Glasgow Outcome Scale (GOS), to examine the potential of apoptosis as a biomarker for a TBI outcome. Patients with severe TBI were recruited at the Department of Neurosurgery, Wujin Hospital Affiliated with Jiangsu University, between January 2018 and December 2019. As a control group, healthy subjects were recruited. The concentrations of caspase-3, cytochrome c, sFas, and caspase-9 in the cerebrospinal fluid (CSF) were analyzed by enzyme-linked immunosorbent assay (ELISA). The association between the GOS and the clinical variables age, sex, initial Glasgow Coma Scale (GCS) score, intracranial pressure (ICP), cerebral perfusion pressure (CPP), initial computed tomography (CT) findings, and apoptotic factors was determined using logistic regression. The area under the receiver operator characteristic (ROC) curve (AUC), and thus the sensitivity and specificity of each risk factor, were obtained.The levels of caspase-3, cytochrome c, sFas, and caspase-9 in the TBI group were significantly higher than those in the control group (P < .05). The logistic regression results showed that ICP and caspase-3 were significant predictors of outcome at 6 months post-TBI (P < .05). The AUC was 0.925 and 0.888 for ICP and caspase-3, respectively. However, the AUC for their combined prediction was 0.978, with a specificity and sensitivity of 96.0% and 95.2%, respectively, showing that the combined prediction was more reliable than that of the 2 separate factors.We demonstrated that caspase-3, cytochrome C, sFas, and caspase-9 were significantly increased in the CSF of patients following severe TBI. Furthermore, we found that ICP and caspase-3 were more reliable for outcome prediction in combination, rather than separately.