Production Of Oxygen Metabolites Research Articles

Apigeninidin chloride disrupts Toxoplasma gondii Mitochondrial membrane potential and induce reactive oxygen species and metabolites production.

Apigeninidin chloride (APi) is a form of 3-deoxyanthrocyanidins (3-DAs) abundantly produced by the red Sorghum bicolor plant. It has been previously reported to be effective against Toxoplasma gondii (T. gondii) tachyzoites grown in vitro with less cytotoxic effect. However, its possible mechanism(s) of action has not been elucidated. Biochemically, we discovered that APi induced high reactive oxygen species (ROS) and mitochondria superoxide (MitoSOX) productions in tachyzoites, leading to mitochondrial membrane potential (MMP) disruption in vitro. To confirm our biochemical results at the molecular level, we performed a liquid chromatography-mass spectrometry (LC-MS) analysis on APi-treated parasites to assess any metabolite and lipid alterations often associated with high ROS/MitoSOX production in cells. Noteworthy is that we detected several important oxidative stress-induced metabolites such as hexanal, aldehydes, methyl undeo10-enoate, butadiynyl phenyl ketone, 16-hydroxyhexadecanoic acid (16-OH, 16:0), 2-hydroxytricosanoic acid (C23:0; O), 3-oxodecanosanoic acid (C22:1; O), 2-hydroxypropylsterate, and furan fatty acids F6 (19FU-FA). These metabolites are associated with lipid, protein, and nucleic acid disruptions. Using atovaquone (Atov) as a control, we observed that it disrupted intracellular tachyzoites' mitochondrial membrane potential, increased ROS and MitoSOX production, and altered metabolite and lipid production similar to what was observed with our experimental compound APi. Overall, our results indicated that APi targets T. gondii tachyzoite growth through inducing oxidative stress, mitochondrial dysfunction, and eventually parasite death.

Effects of antioxidant-rich Lactiplantibacillus plantarum inoculated alfalfa silage on rumen fermentation, antioxidant and immunity status, and mammary gland gene expression in dairy goats

BackgroundMilk synthesis in lactating animals demands high energy metabolism, which results in an increased production of reactive oxygen metabolites (ROM) causing an imbalance between oxidants and antioxidants thereby inducing oxidative stress (OS) on the animals. To mitigate OS and postpartum disorders in dairy goats and gain insight into the impact of dietary choices on redox status during lactation, a feeding trial was conducted using alfalfa silage inoculated with a high-antioxidant strain of Lactiplantibacillus plantarum.MethodsTwenty-four Guanzhong dairy goats (38.1 ± 1.20 kg) were randomly assigned to two dietary treatments: one containing silage inoculated with L. plantarum MTD/1 (RSMTD-1), and the other containing silage inoculated with high antioxidant activity L. plantarum 24-7 (ES24-7).ResultsES24-7-inoculated silage exhibited better fermentation quality and antioxidant activity compared to RSMTD-1. The ES24-7 diet elevated the total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activities in milk, serum, and feces of lactating goats (with the exception of T-AOC in milk). Additionally, the diet containing ES24-7 inoculated silage enhanced casein yield, milk free fatty acid (FFA) content, and vitamin A level in the goats’ milk. Furthermore, an increase of immunoglobulin (Ig)A, IgG, IgM, interleukin (IL)-4, and IL-10 concentrations were observed, coupled with a reduction in IL-1β, IL-2, IL-6, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α concentrations in the serum of lactating goats fed ES24-7. Higher concentrations of total volatile fatty acid (VFA), acetate, and propionate were observed in the rumen fluid of dairy goats fed ES24-7 inoculated silage. Moreover, the diet containing ES24-7 inoculated silage significantly upregulated the expression of nuclear factor erythroid 2 like 2 (NFE2L2), beta-carotene oxygenase 1 (BCO1), SOD1, SOD2, SOD3, GPX2, CAT, glutathione-disulfide reductase (GSR), and heme oxygenase 1 (HMOX1) genes in the mammary gland, while decreased the levels of NADPH oxidase 4 (NOX4), TNF, and interferon gamma (IFNG).ConclusionsThese findings indicated that feeding L. plantarum 24-7 inoculated alfalfa silage not only improved rumen fermentation and milk quality in lactating dairy goats but also boosted their immunity and antioxidant status by modulating the expression of several genes related to antioxidant and inflammation in the mammary gland.

​Assessment of the Levels of Biomarkers of Oxidative Stress during Peri-parturient Period in Gir Cows

Background: Dairy cows during late pregnancy, parturition or early lactation may be subjected to oxidative stress or the production of reactive oxygen metabolites. An imbalance between increased production of ROS and reduced availability of antioxidant defenses near the time of parturition increases oxidative stress and may contribute to peri-parturient disorders in dairy cows therefore in present investigation, of biomarkers of oxidative stress during peri-parturient period in Gir cows were studied. Methods: Total 42 peri-parturient Gir cows were included in the present study. Ten apparently healthy Gir cows were also selected (which were outside the peri-parturient period) to have base line data on levels of biomarkers of oxidative stress for the comparison and analysis. The study was conducted in southern part of Rajasthan. Result: The mean values of malondialdehyde (n mol/g Hb), super oxidase dismutase (U/mg Hb), reduced glutathione (mM) and catalase (µmol of H2O2 consumed /min/mg Hb) during pre-partum period and post-partum period in Gir cows was 227.34±9.48 and 255.85±11.15; 72.97±4.96 and 56.13±4.30; 2.72±0.15 and 2.27±0.15 and 139.85±3.81 and 121.57±4.65, respectively. Determination of levels of biomarkers of oxidative stress revealed that level of malondialdehyde (MDA) was significantly (P less than 0.05) increased during peri-parturient period in Gir cows whereas, levels of superoxide dismutase (SOD) and reduced Glutathione (GSH) were significantly (P less than 0.05) decreased. The level of catalase was also significantly (P less than 0.01) decreased during peri-parturient period in Gir cows.

Lycopene minimizes skin toxicity and oxidative stress in patients treated with panitumumab-containing therapy for metastatic colorectal cancer

• Lycopene reduces ≥ G2 skin toxicity frequency in course of panitumumab therapy. • Lycopene prolongs time to appearance of ≥ G2 skin toxicity ( p = 0.0007 logrank test) • Lycopene protects tissues from oxidative stress due to panitumumab therapy. • Lycopene administration saves antioxidants consumption due to panitumumab therapy. Epidermal Growth Factor Receptor (EGFR) inhibition leads to the production of reactive oxygen metabolites causing skin inflammatory reactions. Anti-EGFR therapies are frequently associated with skin toxicities which often cause treatment delay and impairment to patient quality of life. Lycopene is a compound in the carotenoid group with an extreme antioxidant action which accumulates in the skin due to its hydrophobic structure. In a pilot study, we describe lactolycopene effectiveness in reducing skin toxicity and protecting tissues from oxidative stress in patients treated with panitumumab. Despite the limited number of patients, we show an absolute reduction of skin grade 2–3 toxicity in 41% of patients and 46% of panitumumab cumulative cycles in the experimental group versus placebo; lactolycopene administration was able to abolish malondialdehyde (MDA) production, a biomarker used to measure lipid peroxidation in the organism, and replenish antioxidant consumption in the course of anti-EGFR therapy. Trial registration: Clinicaltrials.gov NCT 03,167,268 (Pasto trial).

The impact of elevated ozone on growth, secondary metabolites, production of reactive oxygen species and antioxidant response in an anti-diabetic plant Costus pictus

Tropospheric ozone (O3) is a global air pollutant that causes deleterious effect to the plants. The present objective was to investigate the growth response, foliar injury, reactive oxygen species (ROS) accumulation and metabolites production in Costus pictus D. Don (insulin plant) at two developmental stages under ambient O3 (AO) and ambient + 20 ppb O3 (EO) using the open-top chambers (OTCs). A significant reduction in leaf area and total biomass was observed under EO as compared with AO. EO induced ROS (.O2- and H2O2) and lipid peroxidation led to more significant foliar injury and solute leakage. Image obtained from the fluorescence microscope and biochemical estimations reflected high levels of ROS under EO. A differential response in flavonoids and anthocyanin content, ascorbic acid, and antioxidative enzymes such as catalase (CAT), superoxide dismutase (SOD) and peroxidase (POX) has been observed with the growth stages of C. pictus plant. EO exposure negatively affected thiols and protein contents at all the growth stages. Secondary metabolites (tannins, lignin, saponins and alkaloids) were increased in both leaves and rhizomes due to EO, whereas phytosterols were induced only in rhizomes. Apart from other metabolites, the key bioactive compound (corosolic acid) showed its synthesis to be stimulated under EO at later growth stage. The study concludes that O3 is a potent stimulating factor for changing the levels of secondary metabolites and antioxidants in an antidiabetic C. pictus plants as it can alter its medicinal properties.

Free radical reactions in socially significant infectious diseases: HIV infection, hepatitis, tuberculosis

The analysis of current literature data on the study of the features of the course of free-radical reactions, as well as the state of the antioxidant defense system at socially significant infectious diseases HIV infection, hepatitis, tuberculosis was carried out. The role of this kind of reaction in the genesis and progression of socially significant infections a long time has been studied. Foreign studies of recent years have been focused on the identification of specific markers of oxidative and carbonyl stress, which make it possible to identify the redox imbalance of the cell under conditions of infection and target affect it to modulate the activity of the main transcription factors of viral proteins and the bacteria pathogenicity. Numerous sources indicate the involvement of active oxygen metabolites in a wide range of events in infected cells and tissues, including neoplastic transformation processes. These biochemical markers can be used as additional criteria for monitoring the progression of infection. At the same time, noticeable gaps in this area there are that may become the goal of future research. The issues of changing free radical reactions depending on gender, age, place of residence of patients remain practically unstudied. There is little data about intensity of oxidative stress in patients of reproductive age with HIV, hepatitis B and C, and pulmonary tuberculosis, as well as the relationship of antioxidant deficiency with reproductive disorders in conditions of infection. These data could serve as the basis for the development of pathogenetically substantiated methods for the correction of socially significant infectious diseases. Modulation of the production of reactive oxygen metabolites and oxidative stress is a potentially new pharmacological approach to reduce the effects of viral and bacterial exposure.

Anti-amebic effects of Chinese rhubarb (Rheum palmatum) leaves' extract, the anthraquinone rhein and related compounds

Entamoeba histolytica infects 50 million people worldwide and causes 55 thousand fatalities every year. Current anti-amebic drugs (e.g. paromomycin) work either at the level of the intestinal lumen (where trophozoites proliferate via cell divisions) or on the invasive trophozoites that have penetrated the gut or colonized internal organs (e.g. metronidazole). Some of these drugs are highly toxic to patients, have generated trophozoite resistance, or caused mutations and cancer in laboratory animals. Thus, alternative anti-amebic compounds need to be identified to minimize the side effects (on patients) or resistance (by amebas) to current treatments. The literature suggests that anthraquinones (chemicals found in medicinal plants) have antibacterial, antiparasitic, anti-inflammatory and antioxidant properties. Here we provide experimental evidence that Chinese rhubarb (Rheum palmatum) leaves' extract (rich in the anthraquinone rhein) inhibits E. histolytica trophozoite growth in vitro. In addition, from a set of ten isolated/synthetic anthraquinones (which we suspected to have anti-amebic properties), four analogs (rhein; AHHDAC = 1-amino-4-hydroxy-9, 10-dioxo-9, 10-dihydro-anthracene-2-carboxylic acid; unisol blue AS; and sennoside B) efficiently inhibited amebic growth at EIC50 concentrations comparable to metronidazole. The mechanism of action of these compounds still needs to be determined, although anthraquinones might enhance the production of toxic oxygen metabolites as it has been suggested for various protists (e.g. Leishmania, Plasmodium, Trypanosoma). Our research is the first to explore anti-amebic effects of Chinese rhubarb leaves' extract and isolated/synthetic anthraquinones on pathogenic Entamoeba.

Lycopene Reduces Skin Toxicity in Patients Treated with Panitumumab-Containing Therapy for Metastatic Colorectal Cancer – A Randomized Double-Blind Placebo-Controlled Pilot Study

Background: EGFR inhibition leads to the production of reactive oxygen metabolites, causing skin inflammatory reactions. Lycopene has an extreme antioxidant activity and, due to its hydrophobic structure, accumulates specifically in the skin. Methods: To assess lycopene effectiveness in reducing skin toxicity induced by panitumumab, 28 patients received lactolycopene or placebo in a phase-II PASTO study. Patients were evaluated at the beginning of each cycle and data were analyzed as the difference in mean grade of skin toxicity, frequency and duration of > G2 skin toxicity and frequency of tetracyclines administration. Lycopene, β-carotene and malondialdehyde plasma concentrations were analyzed to measure antioxidant consumption and oxidative stress. Findings: In the placebo group, ten out of 15 patients (66·67%), developed a > G2 skin toxicity, during 36 out of 92 cumulative cycles (39·1%), versus 3 out of 13 (23·08%), during 8 out of 64 cumulative cycles (12·5%) in the lactolycopene group. The time to the appearance of > G2 skin toxicity was different in the two groups (p= 0.0007 logrank test). Mean value of skin toxicity during treatment was 2·4 and 1·4 for placebo and lactolycopene group, respectively. Tetracyclines were administered for > G2 skin toxicity according to the protocol. Mean plasma concentration of lycopene and β-carotene decreased during treatment compared to day 0 (-141·94% and -157·22% respectively) in the placebo group, with a +109·36% mean malondialdehyde concentration increase. In contrast, in the lactolycopene group, mean plasma lycopene concentration increased during treatment (+94·18%), with a mean malondialdehyde concentration decrease (-68·83%) and a mean β-carotene concentration decrease (-105·5%) smaller than in the placebo group . Interpretation: Despite the limited number of patients, results clearly show lycopene effectiveness in reducing skin toxicity in patients treated with panitumumab. In our series lactolycopene administration was able to protect tissues from oxidative stress and to replenish antioxidants consumption. Trial Registration: registered in ClinicalTrials.gov (PaSTo – Clinicaltrials.gov: NCT 03167268) Funding Statement: AMOlavitaONLUS, Indena Spa. Declaration of Interests: MM1 is the vice-president and MP the treasurer of AMOlavitaONLUS. AMOlavitaONLUS is the sole owner and it holds all the ownership rights on the international patent application entitled “Use of carotenoid derivatives to reduce the toxicity and increase the efficacy of antiEGFR antitumor treatments” (N°PCT/IB2018/050229). MM1 and MP are exclusively inventors of the patent and they have no ownership right on the patent. Indena Spa, who supplied free of charge lactolycopene for the Pasto Clinical Trial, is not the owner of the patent for lactolycopene production. All other authors declare no competing interest. Ethics Approval Statement: The protocol was approved by the ethics committees of all participating institutions. Written informed consent was obtained from all patients.

The Risk Assessment of Oxidative Stress in Postpartum Dairy Cattle

Background: Negative energy balance in cows will induce catabolism especially of lipids and the resulting rapid increase of free fatty acids (NEFA) in the blood stream increases the production of reactive oxygen metabolites (ROMs) and promote oxidative stress. Once the animal body is in an oxidative stress state, many molecules in the body may be peroxidized, accelerating the destruction of cells and causing damage to tissues or organs. This study conducted risk assessment of the oxidative stress caused by negative postpartum energy balance in cows by exploring the relationship between negative energy balance and oxidative stress.Materials, Methods & Results: This experiment randomly selected 120 cows at 14 to 21 days postpartum, from an intensive cattle farm in Heilongjiang province, China. Using a negative energy balance standard of β-hydroxybutyric acid (BHBA) greater than 1.2 mmol/L, nonesterified fatty acid (NEFA) greater than 0.4 mmol/L and glucose (GLU) less than three mmol/L, the cattle were divided into a healthy group of 74 cows and a diseased group of 46 cows. The oxidative stress indices of the experimental cows were measured and analyzed using the independent sample t test. Spearman correlation analysis and regression analysis were performed and by using the binary Logistic regression analysis to predict disease, the receiver-operating characteristic curve (ROC) analysis established diagnosis effect and boundary values. Compared with the healthy group, the levels of glutathione peroxidase (GSH-Px), catalase (CAT), vitamin E (VE), selenium (Se) and total nitric oxide synthetase (T-NOS) in the blood of dairy cows in the diseased group were significantly reduced and the level of malondialdehyde (MDA) was significantly increased. The study concluded that negative energy balance is associated with oxidative stress in cows and the blood levels of GSH-Px, CAT and Se can be used to evaluate the degree of resulting oxidative stress. The early warning levels were determined to be GSH-Px less than 619.28 U/L, CAT less than 7.87 U/ml and Se less than 0.51 μg/L.Discussion: The levels of GSH-Px, CAT, Se, T-NOS and VE in the blood of cows in the disease group were lower than those in the healthy control group. This may be due to the increase in energy demand of cows in the perinatal period and the decrease of DMI in this period, which promotes the NEB of cows. In order to alleviate the NEB, the body forces its metabolic enhancement. Some tissues of the body cannot adjust their metabolism to adapt to NEB and this leads to excessive NEFA, which causes oxidative stress. Excessive NEFA can also affect the expression of GSH-Px in cells, and causing a series of oxidation reactions, increased free radicals and as the antioxidant system is overwhelmed. The MDA content in the blood of the deficient group was significantly increased compared with the healthy group, positively correlated with NEFA and BHBA and negatively correlated with GLU. This may be due to severe oxidative stress in cows during the period of negative energy balance as MDA is one of the important products of membrane lipid peroxidation. Long chain PUFA are an important component of all cell membranes and their oxidation damages the integrity, fluidity and function of cell membranes. In summary MDA causes oxidative damage to cells, damages mitochondrial membranes and changes membrane permeability, resulting in more free radicals produced by mitochondria during biological oxidation.

Skewed X-inactivation in a Female Carrier with X-linked Chronic Granulomatous Disease

Chronic granulomatous disease (CGD) is a primary immunodeficiency caused by defective phagocytic NADPH oxidase, causing a complete lack or significant decrease in the production of microbicidal reactive oxygen metabolites. It mainly affects male children; however, there are scarce reports of adult females diagnosed with X-linked-CGD attributed to an extremely skewed X-chromosome inactivation. This condition is characterized by severe and recurrent infections that usually develop after childhood. In clinical practice, physicians who usually confront these patients should suspect this entity and differentiate it from a secondary immunodeficiency. Here, we report a 38-year-old Mexican female with juvenile-onset X linked-CGD, caused by a de novo mutation and extremely skewed X-inactivation, whose clinical features were similar to those in patients with classic X-linked-CDG.

Dietary Biotechnological Ajuga reptans Extract in Post Weaning Piglets: Effects on Growth Performance, Oxidative Status and Immune Parameters

Abstract The effect of dietary supplementation with a biotechnological extract of Ajuga reptans on growth performance, oxidative status and immune parameters was evaluated in post weaning piglets. At weaning, 120 piglets with an average live weight of 8.1±1.3 kg, were assigned to one of three experimental groups. The first group was fed a control diet (C). The second and third groups were fed the same diet supplemented with 5 mg (T1) and 10 mg (T2) of teupolioside/kg feed from a biotechnological plant extract. Growth performances were recorded and blood samples were collected at the beginning, at 14 days, and at the end of the trial (56 days). Serum biochemical parameters, oxidative status and immunoglobulin titres were determined. Average daily gain tended to be higher (P=0.057) and live weight was higher in piglets (P<0.05) fed with different amounts of plant extract (T1 and T2) than the controls. The production of reactive oxygen metabolites (ROMs) was higher (P<0.05) in the control group than in the groups receiving teupolioside (T1 and T2). Concentration of serum immunoglobulin of class G improved (P<0.001) in piglets fed the T1 and T2 diets than the controls. Overall, the results suggested that the biotechnological extract of Ajuga reptans containing teupolioside has an antioxidant and immunomodulant effect.

Treatment with estrogen receptor agonist ERβ improves torsion-induced oxidative testis injury in rats

AimsThe purpose of the present study was to investigate the potential antioxidant, anti-apoptotic and sperm function-preserving effects of estrogen, estrogen receptor (ER)α and ERβ agonists in a rat model of testis torsion-detorsion (T/D). Main methodsUnder anesthesia, 6–8-week-old male Sprague-Dawley rats underwent sham-operation or testicular torsion by fixing left testis rotated at 720° for 2 h. After detorsion, rats were treated with ERα agonist (1 mg/kg/day, subcutaneously, sc) or ERβ agonist (1 mg/kg/day, sc) or estradiol (E2, 1 mg/kg/day, in drinking water) or vehicle on the following two days. On the third day, testicular blood-flow was recorded and then left testes were extracted for molecular and histochemical analysis. Key findingsThe findings showed that reduced testicular blood-flow following torsion was partially restored on the 3rd day of detorsion, while treatments with either of the ER agonists or E2 returned blood flow fully back to the control levels. When the testis-torsioned rats were given ERβ agonist during the detorsion period, tubular injury was lessened, sperm count and motility were increased, while the production of reactive oxygen metabolites and apoptosis in the testis tissues were totally suppressed. Although a down-regulated expression of androgen receptor (AR) along with a reduction in serum testosterone level was observed in the vehicle-treated T/D group, all three treatments up-regulated the expressions of AR and its mRNA, while ERα agonist and E2 suppressed the testosterone level. SignificanceERβ receptor activation during the post-ischemic period may be beneficial in protection against torsion-related oxidant testicular injury and infertility.

Transplanting gravid lizards to high elevation alters maternal and embryonic oxygen physiology, but not reproductive success or hatchling phenotype.

Increased global temperatures have opened previously inhospitable habitats, such as at higher elevations. However, the reduction of oxygen partial pressure with increase in elevation represents an important physiological constraint that may limit colonization of such habitats, even if the thermal niche is appropriate. To test the mechanisms underlying the response to ecologically relevant levels of hypoxia, we performed a translocation experiment with the common wall lizard (Podarcis muralis), a widespread European lizard amenable to establishing populations outside its natural range. We investigated the impacts of hypoxia on the oxygen physiology and reproductive output of gravid common wall lizards and the subsequent development and morphology of their offspring. Lowland females transplanted to high elevations increased their haematocrit and haemoglobin concentration within days and maintained routine metabolism compared with lizards kept at native elevations. However, transplanted lizards suffered from increased reactive oxygen metabolite production near the oviposition date, suggesting a cost of reproduction at high elevation. Transplanted females and females native to different elevations did not differ in reproductive output (clutch size, egg mass, relative clutch mass or embryonic stage at oviposition) or in post-oviposition body condition. Developing embryos reduced heart rates and prolonged incubation times at high elevations within the native range and at extreme high elevations beyond the current range, but this reduced oxygen availability did not affect metabolic rate, hatching success or hatchling size. These results suggest that this opportunistic colonizer is capable of successfully responding to novel environmental constraints in these important life-history stages.

Therapeutic efficacy of osthole against dinitrobenzene sulphonic acid induced-colitis in rats

Several mediators were associated with the pathogenesis of inflammatory bowel disease such as oxidative stress through the production of reactive oxygen metabolites, neutrophils infiltration and release of pro-inflammatory cytokines. This study was designed to investigate the therapeutic efficacy of osthole against dinitrobenzene sulfonic acid (DNBS) induced-colitis in rats through its anti-oxidant and anti-inflammatory properties. Colitis was induced in rats by single intracolonic instillation of (250 μl DNBS-25 mg/rat). Then 4 days later, rats were received oral administration of either (osthole 50 mg/kg), (sulfasalazine 500 mg/kg) or both in combination for 7 consecutive days. Body weight, some hematological parameters, colonic malondialdehyde (MDA) and myeloperoxidase activity (MPO), antioxidant parameters, colon injury and mucosa architectures were assessed. T helper (Th1)-related cytokines [Tumor necrosis factor alpha (TNF-α) and interferon-gamma (INF-γ)], Th2-relarted cytokines (interleukin-4 [IL-4 and IL-10], and Th-17 related cytokines [IL-17] were determined by ELISA. Osthole significantly improved the loss in body weight. That was accompanied with a remarkable amelioration of the disruption of the colonic architecture as well as a significant improvement in the antioxidant defense system. A reduction in MPO and MDA was observed in flamed colon. Treatment with either osthole or combination therapy showed suppressive activities on pro-inflammatory Th2-related cytokines and upregulation of anti-inflammatory Th2-related cytokines The results of this study suggest that osthole exert beneficial therapeutic effect in experimental colitis and improved the efficacy of the synthetized drugs such as sulfasalazine. Therefore, osthole may have a valuable sound in the treatment of inflammatory bowel disease.

Prognostic relevance of oxidative stress measurement in chronic lymphocytic leukaemia.

To evaluate the prognostic significance of oxidative stress (OS) and antioxidant defence status measurement in patients with chronic lymphocytic leukaemia (CLL). d-ROMs test and BAP test were evaluated at diagnosis of 165 patients with CLL and correlated with clinical-biological features and prognosis. An increased oxidative damage (d-ROMs test) and a reduced antioxidant potential (BAP test) were found in CLL patients than normal controls (P<.0001). CLL patients with higher d-ROMs values had higher number of circulating white blood cells and lymphocytes, and higher values of β2 -microglobulin. Higher d-ROMs values were found in female (P=.0003), in patients with unmutated IgVH (P=.04), unfavourable cytogenetics (P=.002) and more advanced clinical stage (P=.002). Higher BAP test values were found in patients expressing CD49d (P=.01) and with more advanced clinical stage (P=.004). At a median follow-up of 48months, CLL patients with d-ROMs ≥418 CARR U were found to have a shorter time to first treatment (TFT) (P=.0002), and a reduced survival (P=.006) than others. CLL patients with BAP test values ≥2155μmol/L had a shorter TFT (P=.008) and a shorter survival (P=.003). OS can affect CLL patients by concomitantly increasing reactive oxygen metabolites production and decreasing antioxidant defences.

Mitochondria control acute and chronic responses to hypoxia

There are numerous mechanisms by which mammals respond to hypoxia. These include acute changes in pulmonary arterial tone due to smooth muscle cell contraction, acute increases in respiration triggered by the carotid body chemosensory cells, and chronic changes such as induction of red blood cell proliferation and angiogenesis by hypoxia inducible factor targets erythropoietin and vascular endothelial growth factor, respectively. Mitochondria account for the majority of oxygen consumption in the cell and have recently been appreciated to serve as signaling organelles required for the initiation or propagation of numerous homeostatic mechanisms. Mitochondria can influence cell signaling by production of reactive oxygen species and metabolites. Here we review recent evidence that mitochondrial signals can imitate acute and chronic hypoxia responses.

Human Neutrophils Use Different Mechanisms To Kill Aspergillus fumigatus Conidia and Hyphae: Evidence from Phagocyte Defects.

Neutrophils are known to play a pivotal role in the host defense against Aspergillus infections. This is illustrated by the prevalence of Aspergillus infections in patients with neutropenia or phagocyte functional defects, such as chronic granulomatous disease. However, the mechanisms by which human neutrophils recognize and kill Aspergillus are poorly understood. In this work, we have studied in detail which neutrophil functions, including neutrophil extracellular trap (NET) formation, are involved in the killing of Aspergillus fumigatus conidia and hyphae, using neutrophils from patients with well-defined genetic immunodeficiencies. Recognition of conidia involves integrin CD11b/CD18 (and not dectin-1), which triggers a PI3K-dependent nonoxidative intracellular mechanism of killing. When the conidia escape from early killing and germinate, the extracellular destruction of the Aspergillus hyphae needs opsonization by Abs and involves predominantly recognition via Fcγ receptors, signaling via Syk, PI3K, and protein kinase C to trigger the production of toxic reactive oxygen metabolites by the NADPH oxidase and myeloperoxidase. A. fumigatus induces NET formation; however, NETs did not contribute to A. fumigatus killing. Thus, our findings reveal distinct killing mechanisms of Aspergillus conidia and hyphae by human neutrophils, leading to a comprehensive insight in the innate antifungal response.

In vitro interferon γ improves the oxidative burst activity of neutrophils in patients with chronic granulomatous disease with a subtype of gp91phox deficiency.

Aim of this studyChronic granulomatous disease (CGD) is a genetically heterogeneous primary immunodeficiency caused by a defect in phagocyte production of oxygen metabolites, and resulting in infections produced by catalase-positive microorganisms and fungi. Interferon γ (IFN-γ) has a multitude of effects on the immune system. Although preliminary studies with CGD patients on treatment with IFN-γ showed that it enhanced phagocytosis and superoxide production, ongoing studies did not reveal a significant increase of this function. Here we investigated the oxidative capacity of phagocytes in different subtypes of CGD patients on treatment with IFN-γ in vitro.Material and methodsFifty-seven patients with CGD from 14 immunology centres were enrolled to our multi-centre study. Twenty-one patients were studied as controls. Oxidative burst assay with dihydrorhodamine 123 (DHR) was used and the stimulation index (SI) was calculated with respect to CGD subtypes in both neutrophils and monocytes before, and then one and 24 hours after adding IFN-γ.ResultsUpon comparison of the SIs of the patients’ neutrophils before in vitro IFN-γ at hour 0, and after adding IFN-γ at hour 1 and 24 were compared, and the differences were determined between hours 0-24 and hours 1-24. This difference was especially apparent between hours 1-24. In CGD subtypes, particularly in gp91phox subtype, it was seen that, following in vitro IFN-γ, SIs of neutrophils began to increase after hour 1, and that increase became more apparent at hour 24.ConclusionsOur study showed that IFN-γ treatment may increase the oxidative bursting activity by increasing the superoxide production in neutrophils, particularly in gp91phox subtype.

EFFICIENCY OF RECOMBINANT TNF-BINDING PROTEIN FROM VARIOLA VIRUS IN A MODEL OF COLLAGEN-INDUCED ARTHRITIS

This paper presents the results of the research on the effectiveness of recombinant TNF-binding protein of variola virus (VARV-CrmB) in a model of collagen-induced arthritis (CIA) in mice (CBAxC57Bl6) F1. The introduction of VARV-CrmB and polyclonal antibody to recombinant mouse TNF (poly-AbMuTNF) led to an improvement of clinical manifestations of CIA by reducing the swelling and increasing the mobility of mice limbs. The introduction of VARV-CrmB and poly-AbMuTNF reduced the number of neutrophilic granulocytes and granulocytic precursors. The introduction of VARV-CrmB and poly-AbMuTNF into mice decreased collagenolysis in the blood serum and the content of glycosaminoglycans at the early stages of experimentation. Treatment with VARV-CrmB and poly-AbMuTNF of mice with CIA significantly decreased the chemiluminescence response of blood leukocytes. VARV-CrmB exerted more pronounced inhibitory effect on the production of reactive oxygen metabolites by blood leukocytes of mice with CIA than poly-AbMuTNF. Improvement of clinical condition of the mice with CIA has a more prolonged effect following introduction of the VARV-CrmB than after injection of poly-AbMuTNF. The results suggest the recombinant viral protein VARVCrmB to be a new potential TNF-antagonist.

SAT0364 Efficacy and Safety of Intra-Articular Therapy with Methotraxate in Large Glucose Injection Volume on Knee Synovitis in Patients with Ankylosing Spondylitis

BackgroundIntra-articular therapy has a long history in treating inflamed arthritis, especially mono-oligoarthritis. Many different compounds are used. Since 1970's, a number of short-term studies have demonstrated Intra-articular administration of MTX...