Systemic circulatory influences on retinal microvascular function in middle-age individuals with low to moderate cardiovascular risk

Purpose: To investigate the relationship between retinal microvascular reactivity, circulatory markers for CVD risk and systemic antioxidative defence capacity in healthy middle-aged individuals with low to moderate risk of CVD. Methods: Retinal vascular reactivity to flickering light was assessed in 102 healthy participants (46-60 years) by means of dynamic retinal vessel analysis (DVA). Other vascular assessments included carotid intima-media thickness (C-IMT) and blood pressure (BP) measurements. Total cholesterol (CHOL), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG) and blood glutathione levels in its reduced (GSH) and oxidized (GSSG) forms were also determined for each participant, along with Framingham risk scores (FRS). Results: Retinal arterial baseline diameter fluctuation (BDF) was independently, significantly and negatively influenced by LDL-C levels (β = -0.53, p = 0.027). Moreover, the arterial dilation slope (SlopeAD) was independently, significantly and positively associated with redox index (GSH: GSSG ratio, β = 0.28, p = 0.016), while the arterial constriction slope (SlopeAC) was significantly and negatively influenced by blood GSH levels (β = -0.20, p = 0.042), and positively associated with FRS (β = 0.25, p = 0.009). Venous BDF and dilation amplitude (DA) were also negatively influenced by plasma LDL-C levels (β = -0.83, p = 0.013; and β = -0.22, p = 0.028, respectively). Conclusions: In otherwise healthy individuals with low to moderate cardiovascular risk, retinal microvascular dilation and constriction responses to stress levels are influenced by systemic antioxidant capacity, and circulating markers for cardiovascular risk.

Glutaredoxin-1 Is up-regulated in preeclampsia and increases soluble Flt-1 expression

INTRODUCTION: Preeclampsia is a vascular disorder in pregnancyand is biochemical characterization by high soluble Flt-1 and lowplacenta growth factor as well as an imbalance in redox homeostasis.During conditions of high oxidative stress, cysteine residues on keyproteins are reversibly altered by S-glutathionylation, modifying theirfunction. Glutaredoxin-1 (Glrx) enzymatically catalyzes the removal of S-glutathione adducts, conferring reversible signaling dynamics toproteins with redox-sensitive cysteines. The role of Glrx in preeclampsiais unknown.METHODS: Immunohistochemistry and Western blot analysis for Glrx orglutathione were conducted on human placenta samples collected pre-termfrom early onset preeclamptic patients (n=10) or non-preeclamptic induceddeliveries (n=9). Human endothelial cells were infected with adenovirusencoding Glrx or LacZ prior to the cells being exposed to hypoxia (0.1%O2, 24h) to measure changes in soluble Flt-1 (sFlt-1). Quantitative PCRand ELISA were used to measure sFlt-1 at mRNA and protein level.RESULTS: Immunohistochemical staining for GSH revealed lowerS-glutathionylation adducts in preeclampsia placenta in comparison tocontrols. Glrx expression, which catalyses de-glutathionylation wasenhanced in early onset preeclampsia compared to pre-term controlsamples. In contrast, no change was observed in preeclamptic and IUGRplacentas at full term. In endothelial cells overexpressing Glrx, sFlt-1expression was dramatically enhanced at mRNA (3-fold P<0.05) andprotein level (5 fold P>0.01, n=4) after hypoxia andoverexpressing Glrxin mice enhanced levels of circulating sFlt-1 during in vivo ischemia.CONCLUSIONS: Enhanced Glrx expression in preeclamptic placentain line with an apparent decrease in S-glutathionylation may leavekey proteins susceptible to irreversible oxidation in conditions of highoxidative stress.

Interaction between mercury (Hg), arsenic (As) and selenium (Se) affects the activity of glutathione S-transferase in breast milk: possible relationship with fish and shellfish intake

Breast milk is regarded as an ideal source of nutrients for the growth and development of neonates, but it can also be a potential source of pollutants. Mothers can be exposed to different contaminants as a result of their lifestyle and environmental pollution. Mercury (Hg) and arsenic (As) could adversely affect the development of fetal and neonatal nervous system. Some fish and shellfish are rich in selenium (Se), an essential trace element that forms part of several enzymes related to the detoxification process, including glutathione S-transferase (GST). The goal of this study was to determine the interaction between Hg, As and Se and analyze its effect on the activity of GST in breast milk. Milk samples were collected from women between day 7 and 10 postpartum. The GST activity was determined spectrophotometrically; total Hg, As and Se concentrations were measured by atomic absorption spectrometry. To explain the possible association of Hg, As and Se concentrations with GST activity in breast milk, generalized linear models were constructed. The model explained 44% of the GST activity measured in breast milk. The GLM suggests that GST activity was positively correlated with Hg, As and Se concentrations. The activity of the enzyme was also explained by the frequency of consumption of marine fish and shellfish in the diet of the breastfeeding women.

Biofloc contribution to antioxidant defence status, lipid nutrition and reproductive performance of broodstock of the shrimp Litopenaeus stylirostris: Consequences for the quality of eggs and larvae

The aim of this study was to determine biofloc contributions to the antioxidant status and lipid nutrition of broodstock of Litopenaeus stylirostris in relationship with their reproductive performance and the health of larvae produced. Shrimp broodstock reared with Biofloc technology (BFT) compared to Clear water (CW) exhibited a higher health status with (i) a better final survival rate during the reproduction period (52.6% in CW against 79.8% in BFT); (ii) higher glutathione level (GSH) and total antioxidant status (TAS), reduced oxidized/reduced glutathione ratio and a higher spawning rate and frequency as well as higher gonado-somatic index and number of spawned eggs. Finally, larvae from broodstock from BFT exhibited higher survival rates at the Zoe 2 (+ 37%) and Post Larvae 1 (+ 51%) stages when compared with those from females from CW treatment. The improved reproductive performance of the broodstock and higher larvae survival rate resulting from BFT treatment may be linked to the dietary supplement obtained by the shrimp from natural productivity during BFT rearing. Indeed, our study confirms that biofloc particulates represent a potential source of dietary glutathione and a significant source of lipids, particularly essential phospholipids and n-3 highly unsaturated fatty acids (HUFA) for shrimps. Thus, broodstock from BFT treatment accumulated phospholipids, n-3 HUFA and arachidonic acid, which are necessary for vitellogenesis, embryogenesis and pre-feeding larval development. The predominant essential fatty acids, arachidonic acid (ARA), eicopentaeonic acid (EPA) and docosahexaenoic acid (DHA), had levels in the eggs that were, respectively, 2.5, 2.8 and 3 fold higher for BFT compared to the CW treatment. Statement of Relevance Today, the influence of biofloc technology on shrimp broodstock is not enough described and no information was available on the larvae quality. Moreover, two key pieces of new information emerge from the present study. Firstly, biofloc is a source of further dietary lipids that can act as energetic substrates, but also as a source of phospholipids and essential fatty acids necessary to sustain reproduction, embryonic and larval development. Second, improving the reproduction of the broodstock also leads to an improvement in the quality of the larvae. We think that our research is new and important to increase knowledge on biofloc topic. We believe the paper will contribute to the development of more efficient and therefore more sustainable systems.

Influência dos ácidos graxos do tipo ômega-3 ou ômega-6 e o nanomaterial de carbono fulereno (C60) no estado antioxidante em suspensões celulares de cérebro de Cyprinus carpio (Pisces, Cyprinidae)

Os nanomateriais de carbono como o fulereno (C60) apresenta comportamentos bioquímicos distintos, podendo atuar como antioxidante ou pró-oxidante em diferentes sistemas biológicos. Outra evidência ao C60 refere-se a sua característica lipofilica, na qual oferece ação mais direta a diferentes tipos de membranas celulares. Do mesmo modo ácidos graxos poliinsaturados (AGPs) como o ômega-3 (DHA) e o ômega-6 (LA) são importantes para funções celulares da membrana, sendo considerados antioxidantes clássicos. Dessa forma este estudo avaliou em suspensões celulares de cérebro da carpa (Cyprinus carpio, Cyprinidae), o efeito de C60 após um pré-tratamento com DHA ou LA. Para tal avaliação os ensaios consistiram em um pré-tratamento com AGPs (48h) e após exposição a C60 (2h). Como resultados observamos que a viabilidade celular e a capacidade antioxidante total não apresentaram diferença (p> 0.05) entre todos os grupos. Em relação a valores de espécies ativas de oxigênio e dano lipídico foi observado redução nos seus valores nos grupos expostos ao C60 pré – tratados com AGPs (p<0.05). Em termos de cisteína, ocorre uma redução da sua concentração em todos os grupos expostos ao C60. Porém para glutationa a exposição ao C60 provoca um aumento de sua concentração nos grupo controle (sem AGPs) e no grupo pré – tratado com DHA. Dessa forma consideramos que o pré – tratamento com AGPs é benéfico às células, uma vez que um aumento nos níveis de glutationa e uma diminuição na concentração de espécies ativas de oxigênio e peroxidação lipídica foram observados nos grupos expostos ao C60. Sendo assim um bom estado nutritivo em termos da concentração de AGPs foi considerado benéfico na exposição ao fulereno.

3-Dichloroacetyl oxazolidine protect maize from imazethapyr herbicide injury

Safeners are an important tool used to ensure the safe using of herbicide. The objective of this paper was to investigate the protective effect of four 3-dichloroacetyl oxazolidine safeners (3-dichloroacetyl-2,2-dimethyl-1,3-oxazolidine [R-28725], racemate of 3-dichloroacetyl-2,2-dimethyl-4- ethyl-1,3-oxazolidine, and its two chiral stereoisomers) in reducing the injury caused by imazethapyr. Physiological and biochemical tests were conducted under laboratory condition, by using seed treatment with safeners and soil treatment with imazethapyr, respectively. The interaction of two safeners (R-28725 and R-stereoisomer) and imazethapyr reduced the injury of maize significantly, and also increased glutathione content, activity of glutathione S-transferases, and activity of acetolactate synthase in maize. When induced by R-stereoisomer, the GSH content in root and in shoot increased 100.7% and 73.6%, respectively. When induced by R-28725, the GST activity in vivo increased threefold and the GST activity in vitro more than doubled. The kinetic parameter Vmax of GST in the maize treated with R-28725 and R-stereoisomer increased by 102.2% and 81.9%, respectively, compared with the control. The results also showed that R-28725 and R-stereoisomer induced glutathione S-transferases affinity for the substrate of conjugation reaction significantly.

Seasonal variation of acetylcholinesterase, catalase, glutathione s transferase in different size of Pinctada radiata and metal pollution (Pb, Cd, Ni) in Persian Gulf

There are various tools for monitoring the concentration of pollutants on aquatic ecosystems. Today these studies are based on biological monitoring and biomarkers. The aim of this study was to measure the concentration of the acetylcholinesterase (AChE), glutathione S-transferase and catalase as biomarkers of heavy metal contamination in pearl oyster Pinctada radiata and their mechanism in aquatic ecosystems. Heavy metals lead, cadmium and nickel were measured in soft tissue and studied stations in four seasons. Samples were collected seasonally in Lavan stations, Hendurabi and Nakhilo (in the northern Persian Gulf) from spring 2013 to winter of that year by scuba diving. Pearl oysters are divided according to their shells size; shells separated from soft tissues and were transferred to the laboratory for analysis of heavy metals and enzymes. Moopam standard method for were used for measuring the concentration of heavy metals and for analyzing tissue concentrations of glutathione S-transferase in Clam the method recommended by Habig et al in 1974 were used. For measuring acetylcholinesterase Ellman method were used. Catalase contamination in pearl oyster in the supernatant obtained from the study based on the method homogeate soft tissue of mussels (Abei, 1974) was evaluated. The results showed that the concentration of lead has significant difference in sediments station, the concentration of lead in Lavan is significantly higher than the other two stations, This could be due to the movement of tanker, boats and floating refueling and with a considerable amount of wastewater containing oil and Petroleum into the water, and also due to precipitation and industrial discharges the lead in the region is increasing, land-disposed sewage sludge, has large concentrations of lead. Compare the results of this study with standards related and other similar studies at the regional and international level showed that pollutant concentration of heavy metals in all cases significantly less than all the standards and guide values associated. And also compared to other world research results have been far less than others, Being Less of the conclusion given in this research according that nickel is one of the indicators of oil pollution in the study area and emissions have been relatively low of oil. The concentration of acetylcholinesterase at several stations, in large and small sizes and in the seasons had no significant difference. Variations of catalase, and glutathione S-transferase were almost similar to each other and parameters, station and seasons were significantly different in the concentrations of these enzymes. The effects and interaction between various parameters indicate that following parameters has impact on the concentration of catalase and glutathione S-transferase. Stations; Seasonal changes in antioxidant enzymes related to (assuming a constant in salinity and oxygen) to age, reproductive cycle, availability of food and water temperature. With increasing temperature at warm season, antioxidant enzymes were increase, with increasing temperature and abundance of food in the environment the amount of antioxidant enzymes may increase. The presence of the enzyme concentration may indicate that the higher levels of the enzyme to eliminate ROS activities to be any healthier situation. At the time of gonads maturation and spawning season catalase activity increases. This study also indicates that catalase was significantly higher in the warm season. Due to low pollutants of heavy metals in the study area, a lower level of contaminants were observed in shellfish tissue incidents of international standards and strong correlation between the amount of heavy metal contamination in pearl oyster tissue and enzymes was not observed. Therefore, we can say that the pearl oyster remains in a healthy condition and the amount of enzyme is normal.

Refolding of omega conotoxin peptide derived from marine snail, Conus magus, to evaluate its analgesic effects

Oxidative refolding is one of the key challenges hampering the development of peptide based compounds as therapeutics. The correct refolding for three disulfide peptide like w-Conotoxi n MVIIA is difficult and crucial for biological activity. This work advanced knowledge of chemical and biological for improve oxidative refolding of synthetic w-Conotoxi n MVIIA in base of Conus magus venom. The present study aimed to set up an appropriate and effective protocols for refolding of disulfide-rich w-Conotoxin MVIIA. In this study, the crude peptide was protected with Acm group, according to the right amino acid sequences (Synthesized by Australian Company). The crude peptide was purified by H PLC. To prepare the peptide to refolding, innovative deprotection applied molar ratio (AMR) method was performed based on mercury. Accuracy of deprotection was approved by reverse phase chromatography. The deprotected target peptide (omega-conotoxin) was determined by SDS-PAGE. Then the Oxidative refolding of target peptide was performed in six protocol based on Guanidinium chloride and oxidized and reduced Glutathione. Analgesic effect of refolded peptide was surveyed with formalin test in mice Balb/c. Non neurotoxic effects of target peptides were survey with ICV injection in mice model (C57/BL6). The innovative deprotection protocol performed based on the best ratio of mercury/2-mercaptoethanol adjusted to 1mg/10p1 in 90 minute. The results showed the yield and purity of omega-conotoxin MVIIA as 93 and 95%, respectively. Refolding of 40 mg omega Conotoxin with GSSG and GSH on ratio of 10:1 and 20 mM ammonium acetate showed the best analgesic effect compared with the other methods. The result showed 95.5% yield and 98% purity of omega-conotoxin MVIIA in this refolding method. Related refolding method reduced 85% pain in experimented mice using 7 ng of the peptide. That was 71.5 fold stronger than morphine and 2 times than standard Prialt®. And it was not neurotoxic in mice. In this study, refolding method for omega-conotoxin MVIIA was optimized in the fourth factor including: reducing the time, amount and number of reagent and increase the efficiency. We introduced new method for deprotection of omega-conotoxin MVIIA. Effective, economic and applied refolding and deprotecti on method was performed in this research may al so be applied to similar omega conotoxin peptides.

Liver injury from ampicillin-induced intestinal microbiota distresses in rats fed carbohydrate- and protein-rich diets

Purpose: To investigate the effect of ampicillin on rat intestinal microflora and liver in the presence of high carbohydrate and protein diets. Methods: Male Wistar albino rats were divided into four groups. The first group served as the control, the second group was treated with ampicillin (50 mg/kg for 3 weeks) and fed with a standard diet, while the third and fourth groups were treated with the same dose of ampicillin and fed with acarbohydrateand protein-rich diets, respectively, to observe the effect of diet on gut flora and liver. Fecal specimens were collected and used for qualitative determination of gut microbiota composition. Serum hepatospecific markers (AST, ALT and ALP) were estimated. The antioxidant status of liver tissues was estimated for GSH, MDA, GST, LDH and vitamin C l, in addition to sodium and potassium. Results: Administration of orogastric dose of ampicillin for 3 weeks induced inhibition of E.coli, yeasts, total anaerobes, and anaerobic lactobacilli with new growth of P. vulgaris and K. pneumonia. The levels of serum AST, ALT and ALP showed significant (p ˂ 0.05) increase to 163, 112.38 and 115.35 %, respectively in ampicillin-treated animals, compared to control. Also significant (p ˂ 0.05) increase in lipid peroxidation (120 %) and LDH (111 %) coupled with significant (p ˂ 0.05) decrease in glutathione (74.57 %), vitamin C (63.49 %) and glutathione S-transferase (41.51 %) were observed in ampicillintreated groups. No significant variation (p ˂ 0.05) in sodium and potassium levels were found between control and the treated group after 3 weeks of treatment. Conclusion: These results confirm that extended ampicillin therapy disrupts gut flora, which results in liver injury; hence, overuse of antibiotics should be avoid.

Characterization of glutathione S transferases from the plant-parasitic nematode, Bursaphelenchus xylophilus

We have previously identified two secreted glutathione S-transferases (GST) expressed in the pharyngeal gland cell of Bursaphelenchus xylophilus, which are upregulated post infection of the host. This study examines the functional role of GSTs in B. xylophilus biology. We analysed the expression profiles of all predicted GSTs in the genome and the results showed that they belong to kappa and cytosolic subfamilies and the majority are upregulated post infection of the host. A small percentage is potentially secreted and none is downregulated post infection of the host. One secreted protein was confirmed as a functional GST and is within a cluster that showed the highest expression fold change in infection. This enzyme has a protective activity that may involve host defences, namely in the presence of terpenoid compounds and peroxide products. These results suggest that GSTs secreted into the host participate in the detoxification of host-derived defence compounds and enable successful parasitism.

Overexpression Of Ucp1 In Tobacco Induces Mitochondrial Biogenesis And Amplifies A Broad Stress Response.

Uncoupling protein one (UCP1) is a mitochondrial inner membrane protein capable of uncoupling the electrochemical gradient from adenosine-5'-triphosphate (ATP) synthesis, dissipating energy as heat. UCP1 plays a central role in nonshivering thermogenesis in the brown adipose tissue (BAT) of hibernating animals and small rodents. A UCP1 ortholog also occurs in plants, and aside from its role in uncoupling respiration from ATP synthesis, thereby wasting energy, it plays a beneficial role in the plant response to several abiotic stresses, possibly by decreasing the production of reactive oxygen species (ROS) and regulating cellular redox homeostasis. However, the molecular mechanisms by which UCP1 is associated with stress tolerance remain unknown. Here, we report that the overexpression of UCP1 increases mitochondrial biogenesis, increases the uncoupled respiration of isolated mitochondria, and decreases cellular ATP concentration. We observed that the overexpression of UCP1 alters mitochondrial bioenergetics and modulates mitochondrial-nuclear communication, inducing the upregulation of hundreds of nuclear- and mitochondrial-encoded mitochondrial proteins. Electron microscopy analysis showed that these metabolic changes were associated with alterations in mitochondrial number, area and morphology. Surprisingly, UCP1 overexpression also induces the upregulation of hundreds of stress-responsive genes, including some involved in the antioxidant defense system, such as superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione-S-transferase (GST). As a consequence of the increased UCP1 activity and increased expression of oxidative stress-responsive genes, the UCP1-overexpressing plants showed reduced ROS accumulation. These beneficial metabolic effects may be responsible for the better performance of UCP1-overexpressing lines in low pH, high salt, high osmolarity, low temperature, and oxidative stress conditions. Overexpression of UCP1 in the mitochondrial inner membrane induced increased uncoupling respiration, decreased ROS accumulation under abiotic stresses, and diminished cellular ATP content. These events may have triggered the expression of mitochondrial and stress-responsive genes in a coordinated manner. Because these metabolic alterations did not impair plant growth and development, UCP1 overexpression can potentially be used to create crops better adapted to abiotic stress conditions.

Antioxidative responses of cell suspension cultures of two Coffea arabica varieties to low aluminum levels at pH 5.8

The effects of aluminum (Al) on the activities of antioxidant enzymes and ferritin expression were studied in cell suspension cultures of two varieties of Coffea arabica, Mundo Novo and Icatu, in medium with pH at 5.8. The cells were incubated with 300 µM Al3+, and the Al speciation as Al3+ was 1.45% of the mole fraction. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) were increased in Mundo Novo, whereas glutathione reductase (GR) and guaiacol peroxidase (GPOX) activities remained unchanged. SOD, GR, and GST activities were increased in Icatu, while CAT activity was not changed, and GPOX activity decreased. The expression of two ferritin genes (CaFer1 and CaFer2) were analyzed by Real-Time PCR. Al caused a downregulation of CaFER1 expression and no changes of CaFER2 expression in both varieties. The Western blot showed no alteration in ferritin protein levels in Mundo Novo and a decrease in Icatu. The differential enzymes responses indicate that the response to Al is variety-dependent.

Interação entre consumo alimentar e polimorfismos da GSTM1 e GSTT1 no risco para o câncer de cabeça e pescoço: estudo caso-controle em São Paulo, Brasil

O objetivo foi investigar a interação entre fatores dietéticos e polimorfismos de enzimas de metabolização de xenobióticos (GSTM1 e GSTT1) associadas ao câncer de cabeça e pescoço em um estudo caso controle de base hospitalar, no Município de São Paulo, Brasil. Participaram 103 casos incidentes, histologicamente confirmados, e 101 controles. O consumo alimentar foi obtido por um questionário de frequência alimentar validado. Os polimorfismos GSTM1 e GSTT1 foram avaliados pelo método PCR. Observou-se aumento de risco no mais alto tercil de consumo de carne bovina na presença do alelo nulo da GSTM1 (OR = 10,79; IC95%: 2,17-53,64) e GSTT1 (OR = 3,41; IC95%: 0,43-27,21). Considerando-se a razão entre alimentos de origem animal e vegetal, verificou-se para o tercil intermediário a OR = 2,02 (IC95%: 0,24-16,0) e no tercil superior OR = 3,23 (IC95%: 0,40-25,92). Os resultados apontam para uma possível interação entre o consumo de carne e variantes polimórficas dos genes GSTM1 e GSTT1 na modulação do risco para o câncer de cabeça e pescoço, influenciados pelo consumo de alimentos de origem vegetal.