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Almeida Pinheiro de Carvalho, Miguel Angelo

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  • Effects of Melatonin-aided therapy on the Glutathione antioxidant system activity and liver protection
    Publication . Popov, Serguey S.; Shulgin, Konstantin K.; Popova, Tatyana N.; Pashkov, Aleksander N.; Agarkov, Aleksander A.; Carvalho, Miguel A. A. Pinheiro de
    Acute hepatitis results from oxidative stress triggered by hepatotoxic drugs causing liver injury and the activation of caspases cascade. The glutathione antioxidant system protects against reactive oxygen species and mitigates development of these processes. The effectiveness of silymarin, a polyphenolic flavonoid, essenthiale, composed of phosphatidyl choline, and melaxen, a melatonin-correcting drug, as hepatoprotectors has been investigated. The variation of 6-sulfatoxymelatonin (aMT6s), resulting from the biotransformation of melatonin, and GSH has been measured. The activities of caspase-1 and caspase-3, glutathione antioxidant system, and NADPH-generating enzymes were determined. The aMT6s decreases in patients with drug hepatitis and recovers with administration of mexalen. GSH increased in the presence of the studied hepatoprotectors. Pathologically activated caspase-1 and caspase-3 decreased their activities in the presence of hepatoprotectors with melaxen showing the highest effect. The positive effect of melatonin appears to be related to the suppression of decompensation of the glutathione antioxidant system functions, recovery of liver redox status, and the attenuation of inhibition of the NADPH supply.
  • Expression of Glutathione peroxidase and Glutathione reductase and level of free radical processes under toxic hepatitis in rats
    Publication . Iskusnykh, Igor Y.; Popova, Tatyana N.; Agarkov, Aleksander A.; Carvalho, Miguel Â. A. Pinheiro de; Rjevskiy, Stanislav G.
    Correlation between intensity of free radical processes estimated by biochemiluminesce parameters, content of lipoperoxidation products, and changes of glutathione peroxidase (GP, EC 1.11.1.9) and glutathione reductase (GR, EC 1.6.4.2) activities at rats liver injury, after 12, 36, 70, 96, 110, and 125 hours & tetrachloromethane administration have been investigated. The histological examination of the liver sections of rats showed that prominent hepatocytes with marked vacuolisation and inflammatory cells which were arranged around the necrotic tissue are more at 96 h after exposure to CCl4. Moreover maximum increase in GR and GP activities, 2.1 and 2.5 times, respectively, was observed at 96 h after exposure to CCl4, what coincided with the maximum of free radical oxidation processes. Using a combination of reverse transcription and real-time polymerase chain reaction, expression of the glutathione peroxidase and glutathione reductase genes (Gpx1 and Gsr) was analyzed by the determination of their respective mRNAs in the rat liver tissue under toxic hepatitis conditions. The analyses of Gpx1 and Gsr expression revealed that the transcript levels increased in 2.5- and 3.0-folds, respectively. Western blot analysis revealed that the amounts of hepatic Gpx1 and Gsr proteins increased considerably after CCl4 administration. It can be proposed that the overexpression of these enzymes could be a mechanism of enhancement of hepatocytes tolerance to oxidative stress.
  • Identification of aluminum resistant genotypes among Madeiran regional wheats
    Publication . Carvalho, Miguel Â. A. Pinheiro de; Slaski, Jan J.; Santos, Teresa M. M. dos; Ganança, Filipe T.; Abreu, Ivo; Taylor, Gregory J.; Vieira, Maria R. Clemente; Popova, Tatyana N.; Franco, Emanuel
    Forty-eight genotypes representing wheat diversity from the Island of Madeira were screened for resistance to aluminum (Al) in nutrient solution. Seeds of wheat used in the experiments were obtained from local farmers. The soil pH and content of ionic Al of plots cultivated with wheat were analyzed. The pH of topsoils varied between 3.83 and 6.59. The amount of ionic Al in soil samples varied between 0.38 and 1.36 cmol Al3 + per kg of soil and was positively correlated with the altitude of a plot. Eriochrome cyanine staining was used to evaluate the effect of Al ions on the root elongation. Seventy-two hour exposure of 3-day-old seedlings to 100 and 200 mM Al in nutrient solution revealed a high number of Al resistant genotypes among wheat germplasm. After withdrawal of Al stress, survival and root regrowth was observed in 28 and 23 genotypes screened at 100 and 200 mM Al in nutrient solution, respectively. Enhanced resistance to Al among Madeiran genotypes was associated with the amount of ionic Al in the soils. Complexity and various patterns of responses of tested cultivars to Al stress may suggest that Madeiran germplasm could be a valuable source of genes controlling Al resistance for conventional breeding programs and for studies of molecular bases of mechanisms of Al resistance.
  • Citrate and isocitrate in plant metabolism
    Publication . Popova, Tatyana N.; Carvalho, Miguel Â. A. Pinheiro de
    The relevance of citrate and isocitrate metabolism in plants is discussed in connection with the different pathways for their conversions. The routes for citrate and isocitrate conversions are incorporated into the system of cross-linked metabolic processes and may provide carbon skeletons for nitrogen assimilation and reducing equivalents for biosynthetic reactions, support the functioning of the glyoxylate cycle and play an important role in the TCA and energy metabolism as a whole. The possibility of the coupling of citrate and isocitrate metabolism with various electron transport systems is discussed from the point of view of the efficiency of the balancing cellular NAD P H Ž . Ž . rNAD P and ATP q rADP ratios. The role of citrate and isocitrate and their derivations as potent effectors of some enzymes is considered. Special attention is paid to the enzymes associated with citrate and isocitrate metabolism and to the mechanisms which regulate their activity. The possibilities of the coordination of the main processes of energy and biosynthetic metabolism at the level of citrate and isocitrate distribution are discussed.