Antioxidant capacity of Lemna gibba L. exposed to wastewater treatment

In this study, the amounts of antioxidant vitamins (A, E and C), selenium (Se), reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio and malondialdehyde (MDA) that is the indicator of lipid peroxidation were determined in Lemna gibba L. plants placed in the secondary clarifier and grown in natural water. The amounts of antioxidant vitamins (A, E and C), GSH, GSSG and MDA were determined with HPLC (high performance liquid chromatography) and the amounts of Se were determined fluorimetrically. While significant decrease in amounts of antioxidant vitamins (A, E and C) and Se takes place between the first and the second or third day depending on species and insignificant decrease takes place after that day, the amount of GSH/GSSG ratio decreases until the second day (p < 0.05) and insignificantly increases after that day. Amounts of vitamins (A, E and C), Se and GSH/GSSG ratio for the plants placed in the secondary clarifier are much less than that for control group while an opposite trend was observed in MDA level. The MDA for the plants placed in secondary clarifier has maximum value at the second day, which can be considered as the maximum stress occurring at that day. Consequently, the first two days of treatment time can be taken as acclimation time and after the acclimation time the plant lives in the hostile environmental condition with the certain amount of oxidative stress. As a result, it is determined that wastewater decreases the lifetime of plant by causing metabolic stress on it and affects antioxidant capacity. The lifetime of the plants in the secondary clarifier was determined to be five days since fading toward yellow color (necrosis) in the plants was observed at the fifth day.     

Biochemical characterization and DNA repair pathway interactions of Mag1-mediated base excision repair in Schizosaccharomyces pombe

The Schizosaccharomyces pombe mag1 gene encodes a DNA repair enzyme with sequence similarity to the AlkA family of DNA glycosylases, which are essential for the removal of cytotoxic alkylation products, the premutagenic deamination product hypoxanthine and certain cyclic ethenoadducts such as ethenoadenine. In this paper, we have purified the Mag1 protein and characterized its substrate specificity. It appears that the substrate range of Mag1 is limited to the major alkylation products, such as 3-mA, 3-mG and 7-mG, whereas no significant activity was found towards deamination products, ethenoadducts or oxidation products. The efficiency of 3-mA and 3-mG removal was 5–10 times slower for Mag1 than for Escherichia coli AlkA whereas the rate of 7-mG removal was similar to the two enzymes. The relatively low efficiency for the removal of cytotoxic 3-methylpurines is consistent with the moderate sensitivity of the mag1 mutant to methylating agents. Furthermore, we studied the initial steps of Mag1-dependent base excision repair (BER) and genetic interactions with other repair pathways by mutant analysis. The double mutants mag1 nth1, mag1 apn2 and mag1 rad2 displayed increased resistance to methyl methanesulfonate (MMS) compared with the single mutants nth1, apn2 and rad2, respectively, indicating that Mag1 initiates both short-patch (Nth1-dependent) and long-patch (Rad2-dependent) BER of MMS-induced damage. Spontaneous intrachromosomal recombination frequencies increased 3-fold in the mag1 mutant suggesting that Mag1 and recombinational repair (RR) are both involved in repair of alkylated bases. Finally, we show that the deletion of mag1 in the background of rad16, nth1 and rad2 single mutants reduced the total recombination frequencies of all three double mutants, indicating that abasic sites formed as a result of Mag1 removal of spontaneous base lesions are substrates for nucleotide excision repair, long- and short-patch BER and RR.     

Effect of carbon source on biological nutrient removal in a sequencing batch reactor

Sequencing batch operation was used for nutrient (COD, NH4-N, NO3-N, PO4-P) removal from synthetic wastewater by using different carbon sources. Operation consisted of anaerobic, anoxic, oxic, anoxic and oxic (An/Ax/Ox/Ax/Ox) phases with durations of 2/1/4.5/1.5/1.5 h. Glucose, acetate and a mixture of glucose/acetate were used as carbon source to yield a COD/N/P ratio of 100/5/1.5 in the feed. Sludge age was kept constant at 10 days. COD, NH4-N, NO3-N and PO4-P removal efficiencies were maximum at the levels of 96%, 87%, 81% and 90% respectively, when a mixture (50/50) of glucose and acetate was used.     

Synthesis and discovery of potent carbonic anhydrase,acetylcholinesterase, butyrylcholinesterase,and α‐glycosidase enzymes inhibitors: The novel N,N′‐bis‐cyanomethylamine and alkoxymethylamine derivatives

During this investigation, N,N′‐bis‐azidomethylamines, N,N′‐bis‐cyanomethylamine, new alkoxymethylamine and chiral derivatives, which are considered to be a new generation of multifunctional compounds, were synthesized, functional properties were investigated, and anticholinergic and antidiabetic properties of those compounds were studied through the laboratory tests, and it was approved that they contain physiologically active compounds rather than analogues. Novel N‐bis‐cyanomethylamine and alkoxymethylamine derivatives were effective inhibitors of the α‐glycosidase, cytosolic carbonic anhydrase I and II isoforms, butyrylcholinesterase (BChE), and acetylcholinesterase (AChE) with K_i values in the range of 0.15–13.31 nM for α‐glycosidase, 2.77–15.30 nM for human carbonic anhydrase isoenzymes I (hCA I), 3.12–21.90 nM for human carbonic anhydrase isoenzymes II (hCA II), 23.33–73.23 nM for AChE, and 3.84–48.41 nM for BChE, respectively. Indeed, the inhibition of these metabolic enzymes has been considered as a promising factor for pharmacologic intervention in a diversity of disturbances.     

Novel amides of 1,1‐bis‐(carboxymethylthio)‐1‐arylethanes: Synthesis,characterization, acetylcholinesterase,butyrylcholinesterase, and carbonic anhydrase inhibitory properties

The thiolation reaction was carried out in a benzene solution at 80°C and p‐substituted ketones and mercaptoacetic acid in a molar ratio (1:4) of in the presence of a catalytic amount of toluene sulfonic acids. The enzyme inhibition activities of the novel amides of 1,1‐bis‐(carboxymethylthio)‐1‐arylethanes derivatives were investigated. These novel amides of 1,1‐bis‐(carboxymethylthio)‐1‐arylethanes derivatives showed good inhibitory action against acetylcholinesterase (AChE) butyrylcholinesterase (BChE), and human carbonic anhydrase I and II isoforms (hCA I and II). AChE inhibitors, interacting with the enzyme as their primary target, are applied as relevant drugs and toxins. Many clinically established drugs are carbonic anhydrase inhibitors, and it is highly anticipated that many more will eventually find their way into the market. The novel synthesized compounds inhibited AChE and BChE with K_i values in the range of 0.64–1.47 nM and 9.11–48.12 nM, respectively. On the other hand, hCA I and II were effectively inhibited by these compounds, with K_i values between 63.27–132.34 and of 29.63–127.31 nM, respectively.     

Generalized rate equation for single-substrate enzyme catalyzed reactions

The most widely used rate expression for single-substrate enzyme catalyzed reactions, namely the Michaelis-Menten kinetics is based upon the assumption that enzyme concentration is in excess of the substrate in the medium and the rate is mainly limited by the substrate concentration according to saturation kinetics. However, this is only a special case and the actual rate expression varies depending on the initial enzyme/substrate ratio (E₀/S₀). When the substrate concentration exceeds the enzyme concentration the limitation is due to low enzyme concentration and the rate increases with the enzyme concentration according to saturation kinetics. The maximum rate is obtained when the initial concentrations of the enzyme and the substrate are equal. A generalized rate equation was developed in this study and special cases were discussed for enzyme catalyzed reactions.     

K+ channel regulator KCR1 suppresses heart rhythm by modulating the pacemaker current If

Hyperpolarization-activated, cyclic nucleotide sensitive (HCN) channels underlie the pacemaker current I(f), which plays an essential role in spontaneous cardiac activity. HCN channel subunits (HCN1-4) are believed to be modulated by additional regulatory proteins, which still have to be identified. Using biochemistry, molecularbiology and electrophysiology methods we demonstrate a protein-protein interaction between HCN2 and the K(+) channel regulator protein 1, named KCR1. In coimmunoprecipitation experiments we show that KCR1 and HCN2 proteins are able to associate. Heterologously expressed HCN2 whole-cell current density was significantly decreased by KCR1. KCR1 profoundly suppressed I(HCN2) single-channel activity, indicating a functional interaction between KCR1 and the HCN2 channel subunit. Endogenous KCR1 expression could be detected in adult and neonatal rat ventriculocytes. Adenoviral-mediated overexpression of KCR1 in rat cardiomyocytes (i) reduced I(f) whole-cell currents, (ii) suppressed most single-channel gating parameters, (iii) altered the activation kinetics, (iv) suppressed spontaneous action potential activity, and (v) the beating rate. More importantly, siRNA-based knock-down of endogenous KCR1 increased the native I(f) current size and single-channel activity and accelerated spontaneous beating rate, supporting an inhibitory action of endogenous KCR1 on native I(f). Our observations demonstrate for the first time that KCR1 modulates I(HCN2)/I(f) channel gating and indicate that KCR1 serves as a regulator of cardiac automaticity.     

Angiotensin II-induced changes of calcium sparks and ionic currents in human atrial myocytes: potential role for early remodeling in atrial fibrillation

AIMS: Atrial angiotensin II (ANG II) levels have been shown to be increased in atrial fibrillation (AF). The purpose of the study was to evaluate a potential role of ANG II in the early remodeling and susceptibility to chronicization of AF. METHODS AND RESULTS: Isolated human atrial myocytes were incubated in ANG II and/or angiotensin type 1 receptor blocker candesartan. ANG II markedly increased the frequency of spontaneous Ca(2+) sparks, spark full duration, time to peak Ca(2+) fluorescence and decay time measured by confocal imaging. Sarcoplasmic reticulum calcium content estimated by caffeine-evoked calcium release did not differ between ANG II-treated cells and controls. Patch-clamp recordings revealed that ANG II significantly decreased I(to) and increased I(Ca,L) current densities. Candesartan blocked these ANG II-mediated alterations. ANG II exhibited no effect on I(K1), I(Kur) and I(f) current size. Expression of connexin 40 and connexin 43 was not significantly changed by ANG II as assessed by immunohistochemistry and Western blot analysis. CONCLUSION: ANG II-induced alterations of calcium handling and electrophysiological changes in human atrial cells similar to those previously observed in the onset of AF. Prevention of these alterations by candesartan might constitute a useful pharmacological strategy for the treatment of AF.     

Fishery resource use in a subarctic Indian community

Cree Indians of Fort George, James Bay, northern Canada, maintain a large and successful subsistence fishery. Methods used in the fishery, seasons and locations of catch, and yield levels were studied, together with the population biology of two sea-run Coregonus species, cisco and whitefish, that dominate the catch. The fishery was characterized by a high degree of order, social regulation of the fishing effort and the gillnet mesh size, and practices that were identified as adaptations to the subarctic ecosystem. Fishing methods used permit the Cree to control the magnitude of the harvest and the species and size composition of the catch. There is evidence that fishers can alter the scarcity-abundance patterns of the fish stocks, and have a biologically measurable effect on the populations.