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Biomedical subjects

Ahmed M Osman

Publications and source records attributed to Ahmed M Osman.

5 recordsLinked to original sources

Kinetic and stability studies on the chloroperoxidase complexes in presence of tert-butyl hydroperoxide.

The inactivation of native chloroperoxidase (CPO) from Caldariomyces fumago in the presence of tert-butyl hydroperoxide (tert-BuOOH) was investigated. A kinetic analysis was made and the inactivation constants (V(3) and K(3)) were evaluated. In prolonged times, uni-exponential equation describes the enzyme time course inactivation. A method based on the rate of inactivation of the enzyme in the presence of the inactivating molecule tert-BuOOH was also performed. A second group of inactivation constants (j(3) and K) was obtained, which is sufficiently close to the first two, thus verifying that the decreasing of enzyme absorbance corresponds to the decay of activity.

Binding Sites↗

A linearization method for low catalytic activity enzyme kinetic analysis.

A kinetic analysis was made and a linear plot based on the general rate equation derived by Laidler [Can. J. Chem. 33, 1614-1624] is proposed. This linearization method allows determining the kinetic parameters (K(m), k(cat)) and [E](0) for enzymes with low catalytic activity. The method was applied to chloroperoxidase from Caldariomyces fumago [EC 1.11.1.10], whose kinetic parameters K(m)(app), k(cat)(app), and [E](0) with monochlorodimedone as substrate, were obtained by using the linearization plot and the V(max) value (calculated by Eadie-Hofstee plot). This plot could also be useful to the study of abenzyme kinetics provided the concentration of the latter is either higher or equal than K(m) value.

Catalysis↗

Studies on the DT-diaphorase-catalysed reaction employing quinones as substrates: evidence for a covalent modification of DT-diaphorase by tetrachloro-p-benzoquinone.

In this study, the kinetic parameters, V(max) and K(m), of rat liver DT-diaphorase were determined for a series of p-benzoquinones, with methyl, methoxy, cyano, hydroxy and halo substituents. The results show that there is no correlation between the experimentally determined rates of p-benzoquinone reduction by DT-diaphorase and the calculated chemical reactivity of the examined substrates as expressed by the energy of the lowest unoccupied molecular orbital, E(LUMO). However, a reasonable correlation was found between the natural logarithm of V(max)/K(m) and the partition coefficient of the p-benzoquinones (r=0.81). Furthermore, tetrachloro-p-benzoquinone, one of the tested quinones is shown to be an inhibitor of rat DT-diaphorase. The presence of bovine serum albumin (BSA) in the incubation mixture protects DT-diaphorase against the inactivation by tetrachloro-p-benzoquinone, probably by interacting with the quinone. Maldi-Tof analysis of the incubation mixture of the purified DT-diaphorase and tetrachloro-p-benzoquinone showed that every subunit of the enzyme shifted about +414 amu, whereas the dimer shifted about +849 amu relative to control values. This indicates a covalent modification of the rat liver DT-diaphorase by tetrachloro-p-benzoquinone.

Animals↗

Biomphalaria alexandrina: cellular responses of susceptible and resistant snails to Schistosoma mansoni infection.

Distribution pattern of Schistosoma mansoni miracidia level of host susceptility/resistance and the basic cellular responses brought into play during the parasite development. Several snail stocks showed a wide spectrum of host reaction to the parasite. From the present study, a vigorous "resistant-type" cellular response to invading miracidia is seen in the histological sections of non-susceptible snails. In this respect, they were classified in our study as "resistant snails". B. alexandrina experimentally infected with S. mansoni exhibit a wide range of histopathological and immunological changes. The rate of phagocytosis as well as the actual number of hemocytes were determined in different groups of snails during the infection cycle. The significant fluctuation (increase and/or decrease) in circulating hemocyte number was only correlated with a shift in hemocyte populations in the first two days after exposure to the infective stage. The in vitro phagocytic activity of resistant snail hemocytes was found to be higher than that observed in susceptible snails. The observed phagocytic activity indicates that susceptible snail hemocytes were capable of recognizing parasite antigen in vitro, despite of that, they were not able to clear the infection in vivo. Thus suggesting the presence of endogenous factors preventing the immune system of susceptible snails from destroying the developed parasite larvae. Therefore, the mechanism underlying the susceptibility of the snails should be investigated by studying the host-parasite interactions.

Animals↗

A possible role for an enkephalinergic system in the internal defense mechanism of Biomphalaria alexandrina exposed to Schistosoma mansoni.

The present study documents that in Bioniphalaria alexandrina coordinated responses to Schistosoma mansoni infection are modulated by receptor-mediated opioid signals. Rather comprehensive tests in susceptible and resistant snails have demonstrated: I- the presence of an endogenaus opioids in the snail hemolymph (in particular, Leu-enkephalin-like material). II- in vitro treatment of snail hemocytes with synthetic Leu-enkephalin analogue (DADLE) resulted in the modulation of cellular adherence, and phagocytic activity. III- the addition of Naloxone, either alone or in combination whith DADLE, generally reduced hemocyte activity indicating opioid-receptor-mediated mechanism. V- the presence of DADLE or Naloxone modulated the level of IL-2-, TNF-gamma- and FNF-alpha-like molecules in S. mansoni resistant and susceptible snails. Specifically, DADLE and DADLE in combination with Naloxone generally were found to be capable of modulating resistant snail hemocytes at concentrations of 10(-6) and 10(-8) M. Similar actions after incubation with the same concentrations were not detected in the susceptible snails. These observations demonstrate the existence of a complete opioid system in B. alexandrina, associated with susceptibility and resistance to S. mansoni infection, the results suggest the role of such opioid system in molecular signaling within the host and in host-parasite interactions.

Animals↗