Microbial transformation of zearalenone to a zearalenone sulfate.

The conversion of zearalenone by various microorganisms was studied. A new polar metabolite was formed in addition to alpha- and beta-zearalenols. The structure of the new metabolite was determined as zearalenone-4-O-sulfate conjugate on the basis of enzymatic and acid hydrolysis, followed by mass spectrometry, nuclear magnetic resonance, and infrared spectroscopic analysis. The results obtained demonstrate that Rhizopus arrhizus catalyzes sulfation of zearalenone at the C-4 hydroxyl group.     

Effects of ambient temperature on brain acetylcholinesterase activity and protein content in three Egyptian vertebrates

Exposure to cold caused an increase in AChE activity of the different brain regions of both Arvicanthis and Columbia and a decrease in the enzyme activity of Scincus midbrain. Heat exposure provoked variable changes in AChE activity of the various brain regions of the three experimental species. The changes in AChE activities may be one of the mechanisms by which birds and mammals tend to acclimatize themselves to various forms of stress. In reptiles, AChE activity varied with changes in ambient temperature and this is probably due to the adaptive significance of thermally directed changes in enzyme substrate affinity.     

In situ synthesis of oligonucleotide microarrays.

This contribution presents a brief overall look of the methods for the preparation of various types of DNA microarrays and a thorough examination of the methods for in situ synthesis of oligonucleotide microarrays.     

Influence of Dangling Ends and Surface-Proximal Tails of Targets on Probe-Target Duplex Formation in 16S rRNA Gene-Based Diagnostic Arrays

Dangling ends and surface-proximal tails of gene targets influence probe-target duplex formation and affect the signal intensity of probes on diagnostic microarrays. This phenomenon was evaluated using an oligonucleotide microarray containing 18-mer probes corresponding to the 16S rRNA genes of 10 waterborne pathogens and a number of synthetic and PCR-amplified gene targets. Signal intensities for Klenow/random primer-labeled 16S rRNA gene targets were dissimilar from those for 45-mer synthetic targets for nearly 73% of the probes tested. Klenow/random primer-labeled targets resulted in an interaction with a complex mixture of 16S rRNA genes (used as the background) 3.7 times higher than the interaction of 45-mer targets with the same mixture. A 7-base-long dangling end sequence with perfect homology to another single-stranded background DNA sequence was sufficient to produce a cross-hybridization signal that was as strong as the signal obtained by the probe-target duplex itself. Gibbs free energy between the target and a well-defined background was found to be a better indicator of hybridization signal intensity than the sequence or length of the dangling end alone. The dangling end (Gibbs free energy of −7.6 kcal/mol) was found to be significantly more prone to target-background interaction than the surface-proximal tail (Gibbs free energy of −64.5 kcal/mol). This study underlines the need for careful target preparation and evaluation of signal intensities for diagnostic arrays using 16S rRNA and other gene targets due to the potential for target interaction with a complex background.     

Biocatalysis of tyrosinase using catechin as substrate in selected organic solvent media

The enzymatic activity of mushroom tyrosinase was investigated using catechin as substrate in selected organic solvent media. The results showed that optimal tyrosinase activity was obtained at pH 6.2, 6.6, 6.0 and 6.2 in the organic solvent media of heptane, toluene, dichloromethane, and dichloroethane, respectively, and at a temperature between 25°C and 27.5°C. In addition, the kinetic studies showed that the K_m values were 5.38, 1.03, 2.52 and 4.03 mM, for the tyrosinase-catechin biocatalysis in the reaction media of heptane, toluene, dichloromethane, and dichloroethane, respectively, while the corresponding V_max values were 1.22×10⁻³, 0.33×10⁻³, 1.47×10⁻³ and 1.20×10⁻³ δA per μg protein per second, respectively. The use of acetone as co-solvent for the tyrosinase-catechin biocatalysis showed that acetone concentrations ranging from 5% to 30% (v/v) in the heptane reaction medium produced a decrease of 4.3% to 96.7% in tyrosinase activity. The results also indicated that the presence of 12.5% acetone in the reaction medium of dichloromethane, and 22.0% in those of toluene and dichloroethane produced a maximal increase of 42.6%, 92.1% and 71.8%, respectively, in tyrosinase activity. However, the overall findings indicated that additional increases in acetone concentration resulted in an inhibition of tyrosinase activity.     

Triazine inhibits electron transfer in photosystem 2 and induces lipid peroxidation in thylakoid membrane of maize

Triazine treatment of thylakoid membranes isolated from the primary leaves ofZea mays L. showed an 80% inhibition of photosystem (PS) 2 activity. No detectable change of PS 1 activity was found. The inhibited membranes showed a selective reduction of the most unsaturated linolenic acid (C_18∶3) biosynthesis by about 15% coupled with a corresponding increase in stearic (C_18∶0), oleic (C_18∶1) and linoleic (C_18∶2) acids. Thus the inhibition of electron transfer of PS 2 induced by triazine treatment was followed by lipid peroxidation and changes in the thylakoid membrane fluidity.     

Effect of essential fatty acid deficiency on G-proteins, cAMP-dependent protein kinase activity and mucin secretion in the rat submandibular salivary glands

Studies were conducted to determine whether β-adrenergic cell signalling is altered in submandibular salivary glands (SMSG) is essential fatty acid (EFA) deficiency. Three groups of rats were fed diets which were deficient in EFA (EFAD), marginally deficient in EFA (MEFAD) or contained sufficient amount of EFA (Control). Rats were killed after 20 wk on diets, SMSG were dissected out and cyclic AMP-dependent protein kinase (PKA) activity was measured. The specific enzyme activities were higher in the homogenates and supernatant fractions of the gland from EFAD and MEFAD rats compared with the controls. The relative levels of guanine nucleotide-binding regulatory proteins (Gs and Gi) were also measured in the SMSG membranes of rats fed the 3 diets. The levels of Gs were significantly higher in the EFAD and MEFAD groups than in the controls. No significant differences were observed in the secretion of trichloroacetic acid-phosphotungstic acid (TCA-PTA) precipitable glycoproteins from the SMSG slices among the 3 dietary groups.