Food utilization and esterase activity in Pieris brassicae during chronic exposure to lindane-containing food

Pieris brassicae larvae were reared on a meridic diet containing 0.75 ppm lindane. The results show that overal food consumption was lower but the approximate digestibility of food and the efficiency of conversion of ingested material into tissue biomass were higher than in control larvae. Total haemolymph esterase activity of larvae fed the lindane diet was reduced. Disc electrophoresis of midgut soluble proteins revealed eight zones of carboxylesterase activity. Results suggest that the activity of two esterase zones was reduced and that of one enhanced in larvae fed the lindane diet.

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Zusammenfassung Fünfte Larvenstadien von Pieris brassicae wurden für 72 h an einer meridischen Diät gehalten, die 0,75 ppm Lindan enthielt. Wie die Ergebnisse zeigen, war die Gesamtnahrungsaufnahme ca 50% niedriger als die der Kontrollarven, dagegen erreichte das Larvengewicht über 70% der Kontrolle. Angenäherte Verdaulichkeit (AD) betrug 80,4% für die lindanhaltige Diät gegenuber 61,7% für die Kontrolldiät. Die Effizienz des Umsatzes von aufgenommener Nahrung (ECI) war ebenfalls größer für lindanhaltige Nahrung, dagegen war die Effizienz des Umsatzes von verdauter Nahrung (ECD) bei lindanhaltiger und Kontrolldiät vergleichbar. Disc-Elektrophorese von löslichen, larvalen Mitteldarmproteinen ergab acht Zonen von Esterase-Aktivität, die verschiedene Empfindlinchkeit gegenüber in-vitro Organophosphat-Hemmung zeigten. Die Aktivität von zwei Carboxylesterasen war vermindert und die Aktivität einer schnell wandernden Esterase erschien bei lindangefütterten Larven vergrößert. Die Ergebnisse lassen auch vermuten, daß die totale hämolymphesterolytische Aktivität bei lindangehaltenen Larven reduziert war. Der Effekt von reduzierter hydrolytischer Aktivität auf larvalen Lipid-Metabolismus wird diskutiert.

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This study was supported by funds from the Finnish Cultural Foundation.     

Single-site chemical modification at C10 of the baccatin III core of paclitaxel and Taxol C reduces P-glycoprotein interactions in bovine brain microvessel endothelial cells

A single-site modification of paclitaxel analogs at the C10 position on the baccatin III core that reduces interaction with P-glycoprotein in bovine brain microvessel endothelial cells is described. Modification and derivatization of the C10 position were carried out using a substrate controlled hydride addition to a key C9 and C10 diketone intermediate. The analogs were tested for tubulin assembly and cytotoxicity, and were shown to retain potency similar to paclitaxel. P-glycoprotein interaction was examined using a rhodamine assay and it was found that simple hydrolysis or epimerization of the C10 acetate of paclitaxel and Taxol C can reduce interaction with the P-glycoprotein transporter that may allow for increased permeation of taxanes into the brain.     

Paclitaxel succinate analogs: Anionic and amide introduction as a strategy to impart blood-brain barrier permeability

A focused library of TX-67 (C10 hemi-succinate) analogs has been prepared, including C7 regioisomers, esters, amides, and one-carbon homologs. These were prepared to investigate whether the lack of TX-67 interaction with P-glycoprotein (Pgp) is due to the presence of the carboxylic acid moiety and whether this phenomenon was restricted to C10 analogs. Tubulin stabilization ability, cytotoxicity, and Pgp interactions were evaluated. All carboxylic acid analogs and several of the amides had no apparent interactions with Pgp at the concentrations used, whereas the ester variants displayed characteristics of Pgp substrates. Furthermore, it was demonstrated that hydrogen-bonding properties were significant with respect to Pgp interactions. Calculations of logD and cross-sectional areas revealed that these analogs are predicted to partition into the membrane and can compete for Pgp binding sites. The anionic and amide introduction strategy may allow for delivery of paclitaxel into the CNS and may be a potential approach for the delivery of other, structurally complex and lipophilic non-CNS permeable drugs.     

Stabilization of the cyclin-dependent kinase 5 activator,p35, by paclitaxel decreases beta-amyloid toxicity in cortical neurons

One hallmark of Alzheimer's disease (AD) is the formation of neurofibrillary tangles, aggregated paired helical filaments composed of hyperphosphorylated tau. Amyloid-beta (Abeta) induces tau hyperphosphorylation, decreases microtubule (MT) stability and induces neuronal death. MT stabilizing agents have been proposed as potential therapeutics that may minimize Abeta toxicity and here we report that paclitaxel (taxol) prevents cell death induced by Abeta peptides, inhibits Abeta-induced activation of cyclin-dependent kinase 5 (cdk5) and decreases tau hyperphosphorylation. Taxol did not inhibit cdk5 directly but significantly blocked Abeta-induced calpain activation and decreased formation of the cdk5 activator, p25, from p35. Taxol specifically inhibited the Abeta-induced activation of the cytosolic cdk5-p25 complex, but not the membrane-associated cdk5-p35 complex. MT-stabilization was necessary for neuroprotection and inhibition of cdk5 but was not sufficient to prevent cell death induced by overexpression of p25. As taxol is not permeable to the blood-brain barrier, we assessed the potential of taxanes to attenuate Abeta toxicity in adult animals using a succinylated taxol analog (TX67) permeable to the blood-brain barrier. TX67, but not taxol, attenuated the magnitude of both basal and Abeta-induced cdk5 activation in acutely dissociated cortical cultures prepared from drug treated adult mice. These results suggest that MT-stabilizing agents may provide a therapeutic approach to decrease Abeta toxicity and neurofibrillary pathology in AD and other tauopathies.     

Improved xylanase production by<Emphasis Type="Italic"> Trichoderma reesei</Emphasis> grown on <Emphasis Type="SmallCaps">l</Emphasis>-arabinose and lactose or <Emphasis Type="SmallCaps">d</Emphasis>-glucose mixtures

Trichoderma reesei Rut C-30 was grown on eight different natural or rare aldopentoses as the main carbon source and on mixtures of an aldopentose with d-glucose or lactose. The fungal cells consumed all aldopentoses tested, except l-xylose and l-ribose. The highest total xylanase and cellulase activities were achieved when cells were grown on l-arabinose as the main carbon source. The total xylanase activity produced by cells grown on l-arabinose was even higher than that produced by cells grown on an equal amount of lactose. In co-metabolism of d-glucose (15 g l^–1) and l-arabinose (5 g l^–1), the total volumetric and specific xylanase productivities were improved (derepressed) approximately 23- and 18-fold, respectively, compared to a cultivation on only d-glucose (20 g l^–1). In a similar experiment, in which cells were grown on a mixture of lactose and l-arabinose, the xylanase productivity was approximately doubled, compared to a cultivation on only lactose. The cellulase productivities, however, were not improved by the addition of l-arabinose. Compared with a typical industrial fungal enzyme production process with lactose as the main carbon source, better volumetric and specific xylanase productivities were achieved both on a lactose/arabinose mixture and on a glucose/arabinose mixture.     

Influence of peroxisome proliferator-activated receptor alpha on ubiquinone biosynthesis

The control of ubiquinone biosynthesis by peroxisome proliferators was investigated using peroxisome proliferator activated receptor alpha (PPARalpha)-null mice. Administration of 2-(diethylhexyl)phthalate to control mice resulted in elevated ubiquinone levels in the liver, while dolichol, dolichyl-P and cholesterol concentrations remained unchanged. In PPARalpha-null mice, the level of these lipids were similar to control levels and administration of the peroxisome proliferator did not increase the levels of ubiquinone. The increase in ubiquinone levels was the result of increased synthesis. Induction was most pronounced in liver, kidney and heart, which have relatively high levels of PPARalpha. When the tissue concentration of hydrogen peroxide was elevated by inhibition of catalase activity with aminotriazole, the amount of ubiquinone was not increased, suggesting that the induction of ubiquinone synthesis occured through a direct mechanism. The activities of branch-point enzymes FPP-synthase, squalene synthase, cis-prenyltransferase, trans-prenyltransferase and NPHB-transferase were substantially increased in control but not in PPARalpha-null mice after treatment with peroxisome proliferators. These data suggest that the induction of ubiquinone biosynthesis after administration of peroxisome proliferators is dependent on the PPARalpha through regulation of some of the mevalonate pathway enzymes.     

Patterns of nitrogen and sulfur accumulation and retention in ombrotrophic bogs, eastern Canada

Nitrogen (N) and sulfur (S) play important roles in peatlands, through their influence on plant production and peat decomposition rates and on redox reactions, respectively, and peatlands contain substantial stores of these two elements. Using peat N and S concentrations and dry bulk density and ²¹⁰Pb dating, we determined the rates of N and S accumulation over the past 150 years in hummock and hollow profiles from 23 ombrotrophic bogs in eastern Canada. Concentrations of N and S averaged 0.80% and 0.18%, respectively, generally increased with depth in the profile and there was a weak but significant correlation between N and S concentrations. Rates of N and S accumulation over the past 50–150 years ranged from 0.5 to 4.8 g N m^?2 yr^?1 and from 0.1 to 0.9 g S m^?2 yr^?1. There were significant but weak correlations between C, N and S accumulation rates over 50‐, 100‐ and 150‐year periods. Over the last 50 years, rates of S accumulation showed little differentiation between hummocks and hollows, whereas the pattern for N accumulation was more variable (hummock minus hollow rate ranged from ?1 to +1.5 g N m^?2 yr^?1), with hummocks generally having a larger N accumulation rate, correlated with the rate of carbon (C) accumulation. There was a modest but significant positive correlation between 50‐year rates of N accumulation and wet atmospheric deposition of N measured between 1990 and 1996, with accumulation rates about four times that of wet deposition. The difference between deposition and accumulation of N is attributed to organic N deposition, dry deposition and N₂ fixation. A weaker, but still significant, correlation was observed between 50‐year S accumulation and 1990–1996 wet atmospheric S deposition, with about 75% of the deposited S accumulating in the peat. A laboratory experiment with peat cores exposed to varying water table position and simulated N and S deposition, showed that on average 87% and 98% of the deposited NH₄⁺ and NO₃^?, respectively, and 58% of the deposited S were retained in the vegetation and unsaturated zone of the cores, supporting the results from the field study.