Characterization of an acyl-coA thioesterase that functions as a major regulator of peroxisomal lipid metabolism.

Peroxisomes function in beta-oxidation of very long and long-chain fatty acids, dicarboxylic fatty acids, bile acid intermediates, prostaglandins, leukotrienes, thromboxanes, pristanic acid, and xenobiotic carboxylic acids. These lipids are mainly chain-shortened for excretion as the carboxylic acids or transported to mitochondria for further metabolism. Several of these carboxylic acids are slowly oxidized and may therefore sequester coenzyme A (CoASH). To prevent CoASH sequestration and to facilitate excretion of chain-shortened carboxylic acids, acyl-CoA thioesterases, which catalyze the hydrolysis of acyl-CoAs to the free acid and CoASH, may play important roles. Here we have cloned and characterized a peroxisomal acyl-CoA thioesterase from mouse, named PTE-2 (peroxisomal acyl-CoA thioesterase 2). PTE-2 is ubiquitously expressed and induced at mRNA level by treatment with the peroxisome proliferator WY-14,643 and fasting. Induction seen by these treatments was dependent on the peroxisome proliferator-activated receptor alpha. Recombinant PTE-2 showed a broad chain length specificity with acyl-CoAs from short- and medium-, to long-chain acyl-CoAs, and other substrates including trihydroxycoprostanoyl-CoA, hydroxymethylglutaryl-CoA, and branched chain acyl-CoAs, all of which are present in peroxisomes. Highest activities were found with the CoA esters of primary bile acids choloyl-CoA and chenodeoxycholoyl-CoA as substrates. PTE-2 activity is inhibited by free CoASH, suggesting that intraperoxisomal free CoASH levels regulate the activity of this enzyme. The acyl-CoA specificity of recombinant PTE-2 closely resembles that of purified mouse liver peroxisomes, suggesting that PTE-2 is the major acyl-CoA thioesterase in peroxisomes. Addition of recombinant PTE-2 to incubations containing isolated mouse liver peroxisomes strongly inhibited bile acid-CoA:amino acid N-acyltransferase activity, suggesting that this thioesterase can interfere with CoASH-dependent pathways. We propose that PTE-2 functions as a key regulator of peroxisomal lipid metabolism.     

Effects of copper, cadmium and contaminated harbour sediments on recolonisation of soft-bottom communities

An extensive field experiment on benthic colonisation on polluted sediments was carried out at 50 m depth at three locations representative of the pollution gradient in the inner Oslofjord. The aim of the study was to investigate the effect of copper, cadmium and polluted harbour sediment on recolonisation of benthic fauna, and whether the response varied with a varying degree of background pollution. Copper and cadmium were added to the local sediments at the three locations, while the contaminated harbour sediment was taken from the hypoxic Oslo harbour and transplanted to well-oxygenated and less-polluted locations. Oslo harbour has a poor macrofauna, the sediments are heavily contaminated and the oxygen levels in the bottom waters are low. One of the aims of this experiment was to separate effects of contamination from effects of low oxygen. Sediments with high levels of copper (∼400-1500 mg/kg) clearly had a negative effect on colonisation by several taxa. This negative response was more pronounced at the relatively pristine location in the outer fjord compared to the more polluted sites, probably as a result of an increased bioavailability of the metals in sediments with lower concentrations of organic carbon. Sediments with high levels of cadmium (∼9-23 mg/kg), on the other hand, did not have negative effects on colonisation of any taxa. In the boxes with polluted harbour sediments, only the polychaete Raricirrus beryli showed reduced abundance. This lack of community response clearly indicates that hypoxia is more critical than high concentrations of contaminants for the establishment of a normal benthic fauna in the Oslo harbour area. The experiment only lasted for 6 months and the recolonising fauna was dominated by opportunistic and r-selected species. Long-term toxic effects may therefore have occurred if the experiment had lasted longer. The experiment also showed that the time for the development of a community resembling the ambient community was faster on the polluted than on the more pristine location.     

The Surface-Associated and Secreted MopE Protein of Methylococcus capsulatus (Bath) Responds to Changes in the Concentration of Copper in the Growth Medium

Expression of surface-associated and secreted protein MopE of the methanotrophic bacterium Methylococcus capsulatus (Bath) in response to the concentration of copper ions in the growth medium was investigated. The level of protein associated with the cells and secreted to the medium changed when the copper concentration in the medium varied and was highest in cells exposed to copper stress.     

Quantitative analysis of sugar constituents of glycoproteins by capillary electrophoresis

A method for quantitative analysis of monosaccharides including N- acetylneuraminic acid derived from sialic acid-containing oligosaccharides and glycoproteins is presented. The analysis is based on the combination of chemical and enzymatic methods coupled with capillary electrophoretic (CE) separation and laser-induced fluorescence (LIF) detection. The present method utilizes a simplified acid hydrolysis procedure consisting of mild hydrolysis (0.1 M TFA) to release sialic acid and strong acid hydrolysis (2.0 N TFA) to produce amino and neutral sugars. Amino sugars released from strong acid hydrolysis of oligosaccharides and glycoproteins were reacetylated and derivatized with 8-aminopyrene-1,3,6-trisulfonate (APTS) along with neutral sugars in the presence of sodium cyanoborohydride to yield quantitatively the highly stable fluorescent APTS adducts. N- acetylneuraminic acid (Neu5Ac), a major component of most mammalian glycoproteins, was converted in a fast specific reaction by the action of neuraminic acid aldolase (N-acylneuraminate pyruvate-lyase EC 4.1.3.3) to N-acetylmannosamine (ManNAc) and pyruvate. ManNAc was then derivatized with APTS in the same manner as the other monosaccharides. This method was demonstrated for the quantitation of pure Neu5Ac and the species derived from mild acid hydrolysis of 6'-sialyl-N- acetyllactosamine and bovine fetuin glycan. Quantitative recovery of the N-acetylmannosamine was obtained from a known amount of Neu5Ac in a mixture of seven other monosaccharides or from the sialylated oligosaccharides occurring in glycoproteins. The sequence of procedures consists of acid hydrolysis, enzymatic conversion and APTS derivatization which produced quantitative recovery of APTS- monosaccharide adducts. The detection limits for sugars derivatized with APTS and detected by CE-LIF are 100 pmol for Neu5Ac and 50 pmol for the other sugars.      

The interplay of climate and land use change affects the distribution of EU bumblebees

Bumblebees in Europe have been in steady decline since the 1900s. This decline is expected to continue with climate change as the main driver. However, at the local scale, land use and land cover (LULC) change strongly affects the occurrence of bumblebees. At present, LULC change is rarely included in models of future distributions of species. This study's objective is to compare the roles of dynamic LULC change and climate change on the projected distribution patterns of 48 European bumblebee species for three change scenarios until 2100 at the scales of Europe, and Belgium, Netherlands and Luxembourg (BENELUX). We compared three types of models: (1) only climate covariates, (2) climate and static LULC covariates and (3) climate and dynamic LULC covariates. The climate and LULC change scenarios used in the models include, extreme growth applied strategy (GRAS), business as might be usual and sustainable European development goals. We analysed model performance, range gain/loss and the shift in range limits for all bumblebees. Overall, model performance improved with the introduction of LULC covariates. Dynamic models projected less range loss and gain than climate‐only projections, and greater range loss and gain than static models. Overall, there is considerable variation in species responses and effects were most pronounced at the BENELUX scale. The majority of species were predicted to lose considerable range, particularly under the extreme growth scenario (GRAS; overall mean: 64% ± 34). Model simulations project a number of local extinctions and considerable range loss at the BENELUX scale (overall mean: 56% ± 39). Therefore, we recommend species‐specific modelling to understand how LULC and climate interact in future modelling. The efficacy of dynamic LULC change should improve with higher thematic and spatial resolution. Nevertheless, current broad scale representations of change in major land use classes impact modelled future distribution patterns.     

A Strategy for Discovery of Endocrine Interactions with Application to Whole-Body Metabolism

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The effect of trichlorfon and other organophosphates on prenatal brain development in the guinea pig

The organophosphates trichlorfon, dichlorvos, dimethoate, soman, triortho-cresyl phosphate (TOCP), and the diethoxy-analogue of trichlorfon (O,O-diethyl 2,2,2-trichloro-1-hydroxyethylphosphonate, ethyl-trichlorfon), were administrated to guinea pigs between day 42 and 46 of gestation. When the offsprings were examined at birth, there was a severe reduction in brain weight in the case of trichlorfon and dichlorvos, but not after treatment with the other organophosphates. The reduction in weight was most pronounced for cerebellum, medulla oblongata, thalamus/hypothalamus and quadrigemina. The effect was less marked for cerebral cortex and hippocampus. Since soman, a potent anticholinesterase, and TOCP, an inhibitor of neuropathy target esterase, did not show any effects, this excludes that the brain hypoplasia can be caused by inhibition of these two enzymes. Further, the lack of effect with ethyl-trichlorfon has shed some light on the part of the trichlorfon molecule which could be involved in the formation of the hypoplasia. It is suggested that alkylation of DNA may be involved in the development of the lesion. The possible consequences for a teratogenic effect of trichlorfon and dichlorvos on humans are discussed.Special issue dedicated to Dr. Bernard W. Agranoff.     

Auditory prey location in a pause--travel predator: search height, search time, and attack range of Tengmalm's owls (Aegolius funereus)

We studied the nocturnal hunting and diurnal roosting behaviorof 17 radio-equipped Tengmalm's owls (Aegolius funereus), 12males and 5 females, in coniferous forest during their nestingseason. The owls perched lower when hunting than when roosting,probably because hunting perches were selected to minimize thepredator-prey distance or to obtain unobstructed access to theground-dwelling small mammal prey, whereas roosting percheswere selected to minimize the probability of being detectedby an avian predator. There was no difference between perchingheights associated with giving up and prey attack, nor werethere any differences between perching heights, perching times,and attack distances associated with successful and unsuccessfulattacks. There were no sexual differences in perching heightduring hunting or roosting. However, giving-up times tendedto be longer for females than for their mates, which is expectedbecause females are larger than males, and the relative costof flight increases with body mass. The instantaneous attackrate was independent of perching time. The owls gave up theirperches at a constant rate and independently of the amount oftime already spent on the perch in an exponentially decayingpattern. The owls perched longer, however, before launchingan attack than before giving up, probably in order to observedetected prey until the right moment for an attack. Attack distancewas independent of both perching height and perching time. Perchingtime was inversely related to perching height, which fits thetheoretical expectation that the search area will decrease withincreasing height in birds that locate prey auditorily.