Acetyl-CoA Synthesizing Enzymes in Cholinergic Nerve Terminals

The activities of five enzymes involved in acetyl-CoA synthesis, pyruvate dehydrogenase complex, ATP citrate lyase, carnitine acetyltransferase, acetyl-CoA synthetase, and citrate synthase, were determined in normal nucleus interpeduncularis and nucleus interpeduncularis in which cholinergic terminals were removed following lesion of the habenulointerpeduncular tract. The activities of aspartate transaminase, fumarase, and GABA transaminase also were determined to compare the effect of lesion on other mitochondrial enzymes which are not linked to the biosynthesis of ACh. In normal nucleus interpeduncularis the activities of carnitine acetyltransferase and pyruvate dehydrogenase complex were higher than the activity of ChAT (choline acetyltransferase), whereas the activities of acetyl-CoA synthetase and citrate synthase were considerably lower than that of ChAT. The effect of the lesion separated the enzymes into two groups: the activities of pyruvate dehydrogenase complex, carnitine acetyltransferase, fumarase and aspartate transaminase decreased by 30--40%, whereas the activities of the other enzymes descreased 5--15%. ChAT activity was in all cases less than 15% of normal. It could be concluded that none of the acetyl-CoA synthesizing enzymes decreased to the degree that ChAT did. Only pyruvate dehydrogenase complex and carnitine acetyltransferase seem to be localized in cholinergic terminals to a significant degree. ATP citrate lyase as well as acetyl-CoA synthetase seem to have less significance in supporting acetyl-CoA formation in cholinergic nerve terminals.     

Neurotoxic Effects of Prenatal Trichlorfon Administration in Pigs

Pregnant sows were administered trichlorfon (Neguvon®) during gestation. Examination of the offspring revealed clinical symptoms which included ataxia and tremor. The weights of the cerebrum and the celebellum were significantly reduced (67% and 41% of the control values). There was a clear correlation between the decrease in total brain weight and decrease in cerebellar weight (r = 0.94). Histological examination showed a well preserved lamination of the cerebral and cerebellar cortex. Periodical loss of Purkinje cells was the most notable finding in the cerebellum. The molecular layer in such regions was markedly thinned. Neurochemical investigations of the cerebellum showed a significant decrease in specific activity of the transmitter synthesizing enzymes choline acetyltransferase, glutamate decarboxylase and aromatic amino acid decarboxylase (61.6%, 30.3% and 47.1% of the control values, respectively). No effect in high affinity D-aspartate uptake was observed.     

Anthropogenic and natural size-related selection act in concert during brown trout (Salmo trutta) smolt river descent

Hydrobiologia - By hindering migration and inducing direct turbine mortality during downstream migration, hydropower is regarded as one of the most serious threats to anadromous salmonids. Yet,...     

An illustrated key to the cuckoo wasps (Hymenoptera,Chrysididae) of the Nordic and Baltic countries,with description of a new species

The Chrysididae are a group of cleptoparasitic and parasitoid aculeate wasps with a large number of rare and endangered species. The taxonomy of this group has long been confusing due to the similarity of species and extensive intraspecific variation. We present for the first time a comprehensive dichotomous key for all 74 species found in the Nordic and Baltic countries. In addition to diagnostic characters, information on the distribution and biology of each species is also presented. A new species, Chrysis borealis Paukkunen, Ødegaard & Soon, sp. n. is described on the basis of specimens collected from Fennoscandia. Chrysis gracillima Förster, 1853 is recorded as new to the Nordic and Baltic countries.     

Analysing the outer membrane subproteome of Methylococcus capsulatus (Bath) using proteomics and novel biocomputing tools

High-resolution two-dimensional gel electrophoresis and mass spectrometry has been used to identify the outer membrane (OM) subproteome of the Gram-negative bacterium Methylococcus capsulatus (Bath). Twenty-eight unique polypeptide sequences were identified from protein samples enriched in OMs. Only six of these polypeptides had previously been identified. The predictions from novel bioinformatic methods predicting β-barrel outer membrane proteins (OMPs) and OM lipoproteins were compared to proteins identified experimentally. BOMP () predicted 43 β-barrel OMPs (1.45%) from the 2,959 annotated open reading frames. This was a lower percentage than predicted from other Gram-negative proteomes (1.8–3%). More than half of the predicted BOMPs in M. capsulatus were annotated as (conserved) hypothetical proteins with significant similarity to very few sequences in Swiss-Prot or TrEMBL. The experimental data and the computer predictions indicated that the protein composition of the M. capsulatus OM subproteome was different from that of other Gram-negative bacteria studied in a similar manner. A new program, Lipo, was developed that can analyse entire predicted proteomes and give a list of recognised lipoproteins categorised according to their lipo-box similarity to known Gram-negative lipoproteins (). This report is the first using a proteomics and bioinformatics approach to identify the OM subproteome of an obligate methanotroph.     

Specific modification of peptide-bound citrulline residues

Immune reactions to citrulline-containing proteins appear to be central in the immunopathogenesis of rheumatoid arthritis. Citrulline residues are introduced into proteins by deimination of arginine residues, likely by an enzymatic process. There is a need to characterize which proteins in the inflamed joints of rheumatoid patients contain citrulline in situ. The characterization of deiminated proteins will be greatly facilitated by specific modification of peptide-bound citrulline residues that will enable specific enrichment and detection of citrulline-containing peptides. This study presents the details of such a modification method. The chemistry behind the reaction of the ureido group of citrulline with 2,3-butanedione in the presence of antipyrine is unraveled. Parameters for optimization of the reaction with respect to specificity and completeness, including the testing of different acids, reactant concentrations, and reaction time, are presented. This modification reaction is specific for citrulline residues. The modified product shows a characteristic mass shift of +238Da, as demonstrated by mass spectrometry. The product absorbs UV-Vis radiation at 464nm, and it is demonstrated that this can be used to selectively monitor citrulline-containing peptides during the separation of protein digests. Finally, the structure of the product of modified citrulline is solved by nuclear magnetic resonance spectroscopy using N-butylurea as a model substance. The results presented should facilitate the development of tags that can be used for the enrichment and subsequent detection of citrulline-containing protein fragments by mass spectrometry.     

Host Specificity, Alpha- and Beta-Diversity of Phytophagous Beetles in Two Tropical Forests in Panama

Species diversity, host specificity and species turnover among phytophagous beetles were studied in the canopy of two tropical lowland forests in Panama with the use of canopy cranes. A sharp rainfall gradient occurs between the two sites located 80 km apart. The wetter forest is located in San Lorenzo Protected Area on the Caribbean side of the isthmus, and the drier forest is a part of the Parque Natural Metropolitano close to Panama City on the Pacific slope. Host specificity was measured as effective specialization and recorded by probability methods based on abundance categories and feeding records from a total of 102 species of trees and lianas equally distributed between the two sites. The total material collected included more than 65,000 beetles of 2462 species, of which 306 species were shared between the two sites. The wet forest was 37% more species rich than the dry forest due to more saproxylic species and flower visitors. Saproxylic species and flower visitors were also more host-specific in the wet forest. Leaf chewers showed similar levels of species richness and host specificity in both forests. The effective number of specialized species per plant species was higher in the wet forest. Higher levels of local alpha- and beta-diversity as well as host specificity based on present data from a tropical wet forest, suggests higher number of species at regional levels, a result that may have consequences for ecological estimates of global species richness.     

Mechanisms involved in the neurotoxic effects of environmental toxicants such as polychlorinated biphenyls and brominated flame retardants

Many toxic substances have been distributed to the environment, some of which have properties that promote accumulation and biomagnification in living organisms. Approximately 1.2 million metric tons of polychlorinated biphenyls (PCBs) have been produced and about 30% have been discharged to the environment. Approximately 200 000 metric tons of brominated flame retardants (BFRs) are produced annually, of which considerable amounts have been spread globally, even to the Polar Regions. Behavioral testing of animals has shown that these compounds may affect learning, memory and fine motor functions. Animals are most sensitive during early development. Several epidemiological studies have shown that PCBs and BFRs may be responsible for similar effects in humans. Of especially concern are possible effects of PCBs and BFRs in mixtures containing the highly neurotoxic methyl mercury. The compounds affect several targets in the nervous system that seem to be interconnected, and may be responsible for the observed behavioral deficits. It was shown early that PCBs affect dopamine and serotonin levels in the brain. Later studies showed that transport mechanisms of these neurotransmitters appear to be particularly sensitive to PCBs. Furthermore, PCBs affect intracellular calcium levels and induce formation of reactive oxygen species both in vivo and in vitro, and reduce cell viability in vitro. Neuroendocrine functions, particularly the thyroid hormone system, are also sensitive to disruption by PCBs and BFRs. Their metabolites, such as hydroxy‐metabolites, appear to be particularly potent. We conclude that PCBs are particularly toxic during early development and that the toxic effects are a combination of several factors, including disturbance of calcium homeostasis, oxidative stress, and influence on neurotransmitter transport. Monoaminergic cells appear to be particularly vulnerable.     

Fixed Rejection Responses to Single and Multiple Experimental Parasitism in Two Fringilla Hosts of the Common Cuckoo

Previous studies have shown that the tendency to reject parasitic eggs among certain hosts is strongly dependent on the degree of similarity with own eggs, whereas other conditional cues do not affect rejection decisions. This paper examines whether two such hosts, the closely related brambling and chaffinch, show a different tendency to reject parasitic eggs if they are multiply parasitized. Some individuals were experimentally parasitized with both a non-mimetic and a low–intermediate contrasting egg in the same breeding attempt. The non-mimetic egg was rejected almost without exception. In chaffinches, the low–intermediate contrasting egg was introduced shortly after rejection of the non-mimetic egg whereas in bramblings, both eggs were introduced simultaneously. A control group consisted of individuals that were parasitized with one low–intermediate contrasting egg. There was no significant difference in the tendency to reject the low–medium contrasting eggs between the experimental and control group in any of the species, implying that the same acceptance threshold is applied to each parasitic egg independently. Moreover, the rejection rate of non-mimetic eggs was high (>90%) regardless of whether the egg was introduced alone or together with a low–medium contrasting egg. The results are discussed in relation to recent studies with great reed warblers Acrocephalus arundinaceus that obtained contrasting results in similar experimental designs. The different degrees of flexibility displayed by Fringilla and great reed warbler hosts are likely to reflect differences in both the perception and action component of the recognition system.