Effects of restrictions in dietary protein and vitamin A on the responses of rats to infections by Nippostrongylus brasiliensis (Nematoda) and N. brasiliensis plus Eimeria nieschulzi (Coccidia)

The effects of adequate and restricted dietary protein and vitamin A on responses to infections by Nippostrongylus brasiliensis and N. brasiliensis plus Eimeria nieshulzi were determined in growing Sprague-Dawley rats. Infected rats experienced anorexia followed by a rebound in consumption that compensated for weight losses during anorexia. On certain days, reductions in the urinary/fecal nitrogen ratio, fecal and absorbed nitrogen, and apparent nitrogen and dry matter digestibilities occurred with the combined infections but not with those by nematodes alone. Effects of different levels of vitamin A were expressed only as an increase in nitrogen absorption occurring during the post-anoretic increase in appetite found with infected rats and in rats restricted in protein but receiving the higher level of the vitamin. Protein level was the most significant treatment effect: rats on high protein performed significantly better than those on low, regardless of the level of the other experimental variables.     

Direct Evidence That Excitotoxicity in Cultured Neurons Is Mediated via N-Methyl-D-Aspartate (NMDA) as well as Non-NMDA Receptors

Cultured GABAergic cerebral cortex neurons were exposed to the excitatory amino acid (EAA) L-glutamate, kainate (KA), N-methyl-D-aspartate (NMDA), or RS-alpha-amino-3-hydroxy-5-methyl-4-isoxazolopropionate (AMPA). To ensure a constant glutamate concentration in the culture media during the exposure periods, the glutamate uptake inhibitor L-aspartic acid beta-hydroxamate was added at 500 microM to the cultures that were exposed to glutamate. Each of these EAAs was able to induce neurotoxicity. It was not possible to reduce or prevent glutamate-induced cytotoxicity by blocking only one of the glutamate receptor subtypes with either the NMDA receptor antagonist D-(-)-2-amino-5-phosphonopentanoate (APV) or with one of the specific non-NMDA antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX). However, if the cultures were exposed simultaneously to glutamate and the antagonists in combination, i.e., APV plus CNQX or APV plus DNQX, the toxicity was completely prevented. Furthermore, CNQX and DNQX were shown to be selective blockers of cytotoxic phenomena induced by non-NMDA glutamate agonists with no effect on NMDA-induced cell death. Likewise, APV prevented NMDA-induced cell death without affecting the KA- or AMPA-induced neurotoxicity. It is concluded that EAA-dependent neurotoxicity is induced by NMDA as well as non-NMDA receptors.     

AICAR is not an endogenous mutagen in Escherichia coli

A number of observations in the Escherichia coli and Salmonella typhimurium literature could be explained by the hypothesis that a particular purine ribonucleotide precursor can be converted to the corresponding deoxyribonucleotide triphosphate, thereby becoming a base-analogue mutagen. The metabolite in question, AICAR (5-amino-4-carboxamide imidazole riboside 5′-phosphate), is also a by-product of histidine biosynthesis, and its (ribo)triphosphate derivative, ZTP, has been detected in E. coli. We constructed E. coli tester strains that had either a normal AICAR pool (pur ⁺ his ⁺ strains cultivated without purines or histidine) or no AICAR pool (purF hisG mutant strains, lacking the first enzyme of each pathway and cultivated in the presence of adenine and histidine). Using a set of lacZ mutations, each of which can revert to Lac⁺ only by a specific substitution mutation, we found that no base substitution event occurs at a higher frequency in the presence of an AICAR pool. We conclude that the normal AICAR pool in E. coli is not a significant source of spontaneous base substitution mutagenesis.     

Entomopathogenic fungi associated with Ixodes ricinus ticks

The objective of this study was to demonstrate the occurrence of entomopathogenic fungi on Ixodes ricinus ticks in relation to the tick stage, engorgement and season. Ticks were collected from the vegetation, from small rodents and from deer. All entomopathogenic fungi found belonged to the Hyphomycetes. Paecilomyces farinosus and Verticillium lecanii were the predominant species. Other species, found only on engorged females were: Beauveria bassiana, B. brongniartii, P. fumosoroseus and V. aranearum. Eight out of 1833 ticks collected from the vegetation and three out of 269 engorged nymphs were infected with fungi. Thirty-three out of 149 engorged females were infected, whereas males and engorged larvae were not infected. Throughout the season, a significantly higher proportion of ticks collected in autumn were infected. Entomopathogenic fungi may have a significant impact on the size of the I. ricinus population, since females were the most frequently infected stage.     

Schistosoma mansoni and Fasciola hepatica: Cross-resistance in mice with single-sex schistosome infections

Primary Schistosoma mansoni single-sex infections in mice, i.e., either male only or female only, did not stimulate any detectable level of heterologous resistance to challenge with Fasciola hepatica after 22 to 76 days, while statistically significant resistance to a challenge with F. hepatica was demonstrated in the presence of patent mixed-sex S. mansoni infections. Simultaneous infections with S. mansoni and F. hepatica induced a statistically significant reduction in the number of schisto-some worms established, i.e., the burden being reduced by 40.1 and 43.9%, respectively. There was no reduction of the F. hepatica worm burden. Similar features could be observed with a time interval of 48 hr between the S. mansoni infection and the F. hepatica challenge, i.e., the schistosome burden being reduced by 34.2 and 45.6%, respectively. Furthermore, simultaneous infections with S. mansoni and F. hepatica induced a statistically significant reduction of the egg production capacity per paired female schistosome worm as compared with that of the S. mansoni control group. Tissue egg counts of the various intestinal sections were reduced by 92.8–99.6%.     

Ultrastructural localization of the Pasteurella multocida toxin in a toxin-producing strain

Toxigenic strains of Pasteurella multocida produce the 147 kDa protein Pasteurella multocida toxin (PMT) which is responsible for the osteoclastic bone resorption in progressive atrophic rhinitis in pigs and induces such resorption in all experimental animals tested so far. In the present study we have carried out immunocytochemistry on formaldehyde- and glutaraldehyde-fixed ultracryocut P. multocida using a pool of monoclonal antibodies against different epitopes on PMT as the first layer and affinity purified rabbit anti-mouse IgG as the second layer. Goat anti-rabbit IgG conjugated with 5 nm gold particles was used as marker. The gold particles were silver-enhanced prior to examination in the transmission electron microscope. Whole bacteria were also immunostained after fixation and critical point drying and examined by scanning transmission electron microscopy. The results showed that PMT was located in the cytoplasm of P. multocida. PMT could not be detected on intact, undamaged P. multocida by scanning electron microscopy. Neither pili nor flagella could be detected on the surface of the negatively stained P. multocida strains investigated. PMT has a series of characteristics encompassed in the definition of an exotoxin. However, that PMT was not secreted by living intact P. multocida is unexpected for an exotoxin.     

A review of the biology and transmission ecology of African bovine species of the genus Schistosoma

The present paper reviews the information available concerning the biology and transmission ecology of the African bovine species Schistosoma bovis, S. mattheei, S. margrebowiei and S. leiperi. Criteria for species identification (egg morphology, intermediate host spectra, definitive host-parasite relationships, etc.) are listed and the geographical distribution of the four species and factors determining the relative occurrence within their overall distributional ranges are described. S bovis and S. mattheei occur north and south of 10 degrees S, respectively, and S. margrebowiei occurs mainly, and S. leiperi only, in southern central Africa. Definitive host-related factors (susceptibility, water contact pattern, ect.) providing the background for being a primary definitive host and the primary definitive host spectra for the four schistosome species are described. The primary definitive host spectrum for S. margrebowiei and S. leiperi comprise lechwe, puku and waterbuck, for S. mattheei lechwe, puku, waterbuck plus cattle, and for S. bovis cattle and possibly also some of the listed antelope species. In addition, wild bovines and cattle may provide a reservoir of S. mattheei and S. margrebowiei in humans, but wild bovines and domestic stock play no major role in the transmission of other human species of schistosomes. The intermediate snail host spectra of S. mattheei and S. leiperi only comprise members of the Bulinus africanus species complex; S. bovis is transmitted by members of the B. truncatus, B. africanus and B. forskalii species groups, and S. margrebowiei is transmitted by members of the B. forskalii species group and possibly also by members of the B. tropicus and B. truncatus species groups. Factors determining the transmission ecology of the four schistosome species, and thereby the epidemiology of bovine schistosomiasis, are discussed. Influential factors comprise environmental conditions mediated via the effect of these on the size of the snail host population and on the rate of the intramolluscan development, behavioural patterns of the definitive host population and the course of the infection in the definitive host as related to aspects of susceptibility and level of endemicity. The epidemiological pattern (prevalence and intensity of infection, seasonality of transmission, etc.) is described and exemplified, and it is finally concluded that the increasing water conservation and changing methods of husbandry may result in bovine schistosomiasis becoming a major veterinary problem in Africa.     

Proteomic study of skeletal muscle in obesity and type 2 diabetes: progress and potential

ABSTRACT

Introduction: Skeletal muscle is the major site of insulin-stimulated glucose uptake and imparts the beneficial effects of exercise, and hence is an important site of insulin resistance in obesity and type 2 diabetes (T2D). Despite extensive molecular biology-oriented research the molecular mechanisms underlying insulin resistance in skeletal muscle remain to be established.

Areas covered: The proteomic capabilities have greatly improved over the last decades. This review summarizes the technical challenges in skeletal muscle proteomics studies as well as the results of quantitative proteomic studies of skeletal muscle in relation to obesity, T2D, and exercise.

Expert commentary: Current available proteomic studies contribute to the view that insulin resistance in obesity and T2D is associated with increased glycolysis and reduced mitochondrial oxidative metabolism in skeletal muscle, and that the latter can be improved by exercise. Future proteomics studies should be designed to markedly intensify the identification of abnormalities in metabolic and signaling pathways in skeletal muscle of insulin-resistant individuals to increase the understanding of the pathogenesis of T2D, but more importantly to identify multiple novel targets of treatment of which at least some can be safely targeted by novel drugs to treat and prevent T2D and reduce risk of cardiovascular disease.     

Functional Modulation of the Glutamate Transporter Variant GLT1b by the PDZ Domain Protein PICK1

The dominant glutamate transporter isoform in the mammalian brain, GLT1, exists as at least three splice variants, GLT1a, GLT1b, and GLT1c. GLT1b interacts with the scaffold protein PICK1 (protein interacting with kinase C1), which is implicated in glutamatergic neurotransmission via its regulatory effect on trafficking of AMPA-type glutamate receptors. The 11 extreme C-terminal residues specific for the GLT1b variant are essential for its specific interaction with the PICK1 PDZ domain, but a functional consequence of this interaction has remained unresolved. To identify a functional effect of PICK1 on GLT1a or GLT1b separately, we employed the Xenopus laevis expression system. GLT1a and GLT1b displayed similar electrophysiological properties and EC₅₀ for glutamate. Co-expressed PICK1 localized efficiently to the plasma membrane and resulted in a 5-fold enhancement of the leak current in GLT1b-expressing oocytes with only a minor effect on [³H]glutamate uptake. Three different GLT1 substrates all caused a slow TBOA-sensitive decay in the membrane current upon prolonged application, which provides support for the leak current being mediated by GLT1b itself. Leak and glutamate-evoked currents in GLT1a-expressing oocytes were unaffected by PICK1 co-expression. PKC activation down-regulated GLT1a and GLT1b activity to a similar extent, which was not affected by co-expression of PICK1. In conclusion, PICK1 may not only affect glutamatergic neurotransmission by its regulatory effect on glutamate receptors but may also affect neuronal excitability via an increased GLT1b-mediated leak current. This may be particularly relevant in pathological conditions such as amyotrophic lateral sclerosis and cerebral hypoxia, which are associated with neuronal GLT1b up-regulation.