Single Nucleotide Polymorphism Genotyping and Distribution of Coxiella burnetii Strains from Field Samples in Belgium

The genotypic characterization of Coxiella burnetii provides useful information about the strains circulating at the farm, region, or country level and may be used to identify the source of infection for animals and humans. The aim of the present study was to investigate the strains of C. burnetii circulating in caprine and bovine Belgian farms using a single nucleotide polymorphism (SNP) technique. Direct genotyping was applied to different samples (bulk tank milk, individual milk, vaginal swab, fetal product, and air sample). Besides the well-known SNP genotypes, unreported ones were found in bovine and caprine samples, increasing the variability of the strains found in the two species in Belgium. Moreover, multiple genotypes were detected contemporarily in caprine farms at different years of sampling and by using different samples. Interestingly, certain SNP genotypes were detected in both bovine and caprine samples, raising the question of interspecies transmission of the pathogen.     

Function and regulation of the acid and neutral trehalases of Mucor rouxii

Two different trehalose-hydrolysing activities, known as acid or non-regulatory trehalases, and neutral or regulatory trehalases, have been recognised in a number of fungal species. The true role of these apparently redundant hydrolases remained obscure for many years. However, recent evidence suggests that neutral trehalases would be specialised in the mobilisation of cytosolic trehalose, while acid trehalases would only hydrolyse extracellular trehalose. Results obtained with Mucor rouxii, a Zygomycete initially thought to posses only neutral trehalase activity, reinforced this hypothesis. M. rouxii grows efficiently in trehalose as the sole carbon source. Trehalose-grown or carbon-starved cells exhibit a high trehalase activity of optimum pH 4.5, bound to the external surface of the cell wall, in contrast with the neutral (pH 6.5) trehalase, which occurs in the cytosol. Other differences between the neutral and the acid trehalases are the temperature optimum (35°C and 45°C, respectively) and thermal stability (half-life of 2.5 min and 12 min at 45°C, respectively). The neutral trehalase, but not the acid trehalase, is activated in vitro by cAMP-dependent phosphorylation, stimulated by Ca²⁺, and inhibited by EDTA. It shows maximal activity at germination and decreases as growth proceeds. In contrast the activity of the acid trehalase is totally repressed in glucose-grown cultures and increases upon exhaustion of the carbon source, and is strongly induced by extracellular trehalose.     

The effect of different solvents on the ATP/ADP content and growth properties of HeLa cells

Testing of the effects of xenobiotics in cultured cells often requires the use of organic solvents to effect suspension of the test agents in cell culture media. However, the toxic effects of the solvents themselves may introduce artifacts, which obscure interpretation of the experimental results. In this article, the toxicity of different solvents commonly used for solvation of a variety of xenobiotic agents was studied. We show that ethanol, acetone, isooctane, methanol, and hexane were considerably less toxic than the more commonly used solvent, DMSO, when ATP content and growth rates of HeLa cells exposed to these solvents was measured. © 1998 John Wiley & Sons, Inc. J Biochem Toxicol 13: 11–15, 1999     

Atorvastatin and Fluoxetine Prevent Oxidative Stress and Mitochondrial Dysfunction Evoked by Glutamate Toxicity in Hippocampal Slices

Atorvastatin has been shown to exert a neuroprotective action by counteracting glutamatergic toxicity. Recently, we have shown atorvastatin also exerts an antidepressant-like effect that depends on both glutamatergic and serotonergic systems modulation. Excitotoxicity is involved in several brain disorders including depression; thus, it is suggested that antidepressants may target glutamatergic system as a final common pathway. In this study, a comparison of the mechanisms involved in the putative neuroprotective effect of a repetitive atorvastatin or fluoxetine treatment against glutamate toxicity in hippocampal slices was performed. Adult Swiss mice were treated with atorvastatin (10 mg/kg, p.o.) or fluoxetine (10 mg/kg, p.o.), once a day during seven consecutive days. On the eighth day, animals were killed and hippocampal slices were obtained and subjected to an in vitro protocol of glutamate toxicity. An acute treatment of atorvastatin or fluoxetine was not neuroprotective; however, the repeated atorvastatin or fluoxetine treatment prevented the decrease in cellular viability induced by glutamate in hippocampal slices. The loss of cellular viability induced by glutamate was accompanied by increased D-aspartate release, increased reactive oxygen species (ROS) and nitric oxide (NO) production, and impaired mitochondrial membrane potential. Atorvastatin or fluoxetine repeated treatment also presented an antidepressant-like effect in the tail suspension test. Atorvastatin or fluoxetine treatment was effective in protecting mice hippocampal slices from glutamate toxicity by preventing the oxidative stress and mitochondrial dysfunction.     

Effect of temporal and geographical factors on fatty acid composition of M. galloprovincialis from the Adriatic sea

Soft body tissue and gill and mantle microsomal membranes of Mytilus galloprovincialis, were analyzed to assess geographical and temporal effects on fatty acid distribution with special focus on n-3 polyunsaturated fatty acid (PUFA) content. Mussels and plankton samples were collected in April and in October from three locations of the North Adriatic Sea. Plankton fatty acid composition apparently depended both on the sampling site and time and differences referable to temporal features prevailed on the geographical ones. Accordingly, also mussel fatty acid composition appeared to be more effectively modulated by sampling time rather than by sampling location. Membrane fatty acids showed high homeostatic capabilities to reduce effects of exogenous input on cellular organization. Selective capabilities apparently enable mussels to modulate their fatty acid composition combining plankton input, environmental effects on lipid metabolism and physiological requirements. The constantly higher n-3/n-6 ratio in April than in October, shared by fatty acids of plankton, soft tissues and microsomal membranes, confirms the influence of temporal factors in fatty acid modulation. On these bases, late spring is suggested to be the more suitable period for collecting mussels destined for healthy diet of humans or other animals.     

Regeneration in invertebrates

The mechanisms of regeneration are reviewed from a genetic, cytological and molecular biological points of view. Planarians and Hydra have been chosen and illustrated as biological examples.      

Trap-nests used by Centris (Heterocentris) terminata Smith (Hymenoptera: Apidae, Centridini) at secondary Atlantic Forest fragments, in Salvador, Bahia State

Ninety-five nests of Centris (Heterocentris) terminata Smith were collected in trap-nests, during November/2001 and January/2003, at two fragments (PZGV e CFO-UFBA) of secondary Atlantic Forest, in Salvador, Bahia State (13 masculine01 W e 38 masculine30 S). The highest nest frequencies occurred from December to February (summer), with no nests foundations from August to October (winter - early spring). Two-hundred eight adults emerged from 347 brood cells, being 164 males and 116 females (1: 0.42). During the study period sex ratio was male biased (c2 = 9.342; gl = 10; P < 0.05). C. terminata nested in holes with diameters 6, 8, 10 mm, but 84,2% were constructed in 8 and 10 mm. nests had one to seven cells arranged in a linear series with the cells partitions built with a mixture of sand and resin or oil. Male is significantly smaller than female, which emerges from the first cells constructed. Immature mortality occurred in 14.1% of brood cells (n = 49), of which 13.0% were due fail in development and 1.2% due to parasitism of Coelioxys sp. (Hymenoptera: Megachilidae) e Tetraonyx sp. (Coleoptera: Meloidae). In the study site, weather, mainly pluviosity, rather than natural enemies influenced seasonal population abundance. The long period of nesting activity, local abundance and usage of trap nests, suggest the potential of C. terminata for management aiming at pollination of native and cultivated plants.     

Myricitrin as a substrate and inhibitor of myeloperoxidase: implications for the pharmacological effects of flavonoids

Flavonoids are increasingly being ingested by the general population as chemotherapeutic and anti-inflammatory agents. They are potentially toxic because of their conversion to free radicals and reactive quinones by peroxidases. Little detailed information is available on how flavonoids interact with myeloperoxidase, which is the predominant peroxidase present at sites of inflammation. This enzyme uses hydrogen peroxide to oxidize chloride to hypochlorous acid, as well as to produce an array of reactive free radicals from organic substrates. We investigated how the flavonoid myricitrin is oxidized by myeloperoxidase and how it affects the activities of this enzyme. Myricitrin was readily oxidized by myeloperoxidase in the presence of hydrogen peroxide. Its main oxidation product was a dimer that underwent further oxidation. In the presence of glutathione, myricitrin was oxidized to a hydroquinone that was conjugated to glutathione. When myeloperoxidase oxidized myricitrin and related flavonoids it became irreversibly inactivated. The number of hydroxyl groups in the B ring of the flavonoids and the presence of a free hydroxyl m-phenol group in the A ring were important for the inhibitory effects. Less enzyme inactivation occurred in the presence of chloride. Neutrophils also oxidized myricitrin to dimers in a reaction that was partially dependent on myeloperoxidase. Myricitrin did not affect the production of hypochlorous acid by neutrophils. We conclude that myricitrin will be oxidized by neutrophils at sites of inflammation to produce reactive free radicals and quinones. It is unlikely to affect hypochlorous acid production by neutrophils.     

Airborne pollen patterns in Mar del Plata atmosphere (Argentina) and its relationship with meteorological conditions

This work is part of a series of aerobiological researches conducted in the city of Mar del Plata, Argentina. The annual, seasonal and daily features of the pollen cloud were analyzed over 2 years, together with the effects of the meteorological variables. Cupressaceae amounted to 75 and 54% of the annual total in each year, and it was analyzed separately due to its proven overrepresentation. Eighty-five pollen types were registered during the 2 years, bearing a similar annual pattern (more than 50% from August to November, December also being an important month). Arboreal pollen predominated between late winter and spring (61 and 49% of the annual total), while non-arboreal pollen did so from late spring to summer. The observed seasonality was significantly correlated with the monthly mean temperatures and associated with its effect on phenology. No significant differences along the day were detected between years and 50% of the daily total was recorded between 1000 and 1600 hours. The maximum concentration was registered at 1000 hours in spring and summer, but later in autumn and winter. Regarding the hourly scale, the significant correlation of total pollen with temperature and wind speed was positive, albeit negative with relative humidity. These effects are linked to airborne pollen release and transport. The role rainfall plays on airborne pollen is discussed. To interpret the results, it is important to consider the time scale at which the aerobiological phenomenon is analyzed.