The human immunodeficiency virus (HIV) protease inhibitor indinavir directly affects the opportunistic fungal pathogen Cryptococcus neoformans

Highly active antiretroviral therapy (HAART), that includes human immunodeficiency virus (HIV) protease inhibitors (PIs), has been remarkably efficacious including against some opportunistic infections. In this report we investigated the effect(s) of the PI indinavir on protease activity by Cryptococcus neoformans, an opportunistic fungal pathogen responsible for recurrent meningoencephalitis in AIDS patients. Indinavir was also tested for potential effects on other parameters, such as fungal viability, growth ability and susceptibility to immune effector cells. It was found that indinavir impaired cryptococcal protease activity in a time- and dose-dependent fashion. The phenomenon was similarly detectable in ATCC/laboratory strains and clinical isolates. C. neoformans growth rate was also significantly reduced upon exposure to indinavir, while fungal viability was not affected and mitochondrial toxicity not detected. Furthermore, as assessed by an in vitro infection model, indinavir significantly and consistently augmented C. neoformans susceptibility to microglial cell-mediated phagocytosis and killing. Overall, by providing the first evidence that indinavir directly affects C. neoformans, these data add new in vitro insights on the wide-spectrum efficacy of PIs, further arguing for the clinical relevance of HAART against opportunistic infections in AIDS.     

The evaluation of the oxidative state of native-LDL: three methods compared

LDL-oxidation is considered a contributing factor to the development of atherosclerotic lesions. However, to utilise the oxidative state of LDL as a marker of cardiovascular risk, reliable analytical methods for its detection must be defined. We have compared three methods for their capacity to evaluate the difference in the oxidation state of isolated LDL subjected to either dialysis (D-LDL) or gel filtration (F-LDL) to remove EDTA. Their susceptibility to oxidation promoted by Cu(2+) was monitored by following the time course of conjugated diene (CD) and lipid hydroperoxide (ROOH) accumulation. The relative electrophoretic mobility (REM) of the same LDL samples was evaluated by capillary electrophoresis. As measured by all three methods, F-LDL are less prone to oxidation than D-LDL when added with CuSO(4). REM of F-LDL and D-LDL significantly differs already before the addition of the metal catalyst, whereas CD and ROOH contents become significantly different only after it. Besides confirming that a rapid centrifugation followed by gel filtration is a more convenient procedure than dialysis to remove EDTA during LDL isolation, our study suggests the REM of isolated-LDL as the biochemical marker of choice in the evaluation of its oxidative state.     

Production of oligosaccharides by autohydrolysis of brewery's spent grain

Brewery's spent grain was treated with water in a process oriented towards the production of xylo-oligosaccharides (XOS). A wide range of temperatures and reaction times were tested and the effects of these operational variables on hemicellulose solubilization and reaction products were investigated. The maximal XOS yield (61% of the feedstock xylan) was obtained at 190 degrees C after 5 min of reaction. Several oligosaccharide mixtures with different molecular weight distributions were obtained depending on temperature and reaction time. Longer reaction times led to decreased oligosaccharide production and enhanced concentrations of monosaccharides, sugar decomposition products and acetic acid. With reaction times leading to the maximal yields of XOS, little decomposition into organic acids and aldehydes was found at all the temperatures assayed. From the composition of processed solids, it was calculated that 63-77% of the initial xylan was selectively solubilized in autohydrolysis treatments.     

Nonintrusive Field Experiments Show Different Plant Responses to Warming and Drought Among Sites,Seasons, and Species in a North–South European Gradient

We used a novel, nonintrusive experimental system to examine plant responses to warming and drought across a climatic and geographical latitudinal gradient of shrubland ecosystems in four sites from northern to southern Europe (UK, Denmark, The Netherlands, and Spain). In the first two years of experimentation reported here, we measured plant cover and biomass by the pinpoint method, plant ¹⁴C uptake, stem and shoot growth, flowering, leaf chemical concentration, litterfall, and herbivory damage in the dominant plant species of each site. The two years of approximately 1°C experimental warming induced a 15% increase in total aboveground plant biomass growth in the UK site. Both direct and indirect effects of warming, such as longer growth season and increased nutrient availability, are likely to be particularly important in this and the other northern sites which tend to be temperature-limited. In the water-stressed southern site, there was no increase in total aboveground plant biomass growth as expected since warming increases water loss, and temperatures in those ecosystems are already close to the optimum for photosynthesis. The southern site presented instead the most negative response to the drought treatment consisting of a soil moisture reduction at the peak of the growing season ranging from 33% in the Spanish site to 82% in The Netherlands site. In the Spanish site there was a 14% decrease in total aboveground plant biomass growth relative to control. Flowering was decreased by drought (up to 24% in the UK and 40% in Spain). Warming and drought decreased litterfall in The Netherlands site (33% and 37%, respectively) but did not affect it in the Spanish site. The tissue P concentrations generally decreased and the N/P ratio increased with warming and drought except in the UK site, indicating a progressive importance of P limitation as a consequence of warming and drought. The magnitude of the response to warming and drought was thus very sensitive to differences among sites (cold-wet northern sites were more sensitive to warming and the warm-dry southern site was more sensitive to drought), seasons (plant processes were more sensitive to warming during the winter than during the summer), and species. As a result of these multiple plant responses, ecosystem and community level consequences may be expected.     

Restriction fragment alleles of the rabbit IGHG genes with reference to the rabbit IGHGCH2 or e locus polymorphism

Among domesticated mammals, rabbit (Oryctolagus cuniculus) is the only species possessing not more than one subclass of immunoglobulin (IgG) antibodies. The rabbit IGHGCH2 or e locus presents two serologically defined alleles, the e14 and e15 allotypes, which are correlated with amino acid variation at the IgG CH2-CH3 interface. Genetic studies, while revealing the adaptive value of this polymorphism, have relied so far entirely upon allo-antisera. Here we show how these alleles can be distinguished by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. The proposed PCR-RFLP approach allows the monitoring of IGHG locus diversity in rabbit.     

Clear water associated with biomass and nutrient variation during the growth of a Charophyte stand after a drawdown,in a tropical coastal lagoon

The community of Charophytes in the Imboassica coastal lagoon in Brazil (22° 24 S and 42° 42 W) sometimes occupies almost the entire benthic region, and presents a large variation in C:N:P ratio. The effect of drawdown on the regeneration and buildup of biomass and on the nutrient concentration of these macroalgae was studied at three different sampling sites. Drawdown results in a high level of mortality in the macroalgae stands and after the water level later rises, the process of oospore germination begins. The drawdowns occurred in November 96 and January 97, and after March 97 we took samples in order to determine biomass values and the concentration of carbon, nitrogen and phosphorus. The results indicate that the fast growth of Charophytes may absorb a great amount of the nutrients entering the lagoon. The biomass reached maximum values of between 400 and 600 g DW m^–2, and the C:N:P ratio varied from 51:7:1 to 1603:87:1, indicating that this macroalgae may grow in a wide range of nutrient concentration. The presence of this community in the Imboassica lagoon may act as one of the limiting factors controlling phytoplanktonic primary production, decreasing nutrient availability in the water column (`bottom-up' control) and keeping the water clear after drawdowns. Probably through the habitat structure produced by the great biomass reached, they provide substrate and shelter for the structuring of a community with grazing zooplankton, which acts as a `top-down' controlling mechanism on the phytoplankton.     

Increase in therapeutic index of doxorubicin and vinblastine by aptameric oligonucleotide in human T lymphoblastic drug-sensitive and multidrug-resistant cells

Aptameric GT oligomers are a new class of potential anticancer molecules that inhibit the growth of human cancer cell lines by binding to specific nuclear proteins. We demonstrated that an aptameric GT oligonucleotide increased the therapeutic index of doxorubicin and vinblastine in T lymphoblastic drug-sensitive and multidrug-resistant (MDR) cells. The doxorubicin ID50 decreased 6.5-fold by coadministration of 1 microM GT to CCRF-CEM cells and by 24-fold by coadministration of 0.75 microM GT to CEM-VLB300 cells. In CEM-VLB300 cells, the vinblastine ID50 decreased 11-fold by coadministration of 0.5 microM GT. Control CT sequence did not potentiate the drugs in either CCRF-CEM or CEM-VLB300 cells. The ability of GT to bind to specific nuclear proteins in cancer cells related to the increase in the therapeutic index of doxorubicin and vinblastine. No cooperation was detected by the administration of GT oligomer together with doxorubicin to rat differentiated thyroid FRTL-5 cells and to normal human lymphocytes. These cells did not show binding of GT to the specific nuclear proteins, and they were not sensitive to the cytotoxic action of the GT sequence. Drug potentiation by GT not involving normal human lymphocytes might be exploited to develop a more selective treatment of drug-sensitive and MDR tumors.     

Influence of photoperiod on the uptake of nitrogen and phosphorus in the water by Eichhornia crassipes and Salvinia auriculata

The main goal of this research was to quantify the concentrations of total and ammonium N, nitrate, total and soluble P in the water in the presence of Eichhornia crassipes and Salvinia auriculata, which were submitted to two different photoperiods in a 24 hours incubation period in the laboratory. The macrophytes were incubated in plastic vials of approximately 1.5 litters, with a previously prepared solution with NH4NO3, NH4Cl, and KH2PO4. Eichhornia crassipes showed the highest average rate of reduction of all the nutrients analysed in relation to Salvinia auriculata. The largest photoperiod reflected in a higher average rate of reduction of nutrients, in both plants. Therefore, we may expect that in some periods of the year (e.g. summer), the aquatic macrophytes would show higher growth rates and higher rates of nitrogen and phosphorus absorption. These results are important for the implementation and management of wastewater treatment systems in tropical areas using aquatic macrophytes.     

Diurnal variation of rates of denitrification and nitrogen fixation of periphyton associated with Oryza glumaepatula Steud in an Amazonian Lake

The rates of denitrification and biological nitrogen fixation of the periphyton communities associated with the culms and adventitious root of Oryza glumaepatula in Lake Batata, State of Pará, Brazilian Amazonia, were investigated over the course of 24 h. N₂O was not detected during the period, demonstrating that denitrification was not occurring. The low concentration of nitrate in the water column was indicated as the main factor limiting the denitrification process. The rates of biological nitrogen fixation (BNF) in culms were lowest at night and early morning, while the highest values were measured at 1200 h. BNF rates in periphyton associated with adventitious roots were lowest during the morning, while highest values were measured at 1600 h. The difference in the daily patterns of BNF rates in the periphyton of culms and roots of O. glumaepatula in the euphotic zone was attributed to defferences in the availability of organic carbon which supports this process. In the periphyton of the culms, the BNF process was carried out by heterocystous cyanobacteria, which was photoautotrophs. In the periphyton of the adventitious roots, BNF was carried out by periphytic bacteria, and is heterotrophically mediated. This revised version was published online in July 2006 with corrections to the Cover Date.