Long-term spatio-temporal dynamics of the mosquito Aedes aegypti in temperate Argentina.

Buenos Aires city is located near the southern limit of the distribution of Aedes aegypti (Diptera: Culicidae). This study aimed to assess long-term variations in the abundance of Ae. aegypti in Buenos Aires in relation to changes in climatic conditions. Ae. aegypti weekly oviposition activity was analyzed and compared through nine warm seasons from 1998 to 2014, with 200 ovitraps placed across the whole extension of the city. The temporal and spatial dynamics of abundances were compared among seasons, and their relation with climatic variables were analyzed. Results showed a trend to higher peak abundances, a higher number of infested sites, and longer duration of the oviposition season through subsequent years, consistent with a long-term colonization process. In contrast, thermal favorability and rainfall pattern did not show a consistent trend of changes. The long-term increase in abundance, and the recently documented expansion of Ae. aegypti to colder areas of Buenos Aires province suggest that local populations might be adapting to lower temperature conditions. The steadily increasing abundances may have implications on the risk of dengue transmission.

Combined PDK1 and CHK1 inhibition is required to kill glioblastoma stem-like cells in vitro and in vivo.

Glioblastoma (GBM) is the most common and deadly adult brain tumor. Despite aggressive surgery, radiation, and chemotherapy, the life expectancy of patients diagnosed with GBM is ∼14 months. The extremely aggressive nature of GBM results from glioblastoma stem-like cells (GSCs) that sustain GBM growth, survive intensive chemotherapy, and give rise to tumor recurrence. There is accumulating evidence revealing that GSC resilience is because of concomitant activation of multiple survival pathways. In order to decode the signal transduction networks responsible for the malignant properties of GSCs, we analyzed a collection of GSC lines using a dual, but complementary, experimental approach, that is, reverse-phase protein microarrays (RPPMs) and kinase inhibitor library screening. We treated GSCs in vitro with clinically relevant concentrations of temozolomide (TMZ) and performed RPPM to detect changes in phosphorylation patterns that could be associated with resistance. In addition, we screened GSCs in vitro with a library of protein and lipid kinase inhibitors to identify specific targets involved in GSC survival and proliferation. We show that GSCs are relatively insensitive to TMZ treatment in terms of pathway activation and, although displaying heterogeneous individual phospho-proteomic profiles, most GSCs are resistant to specific inhibition of the major signaling pathways involved in cell survival and proliferation. However, simultaneous multipathway inhibition by the staurosporin derivative UCN-01 results in remarkable inhibition of GSC growth in vitro. The activity of UCN-01 on GSCs was confirmed in two in vivo models of GBM growth. Finally, we used RPPM to study the molecular and functional effects of UCN-01 and demonstrated that the sensitivity to UCN-01 correlates with activation of survival signals mediated by PDK1 and the DNA damage response initiated by CHK1. Taken together, our results suggest that a combined inhibition of PDK1 and CHK1 represents a potentially effective therapeutic approach to reduce the growth of human GBM.

Hatching response of Aedes aegypti (Diptera: Culicidae) eggs at low temperatures: effects of hatching media and storage conditions.

In temperate regions, Aedes aegypti (L.) (Diptera: Culicidae) populations remain in the egg stage during the cold season. To ensure the start of a new breeding season, eggs should hatch at the beginning of a favorable period. The aim of the current study was to investigate the hatching response of two Ae. aegypti egg batches collected and stored for 3 mo under different conditions, to different low immersion temperatures. Two different hatching media (water and yeast solution) were used for the first batch and only one (water) for the second egg batch. Eggs were immersed for 8 d, during which the number of hatched eggs was recorded daily. The proportion of hatched eggs, delay of the hatching response, proportion of dead larvae, and proportion of remaining eggs within the first egg batch were compared between the two hatching media at each temperature. These parameters also were compared between the two batches immersed in water. Hatching rates were higher and faster in the yeast solution. The hatching response was lower at lower immersion temperatures and among eggs stored under field conditions at colder temperatures (second batch). Among the eggs stored in the laboratory (first batch), older eggs exhibited lower hatching response. The proportion of dead larvae was higher in the yeast solution and in the eggs stored in the laboratory. The conditions that triggered a lower hatching response led to higher proportions of remaining eggs, allowing the population to maintain an egg bank for future favorable opportunities.

Effects of thermal heterogeneity and egg mortality on differences in the population dynamics of Aedes aegypti (Diptera: Culicidae) over short distances in temperate Argentina.

In temperate regions, the seasonal dynamics of Aedes aegypti (L.) (Diptera: Culicidae) is mainly influenced by temperature, whereas the probability of local extinction depends on the egg mortality during the cold season. The aim of the current study was to assess the importance of temperature and egg mortality in determining the differences in the oviposition dynamics of Ae. aegypti between favorable and less favorable areas in Buenos Aires City (Argentina). Year-round temperature dynamics were monitored, and oviposition dynamics were experimentally studied with ovitraps at two sites. Daily egg mortality values were calculated from a previous study performed at the same sites. The relative contribution of the differences in temperature and egg mortality between sites to the oviposition dynamics was assessed by means of a mathematical stochastic population dynamics model for Ae. aegypti. The results showed higher temperature and lower daily egg mortality at the site where higher oviposition activity was recorded. A larger influence of temperature than of egg mortality on population abundance during most of the activity season was detected in the results of the simulations. Our results showed a temperature gradient that relates to the distance to the Río de la Plata river and contributes to explaining the spatial heterogeneity in Ae. aegypti population abundances previously reported. The hypothesis of local extinctions because of egg mortality during the winter was not supported by the present analysis. The differences between field oviposition dynamics and simulation results suggest that rainfall might also be an important variable under extremely dry conditions.

Use of laparoscopy for the evaluation of the reproductive status of tench (Tinca tinca).

The aim of the study was to evaluate the use of laparoscopy for the assessment of the reproductive tract and stage of gonadal development of the tench (Tinca tinca). Laparoscopy was carried out from February to March 2009, on 30 anesthetized tenchs, 15 males and 15 females, of approximately 250 g bodyweight and 30 cm length, using a 1.9-mm rigid cystoscope. Male and female gonads, attached to the swim bladder, were visualized. Testes were orange with a striped appearance. Mature ovaries were cluster-shaped and whitish. Histological samples were collected, from the reproductive organs of both the sexes, with a forceps and showed the presence of gametes at different stages of maturity. The application of this minimally invasive diagnostic imaging technique could clinically be used as a powerful tool to evaluate the gender and to establish the reproductive status of tench and other fish.

Rosiglitazone reduces the evolution of experimental periodontitis in the rat.

The peroxisome proliferator-activated receptor-gamma (PPAR-gamma) receptor appears to play a pivotal role in the regulation of cellular proliferation and inflammation. Recent evidence also suggests that rosiglitazone, a PPAR-gamma agonist, reduces acute and chronic inflammation. We hypothesized that rosiglitazone would attenuate periodontal inflammation. In the present study, we investigated the effects of rosiglitazone in a rat model of ligature-induced periodontitis. At day 8, ligation significantly induced an increase in neutrophil infiltration, as well as of gingivomucosal tissue expression of iNOS, nitrotyrosine formation, and poly (ADP-ribose) polymerase activation. Ligation significantly increased Evans blue extravasation in gingivomucosal tissue and alveolar bone destruction. Intraperitoneal injection of rosiglitazone (10 mg/kg 10% DMSO daily for 8 days) significantly decreased all of the parameters of inflammation, as described above. Analysis of these data demonstrated that rosiglitazone exerted an anti-inflammatory role during experimental periodontitis, and was able to ameliorate the tissue damage associated with ligature-induced periodontitis.

Efficacy of the treatment of dogs with leishmaniosis with a combination of metronidazole and spiramycin.

Twenty-seven dogs infected naturally with Leishmania infantum were used in a randomised controlled trial to compare the clinical and parasitological efficacy of an oral treatment with a combination of metronidazole and spiramycin (13 dogs) with the efficacy of conventional treatment with meglumine antimonate and allopurinol (14 dogs) as controls. In the test group one dog had to be withdrawn from the treatment because it developed pemphigus foliaceus; 10 of the dogs were clinically responsive but none was cured parasitologically. In the control group four dogs were withdrawn from the treatment because of side effects; eight of the dogs were clinically responsive but none was cured parasitologically. The control group showed signs of improvement after an average of 30 days, whereas the test group did not show signs of improvement until after an average of 45 days.

Effect of aminoguanidine in ligature-induced periodontitis in rats.

The role of nitric oxide and reactive oxygen species is well-demonstrated in inflammation. In this study, we evaluated the effect of aminoguanidine, a nitric oxide synthase inhibitor, in a rat model of periodontitis. We induced periodontitis in rats by placing a piece of 2/0 braided silk around the lower left 1st molar. At day 8, the gingivomucosal tissue encircling the mandibular 1st molar was removed for biochemical and histological analysis. Ligation significantly increased inducible nitric oxide synthase activity and expression, and damaged tissue revealed increased neutrophil infiltration, lipid peroxidation, and positive staining for nitrotyrosine formation and poly (ADP-ribose) polymerase activation. Ligation significantly increased Evans blue extravasation in gingivomucosal tissue and alveolar bone destruction. Aminoguanidine (100 mg/kg i.p., daily for 8 days) treatment significantly reduced all these inflammatory parameters, indicating that it protects against the tissue damage associated with periodontitis by reducing nitric oxide production and oxidative stress.

Early detection of Leishmania promastigotes in dog bone marrow cultures by acridine orange stain.

An acridine orange staining technique was evaluated in comparison with other well-known methods for the laboratory diagnosis of leishmaniasis. A higher number of promastigotes was found in Novy-MacNeal-Nicolle (NNN) cultures inoculated with canine bone marrow, when culture samples were stained with acridine orange vital stain, compared with those detected using either Giemsa staining or unstained wet mount examination. Based on our data the acridine orange stain is a useful and timely technique in reflecting the true numbers of microorganisms present in a culture and also enhances the visualization of the parasites. The present results warrant further studies with human samples from suspected leishmaniasis patients.