Dengue virus 3 clinical isolates show different patterns of virulence in experimental mice infection

Dengue virus (DENV) may cause symptomatic infection with mild, undifferentiated febrile illness called classical dengue fever (DF) or a more severe disease, potentially fatal, known as dengue hemorrhagic fever (DHF) or dengue shock syndrome. The pathogenesis of DHF is based on the virulence of the infecting DENV and depends on the infecting serotypes and genotypes; it is also based on the immunopathogenesis that is mediated by host immune responses, including dengue virus-cross-reactive antibodies that augment the severity of infections. Involvement of central nervous system (CNS) is extensively described. The present study describes the virulence of DENV-3 isolates in a mouse model by intracranial (i.c.) inoculation with genotypes I and III. Our data suggest that, in this experimental model, DENV-3 genotype I may have the propensity to cause neurological disease in mice, whereas the genotype III is associated with asymptomatic infection in mice. Additionally, the symptomatic mice show a decrease of white blood cell count, infectious DENV in the brains and alterations in levels of IFN-gamma, IL-6 and MCP-1. The results confirm the mouse model as a way to study the biology of DENV-3 isolates and to improve the knowledge about the neurovirulence of the different genotypes of DENV.     

Argali Abundance in the Afghan Pamir Using Capture–Recapture Modeling From Fecal DNA

Abstract: Estimating population size in a mark-recapture framework using DNA obtained from remotely collected genetic samples (e.g., feces) has become common in recent years but rarely has been used for ungulates. Using DNA extracted from fecal pellets, we estimated the size of an argali (Ovis ammon) population that was believed to be isolated from others within the Big Pamir Mountains, Afghanistan, an area where access was difficult and expensive. We used closed-capture models to estimate abundance, and Pradel models to examine closure assumptions, both as implemented in Program MARK. We also made visual counts of argali in the Big Pamirs, allowing comparison of count indices of abundance with modeled estimates. Our model-averaged estimate for female argali in the Big Pamir was 172 (95% CI = 117–232), which was about 23% higher than our best assessment using uncorrected visual counts. However, mark-recapture models suggested that males were not a closed population; thus, we were unable to provide a meaningful estimate of overall population size. Males either suffered much higher mortality than females during the sampling period, or, more likely, males moved in and out of the Big Pamir area. Although information from DNA did not provide a clear overall population estimate, it suggested that the Big Pamir was not isolated from other argali populations, which could not have been confirmed with visual observations alone. Estimating argali population size using mark-recapture models and fecal DNA is feasible but may be too expensive for frequent monitoring of large and remote populations. Our study demonstrates the importance of sex identification and separate abundance estimation for each sex, especially if movement ecology differs by sex.     

Biological and Genetic Characterization of Cryptosporidium spp. and Giardia duodenalis Isolates from Five Hydrographical Basins in Northern Portugal

To understand the situation of water contamination with Cryptosporidium spp. and Giardia spp. in the northern region of Portugal, we have established a long-term program aimed at pinpointing the sources of surface water and environmental contamination, working with the water-supply industry. Here, we describe the results obtained with raw water samples collected in rivers of the 5 hydrographical basins. A total of 283 samples were analyzed using the Method 1623 EPA, USA. Genetic characterization was performed by PCR and sequencing of genes 18S rRNA of Cryptosporidium spp. and β-giardin of Giardia spp. Infectious stages of the protozoa were detected in 72.8% (206 of 283) of the water samples, with 15.2% (43 of 283) positive for Giardia duodenalis cysts, 9.5% (27 of 283) positive for Cryptosporidium spp. oocysts, and 48.1% (136 of 283) samples positive for both parasites. The most common zoonotic species found were G. duodenalis assemblages A-I, A-II, B, and E genotypes, and Cryptosporidium parvum, Cryptosporidium andersoni, Cryptosporidium hominis, and Cryptosporidium muris. These results suggest that cryptosporidiosis and giardiasis are important public health issues in northern Portugal. To the authors'' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in raw water samples in the northern region of Portugal.     

Survival and Breeding Transitions for a Reintroduced Bison Population: a Multistate Approach

Abstract: The iconic plains bison (Bison bison) have been reintroduced to many places in their former range, but there are few scientific data evaluating the success of these reintroductions or guiding the continued management of these populations. Relying on mark-recapture data, we used a multistate model to estimate bison survival and breeding transition probabilities while controlling for the recapture process. We tested hypotheses in these demographic parameters associated with age, sex, reproductive state, and environmental variables. We also estimated biological process variation in survival and breeding transition probabilities by factoring out sampling variation. The recapture rate of females and calves was high (0.78 ± 0.15 [SE]) and much lower for males (0.41 ± 0.23), especially older males (0.17 ± 0.15). We found that overall bison survival was high (>0.8) and that males (0.80 ± 0.13) survived at lower rates than females (0.94 ± 0.04), but as females aged survival declined (0.89 ± 0.05 for F ≥15 yr old). Lactating and non-lactating females survived at similar rates. We found that females can conceive early (approx. 1.5 yr of age) and had a high probability (approx. 0.8) of breeding in consecutive years, until age 13.5 years, when females that were non-lactating tended to stay in that state. Our results suggest senescence in reproduction and survival for females. We found little support for the effect of climatic covariates on demographic rates, perhaps because the park's current population management goals were predicated from drought-year conditions. This reintroduction has been successful, but continued culling actions will need to be employed and an adaptive management approach is warranted. Our demographic approach can be applied to other heavily managed large-ungulate systems with few or no natural predators.     

Lipid peroxidation, chloroplastic pigments and antioxidant strategies in Carapa guianensis (Aubl.) subjected to water-deficit and short-term rewetting

The effects of drought on membrane lipids and leaf pigments and the ability of andiroba (Carapa guianensis Aubl.) plants to attenuate oxidative damage through antioxidant enzymes or adjusting carotenoids and glycinebetaine (GB) were examined. Assessments were performed when pre-dawn leaf water potential (Ψ_pd) of water-stressed plants reached −1.35 and −3.21 MPa (15 and 27 days after withholding irrigation) and 12 h after resuming watering (short-term rewetting, day 28). Oxidative damages to lipids were evident on day 15, in which drought caused an increase of 47% in malondialdehyde (MDA) content. On day 27, MDA content did not differ between treatments. The activity of superoxide dismutase remained unchanged over experimental period, while significant increases in the ascorbate peroxidase (APX, 110%) and catalase (CAT, 50%) activities were observed only on day 27. GB content was 62% (day 15) and 112% (day 27) higher in water-stressed plants than in control. Regardless of Ψ_pd, both chlorophyll (Chl) a, Chl b and total carotenoids remained unchanged between well-watered and water-stressed plants, indicating that drought did not result in degradation of leaflet pigments. On day 28, Ψ_pd of water-stressed plants increased near to control plants and both activities of APX and CAT did not differ between treatments. Altogether, adjustments in APX and CAT activity and in the GB content were efficient strategies to prevent expressive oxidative damages in water-stressed andiroba plants.     

Steady‐state level of kit ligand mRNA in goat ovaries and the role of kit ligand in preantral follicle survival and growth in vitro

The aims of this study were to investigate steady‐state level of Kit Ligand (KL) mRNA and its effects on in vitro survival and growth of caprine preantral follicles. RT‐PCR was used to analyze caprine steady‐state level of KL mRNA in primordial, primary, and secondary follicles, and in small (1–3 mm) and large (3–6 mm) antral follicles. Furthermore, ovarian fragments were cultured for 1 or 7 days in Minimal Essential Medium (MEM⁺) supplemented with KL (0, 1, 10, 50, 100, or 200 ng/ml). Noncultured (control) and cultured fragments were processed for histology and transmission electron microscopy (TEM). RT‐PCR demonstrated an increase in steady‐state level of KL mRNA during the transition from primary to secondary follicles. Small antral follicles had higher steady‐state levels of KL mRNA in granulosa and theca cells than large follicles. After 7 days, only 50 ng/ml of KL had maintained the percentage of normal follicles similar to control. After 1 day, all KL concentrations reduced the percentage of primordial follicles and increased the percentage of growing follicles. KL at 10, 50, 100, or 200 ng/ml increased primary follicles, compared to MEM⁺ after 7 days. An increase in oocyte and follicular diameter was observed at 50 ng/ml of KL. TEM confirmed ultrastructural integrity of follicles after 7 days at 50 ng/ml of KL. In conclusion, the KL mRNAs were detected in all follicular categories. Furthermore, 50 ng/ml of KL maintained the integrity of caprine preantral follicle cultured for 7 days and stimulated primordial follicle activation and follicle growth. Mol. Reprod. Dev. 77: 231–240, 2010. © 2009 Wiley‐Liss, Inc.     

Domain III peptides from flavivirus envelope protein are useful antigens for serologic diagnosis and targets for immunization

The Flavivirus genus of the Flaviviridae family includes 70 enveloped single-stranded-RNA positive-sense viruses transmitted by arthropods. Among these viruses, there are a relevant number of human pathogens including the mosquito-borne dengue virus (DENV), yellow fever virus (YFV), Japanese encephalitis virus (JEV) and West Nile virus (WNV), as well as tick-borne viruses such as tick-borne encephalitis virus (TBEV), Langat virus (LGTV) and Omsk hemorrhagic fever (OHFV). The flavivirus envelope (E) protein is a dominant antigen inducing immunologic responses in infected hosts and eliciting virus-neutralizing antibodies. The domain III (DIII) of E protein contains a panel of important epitopes that are recognized by virus-neutralizing monoclonal antibodies. Peptides of the DIII have been used with promising results as antigens for flavivirus serologic diagnosis and as targets for immunization against these viruses. We review here some important aspects of the molecular structure of the DIII as well as its use as antigens for serologic diagnosis and immunization in animal models.     

Building and analyzing protein interactome networks by cross-species comparisons

Background  

A genomic catalogue of protein-protein interactions is a rich source of information, particularly for exploring the relationships between proteins. Numerous systems-wide and small-scale experiments have been conducted to identify interactions; however, our knowledge of all interactions for any one species is incomplete, and alternative means to expand these network maps is needed. We therefore took a comparative biology approach to predict protein-protein interactions across five species (human, mouse, fly, worm, and yeast) and developed InterologFinder for research biologists to easily navigate this data. We also developed a confidence score for interactions based on available experimental evidence and conservation across species.     

Using species distribution models to identify suitable areas for biofuel feedstock production

The 2007 Energy Independence and Security Act mandates a five‐fold increase in US biofuel production by 2022. Given this ambitious policy target, there is a need for spatially explicit estimates of landscape suitability for growing biofuel feedstocks. We developed a suitability modeling approach for two major US biofuel crops, corn (Zea mays) and switchgrass (Panicum virgatum), based upon the use of two presence‐only species distribution models (SDMs): maximum entropy (Maxent) and support vector machines (SVM). SDMs are commonly used for modeling animal and plant distributions in natural environments, but have rarely been used to develop landscape models for cultivated crops. AUC, Kappa, and correlation measures derived from test data indicate that SVM slightly outperformed Maxent in modeling US corn production, although both models produced significantly accurate results. When compared with results from a mechanistic switchgrass model recently developed by Oak Ridge National Laboratory (ORNL), SVM results showed higher correlation than Maxent results with models fit using county‐scale point inputs of switchgrass production derived from expert opinion estimates. However, Maxent results for an alternative switchgrass model developed with point inputs from research trial sites showed higher correlation to the ORNL model than the corresponding results obtained from SVM. Further analysis indicates that both modeling approaches were effective in predicting county‐scale increases in corn production from 2006 to 2007, a time period in which US corn production increased by 24%. We conclude that presence‐only methods are a powerful first‐cut tool for estimating relative land suitability across geographic regions in which candidate biofuel feedstocks can be grown, and may also provide important insight into potential land‐use change patterns likely to be associated with increased biofuel demand.