Indirect Immunofluorescence Assay for the Simultaneous Detection of Antibodies against Clinically Important Old and New World Hantaviruses

In order to detect serum antibodies against clinically important Old and New World hantaviruses simultaneously, multiparametric indirect immunofluorescence assays (IFAs) based on biochip mosaics were developed. Each of the mosaic substrates consisted of cells infected with one of the virus types Hantaan (HTNV), Puumala (PUUV), Seoul (SEOV), Saaremaa (SAAV), Dobrava (DOBV), Sin Nombre (SNV) or Andes (ANDV). For assay evaluation, serum IgG and IgM antibodies were analyzed using 184 laboratory-confirmed hantavirus-positive sera collected at six diagnostic centers from patients actively or previously infected with the following hantavirus serotypes: PUUV (Finland, n = 97); SEOV (China, n = 5); DOBV (Romania, n = 7); SNV (Canada, n = 23); ANDV (Argentina and Chile, n = 52). The control panel comprised 89 sera from healthy blood donors. According to the reference tests, all 184 patient samples were seropositive for hantavirus-specific IgG (n = 177; 96%) and/or IgM (n = 131; 72%), while all control samples were tested negative. In the multiparametric IFA applied in this study, 183 (99%) of the patient sera were IgG and 131 (71%) IgM positive (accordance with the reference tests: IgG, 96%; IgM, 93%). Overall IFA sensitivity for combined IgG and IgM analysis amounted to 100% for all serotypes, except for SNV (96%). Of the 89 control sera, 2 (2%) showed IgG reactivity against the HTNV substrate, but not against any other hantavirus. Due to the high cross-reactivity of hantaviral nucleocapsid proteins, endpoint titrations were conducted, allowing serotype determination in >90% of PUUV- and ANDV-infected patients. Thus, multiparametric IFA enables highly sensitive and specific serological diagnosis of hantavirus infections and can be used to differentiate PUUV and ANDV infection from infections with Murinae-borne hantaviruses (e.g. DOBV and SEOV).     

Identification of Seroreactive Proteins of Leptospira interrogans Serovar Copenhageni Using a High-Density Protein Microarray Approach

Background

Leptospirosis is a widespread zoonotic disease worldwide. The lack of an adequate laboratory test is a major barrier for diagnosis, especially during the early stages of illness, when antibiotic therapy is most effective. Therefore, there is a critical need for an efficient diagnostic test for this life threatening disease.

Methodology

In order to identify new targets that could be used as diagnostic makers for leptopirosis, we constructed a protein microarray chip comprising 61% of Leptospira interrogans proteome and investigated the IgG response from 274 individuals, including 80 acute-phase, 80 convalescent-phase patients and 114 healthy control subjects from regions with endemic, high endemic, and no endemic transmission of leptospirosis. A nitrocellulose line blot assay was performed to validate the accuracy of the protein microarray results.

Principal findings

We found 16 antigens that can discriminate between acute cases and healthy individuals from a region with high endemic transmission of leptospirosis, and 18 antigens that distinguish convalescent cases. Some of the antigens identified in this study, such as LipL32, the non-identical domains of the Lig proteins, GroEL, and Loa22 are already known to be recognized by sera from human patients, thus serving as proof-of-concept for the serodiagnostic antigen discovery approach. Several novel antigens were identified, including the hypothetical protein LIC10215 which showed good sensitivity and specificity rates for both acute- and convalescent-phase patients.

Conclusions

Our study is the first large-scale evaluation of immunodominant antigens associated with naturally acquired leptospiral infection, and novel as well as known serodiagnostic leptospiral antigens that are recognized by antibodies in the sera of leptospirosis cases were identified. The novel antigens identified here may have potential use in both the development of new tests and the improvement of currently available assays for diagnosing this neglected tropical disease. Further research is needed to assess the utility of these antigens in more deployable diagnostic platforms.     

Modelling potential impacts of climate change on the bioclimatic envelope and conservation of the Maned Wolf (Chrysocyon brachyurus)

Forecasting the influence of climatic changes on the distribution of the Maned Wolf (Chrysocyon brachyurus) is important for the conservation of the species. We explored the environmental characteristics than best explain the current distribution of the species, modelled the past and present distribution, projected the niche model into the future, and identified suitable areas for conservation. Niche modelling was performed using Maxent and 21 environmental variables. For past conditions, we considered the Last Glacial Maximum (LGM) and the mid-Holocene (MH) climates. For future conditions, we used the A2a greenhouse gas emission scenario for 2050. Four General Circulation Models (FGOALS 1.0, HADCM3, IPSL-CM4 and MIROC 3.2) were used. The resulting niche model (AUC = 0.89 ± 0.02) predicts maximum probability of presence at precipitation of 106 mm during the coldest quarter, of 396 mm during the warmest quarter, and in totally flat areas. The suitable area for the Maned Wolf currently covers 4,320,364 km². For the LGM, there were inter-model differences in predicted areas (from 819,324 km² to 6,395,886 km²) and in geographic location. The MH models showed drastic changes with respect to the present and considerable inter-model variation. Predictions for 2050 show significant (at least 33%) reductions in distribution. Only a minor fraction (39%) of the current distribution can be considered stable for the period LGM-2050. The FGOALS model was the best option for projecting species occurrence into the future because it included the three localities known for the Maned Wolf from the late Pleistocene and predicts stable areas that coincide with spatial patterns of genetic diversity. The FGOALS projection for 2050 predicts a 33% reduction in suitable habitats, indicating some stable areas (central South America) that will probably be key sites for the conservation of the species.     

Reproductive aspects of American minks (Neovison vison) in central Spain: Testing the effects of prey availability

Some reproductive components of fitness (breeding times, litter size and the proportion of breeding females per year) of two feral populations of American minks in central Spain were investigated by means of direct observation from 2006 to 2010. The effects of prey availability were explored. Mating season was from February to March. Parturition dates did not differ between the two sites and pooled births took place between April (33.3%) and July (2.8%), peaking in May (40.3%). Mean litter size (the number of small cubs following their mother) was 3.43 ± 1.01 cubs (±SD; n = 30) with statistical significant differences between the two study sites. The strong seasonality in births and differences in litter size were related to prey availability (i.e. these are food-limited life-history traits), but in a complex non-linear fashion. A minimum value of prey abundance is necessary to breed. While litter size rises with prey abundance, there is a point where increasing prey did not result in an increase in litter size. Up to 75% of the females in the breeding pool reared cubs each year in both areas, which represents a somewhat high pool of breeding females. Delayed implantation is shorter than a month or does not exist in these populations. Such a short time-span seems to emerge because longer delays would have an important fitness cost to females. The breeding calendar and the litter size were similar to that reported previously from other areas.     

Effects of human activity on the distribution and abundance of an endangered Mediterranean high-mountain plant (Erysimum penyalarense)

The Iberian mountains are important biodiversity hotspots that face great immediate threats like other species-rich regions on Earth. Preservation of natural communities has historically been based on protecting them from physical disturbance. One of the most important causes of disturbance in natural communities today is human activity. Erysimum penyalarense is an endemic and endangered Mediterranean high-mountain herb. In this study, we explore the effect of environmental factors (microhabitat characteristics) and human activity on its abundance and distribution within the only remaining population of the species. Our results indicate that the species is much more abundant than previously thought. The only population known benefited from human disturbance but was negatively affected by animal disturbance and shrub encroachment. Therefore, the net effect of human activity on the species depends on the balance between the different activities carried out within its distribution range, suggesting that intermediate levels of disturbance could help preserve some endangered plants.     

Endogenous Stress Caused by Faulty Oxidation Reactions Fosters Evolution of 2,4-Dinitrotoluene-Degrading Bacteria

Environmental strain Burkholderia sp. DNT mineralizes the xenobiotic compound 2,4-dinitrotoluene (DNT) owing to the catabolic dnt genes borne by plasmid DNT, but the process fails to promote significant growth. To investigate this lack of physiological return of such an otherwise complete metabolic route, cells were exposed to DNT under various growth conditions and the endogenous formation of reactive oxygen species (ROS) monitored in single bacteria. These tests revealed the buildup of a strong oxidative stress in the population exposed to DNT. By either curing the DNT plasmid or by overproducing the second activity of the biodegradation route (DntB) we could trace a large share of ROS production to the first reaction of the route, which is executed by the multicomponent dioxygenase encoded by the dntA gene cluster. Naphthalene, the ancestral substrate of the dioxygenase from which DntA has evolved, also caused significant ROS formation. That both the old and the new substrate brought about a considerable cellular stress was indicative of a still-evolving DntA enzyme which is neither optimal any longer for naphthalene nor entirely advantageous yet for growth of the host strain on DNT. We could associate endogenous production of ROS with likely error-prone repair mechanisms of DNA damage, and the ensuing stress-induced mutagenesis in cells exposed to DNT. It is thus plausible that the evolutionary roadmap for biodegradation of xenobiotic compounds like DNT was largely elicited by mutagenic oxidative stress caused by faulty reactions of precursor enzymes with novel but structurally related substrates-to-be.     

A Systematic Review of the Relationship between Blood Loss and Clinical Signs

Introduction

This systematic review examines the relationship between blood loss and clinical signs and explores its use to trigger clinical interventions in the management of obstetric haemorrhage.

Methods

A systematic review of the literature was carried out using a comprehensive search strategy to identify studies presenting data on the relationship of clinical signs & symptoms and blood loss. Methodological quality was assessed using the STROBE checklist and the general guidelines of MOOSE.

Results

30 studies were included and five were performed in women with pregnancy-related haemorrhage (other studies were carried in non-obstetric populations). Heart rate (HR), systolic blood pressure (SBP) and shock index were the parameters most frequently studied. An association between blood loss and HR changes was observed in 22 out of 24 studies, and between blood loss and SBP was observed in 17 out of 23 studies. An association was found in all papers reporting on the relationship of shock index and blood loss. Seven studies have used Receiver Operating Characteristic Curves to determine the accuracy of clinical signs in predicting blood loss. In those studies the AUC ranged from 0.56 to 0.74 for HR, from 0.56 to 0.79 for SBP and from 0.77 to 0.84 for shock index. In some studies, HR, SBP and shock index were associated with increased mortality.

Conclusion

We found a substantial variability in the relationship between blood loss and clinical signs, making it difficult to establish specific cut-off points for clinical signs that could be used as triggers for clinical interventions. However, the shock index can be an accurate indicator of compensatory changes in the cardiovascular system due to blood loss. Considering that most of the evidence included in this systematic review is derived from studies in non-obstetric populations, further research on the use of the shock index in obstetric populations is needed.     

Major Cardiovascular Risk Factors in Latin America: A Comparison with the United States. The Latin American Consortium of Studies in Obesity (LASO)

Background

Limited knowledge on the prevalence and distribution of risk factors impairs the planning and implementation of cardiovascular prevention programs in the Latin American and Caribbean (LAC) region.

Methods and Findings

Prevalence of hypertension, diabetes mellitus, abnormal lipoprotein levels, obesity, and smoking were estimated from individual-level patient data pooled from population-based surveys (1998–2007, n = 31,009) from eight LAC countries and from a national survey of the United States (US) population (1999–2004) Age and gender specific prevalence were estimated and age-gender adjusted comparisons between both populations were conducted. Prevalence of diabetes mellitus, hypertension, and low high-density lipoprotein (HDL)-cholesterol in LAC were 5% (95% confidence interval [95% CI]: 3.4, 7.9), 20.2% (95% CI: 12.5, 31), and 53.3% (95% CI: 47, 63.4), respectively. Compared to LAC region’s average, the prevalence of each risk factor tended to be lower in Peru and higher in Chile. LAC women had higher prevalence of obesity and low HDL-cholesterol than men. Obesity, hypercholesterolemia, and hypertriglyceridemia were more prevalent in the US population than in LAC population (31 vs. 16.1%, 16.8 vs. 8.9%, and 36.2 vs. 26.5%, respectively). However, the prevalence of low HDL-cholesterol was higher in LAC than in the US (53.3 vs. 33.7%).

Conclusions

Major cardiovascular risk factors are highly prevalent in LAC region, in particular low HDL-cholesterol. In addition, marked differences do exist in this prevalence profile between LAC and the US. The observed patterns of obesity-related risk factors and their current and future impact on the burden of cardiovascular diseases remain to be explained.