Comparison of DAS-ELISA and qRT-PCR for the detection of cucurbit viruses in seeds

The importance of seeds as virus vehicles for long-distance dissemination makes essential the availability of adequate methods of analysis to guarantee the quality of seed lots. To improve the repertoire of sensitive methods for seed diagnosis, we have developed quantitative real-time RT-PCR assays (RT-qPCR) based on the TaqMan technology to detect three viruses which are seed transmitted in cucurbits, namely, cucumber green mottle mosaic virus (CGMMV), squash mosaic virus (SqMV) and melon necrotic spot virus (MNSV), and compared these assays with DAS-ELISA, the main method used for virus detection in seeds. The estimated RT-qPCR limits of detection were 96, 97 and 740 RNA target molecules for CGMMV, SqMV and MNSV, respectively. The estimated RT-qPCR analytical sensitivity (highest dilution capable of generating a detectable amplification signal) ranged between 10 and 1 pg/μL for the three viruses. Using RT-qPCR, we could reliably detect a single SqMV- or CGMMV-contaminated seed among 999 uncontaminated seeds in a seed lot, and sensitivities were 1,000 and 10,000 times of those provided by DAS-ELISA for SqMV and CGMMV, respectively. Our RT-qPCR assays have proved to be highly suitable for the analysis of seed lots, and the possibility of their implementation into certification programme should be taken into consideration.     

Q Fever Outbreak among Workers at a Waste-Sorting Plant

An outbreak of Q fever occurred in February–April 2014 among workers at a waste-sorting plant in Bilbao (Spain). The outbreak affected 58.5% of investigated employees, 47.2% as confirmed cases (PCR and/or serology) and 11.3% as probable cases (symptoms without laboratory confirmation). Only employees who had no-access to the waste processing areas of the plant were not affected and incidence of infection was significantly higher among workers not using respiratory protection masks. Detection by qPCR of Coxiella burnetii in dust collected from surfaces of the plant facilities confirmed exposure of workers inside the plant. Animal remains sporadically detected among the residues received for waste-sorting were the most probable source of infection. After cleaning and disinfection, all environmental samples tested negative. Personal protection measures were reinforced and made compulsory for the staff and actions were taken to raise farmers’ awareness of the biological risk of discharging animal carcasses as urban waste.     

Using multiple sources of characters to delimit species in the genus Crataegus (Rosaceae): the case of the Crataegus rosei complex

The aim of this study was to delimit the taxa of the Crataegus rosei complex using an integrative approach that incorporates a suite of molecular (cpDNA and nuclear microsatellite markers), morphological, and geometric morphometric characters. One hundred and ten plants from 19 populations that encompass the entire distribution range of the species complex were collected and examined along with herbarium specimens. Parsimony and Bayesian inference analyses were run using morphological, molecular, and both the morphological and molecular data sets combined. Analyses to determine genetic structure based on microsatellite data and multivariate analyses incorporating geometric morphometrics were also done to identify differences in leaf shape. The results supported the recognition of two taxa: C. rosei with high levels of gene flow among its populations, remarkable morphological variation and a wide distribution range and C. rosei var. amoena, composed of a few isolated populations in the high elevation location of Cerro Potosí; a new specific epithet will be decided for the latter in accordance with the International Code of Nomenclature for algae, fungi, and plants.     

Coagulation activation in an experimental pneumonia model in malnourished mice

Malnutrition induces a decrease in immunity that affects the ability of the organism to deal with an infectious challenge. The clotting system is considered a branch of immunity and its activation is important in the pathogenesis of an infectious disease. This work was conducted to determine coagulation modifications in malnourished hosts before and during infection. Weaned mice were malnourished via a protein-free diet. Well-nourished control mice (WNC) consumed a balanced conventional diet. Malnourished mice (MN) and WNC were challenged intranasally with Streptococcus pneumoniae. Blood, bronchoalveolar lavages (BAL), and lung samples were taken at different times post infection. The results were that MN showed altered hemostatic tests and fibrin(ogen) deposits in the lung. Thus, an increase in thrombin-antithrombin complexes (TATc) in plasma and BAL was observed. In the MN group, infection induced a rise in TATc in plasma and BAL and increased plasma fibrinogen and fibrin(ogen) deposits in the lung. A decrease in activated protein C and antithrombin in BAL and an early decrease followed by an increase in plasma Factor VIII were also observed. Thus, malnourishment induced a procoagulant state increased by infection. This is the first work that presents results of an exhaustive study of coagulation in malnourished hosts before and during an infection.     

Synergistic and additive interactions among components of food-based baits underlie female fruit fly attraction

Attraction of tephritid fruit flies to some food sources can be enhanced by the presence of ammonia derivatives, compounds that are perceived as volatile cues for protein-rich food sources. Using a comparative approach, we (1) evaluated the behavioral responses of females of three invasive fruit fly species, Bactrocera (Zeugodacus) cucurbitae (Coquillett), Bactrocera dorsalis (Hendel), and Ceratitis capitata (Wiedemann) (all Diptera: Tephritidae) to trub (a protein-rich waste brewer’s yeast product generated during the production of beer), Concord grape juice (a protein-deficient material), and ammonium acetate, and (2) identified synergistic and additive interactions between low- and high-attractiveness materials and ammonium acetate. We established the attractiveness of fresh trub, grape juice, and ammonium acetate when tested singly to females of all three fly species. Although ammonium acetate did not enhance significantly the response of females of any species to fresh trub, the most attractive material, ammonium acetate, did significantly enhance females’ level of response to aged trub (a comparatively less attractive material) and to grape juice. Our research found a synergistic interaction between diluted grape juice and ammonium acetate for B. cucurbitae, as well as between aged trub and ammonium acetate for B. dorsalis. For C. capitata, additive effects among food attractants and ammonium acetate were identified. Our findings increase our understanding of fruit fly female olfactory-driven behavior in response to food-based materials and the extent to which ammonium acetate modulates female response to protein-rich and protein-deficient materials.     

Medicinal plants: active compounds,properties and antiproliferative effects in colorectal cancer

Cancer development consists of a multitude of steps that occur progressively starting with initial driver mutation(s), to tumorigenesis, and ultimately metastasis. However, relatively little is known about the key epigenetic events that lead to tumour progression and metastasis. On the other hand, it is widely known that cancer cells acquire instruments to circumvent programmed cell death. Epidemiological studies suggest that medicinal plants can act as epigenetic compounds promoting several strategies to override apoptosis, including the induction of programmed cell death, the arrest of cell proliferation, or both. Interestingly, around half of current pharmaceutical drugs have been derived, directly or indirectly, from natural sources. Many species have been recognized to have medicinal properties and beneficial impact on health, such as, antioxidant or anti-inflammatory activities, anti-mutagenic effects and anti-carcinogenic potential. This review thoroughly discusses about the chemical composition and biochemical properties of four medicinal and Mediterranean plants such as Aloe vera, Salvia officinalis L. (Sage), Rosmarinus officinalis L. (Rosemary) and Urtica dioica (Nettle), and compares the potential of these plants to modify proliferation, migration, angiogenesis and apoptosis in colorectal cancer.

     

A Novel Electrocardiographic Index for the Diagnosis of Diastolic Dysfunction

Background

Although the assessment of diastolic dysfunction (DD) is an integral part of routine cardiologic examinations, little is known about associated electrocardiographic (ECG) changes. Our aim was to investigate a potential role of ECG indices for the recognition of patients with DD.

Methods and Results

ECG parameters correlating with echocardiographic findings of DD were retrospectively assessed in a derivation group of 172 individuals (83 controls with normal diastolic function, 89 patients with DD) and their diagnostic performance was tested in a validation group of 50 controls and 50 patients. The patient group with a DD Grade 1 and 2 showed longer QTc (422±24ms and 434±32ms vs. 409±25ms, p<0.0005) and shorter Tend–P and Tend–Q intervals, reflecting the electrical and mechanical diastole (240±78ms and 276±108ms vs. 373±110ms, p<0.0001; 409±85ms and 447±115ms vs. 526±119ms, p<0.0001). The PQ–interval was significantly longer in the patient group (169±28ms and 171±38ms vs. 153±22ms, p<0.005). After adjusting for possible confounders, a novel index (Tend–P/[PQxAge]) showed a high performance for the recognition of DD, stayed robust in the validation group (sensitivity 82%, specificity 93%, positive predictive value 93%, negative predictive value 82%, accuracy 88%) and proved a substantial added value when combined with the indexed left atrial volume (LAESVI, sensitivity 90%, specificity 92%, positive predictive value 95%, negative predictive value 86%, accuracy 91%).

Conclusions

A novel electrocardiographic index Tend–P/(PQxAge) demonstrates a high diagnostic accuracy for the diagnosis of DD and yields a substantial added value when combined with the LAESVI.     

Impact of a Partial Smoke-Free Legislation on Myocardial Infarction Incidence,Mortality and Case-Fatality in a Population-Based Registry: The REGICOR Study

Background and Objective

Coronary heart disease (CHD) is the leading cause of death, and smoking its strongest modifiable risk factor. Our aim was to determine the impact of the Spanish 2006 partial smoke-free legislation on acute myocardial infarction (AMI) incidence, hospitalization and mortality rates, and 28-day case-fatality in Girona, Spain.

Methods

Using a population-based registry (the REGICOR Study), we compared population incidence, hospitalization, and mortality rates, and 28-day case-fatality in the pre- and post-ban periods (2002–2005 and 2006–2008, respectively) by binomial regression analysis adjusted for confounding factors. We also analyzed the ban''s impact on the outcomes of interest using the AMI definitions of the American Heart Association (AHA)/European Society of Cardiology (ESC) and the World Health Organization (WHO)-Monitoring trends and determinants in cardiovascular diseases (MONICA).

Results

In the post-ban period, AMI incidence and mortality rates significantly decreased (relative risk [RR] = 0.89; 95% confidence interval [CI] = 0.81–0.97 and RR = 0.82; 95% CI = 0.71–0.94, respectively). Incidence and mortality rates decreased in both sexes, especially in women, and in people aged 65–74 years. Former and non-smokers (passive smokers) showed diminished incidence rates. Implementation of the ban was not associated with AMI case-fatality. Models tended to be more significant with the WHO-MONICA than with the AHA/ESC definition.

Conclusions

The 2006 Spanish partial smoke-free legislation was associated with a decrease in population AMI incidence and mortality, particularly in women, in people aged 65–74 years, and in passive smokers. These results clarify the association between AMI mortality and the enactment of a partial smoke-free legislation and reinforce the effectiveness of smoking regulations in preventing CHD.     

Phylogeography of Pinus subsection Australes in the Caribbean Basin

Background and Aims

Four species of Pinus subsection Australes occur in the Caribbean Basin: P. caribaea, P. cubensis, P. maestrensis and P. occidentalis. This study analyses the phylogeography of these species to assess possible colonization events from Central America to the islands and subsequent population expansions during glacial periods driven by both drier climate and larger emerged land areas.

Methods

Allele size data were obtained for plastid microsatellites for 314 individuals from 24 populations, covering the distribution range of subsection Australes in the Caribbean Basin.

Key Results

In total, 113 plastid haplotypes were identified. The highest genetic diversity was found in populations of P. caribaea. Overall, Caribbean Basin populations fit the isolation by distance model. Significant phylogeographical structure was found (R_ST = 0·671 > permuted R_ST = 0·101; P < 0·0001). The haplotype network and a Bayesian analysis of population structure (BAPS) indicated different Central American origins for P. caribaea var. bahamensis and P. caribaea var. caribaea plastids, with Central America populations in northern and south-eastern groups. Sudden expansion times for BAPS clusters were close to three glacial maxima.

Conclusions

Central America contains ancestral plastid haplotypes. Population expansion has played a major role in the distribution of genetic diversity in P. caribaea var. hondurensis. Two colonization events gave rise to the P. caribaea var. bahamensis and P. caribaea var. caribaea lineages. Plastid variation in the eastern species (P. cubensis, P. maestrensis and P. occidentalis) evolved independently from that in P. caribaea var. caribaea. Incomplete lineage sorting between P. cubensis and P. maestrensis is apparent. Inferred expansion times for P. caribaea var. bahamensis and for the eastern lineages correspond to glacial maxima, whereas those for P. caribaea var. hondurensis correspond to the beginning of the temperature decrease that led to Marine Isotope Stage 8.