Starvation at the larval stage increases the vector competence of Aedes aegypti females for Zika virus

Aedes aegypti is the primary vector of Zika virus (ZIKV), a flavivirus which typically presents itself as febrile-like symptoms in humans but can also cause neurological and pregnancy complications. The transmission cycle of mosquito-borne arboviruses such as ZIKV requires that various key tissues in the female mosquito get productively infected with the virus before the mosquito can transmit the virus to another vertebrate host. Following ingestion of a viremic blood-meal from a vertebrate, ZIKV initially infects the midgut epithelium before exiting the midgut after blood-meal digestion to disseminate to secondary tissues including the salivary glands. Here we investigated whether smaller Ae. aegypti females resulting from food deprivation as larvae exhibited an altered vector competence for blood-meal acquired ZIKV relative to larger mosquitoes. Midguts from small ‘Starve’ and large ‘Control’ Ae. aegypti were dissected to visualize by transmission electron microscopy (TEM) the midgut basal lamina (BL) as physical evidence for the midgut escape barrier showing Starve mosquitoes with a significantly thinner midgut BL than Control mosquitoes at two timepoints. ZIKV replication was inhibited in Starve mosquitoes following intrathoracic injection of virus, however, Starve mosquitoes exhibited a significantly higher midgut escape and population dissemination rate at 9 days post-infection (dpi) via blood-meal, with more virus present in saliva and head tissue than Control by 10 dpi and 14 dpi, respectively. These results indicate that Ae. aegypti developing under stressful conditions potentially exhibit higher midgut infection and dissemination rates for ZIKV as adults, Thus, variation in food intake as larvae is potentially a source for variable vector competence levels of the emerged adults for the virus.     

Interspecific hybridization causes long‐term phylogenetic discordance between nuclear and mitochondrial genomes in freshwater fishes

Classification, phylogeography and the testing of evolutionary hypotheses rely on correct estimation of species phylogeny. Early molecular phylogenies often relied on mtDNA alone, which acts as a single linkage group with one history. Over the last decade, the use of multiple nuclear sequences has often revealed conflict among gene trees. This observation can be attributed to hybridization, lineage sorting, paralogy or selection. Here, we use 54 groups of fishes from 48 studies to estimate the degree of concordance between mitochondrial and nuclear gene trees in two ecological grades of fishes: marine and freshwater. We test the hypothesis that freshwater fish phylogenies should, on average, show more discordance because of their higher propensity for hybridization in the past. In keeping with this idea, concordance between mitochondrial and nuclear gene trees (as measured by proportion of components shared) is on average 50% higher in marine fishes. We discuss why this difference almost certainly results from introgression caused by greater historical hybridization among lineages in freshwater groups, and further emphasize the need to use multiple nuclear genes, and identify conflict among them, in estimation of species phylogeny.     

Species richness in macroalgae and macrofauna assemblages on Fucus serratus L. (Phaeophyceae) and Zostera marina L. (Angiospermae) in Skagerrak, Norway

The macrophytes Fucus serratus and Zostera marina form similar substrates for associated flora and fauna in shallow waters in Norway. While F. serratus forms a more or less continuous belt on rocky substrate along the coast, Z. marina forms disjunct populations on sandy or muddy bottoms. This study focused on the organisms associated with these two macrophytes in two localities in the Skagerrak region. In total, 130 taxa of epiphytic organisms were identified: 22 green algae, 41 red algae, 32 brown algae, eight diatoms and 27 sessile animals. One hundred and twenty-seven taxa of mobile macrofauna were registered. The dominant group was crustaceans, with amphipods as the order containing most species. Many species of both plants and animals preferred one or the other habitat. It is concluded that coastal macrophyte systems have high species diversity.     

Crop Yields, Internal Nutrient Efficiency, and Changes in Soil Properties in Rice–Wheat Rotations Under Non-Flooded Mulching Cultivation

A field experiment was conducted for 5 years to examine the effects of non-flooded mulching cultivation on crop yield, internal nutrient efficiency and soil properties in rice–wheat (R–W) rotations of the Chengdu Plain, southwest China. Compared with traditional flooding (TF), non-flooded plastic film mulching (PM) resulted in 12 and 11% higher average rice (Oryza sativa L.) yield and system productivity (combined rice and wheat yields), and the trends in rice and wheat (Triticum aestivum L.) yields under PM were stable over time. However, non-flooded wheat straw mulching (SM) decreased average rice yield by 11% compared with TF, although no significant difference in system productivity was found between SM and TF. Uptakes of N and K by rice under PM were higher than those under TF and SM, but internal nutrient efficiency was significantly lower (N) or similar (K) under PM compared to SM and TF. This implies that more N and K accumulated in rice straw under PM. After 5-year rice–wheat rotation, apparent P balances (112–160 kg ha⁻¹) were positive under all three cultivation systems. However, the K balances were negative under PM (−419 kg ha⁻¹) and TF (−90 kg ha⁻¹) compared with SM (45 kg ha⁻¹). This suggests that higher K inputs from fertilizer, straw or manure may be necessary, especially under PM. After five rice seasons and four wheat seasons, non-flooded mulching cultivation led to similar (PM) or higher (SM) soil organic carbon (SOC), total N (TN) and alkali hydrolyzable N (AH-N) in the top 0–5 and 5–12 cm layers compared with TF. SOC, TN, AH-N and Olsen-P (OP) in the sub-surface layer (12–24 cm) were significantly higher under PM or SM than under TF, indicating that rice under non-flooded mulching conditions may fail to make use of nutrients from the subsoil. Thus, the risk of decline in soil fertility under non-flooded mulching cultivation could be very low if input levels match crop requirements. Our data indicate that PM and SM may be alternative options for farmers using R–W rotations for enhancement or maintenance of system productivity and soil fertility.     

Combining MALDI-FTMS and bioinformatics for rapid peptidomic comparisons

Increasing research efforts in large-scale mass spectral analyses of peptides and proteins have led to many advances in technology and method development for collecting data and improving the quality of data. However, the resultant large data sets often pose significant challenges in extracting useful information in a high-throughput manner. Here, we describe one such method where we analyzed a large mass spectral data set collected using decapod crustacean nervous tissue extracts separated via high-performance liquid chromatography (HPLC) coupled to high-resolution matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS). Following their acquisition, the data collected from discrete LC fractions was compiled and analyzed using an in-house developed software package that deisotoped, compressed, calibrated, and matched peaks to a list of known crustacean neuropeptides. By processing these data via bioinformatics tools such as hierarchical clustering, more than 110 neuropeptides that belong to 14 peptide families were mapped in five crustacean species. Overall, we demonstrate the utility of MALDI-FTMS in combination with a bioinformatics software package for the elucidation and comparison of peptidomes of varying crustacean species. This study established an effective methodology and will provide the basis for future investigations into more comprehensive comparative peptidomics with larger collection of species and phyla in order to gain a deeper understanding of the evolution and diversification of peptide families.     

Coupled global and targeted proteomics of human embryonic stem cells during induced differentiation

Elucidating the complex combinations of growth factors and signaling molecules that maintain pluripotency or, alternatively, promote the controlled differentiation of human embryonic stem cells (hESCs) has important implications for the fundamental understanding of human development, devising cell replacement therapies, and cancer cell biology. hESCs are commonly grown on irradiated mouse embryonic fibroblasts (MEFs) or in conditioned medium from MEFs. These culture conditions interfere with many experimental conclusions and limit the ability to perform conclusive proteomics studies. The current investigation avoided the use of MEFs or MEF-conditioned medium for hESC culture, allowing global proteomics analysis without these confounding conditions, and elucidated neural cell-specific signaling pathways involved in noggin-induced hESC differentiation. Based on these analyses, we propose the following early markers of hESC neural differentiation: collapsin response mediator proteins 2 and 4 and the nuclear autoantigenic sperm protein as a marker of pluripotent hESCs. We then developed a directed mass spectrometry assay using multiple reaction monitoring (MRM) to identify and quantify these markers and in addition the epidermal ectoderm marker cytokeratin-8. Analysis of global proteomics, quantitative RT-PCR, and MRM data led to testing the isoform interference hypothesis where redundant peptides dilute quantification measurements of homologous proteins. These results show that targeted MRM analysis on non-redundant peptides provides more exact quantification of homologous proteins. This study describes the facile transition from discovery proteomics to targeted MRM analysis and allowed us to identify and verify several potential biomarkers for hESCs during noggin-induced neural and BMP4-induced epidermal ectoderm differentiation.     

Thioredoxin 2 haploinsufficiency in mice results in impaired mitochondrial function and increased oxidative stress

The mitochondrial form of thioredoxin, thioredoxin 2 (Txn2), plays an important role in redox control and protection against ROS-induced mitochondrial damage. To evaluate the effect of reduced levels of Txn2 in vivo, we measured oxidative damage and mitochondrial function using mice heterozygous for the Txn2 gene (Txn2(+/-)). The Txn2(+/-) mice showed approximately 50% decrease in Trx-2 protein expression in all tissues without upregulating the other major components of the antioxidant defense system. Reduced levels of Txn2 resulted in decreased mitochondrial function as shown by reduced ATP production by isolated mitochondria and reduced activity of electron transport chain complexes (ETCs). Mitochondria isolated from Txn2(+/-) mice also showed increased ROS production compared to wild type mice. The Txn2(+/-) mice showed increased oxidative damage to nuclear DNA, lipids, and proteins in liver. In addition, we observed an increase in apoptosis in liver from Txn2(+/-) mice compared with wild type mice after diquat treatment. Our results suggest that Txn2 plays an important role in protecting the mitochondria against oxidative stress and in sensitizing the cells to ROS-induced apoptosis.     

Influence of root zone nitrogen management and a summer catch crop on cucumber yield and soil mineral nitrogen dynamics in intensive production systems

Nutrient and water management is crucially important in shallow-rooted vegetable production systems characterized by high input and high environmental risk. A 2-year field experiment on greenhouse cucumber double-cropping systems examined the effects of root zone nitrogen management and planting of sweet corn as a catch crop in the summer fallow period on cucumber yield and soil N_min dynamics compared to conventional practices. Cucumber fruit yields were not significantly affected by root zone N management and catch crop planting despite a decrease in N fertilizer application of 53% compared to conventional N management. Soil N_min content to a depth of 0.9 m decreased markedly and root zone (0–0.3 m) soil N_min content was maintained at about 200 kg N ha^?1. Root zone N management efficiently and directly reduced apparent N losses by 44% and 45% in 2005 and 2006, respectively. Sweet corn, the summer catch crop, depleted N_min residue in the soil profile of 1.8 m at harvest of winter–spring season cucumber by 304–333 kg N ha^?1, which contributed 19–22% reduction in N loss. Compared to conventional N management, N loss was reduced by 56% under root zone N management and catch crop planting.