Protistan Communities Within the Galápagos Archipelago With an Emphasis on Micrograzers

The Galápagos Archipelago is a globally significant biodiversity hotspot. However, compared to the relatively well-known megafauna, the distribution and ecological significance of marine protists in this system are poorly understood. To gain an understanding of the protistan assemblages across trophic modes, an intensive oceanographic survey was conducted in the Galápagos Marine Reserve (GMR) in October of 2018. The Equatorial Undercurrent (EUC)-influenced region had higher chlorophyll-a (Chl-a) concentrations than those of the eastern regions of the archipelago, along with higher abundances of protistan grazers. Specifically, proportions of autotrophic and potentially mixotrophic dinoflagellates were higher in the EUC, whereas in the eastern regions, heterotrophic dinoflagellates and chlorophytes dominated. Taxonomic composition and biochemical indicators suggested proportions of micrograzers and their associated heterotrophic biomass was higher in the oligotrophic, low Chl-a regions in the east. We also report observations from a dinoflagellate bloom in the western archipelago, which was heavily influenced by upwelling of the EUC. The red tide-forming dinoflagellate Scrippsiella lachrymosa was highly detected through light microscopy and DNA amplicon sequencing. In addition, the heterotrophic dinoflagellate Polykrikos kofoidii was detected and, based on cell densities observed in this study and grazing rates obtained from the literature, estimated to potentially graze up to 62% of S. lachrymosa bloom population. Our findings thus provide new insights into the composition of micrograzers and their potential roles in structuring protistan communities in the Galápagos Archipelago

Protistan plankton communities in the Galápagos Archipelago respond to changes in deep water masses resulting from the 2015/16 El Niño

The Galápagos Archipelago lies within the Eastern Equatorial Pacific Ocean at the convergence of major ocean currents that are subject to changes in circulation. The nutrient-rich Equatorial Undercurrent upwells from the west onto the Galápagos platform, stimulating primary production, but this source of deep water weakens during El Niño events. Based on measurements from repeat cruises, the 2015/16 El Niño was associated with declines in phytoplankton biomass at most sites throughout the archipelago and reduced utilization of nitrate, particularly in large-sized phytoplankton in the western region. Protistan assemblages were identified by sequencing the V4 region of the 18S rRNA gene. Dinoflagellates, chlorophytes and diatoms dominated most sites. Shifts in dinoflagellate communities were most apparent between the years; parasitic dinoflagellates, Syndiniales, were highly detected during the El Niño (2015) while the dinoflagellate genus, Gyrodinium, increased at many sites during the neutral period (2016). Variations in protistan communities were most strongly correlated with changes in subthermocline water density. These findings indicate that marine protistan communities in this region are regimented by deep water mass sources and thus could be profoundly affected by altered ocean circulation

LINE-1 methylation in leukocyte DNA, interaction with phosphatidylethanolamine N-methyltransferase variants and bladder cancer risk

BACKGROUND: Aberrant global DNA methylation is shown to increase cancer risk. LINE-1 has been proven a measure of global DNA methylation. The objectives of this study were to assess the association between LINE-1 methylation level and bladder cancer risk and to evaluate effect modification by environmental and genetic factors. METHODS: Bisulphite-treated leukocyte DNA from 952 cases and 892 hospital controls was used to measure LINE-1 methylation level at four CpG sites by pyrosequencing. Logistic regression model was fitted to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs). Interactions between LINE-1 methylation levels and environmental and genetic factors were assessed. RESULTS: The risk of bladder cancer followed a nonlinear association with LINE-1 methylation. Compared with subjects in the middle tertile, the adjusted OR for subjects in the lower and the higher tertiles were 1.26 (95% CI 0.99–1.60, P=0.06) and 1.33 (95% CI 1.05–1.69, P=0.02), respectively. This association significantly increased among individuals homozygous for the major allele of five single-nucleotide polymorphisms located in the phosphatidylethanolamine N-methyltransferase gene (corrected P-interaction<0.05). CONCLUSIONS: The findings from this large-scale study suggest that both low and high levels of global DNA methylation are associated with the risk of bladder cancer

Immunogenicity of Self-Associated Aggregates and Chemically Cross-Linked Conjugates of the 42 kDa Plasmodium falciparum Merozoite Surface Protein-1

Self-associated protein aggregates or cross-linked protein conjugates are, in general, more immunogenic than oligomeric or monomeric forms. In particular, the immunogenicity in mice of a recombinant malaria transmission blocking vaccine candidate, the ookinete specific Plasmodium falciparum 25 kDa protein (Pfs25), was increased more than 1000-fold when evaluated as a chemical cross-linked protein-protein conjugate as compared to a formulated monomer. Whether alternative approaches using protein complexes improve the immunogenicity of other recombinant malaria vaccine candidates is worth assessing. In this work, the immunogenicity of the recombinant 42 kDa processed form of the P. falciparum merozoite surface protein 1 (MSP142) was evaluated as a self-associated, non-covalent aggregate and as a chemical cross-linked protein-protein conjugate to ExoProtein A, which is a recombinant detoxified form of Pseudomonas aeruginosa exotoxin A. MSP142 conjugates were prepared and characterized biochemically and biophysically to determine their molar mass in solution and stoichiometry, when relevant. The immunogenicity of the MSP142 self-associated aggregates, cross-linked chemical conjugates and monomers were compared in BALB/c mice after adsorption to aluminum hydroxide adjuvant, and in one instance in association with the TLR9 agonist CPG7909 with an aluminum hydroxide formulation. Antibody titers were assessed by ELISA. Unlike observations made for Pfs25, no significant enhancement in MSP142 specific antibody titers was observed for any conjugate as compared to the formulated monomer or dimer, except for the addition of the TLR9 agonist CPG7909. Clearly, enhancing the immunogenicity of a recombinant protein vaccine candidate by the formation of protein complexes must be established on an empirical basis

H3K4me1 marks DNA regions hypomethylated during aging in human stem and differentiated cells

In differentiated cells, aging is associated with hypermethylation of DNA regions enriched in repressive histone post-translational modifications. However, the chromatin marks associated with changes in DNA methylation in adult stem cells during lifetime are still largely unknown. Here, DNA methylation profiling of mesenchymal stem cells (MSCs) obtained from individuals aged 2 to 92 yr identified 18,735 hypermethylated and 45,407 hypomethylated CpG sites associated with aging. As in differentiated cells, hypermethylated sequences were enriched in chromatin repressive marks. Most importantly, hypomethylated CpG sites were strongly enriched in the active chromatin mark H3K4me1 in stem and differentiated cells, suggesting this is a cell type–independent chromatin signature of DNA hypomethylation during aging. Analysis of scedasticity showed that interindividual variability of DNA methylation increased during aging in MSCs and differentiated cells, providing a new avenue for the identification of DNA methylation changes over time. DNA methylation profiling of genetically identical individuals showed that both the tendency of DNA methylation changes and scedasticity depended on nongenetic as well as genetic factors. Our results indicate that the dynamics of DNA methylation during aging depend on a complex mixture of factors that include the DNA sequence, cell type, and chromatin context involved and that, depending on the locus, the changes can be modulated by genetic and/or external factors

A virtual reality collaborative planning simulator and its method for three machines in a fully mechanized coal mining face

The existing automatic control program and its parameters for three machines in a fully mechanized Coal Mining face are static and simplex and are therefore inadequate for satisfying the complex and dynamic environment of underground coal mines. To overcome this problem, a collaborative mathematical model is established that includes the effects of a dynamic environment. A virtual reality collaborative planning simulator with methods for the three machines is also proposed based on a multi-agent system. According to the dynamic characteristics of the environment, equipment, and technologies, a fully mechanized Unity3D simulator (FMUnitySim) is designed in terms of multiple factors and multiple dimensions. The factors affecting the coordinated operation of the three machines are analyzed and modeled. The communication modes, coordination, and redundant sensing process among multiple agents, which include the shearer agent and the scraper conveyor agent, are also investigated in detail. Using this system, the key parameters of the three machines can be planned and adjusted online to design and distinctly observe the corresponding collaborative simulations of coordinated operation with multiple perspectives and in real time. Tests of different maximum shearer haulage speeds for regular or reverse transporting coal are designed; their key parameters, including the average shearer haulage speed, average follower distance, and average scraper conveyor load, are planned and simulated using FMUnitySim. The optimal parameter combination is obtained by analyzing and comparing the simulation results. The proposed FMUnitySim offers an effective means and theoretical basis for the rapid planning and safe automatic production of a fully mechanized Coal Mining face