Rewiring of the Host Cell Metabolome and Lipidome during Lytic Gammaherpesvirus Infection Is Essential for Infectious-Virus Production.

Oncogenic virus infections are estimated to cause ~15% of all cancers. Two prevalent human oncogenic viruses are members of the gammaherpesvirus family: Epstein-Barr virus (EBV) and Kaposi's sarcoma herpesvirus (KSHV). We use murine herpesvirus 68 (MHV-68), which shares significant homology with KSHV and EBV, as a model system to study gammaherpesvirus lytic replication. Viruses implement distinct metabolic programs to support their life cycle, such as increasing the supply of lipids, amino acids, and nucleotide materials necessary to replicate. Our data define the global changes in the host cell metabolome and lipidome during gammaherpesvirus lytic replication. Our metabolomics analysis found that MHV-68 lytic infection induces glycolysis, glutaminolysis, lipid metabolism, and nucleotide metabolism. We additionally observed an increase in glutamine consumption and glutamine dehydrogenase protein expression. While both glucose and glutamine starvation of host cells decreased viral titers, glutamine starvation led to a greater loss in virion production. Our lipidomics analysis revealed a peak in triacylglycerides early during infection and an increase in free fatty acids and diacylglyceride later in the viral life cycle. Furthermore, we observed an increase in the protein expression of multiple lipogenic enzymes during infection. Interestingly, pharmacological inhibitors of glycolysis or lipogenesis resulted in decreased infectious virus production. Taken together, these results illustrate the global alterations in host cell metabolism during lytic gammaherpesvirus infection, establish essential pathways for viral production, and recommend targeted mechanisms to block viral spread and treat viral induced tumors. IMPORTANCE Viruses are intracellular parasites which lack their own metabolism, so they must hijack host cell metabolic machinery in order to increase the production of energy, proteins, fats, and genetic material necessary to replicate. Using murine herpesvirus 68 (MHV-68) as a model system to understand how similar human gammaherpesviruses cause cancer, we profiled the metabolic changes that occur during lytic MHV-68 infection and replication. We found that MHV-68 infection of host cells increases glucose, glutamine, lipid, and nucleotide metabolic pathways. We also showed inhibition or starvation of glucose, glutamine, or lipid metabolic pathways results in an inhibition of virus production. Ultimately, targeting changes in host cell metabolism due to viral infection can be used to treat gammaherpesvirus-induced cancers and infections in humans.

Unacylated ghrelin promotes adipogenesis in rodent bone marrow via ghrelin O-acyl transferase and GHS-R1a activity: evidence for target cell-induced acylation.

Despite being unable to activate the cognate ghrelin receptor (GHS-R), unacylated ghrelin (UAG) possesses a unique activity spectrum that includes promoting bone marrow adipogenesis. Since a receptor mediating this action has not been identified, we re-appraised the potential interaction of UAG with GHS-R in the regulation of bone marrow adiposity. Surprisingly, the adipogenic effects of intra-bone marrow (ibm)-infused acylated ghrelin (AG) and UAG were abolished in male GHS-R-null mice. Gas chromatography showed that isolated tibial marrow adipocytes contain the medium-chain fatty acids utilised in the acylation of UAG, including octanoic acid. Additionally, immunohistochemistry and immunogold electron microscopy revealed that tibial marrow adipocytes show prominent expression of the UAG-activating enzyme ghrelin O-acyl transferase (GOAT), which is located in the membranes of lipid trafficking vesicles and in the plasma membrane. Finally, the adipogenic effect of ibm-infused UAG was completely abolished in GOAT-KO mice. Thus, the adipogenic action of exogenous UAG in tibial marrow is dependent upon acylation by GOAT and activation of GHS-R. This suggests that UAG is subject to target cell-mediated activation - a novel mechanism for manipulating hormone activity.

Environmental Chemistry and Chemical Ecology of "Green Tide" Seaweed Blooms.

Green tides are large growths or accumulations of green seaweeds that have been increasing in magnitude and frequency around the world. Because green tides consist of vast biomasses of algae in a limited area and are often seasonal or episodic, they go through periods of rapid growth in which they take up large amounts of nutrients and dissolved gases and generate bioactive natural products that may be stored in the plants, released into the environment, or broken down during decomposition. As a result of the use and production of inorganic and organic compounds, the algae in these blooms can have detrimental impacts on other organisms. Here, we review some of the effects that green tides have on the chemistry of seawater and the effects of the natural products that they produce. As blooms are developing and expanding, algae in green tides take up inorganic nutrients, such as nitrate and ortho-phosphate, which can limit their availability to other photosynthetic organisms. Their uptake of dissolved inorganic carbon for use in photosynthesis can cause localized spikes in the pH of seawater during the day with concomitant drops in the pH at night when the algae are respiring. Many of the algae that form green-tide blooms produce allelopathic compounds, which are metabolites that affect other species. The best documented allelopathic compounds include dimethylsulfoniopropionate (DMSP), dopamine, and reactive oxygen species (ROS) and their breakdown products. DMSP and dopamine are involved in defenses against herbivores. Dopamine and ROS are released into seawater where they can be allelopathic or toxic to other organisms. Thus, these macroalgal blooms can have harmful effects on nearby organisms by altering concentrations of nutrients and dissolved gas in seawater and by producing and releasing allelopathic or toxic compounds.

Wastewater polishing by a channelized macrophyte-dominated wetland and anaerobic digestion of the harvested phytomass.

Constructed wetlands (CW) offer a mechanism to meet increasingly stringent regulatory standards for wastewater treatment while minimizing energy inputs. Additionally, harvested wetland phytomass subjected to anaerobic digestion can serve as a source of biogas methane. To investigate CW wastewater polishing activities and potential energy yield we constructed a pair of secondary wastewater-fed channelized CW modules designed to retain easily harvestable floating aquatic vegetation and maximize exposure of water to roots and sediment. Modules that were regularly harvested averaged a nitrate removal rate of 1.1 g N m(-2) d(-1); harvesting, sedimentation and gasification were responsible for 30.5%, 8.0% and 61.5% of the N losses, respectively. Selective harvesting of a module to maintain dominance of filamentous algae had no effect on nitrate removal rate but lowered productivity by one-half. The average monthly productivity for unselectively harvested modules was 9.3 ± 1.7 g dry wt. m(-2) d(-1) (±SE). Cessation of harvesting in one module resulted in a significant increase in nitrate removal rate and decrease in phosphate removal rate. Compared to the influent, the effluent of the harvested module had significantly lower levels of estrogenic activity, as determined by a quantitative PCR-based juvenile trout bioassay, and significantly lower densities of E. coli. In mixed vertical-flow reactors anaerobic co-digestion of equal dry weight proportions of harvested aquatic vegetation, wine yeast lees and dairy manure was greatly improved when the manure was replaced with the crude glycerol by-product of biodiesel production. Remaining solids were vermicomposted for use as a soil amendment. Our results indicate that incorporation of constructed wetlands into an integrated treatment system can simultaneously enhance the economic and energetic feasibility of wastewater and organic waste treatment processes.

Ecological and physiological controls of species composition in green macroalgal blooms.

Green macroalgal blooms have substantially altered marine community structure and function, specifically by smothering seagrasses and other primary producers that are critical to commercial fisheries and by creating anoxic conditions in enclosed embayments. Bottom-up factors are viewed as the primary drivers of these blooms, but increasing attention has been paid to biotic controls of species composition. In Washington State, USA, blooms are often dominated by Ulva spp. intertidally and Ulvaria obscura subtidally. Factors that could cause this spatial difference were examined, including competition, grazer preferences, salinity, photoacclimation, nutrient requirements, and responses to nutrient enrichment. Ulva specimens grew faster than Ulvaria in intertidal chambers but not significantly faster in subtidal chambers. Ulva was better able to acclimate to a high-light environment and was more tolerant of low salinity than Ulvaria. Ulvaria had higher tissue N content, chlorophyll, chlorophyll b: chlorophyll a, and protein content than Ulva. These differences suggest that nitrogen availability could affect species composition. A suite of five grazers preferred Ulva to Ulvaria in choice experiments. Thus, bottom-up factors allow Ulva to dominate the intertidal zone while resistance to grazers appears to allow Ulvaria to dominate the subtidal zone. While ulvoid algae are in the same functional-form group, they are not functionally redundant.