Temperature and salinity effects on respiration of Mnemiopsis leidyi, the new invader of the Baltic Sea

Temperature and salinity effects on respiration of Mnemiopsis leidyi, the new invader of the Baltic Sea 1 2 Abstract 3 To evaluate the influence of temperature along salinity gradients on metabolic performance, we measured 4 respiration rate of fully acclimatized adults of invasive ctenophore Mnemiopsis leidyi under similar field 5 conditions representative for the Baltic Sea. Three levels of temperature (3, 8 and 17°C) and three levels of 6 salinity (12, 17 and 22‰) were considered. Oxygen consumption was temperature and not salinity dependent, 7 albeit combinations of temperature and salinity showed a significant effect on carbon specific respiration rate. 8 Q 10 value of 3.67 indicates a strong impact of temperature on metabolic regulation. The role of combined effect 9 of temperature and salinity on over-wintering population in higher latitudes was discussed

Innovative metabolic data integration applicable for Therapeutic Protein Development 2.0

Therapeutic proteins development becomes more challenging due to the complexity of the diverse molecule formats. In-depth characterization of high producer cell lines and bioprocesses is essential to ensure robust and consistent production of recombinant therapeutic proteins in high quantity and quality for clinical applications. Controlling the environmental stress present during the cultivation of cells is a key for the successful production of an intended bio-therapeutic protein. The captured data is applied in a metabolic network model for the analysis of intracellular metabolic fluxes of Roche’s working horse of therapeutic protein production - the Chinese Hamster Ovary cell. The generated metabolic information has the potential to set a new standard for efficient and innovative process development bridging from research to market. Innovative approach of analyzing the stored data is key towards process development of therapeutic proteins 2.0. In conclusion, the combination of quantitative metabolite profiling, multivariate data analysis, and mechanistic network model simulations can identify metabolic traits characteristic of high-performance clones and empowers the scientists to develop efficient processes. New approach in metabolic/process modeling and results will be presented

Facets of diazotrophy in the oxygen minimum zone waters off Peru

Nitrogen fixation, the biological reduction of dinitrogen gas (N2) to ammonium (NH4+), is quantitatively the most important external source of new nitrogen (N) to the open ocean. Classically, the ecological niche of oceanic N2 fixers (diazotrophs) is ascribed to tropical oligotrophic surface waters, often depleted in fixed N, with a diazotrophic community dominated by cyanobacteria. Although this applies for large areas of the ocean, biogeochemical models and phylogenetic studies suggest that the oceanic diazotrophic niche may be much broader than previously considered, resulting in major implications for the global N-budget. Here, we report on the composition, distribution and abundance of nifH, the functional gene marker for N2 fixation. Our results show the presence of eight clades of diazotrophs in the oxygen minimum zone (OMZ) off Peru. Although proteobacterial clades dominated overall, two clusters affiliated to spirochaeta and archaea were identified. N2 fixation was detected within OMZ waters and was stimulated by the addition of organic carbon sources supporting the view that non-phototrophic diazotrophs were actively fixing dinitrogen. The observed co-occurrence of key functional genes for N2 fixation, nitrification, anammox and denitrification suggests that a close spatial coupling of N-input and N-loss processes exists in the OMZ off Peru. The wide distribution of diazotrophs throughout the water column adds to the emerging view that the habitat of marine diazotrophs can be extended to low oxygen/high nitrate areas. Furthermore, our statistical analysis suggests that NO2− and PO43− are the major factors affecting diazotrophic distribution throughout the OMZ. In view of the predicted increase in ocean deoxygenation resulting from global warming, our findings indicate that the importance of OMZs as niches for N2 fixation may increase in the futu

Albiglutide and cardiovascular outcomes in patients with type 2 diabetes and cardiovascular disease (Harmony Outcomes): a double-blind, randomised placebo-controlled trial

Background: Glucagon-like peptide 1 receptor agonists differ in chemical structure, duration of action, and in their effects on clinical outcomes. The cardiovascular effects of once-weekly albiglutide in type 2 diabetes are unknown. We aimed to determine the safety and efficacy of albiglutide in preventing cardiovascular death, myocardial infarction, or stroke. Methods: We did a double-blind, randomised, placebo-controlled trial in 610 sites across 28 countries. We randomly assigned patients aged 40 years and older with type 2 diabetes and cardiovascular disease (at a 1:1 ratio) to groups that either received a subcutaneous injection of albiglutide (30–50 mg, based on glycaemic response and tolerability) or of a matched volume of placebo once a week, in addition to their standard care. Investigators used an interactive voice or web response system to obtain treatment assignment, and patients and all study investigators were masked to their treatment allocation. We hypothesised that albiglutide would be non-inferior to placebo for the primary outcome of the first occurrence of cardiovascular death, myocardial infarction, or stroke, which was assessed in the intention-to-treat population. If non-inferiority was confirmed by an upper limit of the 95% CI for a hazard ratio of less than 1·30, closed testing for superiority was prespecified. This study is registered with ClinicalTrials.gov, number NCT02465515. Findings: Patients were screened between July 1, 2015, and Nov 24, 2016. 10 793 patients were screened and 9463 participants were enrolled and randomly assigned to groups: 4731 patients were assigned to receive albiglutide and 4732 patients to receive placebo. On Nov 8, 2017, it was determined that 611 primary endpoints and a median follow-up of at least 1·5 years had accrued, and participants returned for a final visit and discontinuation from study treatment; the last patient visit was on March 12, 2018. These 9463 patients, the intention-to-treat population, were evaluated for a median duration of 1·6 years and were assessed for the primary outcome. The primary composite outcome occurred in 338 (7%) of 4731 patients at an incidence rate of 4·6 events per 100 person-years in the albiglutide group and in 428 (9%) of 4732 patients at an incidence rate of 5·9 events per 100 person-years in the placebo group (hazard ratio 0·78, 95% CI 0·68–0·90), which indicated that albiglutide was superior to placebo (p<0·0001 for non-inferiority; p=0·0006 for superiority). The incidence of acute pancreatitis (ten patients in the albiglutide group and seven patients in the placebo group), pancreatic cancer (six patients in the albiglutide group and five patients in the placebo group), medullary thyroid carcinoma (zero patients in both groups), and other serious adverse events did not differ between the two groups. There were three (<1%) deaths in the placebo group that were assessed by investigators, who were masked to study drug assignment, to be treatment-related and two (<1%) deaths in the albiglutide group. Interpretation: In patients with type 2 diabetes and cardiovascular disease, albiglutide was superior to placebo with respect to major adverse cardiovascular events. Evidence-based glucagon-like peptide 1 receptor agonists should therefore be considered as part of a comprehensive strategy to reduce the risk of cardiovascular events in patients with type 2 diabetes. Funding: GlaxoSmithKline

Predicting the structure and function of coalesced microbial communities: Most efficient community dominates community mix

Immigration has major impacts on both the structure and function of communities and evolutionary dynamics of populations. While most work on immigration deals with relatively low numbers and diversity of immigrants, this does not capture microbial community dynamics, which frequently involve the coalescence of entire communities. The general consequences, if any, of such community coalescence are unclear, although existing theoretical and empirical studies suggest coalescence can result in single communities dominating resulting communities. A recent extension8 of classical ecological theory may provide a simple explanation: communities that exploit niches more fully and efficiently prevent species from other communities invading. Here, we test this prediction using complex anaerobic microbial communities, for which methane production provides a measure of resource use efficiency at community scale. We found that communities producing the most methane when grown in isolation dominated in mixtures of communities. As a consequence, the total methane production increased with the number of communities used as an inoculum. In addition to providing a practical method for enhancing biogas production during anaerobic digestion, these results are likely to be relevant to many other microbial communities. As such, it may be possible to predictably manipulate microbial community function for other biotechnological processes, health and agriculture

Toward in silico CMC: An industrial collaborative approach to model‐based process development

The Third Modeling Workshop focusing on bioprocess modeling was held in Kenilworth, NJ in May 2019. A summary of these Workshop proceedings is captured in this manuscript. Modeling is an active area of research within the biotechnology community, and there is a critical need to assess the current state and opportunities for continued investment to realize the full potential of models, including resource and time savings. Beyond individual presentations and topics of novel interest, a substantial portion of the Workshop was devoted toward group discussions of current states and future directions in modeling fields. All scales of modeling, from biophysical models at the molecular level and up through large scale facility and plant modeling, were considered in these discussions and are summarized in the manuscript. Model life cycle management from model development to implementation and sustainment are also considered for different stages of clinical development and commercial production. The manuscript provides a comprehensive overview of bioprocess modeling while suggesting an ideal future state with standardized approaches aligned across the industry

Toward in silico CMC: An industrial collaborative approach to model‐based process development

The Third Modeling Workshop focusing on bioprocess modeling was held in Kenilworth, NJ in May 2019. A summary of these Workshop proceedings is captured in this manuscript. Modeling is an active area of research within the biotechnology community, and there is a critical need to assess the current state and opportunities for continued investment to realize the full potential of models, including resource and time savings. Beyond individual presentations and topics of novel interest, a substantial portion of the Workshop was devoted toward group discussions of current states and future directions in modeling fields. All scales of modeling, from biophysical models at the molecular level and up through large scale facility and plant modeling, were considered in these discussions and are summarized in the manuscript. Model life cycle management from model development to implementation and sustainment are also considered for different stages of clinical development and commercial production. The manuscript provides a comprehensive overview of bioprocess modeling while suggesting an ideal future state with standardized approaches aligned across the industry

The Contamination of Commercial 15N2 Gas Stocks with 15N–Labeled Nitrate and Ammonium and Consequences for Nitrogen Fixation Measurements

We report on the contamination of commercial 15-nitrogen (N-15) N-2 gas stocks with N-15-enriched ammonium, nitrate and/or nitrite, and nitrous oxide. N-15(2) gas is used to estimate N-2 fixation rates from incubations of environmental samples by monitoring the incorporation of isotopically labeled N-15(2) into organic matter. However, the microbial assimilation of bioavailable N-15-labeled N-2 gas contaminants, nitrate, nitrite, and ammonium, is liable to lead to the inflation or false detection of N-2 fixation rates. N-15(2) gas procured from three major suppliers was analyzed for the presence of these N-15-contaminants. Substantial concentrations of N-15-contaminants were detected in four Sigma-Aldrich N-15(2) lecture bottles from two discrete batch syntheses. Per mole of N-15(2) gas, 34 to 1900 mmoles of N-15-ammonium, 1.8 to 420 mmoles of (15)Nnitrate/nitrite, and 0.01 nmoles N L-1 d(-1), to 530 nmoles N L-1 d(-1), contingent on experimental conditions. These rates are comparable to, or greater than, N-2 fixation rates commonly detected in field assays. These results indicate that past reports of N-2 fixation should be interpreted with caution, and demonstrate that the purity of commercial N-15(2) gas must be ensured prior to use in future N-2 fixation rate determinations