Production and end-of-life scenarios

Funding Information: The studied PHBV pellet production was carried out at a pilot scale (FCT-UNL, Portugal and IVV Fraunhofer, Germany) and subsequently simulated to an industrial scale (i.e. upscaling of 50 times) within the context of the Horizon project “Granting society with low environmental impact innovative packaging” (GLOPACK). The production at these two scales was assumed to obtain a similar function (or the same functional unit (FU), i.e. 1 kg of PHBV pellets formulated for food packaging to be delivered at the gate, including two main steps. PHBV powder was first produced from fruit residues via microbial synthesis, followed by extraction and purification, namely “PHBV powder production” hereafter. Secondly, the additive (i.e. boron nitride as a nucleating agent) was added to the PHBV powder (0.5 wt%) to increase the PHBV crystallinity, followed by compounding into pellets, referred to as “material processing”. Lignocellulosic fibres, e.g. milled wheat straws) were also considered as an optional filler (up to 20 wt%) (Fig. 1). More details regarding microbial synthesis can be found in Matos et al. (2021). Funding Information: We thank GLOPACK's Consortium partners for providing data and sharing expertise, in particular the colleagues from Instituto de Biologia Experimental e Tecnológica (iBET), University of Montpellier, Fraunhofer Institute for Process Engineering and Packaging IVV, and InnovEn, for assisting in acquiring the primary data and modelling the industrial production (upscaling). Publisher Copyright: © 2024 The AuthorsIn the context of a circular bio-based economy, more public attention has been paid to the environmental sustainability of biodegradable bio-based plastics, particularly plastics produced using emerging biotechnologies, e.g. poly(3-hydroxybutyrate-co-3-hydroxyvalerate) or PHBV. However, this has not been thoroughly investigated in the literature. Therefore, this study aimed to address three aspects regarding the environmental impact of PHBV-based plastic: (i) the potential environmental benefits of scaling up pellet production from pilot to industrial scale and the environmental hotspots at each scale, (ii) the most favourable end-of-life (EOL) scenario for PHBV, and (iii) the environmental performance of PHBV compared to benchmark materials considering both the pellet production and EOL stages. Life cycle assessment (LCA) was implemented using Cumulative Exergy Extraction from the Natural Environment (CEENE) and Environmental Footprint (EF) methods. The results show that, firstly, when upscaling the PHBV pellet production from pilot to industrial scale, a significant environmental benefit can be achieved by reducing electricity and nutrient usage, together with the implementation of better practices such as recycling effluent for diluting feedstock. Moreover, from the circularity perspective, mechanical recycling might be the most favourable EOL scenario for short-life PHBV-based products, using the carbon neutrality approach, as the material remains recycled and hence environmental credits are achieved by substituting recyclates for virgin raw materials. Lastly, PHBV can be environmentally beneficial equal to or even to some extent greater than common bio- and fossil-based plastics produced with well-established technologies. Besides methodological choices, feedstock source and technology specifications (e.g. pure or mixed microbial cultures) were also identified as significant factors contributing to the variations in LCA of (bio)plastics; therefore, transparency in reporting these factors, along with consistency in implementing the methodologies, is crucial for conducting a meaningful comparative LCA.publishersversionpublishe

Development and validation of a rabbit model of Pseudomonas aeruginosa non-ventilated pneumonia for preclinical drug development

BackgroundNew drugs targeting antimicrobial resistant pathogens, including Pseudomonas aeruginosa, have been challenging to evaluate in clinical trials, particularly for the non-ventilated hospital-acquired pneumonia and ventilator-associated pneumonia indications. Development of new antibacterial drugs is facilitated by preclinical animal models that could predict clinical efficacy in patients with these infections.MethodsWe report here an FDA-funded study to develop a rabbit model of non-ventilated pneumonia with Pseudomonas aeruginosa by determining the extent to which the natural history of animal disease reproduced human pathophysiology and conducting validation studies to evaluate whether humanized dosing regimens of two antibiotics, meropenem and tobramycin, can halt or reverse disease progression.ResultsIn a rabbit model of non-ventilated pneumonia, endobronchial challenge with live P. aeruginosa strain 6206, but not with UV-killed Pa6206, caused acute respiratory distress syndrome, as evidenced by acute lung inflammation, pulmonary edema, hemorrhage, severe hypoxemia, hyperlactatemia, neutropenia, thrombocytopenia, and hypoglycemia, which preceded respiratory failure and death. Pa6206 increased >100-fold in the lungs and then disseminated from there to infect distal organs, including spleen and kidneys. At 5 h post-infection, 67% of Pa6206-challenged rabbits had PaO2 <60 mmHg, corresponding to a clinical cut-off when oxygen therapy would be required. When administered at 5 h post-infection, humanized dosing regimens of tobramycin and meropenem reduced mortality to 17-33%, compared to 100% for saline-treated rabbits (P<0.001 by log-rank tests). For meropenem which exhibits time-dependent bactericidal activity, rabbits treated with a humanized meropenem dosing regimen of 80 mg/kg q2h for 24 h achieved 100% T>MIC, resulting in 75% microbiological clearance rate of Pa6206 from the lungs. For tobramycin which exhibits concentration-dependent killing, rabbits treated with a humanized tobramycin dosing regimen of 8 mg/kg q8h for 24 h achieved Cmax/MIC of 9.8 ± 1.4 at 60 min post-dose, resulting in 50% lung microbiological clearance rate. In contrast, rabbits treated with a single tobramycin dose of 2.5 mg/kg had Cmax/MIC of 7.8 ± 0.8 and 8% (1/12) microbiological clearance rate, indicating that this rabbit model can detect dose-response effects.ConclusionThe rabbit model may be used to help predict clinical efficacy of new antibacterial drugs for the treatment of non-ventilated P. aeruginosa pneumonia

Global investments in pandemic preparedness and COVID-19: development assistance and domestic spending on health between 1990 and 2026

Background The COVID-19 pandemic highlighted gaps in health surveillance systems, disease prevention, and treatment globally. Among the many factors that might have led to these gaps is the issue of the financing of national health systems, especially in low-income and middle-income countries (LMICs), as well as a robust global system for pandemic preparedness. We aimed to provide a comparative assessment of global health spending at the onset of the pandemic; characterise the amount of development assistance for pandemic preparedness and response disbursed in the first 2 years of the COVID-19 pandemic; and examine expectations for future health spending and put into context the expected need for investment in pandemic preparedness. Methods In this analysis of global health spending between 1990 and 2021, and prediction from 2021 to 2026, we estimated four sources of health spending: development assistance for health (DAH), government spending, out-of-pocket spending, and prepaid private spending across 204 countries and territories. We used the Organisation for Economic Co-operation and Development (OECD)'s Creditor Reporting System (CRS) and the WHO Global Health Expenditure Database (GHED) to estimate spending. We estimated development assistance for general health, COVID-19 response, and pandemic preparedness and response using a keyword search. Health spending estimates were combined with estimates of resources needed for pandemic prevention and preparedness to analyse future health spending patterns, relative to need. Findings In 2019, at the onset of the COVID-19 pandemic, US$9·2 trillion (95$7·3 trillion (95% UI 7·2–7·4) in 2019; 293·7 times the $24·8 billion (95$43·1 billion in development assistance was provided to maintain or improve health. The pandemic led to an unprecedented increase in development assistance targeted towards health; in 2020 and 2021, $1·8 billion in DAH contributions was provided towards pandemic preparedness in LMICs, and$37·8 billion was provided for the health-related COVID-19 response. Although the support for pandemic preparedness is 12·2% of the recommended target by the High-Level Independent Panel (HLIP), the support provided for the health-related COVID-19 response is 252·2% of the recommended target. Additionally, projected spending estimates suggest that between 2022 and 2026, governments in 17 (95% UI 11–21) of the 137 LMICs will observe an increase in national government health spending equivalent to an addition of 1% of GDP, as recommended by the HLIP. Interpretation There was an unprecedented scale-up in DAH in 2020 and 2021. We have a unique opportunity at this time to sustain funding for crucial global health functions, including pandemic preparedness. However, historical patterns of underfunding of pandemic preparedness suggest that deliberate effort must be made to ensure funding is maintained

Changes in soil characteristics and C dynamics after mangrove clearing (Vietnam)

Of the blue carbon sinks, mangroves have one of the highest organic matter (OM) storage capacities in their soil due to low mineralization processes resulting from waterlogging. However, mangroves are disappearing worldwide because of demographic increases. In addition to the loss of CO2 fixation, mangrove clearing can strongly affect soil characteristics and C storage. The objectives of the present study were to quantify the evolution of soil quality, carbon stocks and carbon fluxes after mangrove clearing. Sediment cores to assess physico-chemical properties were collected and in situ CO2 fluxes were measured at the soil-air interface in a mangrove of Northern Vietnam. We compared a Kandelia candel mangrove forest with a nearby zone that had been cleared two years before the study. Significant decrease of clay content and an increase in bulk density for the upper 35 cm in the cleared zone were observed. Soil organic carbon (OC) content in the upper 35 cm decreased by >65% two years after clearing. The quantity and the quality of the carbon changed, with lower carbon to nitrogen ratios, indicating a more decomposed OM, a higher content of dissolved organic carbon, and a higher content of inorganic carbon (three times higher). This highlights the efficiency of mineralization processes following clearing. Due to the rapid decrease in the soil carbon content, CO2 fluxes at sediment interface were >50% lower in the cleared zone. Taking into account carbonate precipitation after OC mineralization, the mangrove soil lost similar to 10 MgOC ha(-1)yr(-1) mostly as CO2 to the atmosphere and possibly as dissolved forms towards adjacent ecosystems. The impacts on the carbon cycle of mangrove clearing as shown by the switch from a C sink to a C source highlight the importance of maintaining these ecosystems, particularly in a context of climate change

Stabilizing additives added during cell lysis aid in the solubilization of recombinant proteins.

Insoluble recombinant proteins are a major issue for both structural genomics and enzymology research. Greater than 30% of recombinant proteins expressed in Escherichia coli (E. coli) appear to be insoluble. The prevailing view is that insolubly expressed proteins cannot be easily solubilized, and are usually sequestered into inclusion bodies. However, we hypothesize that small molecules added during the cell lysis stage can yield soluble protein from insoluble protein previously screened without additives or ligands. We present a novel screening method that utilized 144 additive conditions to increase the solubility of recombinant proteins expressed in E. coli. These selected additives are natural ligands, detergents, salts, buffers, and chemicals that have been shown to increase the stability of proteins in vivo. We present the methods used for this additive solubility screen and detailed results for 41 potential drug target recombinant proteins from infectious organisms. Increased solubility was observed for 80% of the recombinant proteins during the primary and secondary screening of lysis with the additives; that is 33 of 41 target proteins had increased solubility compared with no additive controls. Eleven additives (trehalose, glycine betaine, mannitol, L-Arginine, potassium citrate, CuCl(2), proline, xylitol, NDSB 201, CTAB and K(2)PO(4)) solubilized more than one of the 41 proteins; these additives can be easily screened to increase protein solubility. Large-scale purifications were attempted for 15 of the proteins using the additives identified and eight (40%) were prepared for crystallization trials during the first purification attempt. Thus, this protocol allowed us to recover about a third of seemingly insoluble proteins for crystallography and structure determination. If recombinant proteins are required in smaller quantities or less purity, the final success rate may be even higher

Allelopathic Potential of Rice and Identification of Published Allelochemicals by Cloud-Based Metabolomics Platform

The methanol extracts of nine popular cultivated Vietnamese rice cultivars (Oryza sativa L.cv. OM 2395, 5451, 6976, 380, 5930, 4498, 3536, N406, and 7347) were used to explore their allelopathic potential on barnyardgrass (Echinochola crus-galli L.). At 0.1 g mL−1, OM 5930, OM 4498, and OM 6976 correlatively possessed greatest phytotoxicity on barnyardgrass shoot (98.77%, 90.75%, and 87.17%) and root (99.39%, 92.83%, and 86.56%) growth. The following study aimed to detect previously-known allelochemicals in those rice using XCMS online cloud-based metabolomics platform. Twenty allelochemicals were semi-quantified and seven of them were detected predominantly and five was putatively confirmed in OM 5930 (mg/ 100g fresh rice) as salicylic acid (5.0076), vanillic acid (0.1246), p-coumaric acid (0.1590), 2,4-dimethoxybenzoic acid (0.1045), and cinnamic acid (3.3230). These compounds were active at concentrations greater than 0.5 mM and the average EC50 were 1.24 mM. The results indicated that OM 5930 may use as promising candidates in weed biological control for rice production

Proposed mechanism for rescue of recombinant protein solubility.

<p>We hypothesize that up to 80% of the seemingly insoluble recombinant proteins are in a partially folded state and reside in the <i>E. coli</i> cytosol. If lysed in a non-ideal buffer, the proteins unfold, resulting in aggregates of insoluble protein. When the sample is centrifuged to separate the soluble fraction, the protein aggregates are present in the insoluble cell pellet. If the additives are present during cell lysis, they can either stabilize the proteins from partially unfolding, preventing protein-protein interactions, or aid as chemical chaperones, leading to the properly folded and non-aggregated state. When centrifuged there are minimal protein aggregates and the recombinant protein remains in the soluble fraction.</p

Complete additive and buffer list at final concentrations.

<p>Complete additive and buffer list at final concentrations.</p

The top performing additive molecular structures.

<p>Presented are the molecular structures of the top performing additives. The number of proteins soluble with each additive is presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052482#pone-0052482-t004" target="_blank">table 4</a>.</p