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Severe plastics pollution derived from the wide use and the past incorrect dispersion of petroleum-based plastics. The development of bioplastics/biopolymers with the same chemical/physical characteristics of conventional plastics can be a solution to the problems of their degradability. Polybutylene succinate was among the most investigated biopolymers for these characteristics that can be produced from bio-based succinic acid and butan-1,4-diol. Succinic acid can be produced through fermentation process using microorganisms as Actinobacillus succinogenes that were able to utilise glucose from several sources as lignocellulosic biomasses. Lignocellulosic biomass can be ideal candidates for glucose supply but the processes to release fermentable sugars can produce inhibitors (acids and furans) of the biological processes for succinic acid production. The aim of this paper was to evaluate the growth of A. succinogenes for the production of succinic acid under acetic acid and furfural at different concentration on inoculum with pre-adaptation and without adaptation. The results highlighted that pre-inoculum adaptation was essential for the growth of the strain and succinic acid production that decreased strongly under the synergistic effect of acetic acid and furfural in the broth (63% inhibition rate of growth) respect to inoculum with pre-inoculum adaptation (29% inhibition rate of growth).

Succinic acid was a building block chemical of many industrial products, among these, one of the most interesting applications concerns the bioplastics market. This compound can be produced through the biotechnological routes, but several challenges still need to be addressed. One of the greatest challenges has been the downstream process to recover and purify succinic acid from the fermentative broth. Succinic acid must be taken from the fermentation broth, that was composed of various compounds, such as cellular residues, proteins, and other acids, to reach a recovery and a purity >90%, in order to be marketed. Generally, the downstream process can be divided into three phases: pretreatment, separation and purification. Several studies have been carried out on different technologies with the aim to improve recovery and purity. To date, all these methods were applied only on laboratory-scale, due to various critical issues that emerged, such as high costs, which prevented their application on an industrial scale. This review highlighted the best obtained results from the recent studies and discussed on critical aspects and future perspectives on the topic.

Abstract Background Severe asthma can cause poor health status, poor health-related quality of life (HRQoL) and an impaired functioning at work. However, to date, limited data are available on the impact of the biological therapies on such outcomes. Therefore, aim of the present study was to prospectively assess the clinical, quality of life and work functionality issues in severe asthma patients both at baseline and after 6 months of biological therapies and determine which individual, pathological and occupational factors can influence such parameters. Methods Fifty-two patients were enrolled between December 2022 and June 2023. Patients’ personal, clinical, functional and occupational features were assessed. The Short Form Health Survey (SF-12), the Work Productivity and Activity Impairment (WPAI) questionnaire and the Work Ability Index (WAI) were employed to assess HRQoL, the employee’s productivity and perception of work ability, respectively. Results Among the enrolled patients, 30 (57.70%) were employed. Biological therapy induced a significant improvement in clinical and functional parameters, e.g., FEV1% (72 ± 12 vs.87 ± 13%; 72 ± 14 vs. 86 ± 14%), FVC% (92 ± 11 vs. 101 ± 11%; 90 ± 13 vs. 98 ± 14%) and FEV1/FVC (62 ± 11 vs. 71 ± 8%; 64 ± 9 vs. 70 ± 8%) in workers and non-workers, respectively (P

Succinic acid is one of the most used building blocks in different production chains. Conventionally, this carboxylic acid is produced through petrochemical routes, but can be also produced via biological routes. Many microorganisms, such as Actinobacillus succinogenes, synthesized succinic acid as an intermediate of many biochemical pathways, as the Krebs cycle. Nowadays, the industrial production of bio-succinic acid has been limited due to different critical issues, like the low productivity and yield and downstreaming processes. To improve the bio-based production, the operative conditions of the fermentation process needed to be optimized especially when agro-industrial residues were used as sugars feedstock. To this aim, in this study fermentation batch tests were performed by using different concentrations of A. succinogenes at a given glucose concentration. The results suggested that glucose consumption and succinic acid production were not directly influenced by the initial bacterial concentration respect to glucose concentration. But the best results were obtained at the ratio of bacteria:glucose equal to 1:1 and further tests will be necessary to implement succinic acid yield and productivity.

Bio-based succinic acid production holds great promise for the process sustainability through the use of renewable resources such as lignocellulosic biomass and the use of microorganisms that can utilize CO2 in fermentation processes. A critical review of the latest findings on succinic acid production from lignocellulosic biomass was discussed, by highlighting all the key aspects of optimization processes at different levels of the process from pre-treatment to succinic acid production. Chemical-physical pretreatments seemed to be the most appropriate for the pretreatment of lignocellulosic biomass, but many strategies are still needed to solve the problem of variability in initial composition, the presence of post-treatment inhibitory compounds and the use of green solvent. In addition, the saccharization-fermentation process seemed to be the most suitable for the fermentative process of hydrolysates obtained from pretreatment to produce bio-based succinic acid. Certainly new studies and strategies are still needed to overcome the weaknesses of the whole process of succinic acid production from lignocellulosic biomass.

Abstract Background We evaluated the safety and feasibility of one‐lung ventilation in obese patients undergoing thoracoscopic lobectomy and whether obesity affected peri‐ and postoperative outcomes. Methods This was a retrospective single center study including consecutive patients undergoing thoracoscopic lobectomy between October 2019 and February 2022. Obese patients were statistically compared to a control group to evaluate any differences in relation to one‐lung ventilation and peri‐ and postoperative outcomes. Results Our study population included 111 patients; of these, 26 (23%) were included in the obese group, while 85 (77%) were included within the nonobese group. To obtain one‐lung ventilation in nonobese patients, a double‐lumen tube was more frequently used than a single‐lumen tube with bronchial blocker (61% vs. 39%; p = 0.02), while in obese patients a single‐lumen tube with bronchial blocker was used more than a double‐lumen tube (81% vs. 19%, p = 0.001). Intergroup comparison showed that a double‐lumen tube was the preferred method in nonobese patients, while a single‐lumen tube with bronchial blockers was the strategy of choice in obese patients (p = 0.0002). Intubation time was longer in the obese group than in the nonobese group (94.0 ± 6.1 vs. 85.0 ± 7.0 s; p = 0.0004) and failure rate of first attempt at intubation was higher in the obese group (23% vs. 5%; p = 0.01). Obesity was not associated with increased intra‐, peri‐ and postoperative complications and/or mortality. Conclusions One‐lung ventilation is a feasible and safe procedure also in obese patients and obesity did not negatively affect peri‐ and postoperative outcomes after lung resection.

Background Endothelial dysfunction has been proposed to play a key role in the pathogenesis of coronavirus disease 2019 (COVID-19) and its post-acute sequelae. Flow-mediated dilation (FMD) is recognized as an accurate clinical method to assess endothelial function. Thus, we performed a meta-analysis of the studies evaluating FMD in convalescent COVID-19 patients and controls with no history of COVID-19.Methods A systematic literature search was conducted in the main scientific databases according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Using the random effects method, differences between cases and controls were expressed as mean difference (MD) with 95% confidence intervals (95% CI). The protocol was registered on PROSPERO with reference number CRD42021289684.Results Twelve studies were included in the final analysis. A total of 644 convalescent COVID-19 patients showed significantly lower FMD values as compared to 662 controls (MD: −2.31%; 95% CI: −3.19, −1.44; p

Severe Asthma (SA) is characterized by inadequate disease control despite maximal inhalation therapy. In November 2021, a 68-years old female patient presented at our facility referring a worsening in her asthma-related symptoms and a high exacerbations rate. She reported a liver transplantation in 2015 and was in treatment with tacrolimus. We started treatment with Mepolizumab 100 mg once every 28 days and monitored her lung function as well as her lymphocytes subsets. After one year follow-up, the patient had a substantial improvement in lung function, exacerbation rate, daily OCS intake dose and no variation in the blood concentration of tacrolimus.

BackgroundLung involvement in primary Sjögren’s syndrome (pSS) may vary from 9 to 90%. Interstitial lung disease and tracheobronchial alterations are the most typical findings. The evidence of primarily emphysematous changes at computed tomography of the chest of pSS patients has occasionally been described but poorly characterized. This study aims to assess pulmonary involvement and the impact on respiratory function in a cohort of pSS patients.Materials and methodsA total of 22 consecutive patients diagnosed with pSS underwent pulmonary function tests to investigate the presence of ventilatory impairment and evaluate the exchanges of alveolar gases. All patients underwent a chest high-resolution computed tomography (HRTC).ResultsDynamic volumes were within the normal range in 21 patients (95.4%). A reduction in the diffusing capacity of the lung for carbon monoxide (DLCO) was observed in 18 patients (81.8%). Ten (45.5%) patients showed a mild degree deficit, while 8 patients (36%) showed a moderate degree deficit. Analysis of DLCO revealed a significant difference between pSS patients and controls [t(30.98) = −10.77; p < 0.001], showing a higher DLCO value for the healthy controls (mean ± SE; 101.27 ± 6.08) compared to pSS patients (mean ± SE; 65.95 ± 12.78). Emphysema was found in 21 (94.5%) patients and was the most widespread pulmonary injury. Tracheal thickness was reduced in 15 (67%) patients. Micronodules were observed in 10 (45%) patients in all the pulmonary fields. Bronchial wall thickening and bronchiectasis were observed in 8 (36%) patients, mainly in the lower lobes. Ground glass was found in 5 (22.5%) patients in lower and higher lobes. Cysts were observed in two patients (9%).ConclusionThe reduction of the DLCO could be related to early emphysematous alterations in the absence of spirometric alterations and relevant respiratory symptoms. In conclusion, emphysema might be seen as an early pulmonary involvement mark in patients suffering from pSS.

The concept of fastidious microorganisms currently found in scientific literature is mainly related to the difficulty of isolating/culturing/preserving bacteria. Eukaryotes are investigated much less in this respect, although they represent a fundamental part of the microbial world. Furthermore, not only isolation, but also identification and culturing (in the perspective of long-term preservation) should be considered key aspects often impacting on the study of fastidious microorganisms, especially in terms of preservation in culture collections and biotechnological exploitation. The present review aimed to investigate the current state of the art on fastidious eukaryotes, with special emphasis on the efforts to improve their isolation, identification, culturing and long-term preservation in culture collections practices. A few case studies focused on some fastidious eukaryotic microorganisms (including possible customized solutions to overcome specific issues) are also presented: isolation and preservation of slow-growing fungi, culturing of Haematococcus lacustris, isolation of unialgal strains of Cyanidiophytina (Rhodophyta), identification of Metschnikowia pulcherrima clade yeasts, isolation and preservation of Pyricularia species, preservation of Halophytophtora spp.

Severe acute respiratory syndrome-coronavirus-2 (SARS‐CoV‐2), responsible for COVID-19, has caused a global pandemic. Observational studies revealed a condition, herein called as Long-COVID syndrome (PC), that affects both moderately and severely infected patients, reducing quality-of-life. The mechanism/s underlying the onset of fibrotic-like changes in PC are still not well defined. The goal of this study was to understand the involvement of the Absent in melanoma-2 (AIM2) inflammasome in PC-associated lung fibrosis-like changes revealed by chest CT scans. Peripheral blood mononuclear cells (PBMCs) obtained from PC patients who did not develop signs of lung fibrosis were not responsive to AIM2 activation by Poly dA:dT. In sharp contrast, PBMCs from PC patients with signs of lung fibrosis were highly responsive to AIM2 activation, which induced the release of IL-1α, IFN-α and TGF-β. The recognition of Poly dA:dT was not due to the activation of cyclic GMP-AMP (cGAMP) synthase, a stimulator of interferon response (cGAS-STING) pathways, implying a role for AIM2 in PC conditions. The release of IFN-α was caspase-1- and caspase-4-dependent when AIM2 was triggered. Instead, the release of pro-inflammatory IL-1α and pro-fibrogenic TGF-β were inflammasome independent because the inhibition of caspase-1 and caspase-4 did not alter the levels of the two cytokines. Moreover, the responsiveness of AIM2 correlated with higher expression of the receptor in circulating CD14+ cells in PBMCs from patients with signs of lung fibrosis.

Lutein is a powerful carotenoid that is used as a feed additive for the colouring properties and as supplement in nutraceutical products. Its principal healthy properties are antioxidant, anti-inflammatory and protective against age-related macular degeneration. Lutein is indeed a macular pigment in human eyes between zeaxanthin and their function is very important for the eyes health. Lutein is found in some vegetables as spinach, parsley, and kale with a content of around 1-10 mg/100 g. The commercial and recognized source of lutein is the plant marigold (Tagetes erecta L.). Lutein is extracted from marigold petals as oleoresin and the lutein content can be around 0.03% (based on dry biomass weight). Since the costs for harvesting are expensive, land and water use are requested for marigold cultivation and due to the seasonality of the growth, an alternative source of lutein has been searched among microalgae. Scenedesmus almeriensis is considered a promising source for lutein production whose content is around 4.5 mg/g dry weight. The aim of this paper is to improve lutein extraction from Scenedesmus almeriensis by using pressurized liquid extraction through accelerated solvent extractor (ASE 200©Dionex). Extractions were performed using different solvent as the mixture chloroform:methanol (1:1 v/v), ethanol, hexane, acetone. Several temperatures were tested from 20° to 80°C and biomass was mechanically pre-treated. Each extraction cycle was repeated until to complete biomass decolouring.

Microalgae have been long recognized as potential food and feed solution, since they are able to meet the population growth on demand for a more sustainable food and feed, especially with respect to protein production. In addition, bioactive components, such as eicosapentaenoic acid (EPA), with well-known antioxidant and anti-inflammatory properties, can be extracted and incorporated in food supplements. Microalgae cultivation and processing becomes even more sustainable if simultaneously coupled to green technologies for the environmental protection. One of the most promising techniques is the supercritical fluid extraction which allows to extract bioactive compounds without loss of their activity and free from extraction solvents. In this work, a pilot scale supercritical CO2 (ScCO2) extraction plant was exploited for performing the extraction of active components from microalgae biomass potentially usable in the food and the feed sectors. Nannochloropsis gaditana microalga was selected as natural producer of EPA. The extract obtained after ScCO2 tests was enriched of EPA and protein, therefore suitable for food applications. The exhausted biomass, having a high content of carbohydrates and total dietary fiber, might be proposed as feed suppl

The downstream processing of natural active molecules remains the most significant cost in the production pipeline. This considerable cost is largely attributed to rigorous chromatographic purification protocols. In an ongoing effort to abate the dependence on chromatography in downstream processing, alternative affinity matrices in the form of magnetic particles (e.g., iron oxide) have emerged as viable candidates. Nevertheless, biotechnological applications of iron oxide particles are still confined to the research level or for low-throughput clinical applications. Herein, we describe an efficient, quick, and environmentally friendly method for the isolation of astaxanthin and lutein, two carotenoids with very similar chemical structure, from extracts of the microalga Haematococcus pluvialis. The technology proposed, named Selective Magnetic Separation (SMS), is based on the use of magnetic materials carrying affinity ligands that bind carotenoids and is applied as second step of purification. The method, thanks to functionalized magnetic nanoparticles, reduces the use of organic or toxic solvents. In the present work, we examined the most efficient binding conditions such as temperature, magnetic nanoparticles concentration, and elution time, as well as their effects on carotenoids recovery, with the aim to improve the non-covalent binding between the ligand (amines) and astaxanthin/lutein. Our initial results clearly showed that it is possible to use magnetic separation as an alternative to chromatography to isolate important and valuable compounds.

This work presents a risk analysis of a plant that uses supercritical fluid extraction technology with carbon dioxide as a solvent to obtain bioproducts from microalgae. Given the high pressure at which it operates, the extraction plant called "Luwar" could be affected by dangerous events, such as the gas release in concentrations that are harmful to human health. In order to limit the damage, the risk analysis is of primary importance. The gas release could occur following the breakage of a pipe, which was the scenario being considered. It was analyzed using the commercial software "Phast & Safeti", an alternative tool to the methods traditionally used in the literature in hazard events of this type, such as the Threshold Limit Value-Time Weighted Average (TLV-TWA) and the Threshold Limit Value-Short Term Exposure Limit (TLV-STEL), that are defined on the basis of the characteristics of the substance dispersed in the air. Since the pressure of the extraction vessel is the highest in the system, the risk analysis was carried out on the breakage of a pipe coming out of the extractor. The results obtained made it possible to identify the risk parameters on the basis of which to make the best choices in terms of safety to cope with the type of hazard analyzed.

In this work, a risk analysis of a supercritical fluid extraction plant is presented. The plant aims to extract valuable compounds from microalgae and uses carbon dioxide as a solvent at high temperature and pressure. Hazard events that could occur are related to the high pressure at which equipment operate and they, in addition to causing damage to the plant structure, could cause the release of CO2, which in high concentrations endangers the health of workers involved in the activities, therefore, an assessment of this type is crucial. The hazard scenario considered in this work is the equipment rupture. Generally, the rupture is analysed through methods based on indices determination such as the Fire and Explosion Index, the Probit analysis and the Safety Weighted Hazard Index. In this case, the commercial software “Phast & Safeti” of DNV GL Company was used, based on the modelling of the plant together with the process conditions definition. Among the equipment present in the plant, the extraction vessel was chosen since the extraction pressure of bioproducts is the highest in the plant and is equal to 500 bar. Risk parameters obtained after the analysis allow to select the most appropriate safety system able to limit the damage to structures and people.

The growing consciousness for environmental issues have stimulated the search for more sustainable biochemical processes. One of the most attracting chemicals with enormous potential in a biobased economy is succinic acid (SA, butanedioic acid or 1,2-ethanedicarboxylic acid). This compound represents the chemical building block for various high value-added substances which find applications in the detergent/surfactant, food, ion chelator and pharmaceutical markets. Nowadays, the SA market is estimated to be about 20000–30000 ton per year worldwide and its industrial manufacture is mainly based on the catalytic hydrogenation of petrochemically derived maleic acid or maleic anhydride. However, since SA is fermentative product and an intermediate of several biochemical pathways, including the tricarboxylic acid cycle, it can be produced by many microorganisms. The biological SA production at commercial scale and potentially for the commodity chemical market is still a challenging target and requires the development and the optimization of microorganisms-based processes able to guarantee high product concentrations to justify economically feasible recovery. The aim of this paper is the conversion of a mixture of sugars, mainly containing glucose, into SA by SA-producing strain Actinobacillussuccinogens, that was isolated from the bovine rumen. It is a facultative anaerobic, pleomorphic, Gram-negative rod. Experimental tests were carried out by using Actinobacillussuccinogens bought by Culture Collection of Goteborg and by using TBS (Tryptic Soy broth) by changing the concentration of glucose as well as that of other sugars as fructose, xylose, sucrose with the main focus to evaluate the effect of the fermentation for the Actinobacillussuccinogens with the single sugars in a concentration range from 1 to 15 g/l. All the experiments were carried out in anaerobic conditions at 37°C and in a pH range 7-8 for several days. For each test, carboxylic acids and sugars were analyzed by using u-HPLC while the growth of the selected microorganism was monitored by means of a spectrophotometer at 600nm for OD tests.

Microalgae are among the most interesting eukaryotic photosynthethic microorganisms able to use solar energy, nutrients and carbon dioxide to convert them into proteins, carbohydrates, lipids and other valuable organic compounds including carotenoids. Astaxanthin is one of the most interesting antioxidant molecules which has attracted crescent interest due to its positive effects on health and the numerous applications in different sectors, from nutraceutical to cosmetic and aquaculture. Even though the astaxanthin properties are well-known, its price still remains high if associated to the algal form, exceeding ~6000 Eur/Kg. This can be explained by considering the process expenses related to the extraction and purification steps of microalga intracellular metabolites. In fact, the downstream stage of this biotechnological process often accounts for more than 60–70% of total production costs. Optimized extraction and purification operations might contribute to microalgae market with the advantage to commercialize a natural existing astaxanthin form. The aim of this paper is the evaluation of the use of commercial activated carbon Darco™ G-60 for the purification of astaxanthin from an extraction broth. Astaxanthin was firstly extracted from Haematococcus pluvialis red phase supplied by Micoperi Blue Growth, an Italian Company that is working for a long time and it is specialized in the microalgae growth. Extraction was performed by Accelerated Solvent Extractor (ASE®200 DIONEX) at 100 bar and 67°C by using ethanol as green solvent with the main advantage to separate all the unipolar fractions as well as insoluble fractions from astaxanthin extracts (fibers, carbohydrate, ashes). In the second step, astaxanthin was purified with a column filled with activated carbon. Experimental tests by changing the mass of activated carbon were carried out (50mg, 100mg and 200mg)and with a flow rate in the range 0.9-1.0ml/min. All the experimental tests were carried out at room temperature (20°C). Results showed that by using activated carbon, it is possible to obtain an adsorption capacity of DARCO G60 in the range 21,9-23,9 mg/g.

Microalgae represent one of the most interesting biological source, since they are relatively simple microorganisms containing high-value compounds with potential applications in numerous industrial sectors. The global nutraceutical market is well developed and with an enormous potential for growth: suffice to say that it has shown considerable growth with a value in 2016 between 168 and 174 billion euros, with a growth forecast that will allow it to reach between 285 and 313 billion euros in 2024. In accordance with the National Center for Biotechnology Information, by 2050, microalgae will constitute ~18% of protein source among the more diverse market. Healthy properties and excellent nutritional values have been conferred to microalgae, with particular regard to the high content of carbohydrates, vitamins, carotenoids and polyunsaturated fatty acids (PUFAs). The omega-3 PUFAs are extremely important throughout life and are a dietary need found predominantly in Nannochloropsis gaditana oleaginous microalga. In fact, although fish contain long-chain omega-3, they acquire PUFAs mainly from their microalgal diet. Additionally, considering depleting fish supplies, the use of microalgae as alternative raw material might be considered of increasing importance in the near future. Herein we report the assessment of the economic feasibility of an integrated process which allows to extract PUFAs from Nannochloropsis gaditana, by taking into account the main operational parameters able to influence the final products yield.

The global cosmetic market is promptly growing, showing a strong boost with the growth of economic well-being. In this context, the demand of innovative, more and more specific cosmetic ingredients has been the key for searching alternative, preferably naturally-based, active components. Microalgae represent one of the most attracting microorganisms and natural deposit for bioactive compounds for their peculiar compositions and properties. In the cosmetic sector, their efficient application has recently been highlighted by the placing on the market of different cosmetic preparations. A powerful antioxidant, 550 times more effective than vitamin E, with enormous potential for cosmetic and pharmaceutical applications, is astaxanthin. Although high-quality astaxanthin can be obtained from microalgae, particularly from H. pluvialis specie, actually the major part of the market is occupied by the synthetic form (99%), conferring substantial differences between their price. In fact, algal-based astaxanthin costs ?6000 €/Kg, while the synthetic form 600 €/kg. Thus, along with novel natural product launch, the development of cost-effective technologies able to match the existing ones, represents the major challenge for the microalgae application. This study aims to determine the feasibility of microalgal-based astaxanthin production, by exploring both procedural issues and costs evaluation.

Nutraceuticals have gained growing attention during the last year, becoming one of the most relevant areas of investigation. Among the existing natural sources of high value bioactive compounds, microalgae have assumed paramount importance. Indeed, they are a natural source of carotenoids, essential omega-3 fatty acids, proteins, carbohydrates, pigments, vitamins and minerals. Among these components, beta-carotene, a potent antioxidant pigment, is particularly attractive to the nutraceutical and cosmetic sector, being an ingredient in many supplements, functional, cosmetic and pharmaceutical products. Moreover, beta-carotene is one of the forms (precursor) of Vitamin A, which may be used in the manufacture of food supplements according to Directive 2002/46/EC. Even though some cyanobacteria produce beta-carotene (80% of total carotenoids) as the major components followed by zeaxanthin, the commercial biological source of beta-carotene is Dunaliella salina. This microalga lives in very saline environment and has the capacity to produce beta-carotene up to 10% of the dry weight. In this work, beta-carotenewas extracted by using supercritical CO2 under optimized operational conditions, resulting in increased amount of carotenoids (from 3.46% to 18.62%), proteins (from 10.03% to 43.08%), and lipids (from 3.49% to 7.93%). Then, the investigation on the main operational parameters affecting extraction from Dunaliella salina and a cost-effectiveness evaluation of the process are presented; the results evidence as 194.3-366.6 kWh/day reflecting a cost of 26.89-52.60euros/day are required for natural based beta-carotene recovery(59.3%) from Dunaliella salina. Keywords: nutraceutical, microalgae, beta-carotene, Dunaliella salina, supercritical extraction.

The Supercritical Fluid Extraction (SFE) technology is currently used in many industrial fields thanks to the advantages that it can offer. Experimental tests are carried out for different purposes such as decaffeination of coffee/tea, extraction of aromas and herbal flavours as well as spices, extraction of fats and oils, extraction of cholesterol and extraction of alcohol from beverages. SFE plants safety is closely linked to the high pressure they operate, a condition that can lead to a risk status for plant operations and above all workers. For this reason it is necessary to adopt preventive safety measures and a risk analysis is the means that allows to define and examine hazardous scenarios in order to identify safety devices. Risk scenarios related to an extraction plant as explosions are analysed through methods based on indices determination such as the Fire and Explosion Index (F&EI), the Safety Weighted Hazard Index (SW&HI) and the Probit method. This work presents the risk analysis of a SFE plant named “Luwar” using the commercial software “Phast & Safeti” provided by the DNV GL Company. In the plant is used CO2 as extraction fluid and the process purpose is valuable compounds extraction from microalgae. The risk analysis is performed on the extraction vessel considering the plant placed inside a structure (indoor condition). The results allow to identify two risk parameters, i.e. the distance and the area within which CO2 concentration in the air assumes a value considered unsafe for human health. On the basis of the outputs it is possible to identify the safety system to be adopted in order to limit the damage deriving from the occurrence of a risk situation.

The main purpose of this study is to investigate the effects of operative parameters and bioprocess strategies on the photo-autotrophic cultivation of the microalgae Scenedesmus almeriensis for lutein production. S. almeriensis was cultivated in a vertical bubble column photobioreactor (VBC-PBR) in batch mode and the bioactive compounds were extracted by accelerated solvent extraction with ethanol at 67°C and 10 MPa. The cultivation with a volume fraction of CO2 in the range 0–3.0%v/v showed that the highest biomass and lutein concentrations – 3.7 g/L and 5.71 mg/g, respectively – were measured at the highest CO2 concentration and using fresh growth medium. Recycling the cultivation medium from harvested microalgae resulted in decreased biomass and lutein content. The nutrient chemical composition analysis showed the highest consumption rates for nitrogen and phosphorus, with values higher than 80%, while sulfate and chloride were less consumed.

Abstract The increasing use, and sometimes the abuse, particularly in industrialized countries of air conditioning at home, in car, hotel and shopping centres has highlighted new emerging public health issues, resulting from exposure of the airways to cool air or, more properly, resulting from sudden temperature changes. This is part of a wider problem, relating to air quality in indoor environment, such as homes or offices, where people spend more than 90% of their time. In particular, if indoor exposure occurs quickly and without any gradual adaptation to a temperature 2°–3° lower than the external temperature and especially with a 5° difference (avoiding indoor temperature below 24°) and an humidity between 40 and 60%, there is a risk of negative consequences on the respiratory tract and the patient risks to be in a clinical condition characterized by an exacerbation of the respiratory symptoms of his chronic respiratory disease (asthma and COPD) within a few hours or days. Surprisingly, these effects of cold climate remain out of the focus of the media unless spells of unusually cold weather sweep through a local area or unstable weather conditions associated with extremely cold periods of increasing frequency and duration. Moreover, the energy consumed by air conditioning induces an increase of CO2 in atmosphere with increase of global warming. There is a need to better define the consequences of repeated exposure to cold air and the mechanisms by which such exposure could modify airway function and affect the outcomes of patients with pre-existing airway disease. This could help to promote adequate policy and public health actions to face the incoming challenges induced by climate change and global warming.

Purpose: SARS-CoV-2 infection induces in some patients a condition called long-COVID-19, herein post-COVID-19 (PC), which persists for longer than the negative oral-pharyngeal swab. One of the complications of PC is pulmonary fibrosis. The purpose of this study was to identify blood biomarkers to predict PC patients undergoing pulmonary fibrosis. Patients and Methods: We analyzed blood samples of healthy, anti-SARS-CoV-2 vaccinated (VAX) subjects and PC patients who were stratified according to the severity of the disease and chest computed tomography (CT) scan data. Results: The inflammatory C reactive protein (CRP), complement complex C5b-9, LDH, but not IL-6, were higher in PC patients, independent of the severity of the disease and lung fibrotic areas. Interestingly, PC patients with ground-glass opacities (as revealed by chest CT scan) were characterized by higher plasma levels of IL-1α, CXCL-10, TGF-β, but not of IFN-β, compared to healthy and VAX subjects. In particular, 19 out of 23 (82.6%) severe PC and 8 out of 29 (27.6%) moderate PC patients presented signs of lung fibrosis, associated to lower levels of IFN-β, but higher IL-1α and TGF-β. Conclusions: We found that higher IL-1α and TGF-β and lower plasma levels of IFN-β could predict an increased relative risk (RR = 2.8) of lung fibrosis-like changes in PC patients.

Severe eosinophilic asthma (SEA) is associated with high peripheral blood and airway eosinophilia, recurrent disease exacerbations and severe airflow limitation. Eosinophilic inflammation is also responsible for small airway disease (SAD) development. SEA patients experience poor disease control and response to standard therapy and are prime candidates for anti-IL5 biologicals, such as mepolizumab, but the effect of treatment on SAD is unclear. We investigated the effect of mepolizumab on lung function in SEA patients, focusing on SAD parameters, and searched for an association between patients’ phenotypic characteristics and changes in small airways function. In this real-life study, data from 105 patients with SEA were collected at baseline and after 6, 12 and 18 months of mepolizumab treatment. Along with expected improvements in clinical and lung function parameters brought by Mepolizumab treatment, FEF2525-75% values showed a highly significant, gradual and persistent increase (from 32.7 ± 18.2% at baseline to 48.6 ± 18.4% after 18 months) and correlated with ACT scores at 18 months (r = 0.566; p ≤ 0.0001). A patient subgroup analysis showed that changes in FEF25-75% values were higher in patients with a baseline peripheral blood eosinophil count ≥400 cells/μL and oral corticosteroid use. Mepolizumab significantly improves small airway function. This effect correlates with clinical benefits and may represent an accessible parameter through which to evaluate therapeutic response. This study provides novel insights into the phenotypic characteristics associated with the improved functional outcome provided by mepolizumab treatment.

Background: Endothelial dysfunction has a key role in the pathogenesis of coronavirus disease 2019 (COVID-19) and its disabling complications. We designed a case-control study to assess the alterations of endothelium-dependent flow-mediated dilation (FMD) among convalescent COVID-19 patients. Methods: COVID-19 patients referred to a Pulmonary Rehabilitation Unit within 2 months from swab test negativization were consecutively evaluated for inclusion and compared to controls matched for age, gender, and cardiovascular risk factors. Results: A total of 133 convalescent COVID-19 patients (81.2% males, mean age 61.6 years) and 133 matched controls (80.5% males, mean age 60.4 years) were included. A significantly lower FMD was documented in convalescent COVID-19 patients as compared to controls (3.2% ± 2.6 vs. 6.4% ± 4.1 p < 0.001), confirmed when stratifying the study population according to age and major clinical variables. Among cases, females exhibited significantly higher FMD values as compared to males (6.1% ± 2.9 vs. 2.5% ± 1.9, p < 0.001). Thus, no significant difference was observed between cases and controls in the subgroup analysis on females (6.1% ± 2.9 vs. 5.3% ± 3.4, p = 0.362). Among convalescent COVID-19 patients, FMD showed a direct correlation with arterial oxygen tension (rho = 0.247, p = 0.004), forced expiratory volume in 1 s (rho = 0.436, p < 0.001), forced vital capacity (rho = 0.406, p < 0.001), and diffusing capacity for carbon monoxide (rho = 0.280, p = 0.008). Overall, after adjusting for major confounders, a recent COVID-19 was a major and independent predictor of FMD values (β = −0.427, p < 0.001). Conclusions: Post-acute COVID-19 syndrome is associated with a persistent and sex-biased endothelial dysfunction, directly correlated with the severity of pulmonary impairment.

Abstract Background Small airways are considered the major site of airflow limitation in COPD. Impulse oscillometry (IOS) is a forced oscillation technique, which provides passive measurement of lung mechanics. It can differentiate small airway from large airway obstruction and is more sensitive than spirometry for peripheral airway disease. In this study the efficacy of the combination of Indacaterol/Glycopirronium (IND/GLY) versus Tiotropium on airway resistance (R5, R20, R5–20), lung reactance (X) and resonant frequency in moderate to severe COPD patients has been evaluated. We also evaluated inspiratory capacity (IC), forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC), exacerbation rate and quality of life. Methods Forty patients were monitored with forced oscillation technique and spirometry. Patients were randomized in 2 groups: 20 received fixed dose once daily Indacaterol/Glycopyrronium (Group A) and 20 received single Tiotropium (Group B). The oscillometry parameters were the measure of resistance in the airways at 5 Hz (R5), at 20 Hz (R20) and the lung reactance (X). Results There was a statistically significant difference between pre-dosing at V1 and at follow up visits in R5, R20 and X values in patients receiving dual bronchodilation but not in control group. Pre-dosing IC value at follow up visits in patients receiving dual bronchodilation had a statistical significant variation. Conclusions The “new” bronchodilator combination LABA/LAMA significantly reduces bronchial obstruction in small airways too. The oscillometry demonstrated greater sensitivity compared with spirometry for monitoring outcome measures of airway obstruction and the effect of long-term therapy.

Alpha-1 antitrypsin deficiency is a rare and often underdiagnosed hereditary disorder, which mainly affects the Caucasian population. We report a case of a noncystic fibrosis bronchiectasis patient in the absence of emphysema associated with low serum alpha-1-antitrypsin (AAT) level, in the absence of the most common defective alleles associated with AAT deficiency (PI*S and PI*Z) but with a new mutation in heterozygosis. This mutation is characterized by the substitution in the coding region of exon 3, of a guanine (G) for a thymine (T), generating the replacement of a glutamine (Gln) by a histidine (His) in codon 212 (cod 212 GlnCAG > HisCAT), corresponds to a new S allelic variant. This mutation, never identified before, is called S-Napoli.

Introduction: Pidotimod, a synthetic dipeptide molecule with biological and immunological activities, is used to reduce the number of exacerbations or pneumonitis in patients with inflammatory diseases.In the present study, we investigated whether Pidotimod modifies the metabolomic pathways measured in the exhaled breath condensate (EBC) of non-cystic fibrosis bronchiectatic patients (NCFB).Materials and Methods: We analyzed 40 adult patients affected by NCFB. They were randomly selected to receive Pidotimod 800 mg b/d for 21 consecutive days (3 weeks) per month for 6 months (20 patients, V1 group) or no drug (20 patients, V0 group), with a 1:1 criterion and then followed as outpatients.Results: EBC samples were collected from all patients at baseline and after 6 months. They were investigated by combined nuclear magnetic resonance (NMR) spectroscopy and multivariate statistical analysis to uncover metabolic differences between EBC from NCFB patients before and after therapy with Pidotimod. Pulmonary function test and pulmonary exacerbations were analyzed at baseline and at the end of Pidotimod therapy.The EBC metabolites were all identified, and through statistical evaluation, we were able to discriminate the two samples’ classes, with acetate, acetoin, lactate, and citrate as statistically significant discriminatory metabolites. The model vas validated by using a blind set of 20 NCFB samples, not included in the primary analysis.No differences were observed in PFT after 6 months. At the end of the study, there was a significant decrease of exacerbation rate in V1 group as compared with V0 group, with a substantial reduction of the number of mild or severe exacerbations (p < 0.001).Discussion: Pidotimod modifies the respiratory metabolic phenotype (“metabotype”) of NCFB patients and reduces the number of exacerbations.

Chronic obstructive pulmonary disease (COPD) is a lung disorder characterized by persistent respiratory symptoms and progressive airflow limitation as a consequence of a chronic inflammatory response. Corticosteroids are the main treatment for COPD patients with a history of exacerbation, in that they attenuate exacerbation and dyspnea, and improve the response to bronchodilators. Nevertheless, despite corticosteroid administration, COPD patients still undergo exacerbation phases. In this context, the aim of this study was to evaluate the activity of Absent in melanoma 2 (AIM2) inflammasome-dependent pathways under corticosteroid treatment during COPD exacerbation. Stable and exacerbated COPD-derived Peripheral Blood Mononuclear Cells (PBMCs) were treated with a well-known anti-inflammatory agent, Dexamethasone (DEX), in the presence or not of Poly (deoxyadenylic-deoxythymidylate) acid (Poly dA:dT), an AIM2 ligand. We found that IL-1α was highly increased when AIM2 was activated from Poly dA:dT in exacerbated, but not in stable, COPD-derived PBMCs. To note, the release of IL-1α after the stimulation of AIM2 in PBMCs obtained from stable (hospitalized) COPD patients was not higher from the basal conditions, though it was still as high as that observed for Poly dA:dT-stimulated PBMCs obtained from exacerbated patients. This effect was associated with a higher expression of AIM2 in pair-matched circulating CD14+ cells obtained from hospitalized patients who passed from the exacerbation to stable status. Because the difference between stable and exacerbated COPD patients relies on the treatment with corticosteroids, exacerbated and stable COPD-derived PBMCs were treated with DEX. Indeed, the release of IL-1α and TGF-β was not altered after DEX treatment. In conclusion, we found that the administration of DEX in vitro on exacerbated COPD-derived PBMCs was not able to revert the detrimental inflammatory mechanism associated with AIM2 activation responsible for the release of IL-1α and the ensuing TGF-β, contributing to the severity of disease.

In this work, a membrane-based separation was investigate for Scenedesmus almeriensis solutions dewatering. A commercial polyvinylidene fluoride (PVDF) membrane, having a pore size of 3 ?m was used in order to allow the water passage through it (permeate), retaining, at the same time, algae biomass (retentate). The possibility to reuse the permeate for a second Scenedesmus almeriensis growth step, was also studied. The registered data evidenced the feasibility of the membrane-based dewatering as an alternative competitive technology, even though the recovery of water should need further investigations to be optimized.

The request for natural products such as antioxidant pigments derived from microalgae, i.e. ßbeta-carotene, lutein and astaxanthin, is growing. In this context, astaxanthin, a powerful antioxidant produced by Haematococcus pluvialis, used as an additive in animal feed and as a food supplement, has been extracted by acceleratedor solvent extraction using acetone and ethanol as green and safe solvents, and hexane and chloroform:methanol (1:1) performing the best operative operating conditions. The obtained extracts showed not only the recovery of mainly astaxanthin but also other carotenoids, such as lutein and in lesser part of ßbeta-carotene. In addition, the composition of the extracts was analyzed by highlighting the content of other valuable bio-products such as proteins, carbohydrates, lipids and Total Dietary Fibers. The best extraction performance was finding found using acetone and ethanol as solvent.

The omega-3 group includes substances as eicosapentaenoic acid and docosapentaenoic acid that can be partially synthesized by human body and substances as alpha-linolenic acid that have to be necessarily introduced into the body from dietary intake. Omega-3 are important nutrients thanks to their anti-inflammatory properties and healthy properties in the reduction of cardiovascular diseases. The growth forecasts of omega-3 market will lead to an increase in the demand for EPA and DHA and therefore finding new potential sources, such as microalgae which are the first EPA and DHA producers in the marine environment, is very important. The aim of this work is to evaluate the feasibility of mechanical pretreatment on Nannochloropsis gaditana as source of EPA and at the same time evaluating the effect on the extraction yield of two solvents: a mixture of chloroform/methanol/water (Bligh & Dyer methods) and hexane that is GRAS (generally recognized as safe solvent) by using accelerated solvent extraction technology.

Anaerobic digestion is a technology used to biologically convert organic substrates into biogas in the absence of oxygen. The resulting biogas is a renewable energy source mainly consisting of a mixture of methane (60÷70% v/v) and carbon dioxide (30÷40% v/v), with traces of some minor compounds, such as H2S and NH3. Anaerobic digestion takes place through a sequence of four biological phases - hydrolysis, acidogenesis, acetogenesis, and methanogenesis - performed by the action of particular species of bacteria. Operating parameters such as temperature, pH, pressure and organic substrates govern the process and affect the starting biomass transformation and the content of methane into the biogas. The biogas from anaerobic digestion can be upgraded to biomethane by removing CO2 and the minor compounds. The techniques commonly used for this purpose, like pressure swing adsorption and membrane separation, are energy-intensive as they require the compression of biogas. In this paper, an innovative energy-saving approach for biogas production and its upgrading to biomethane is proposed. The concept is based on anaerobic digestion carried out at a pressure higher than the atmospheric one, called pressured anaerobic digestion (PAD), in order to directly produce high pressure biogas that can be upgraded to high pure biomethane (CH4 = 95% v/v) avoiding the compression phase during the upgrading. The variation of the main operating parameters has been simulated in order to investigate their effect on biomethane production and composition and to define the best operating conditions. The simulation of the process has been carried out by using Aspen Plus®.

Immobilization of living cells is a useful technique that allows to enhance the biotechnological properties of many organisms. Currently, cell-immobilization is obtained via carrier adsorption, self-aggregation and entrapment. We are currently investigating the possibility to immobilize microalgae cells using magnetic nanoparticles on a magnetic surface. The principle is based on the introduction, inside the cell-body of microalgae, superparamagnetic nanoparticles through electroporation. Once microalgae have absorbed enough magnetic nanoparticles they will be responding to an external magnetic field. By using a flat magnetic surface, it will be possible to trap them in a specific position by means of magnetic force. Importantly, immobilized cells will be covered with a thin layer of water and during cell division only one of the daughter cells will keep the magnetic nanoparticles since they will face permanently the magnetic surface. The first step towards this goal is the introduction of magnetic nanoparticles in the microalgae cells. This work will present preliminary results of magnetic cells preparation. Magnetic nanoparticles covered with lipid, to make them biocompatible, and electroporation method to inset nanoparticles in the cells have been used. The protocol to obtain magnetic cells and viability assay that proved the feasibility of the method will be described.

Chronic obstructive pulmonary disease (COPD) is now the fourth-leading cause of death worldwide and its prevalence is increasing. The progressive decline of lung function and airway remodelling are a consequence of chronic inflammatory responses. It was recently postulated the involvement of the inflammasome in COPD, although the underlying mechanism/s still need to be elucidated. Therefore, we isolated peripheral blood mononuclear cells (PBMCs) from exacerbated/unstable COPD patients. The stimulation of PBMCs with an AIM2 inflammasome activator, Poly dA:dT, led to IL-1α, but not IL-1β, release. The release of this cytokine was caspase-1- and caspase-4-dependent and correlated to higher levels of 8-OH-dG in COPD compared to non-smoker and smoker-derived PBMCs. Interestingly, AIM2-depedent IL-1α release was responsible for higher TGF-β levels, crucial mediator during pro-fibrotic processes associated to COPD progression. In conclusion, our data highlight the involvement of AIM2/caspase-1/caspase-4 in IL-1α-induced TGF-β release in unstable COPD-derived PBMCs, opening new therapeutic perspectives for unstable COPD patients.

Background: Growing evidence points to a key role of endothelial dysfunction in the pathogenesis of COVID-19. In this study, we evaluated changes in endothelium-dependent flow-mediated dilation (FMD) in a cohort of convalescent COVID-19 patients undergoing pulmonary rehabilitation (PR). Methods: After swab test negativization, convalescent COVID-19 patients referring to a post-acute care facility for PR were consecutively screened for inclusion. Study procedures were performed at the time of hospitalization and discharge. Results: We enrolled 82 convalescent COVID-19 patients (85.4% males, mean age 60.4 years). After PR, a significant improvement in most pulmonary function tests and exercise capacity was documented. FMD changed from 2.48% ± 2.01 to 4.24% ± 2.81 (p < 0.001), corresponding to a 70.9% increase. Significantly higher changes in FMD were found in patients without a history of vascular events as compared to those with (+2.04% ± 2.30 vs. +0.61% ± 1.83, p = 0.013). Values of forced expiratory volume in 1 s (FEV1%), forced vital capacity (FVC%) and diffusion capacity for carbon monoxide (DLCO%) significantly and directly correlated with FMD both at baseline and after PR. Patients with normal FEV1% (≥80% predicted) during the overall study period or those normalizing FEV1% after PR showed a more significant FMD change as compared to patients with persistently impaired FEV1% (p for trend = 0.029). This finding was confirmed in a multivariate analysis. Conclusions: Clinically evaluated endothelial function improves after PR in convalescent COVID-19 patients. A direct and persistent association between the severity of pulmonary and vascular disease can be hypothesized. Endothelial function testing may be useful in the follow-up of convalescent COVID-19 patients.

The aim of this study was to develop an effective integrated cultivation system for Haematococcus pluvialis as a source of bioactive compounds such as astaxanthin, lutein, proteins, and fatty acids (FAs). The Chlorophyta H. pluvialis was cultivated in a vertical bubble column photobioreactor (VBC-PBR) under batch mode, allowing switching from green to red phase for astaxanthin induction. The combined effect of light intensity and nutrients on bioactive compound formation was investigated. Results showed that growth under lower nutrients availability and light intensity led to a higher concentration of biomass. Growth under high light intensity with an appropriate concentration of nitrate, sulfate, phosphate and magnesium led to ~85% and ~58% higher production of total carotenoids and fatty acids, respectively. Under high stress conditions, ~90% nitrate and phosphate consumption were observed.

Several bacteria pathogens are responsible for plant diseases causing significant economic losses. The antibacterial activity of Dunaliella salina microalgae extracts were investigated in vitro and in vivo. First, biomass composition was chemically characterized and subjected to extraction using polar/non-polar solvents. The highest extraction yield was obtained using chloroform:methanol (1:1 v/v) equal to 170 mg g−1 followed by ethanol (88 mg g−1) and hexane (61 mg g−1). In vitro examination of hexane extracts of Dunaliella salina demonstrated antibacterial activity against all tested bacteria. The hexane extract showed the highest amount of β-carotene with respect to the others, so it was selected for subsequent analyses. In vivo studies were also carried out using hexane extracts of D. salina against Pseudomonas syringae pv. tomato and Pectobacterium carotovorum subsp. carotovorum on young tomato plants and fruits of tomato and zucchini, respectively. The treated young tomato plants exhibited a reduction of 65.7% incidence and 77.0% severity of bacterial speck spot disease. Similarly, a reduction of soft rot symptoms was observed in treated tomato and zucchini fruits with a disease incidence of 5.3% and 12.6% with respect to 90.6% and 100%, respectively, for the positive control.

Haematococcus pluvialis microalgae is a promising source of astaxanthin, an excellent antioxidant carotenoid. H. pluvialis, as well as other species, could find more extensive applications as healthy food for a variegated carotenoids composition in addition to astaxanthin. Official method has not currently been used for this purpose. The objective of this work was to propose a method to characterize carotenoids in H. pluvialis after the comparison between spectrophotometric and liquid chromatography analysis. In addition, in order to improve the use of astaxanthin in the food industry, thermal stability was investigated. In this context, the effect of temperature at 40–80 °C, over a 16 h storage period was tested on astaxanthin produced by H. pluvialis. A further test was carried out at room temperature (20 °C) for seven days. A decrease in the astaxanthin concentration was observed at all tested temperatures with a decrease >50% of all-trans isomer at 80 °C after 16 h and an increase of 9-cis and 13-cis isomers. In conclusion, the obtained results showed the importance of evaluating the degradation effect of temperature on astaxanthin used as a food additive for a future greater enhancement of this bioproduct in the food field.

Chronic obstructive pulmonary disease (COPD) is characterised by a progressive loss of pulmonary function. Often patients do not adhere to inhaled therapies and this leads clinicians to switch treatments in order to improve control of the symptoms. Narrative medicine is a useful approach that helps healthcare professionals to think over the doctor–patient relationship and how patients live with their disease. The aim of this training project was to teach pulmonologists the basics of narrative medicine: to carefully listen to patients and to practice reflective writing in their relationship with them. Training on narrative medicine and parallel charts was provided through a webinar and a weekly newsletter. Across 362 narratives, written by 74 Italian pulmonologists, 92% of patients had activity limitations at their first visit. The main factor influencing the effectiveness and adherence to therapy was a positive doctor–patient relationship; indeed, if such relationship is difficult, only 21% of patients are able to resume all their activities. After learning the narrative approach, clinicians became aware of the need to spend more time listening to patients, to reflect through writing and to understand more deeply the motivations that lead people towards adherence to new therapies.

Idiopathic pulmonary fibrosis (IPF) is a chronic fibro-proliferative disease characterized by poor prognosis, with a mean survival of ~2–3 years after definite diagnosis. The cause of IPF is still unknown but it is a heterogeneous condition in which the aberrant deposition of extracellular matrix leads to extensive lung remodeling. This remodeling is a consequence of inflammatory responses, but the mechanisms involved are poorly understood. In this study, we first analyzed a bleomycin-induced mouse model, which showed that higher expression of IL-1β, but not IL-18, was correlated to pulmonary cell infiltration and fibrosis. Then, we found that peripheral blood mononuclear cells (PBMCs) from IPF patients released IL-1α and IL-18 in a NLRP3- and calpain-independent manner after LPS ± ATP stimulation. Instead, the activation of the absent in melanoma 2 (AIM2) inflammasome induced the release of IL-1α in a caspase-1-/caspase-8-independent manner; whereas IL-18 release was caspase-1 dependent. These effects correlated with the release of the pro-fibrotic TGF-β, which was induced by AIM2 activation in a caspase-1- and TLR4-independent manner, but dependent on IL-1α. In this context, the activation of AIM2 induced the release of caspase-4 from IPF-derived PBMCs, which correlated with the mRNA levels of this caspase that was higher in IPF than in healthy PBMCs. In conclusion, our findings identify a novel molecular mechanism whereby the activation of AIM2 could lead to the activation of the non-canonical inflammasome (caspase-4 dependent) that induces the release of IL-1α responsible for the release of TGF-β from PBMCs of IPF patients.

Chronic obstructive pulmonary disease (COPD) is considered the fourth-leading causes of death worldwide; COPD is caused by inhalation of noxious indoor and outdoor particles, especially cigarette smoke that represents the first risk factor for this respiratory disorder. To mimic the effects of particulate matter on COPD, we isolated peripheral blood mononuclear cells (PBMCs) and treated them with combustion-generated ultrafine particles (UFPs) obtained from two different fuel mixtures, namely, pure ethylene and a mixture of ethylene and dimethylfuran (the latter mimicking the combustion of biofuels). UFPs were separated in two fractions: (1) sub-10 nm particles, named nano organic carbon (NOC) particles and (2) primarily soot particles of 20–40 nm and their agglomerates (200 nm). We found that both NOC and soot UFPs induced the release of IL-18 and IL-33 from unstable/exacerbated COPD-derived PBMCs. This effect was associated with higher levels of mitochondrial dysfunction and derived reactive oxygen species, which were higher in PBMCs from unstable COPD patients after combustion-generated UFP exposure. Moreover, lower mRNA expression of the repairing enzyme OGG1 was associated with the higher levels of 8-OH-dG compared with non-smoker and smokers. It was interesting that IL-18 and IL-33 release from PBMCs of unstable COPD patients was not NOD-like receptor 3/caspase-1 or caspase-8-dependent, but rather correlated to caspase-4 release. This effect was not evident in stable COPD-derived PBMCs. Our data suggest that combustion-generated UFPs induce the release of caspase-4-dependent inflammasome from PBMCs of COPD patients compared with healthy subjects, shedding new light into the biology of this key complex in COPD.

In this study, Scenedesmus almeriensis as green microalga was cultivated on bench-scale for carbon dioxide (CO2) capture and lutein production. The autotrophic cultivation of S. almeriensis was carried out by using a vertical bubble column photo-bioreactor (VBC-PBR) with a continuous flow of a gaseous mixture of oxygen (O2), nitrogen (N2), and CO2, the latter in content of 0.0–3.0 %v/v. The liquid phase was batch. S. almeriensis growth was optimized. In addition, lutein extraction was carried out by using accelerated solvent extraction with ethanol as Generally Recognized as Safe (GRAS) solvent at 67 °C and 10 MPa. Upon optimization of CO2 concentration, the maximum biomass productivity, equal to 129.24 mg·L−1·d−1, was achieved during the cultivation by using a content of CO2 equal to 3.0 %v/v and it allowed to obtain a lutein content of 8.54 mg·g−1, which was 5.6-fold higher in comparison to the analogous process carried out without CO2 addition. The ion chemical analysis in the growth medium showed that by gradually increasing CO2 content, the nutrient consumption during the growth phase also increased. This study may be of potential interest for lutein extraction at industrial scale, since it is focused on pigment production from a natural source with a concomitantly CO2 capture.

In this article, microalgae Nannochloropsis sp. was used for fatty acid (FA) extraction, using a supercritical fluid-carbon dioxide (SF-CO2) extraction method. This study investigated the influence of different pre-treatment conditions by varying the grinding speed (200−600 rpm), pre-treatment time (2.5−10 min), and mixing ratio of diatomaceous earth (DE) and Nannochloropsis sp. biomass (0.5−2.0 DE/biomass) on FAs extraction. In addition, the effect of different operating conditions, such as pressure (100−550 bar), temperature (50−75 °C), and CO2 flow rate (7.24 and 14.48 g/min) on eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) recovery, was analyzed. Experimental data evidenced that, keeping constant the extraction conditions, the pre-treatment step enhanced the FAs extraction yield up to 3.4 fold, thereby the maximum extracted amount of FAs (61.19 mg/g) was attained with the pre-treatment with a ratio of DE/biomass of 1 at 600 rpm for 5 min. Moreover, by increasing both SF-CO2 pressure and temperature, the selectivity towards EPA was enhanced, while intermediate pressure and lower pressure promoted DHA recovery. The highest amount of extracted EPA, i.e., 5.69 mg/g, corresponding to 15.59%, was obtained at 75 °C and 550 bar with a CO2 flow rate of 14.48 g/min, while the maximum amount of extracted DHA, i.e., ~0.12 mg/g, equal to 79.63%, was registered at 50 °C and 400 bar with a CO2 flow rate of 14.48 g/min. Moreover, the increased CO2 flow rate from 7.24 to 14.48 g/min enhanced both EPA and DHA recovery.

Lutein has several benefits for human health, playing an important role in the prevention of age-related macular degeneration (AMD), cataracts, amelioration of the first stages of atherosclerosis, and some types of cancer. In this work, the Scenedesmus almeriensis microalga was used as a natural source for the supercritical fluid (SF) extraction of lutein. For this purpose, the optimization of the main parameters affecting the extraction, such as biomass pre-treatment, temperature, pressure, and carbon dioxide (CO2) flow rate, was performed. In the first stage, the effect of mechanical pre-treatment (diatomaceous earth (DE) and biomass mixing in the range 0.25–1 DE/biomass; grinding speed varying between 0 and 600 rpm, and pre-treatment time changing from 2.5 to 10 min), was evaluated on lutein extraction efficiency. In the second stage, the influence of SF-CO2 extraction parameters such as pressure (25–55 MPa), temperature (50 and 65 °C), and CO2 flow rate (7.24 and 14.48 g/min) on lutein recovery and purity was investigated. The results demonstrated that by increasing temperature, pressure, and CO2 flow rate lutein recovery and purity were improved. The maximum lutein recovery (~98%) with purity of ~34% was achieved operating at 65 °C and 55 MPa with a CO2 flow rate of 14.48 g/min. Therefore, optimum conditions could be useful in food industries for lutein supplementation in food products.

Microalgae Dunaliella salina contains useful molecules such as β-carotene and fatty acids (FAs), which are considered high value-added compounds. To extract these molecules, supercritical carbon dioxide was used at different operative conditions. The effects of mechanical pre-treatment (grinding speed at 0⁻600 rpm; pre-treatment time of 2.5⁻7.5 min) and operating parameters for extraction, such as biomass loading (2.45 and 7.53 g), pressure (100⁻550 bars), temperature (50⁻75 °C) and CO2 flow rate (7.24 and 14.48 g/min) by varying the extraction times (30⁻110 min) were evaluated. Results showed that the maximum cumulative recovery (25.48%) of β-carotene was achieved at 400 bars and 65 °C with a CO2 flow rate of 14.48 g/min, while the highest purity for stage (55.40%) was attained at 550 bars and 65 °C with a CO2 flow rate of 14.48 g/min. The maximum recovery of FAs, equal to 8.47 mg/g, was achieved at 550 bars and 75 °C with a CO2 flow rate of 14.48 g/min. Moreover, the lowest biomass loading (2.45 g) and the first extraction cycle (30 min) allowed the maximum extraction of β-carotene and FAs.

This research shows that carbon dioxide supercritical fluid (CO2-SF) is an emerging technology for the extraction of high interest compounds for applications in the manufacturing of pharmaceuticals, nutraceuticals, and cosmetics from microalgae. The purpose of this study is to recover fatty acids (FAs) and, more precisely, eicosapentaenoic acid (EPA) from Nannochloropsis gaditana biomass by CO2-SF extraction. In the paper, the effect of mechanical pre-treatment was evaluated with the aim of increasing FAs recovery. Extraction was performed at a pressure range of 250⁻550 bars and a CO2 flow rate of 7.24 and 14.48 g/min, while temperature was fixed at 50 or 65 °C. The effect of these parameters on the extraction yield was assessed at each extraction cycle, 20 min each, for a total extraction time of 100 min. Furthermore, the effect of biomass loading on EPA recovery was evaluated. The highest EPA extraction yield, i.e., 11.50 mg/g, corresponding to 27.4% EPA recovery, was obtained at 65 °C and 250 bars with a CO2 flow rate of 7.24 g/min and 1.0 g biomass loading. The increased CO2 flow rate from 7.24 to 14.48 g/min enhanced the cumulative EPA recovery at 250 bars. The purity of EPA could be improved by biomass loading of 2.01 g, even if recovery was reduced.