High-Pressure Processing Effects on Microbiological Stability, Physicochemical Properties, and Volatile Profile of a Fruit Salad.

Nowadays, consumers are more aware of the effects of their diet on their health, and thus demand natural or minimally processed food products. Therefore, research has focused on processes that assure safe products without jeopardizing their nutritional properties. In this context, this work aimed to evaluate the effects of high-pressure processing (550 MPa/3 min/15 °C, HPP) on a fruit salad (composed of melon juice and pieces of Golden apple and Rocha pear) throughout 35 days of storage at 4 °C. For the physicochemical properties analysed (browning degree, polyphenol oxidase activity, antioxidant activity (ABTS assay), and volatile profile), a freshly made fruit salad was used, while for the microbiological tests (total aerobic mesophiles, and yeast and moulds) spoiled melon juice was added to the fruit salad to increase the microbial load and mimic a challenge test with a high initial microbial load. It was determined that processed samples were more microbiologically stable than raw samples, as HPP enabled a reduction of almost 4-log units of both total aerobic mesophiles and yeasts and moulds, as well as an almost 1.5-fold increase in titratable acidity of the unprocessed samples compared to HPP samples. Regarding browning degree, a significant increase (p < 0.05) was observed in processed versus unprocessed samples (roughly/maximum 68%), while the addition of ascorbic acid decreased the browning of the samples by 29%. For antioxidant activity, there were no significant differences between raw and processed samples during the 35 days of storage. An increase in the activity of polyphenol oxidase immediately after processing (about 150%) was confirmed, which was generally similar or higher during storage compared with the raw samples. Regarding the volatile profile of the product, it was seen that the compounds associated with melon represented the biggest relative percentage and processed samples revealed a decrease in the relative quantity of these compounds compared to unprocessed. Broadly speaking, HPP was shown to be efficient in maintaining the stability and overall quality of the product while assuring microbial safety (by inactivating purposely inoculated microorganisms), which allows for longer shelf life (7 versus 28 days for unprocessed and processed fruit salad, respectively).

The High Pressure Preservation of Honey: A Comparative Study on Quality Changes during Storage.

In commercially available honey, the application of a heat treatment to prevent spoilage can potentially compromise its beneficial properties and quality, and these effects worsen with extended storage. The high-pressure processing (HPP) of honey is being explored, but its long-term impact on honey quality has not been characterised yet. This study evaluated the effects of HPP and thermal processing on the microbial load, physicochemical quality (i.e., hydroxymethylfurfural content and diastase activity), and antioxidant capacity of honey after treatment and following extended storage (6, 12, and 24 months) at 20 °C. Pasteurization (78 °C/6 min) effectively eliminated the microorganisms in honey but compromised its physicochemical quality and antioxidant activity. HPP initially showed sublethal inactivation, but storage accelerated the decrease in yeasts/moulds and aerobic mesophiles in honey (being <1 log CFU/g after 24 months of storage) compared to unprocessed honey and honey thermally treated under mild conditions (55 °C/15 min). The physicochemical characteristics of the quality of HPP-treated honey and raw unprocessed honey did change after long-term storage (24 months) but remained within regulatory standards. In conclusion, HPP emerged as a more suitable and safe preservation method for Apis mellifera honey, with a minimal risk of a loss of antioxidant activity compared to traditional industrial honey pasteurization.

Blood Concentration of Macro- and Microelements in Women Who Are Overweight/Obesity and Their Associations with Serum Biochemistry.

Risk elements in blood matrices can affect human health status through associations with biomarkers at multiple levels. The aim of this study was to analyze 15 macro- and microelements in the blood serum of women with overweight (BMI of ≥25 kg/m2) and obesity (BMI of ≥30 kg/m2) and to examine possible associations with biochemical, liver enzymatic parameters, and markers of oxidative stress. Based on the power calculation, the study involved women (in the postmenopausal stage) with overweight (n = 26) and obesity (n = 22), aged between 50-65 years. Multifrequency bioelectrical impedance analysis was used to measure body composition parameters. Concentrations of elements were determined by inductively coupled plasma optical emission spectrometry, and Hg was measured using cold-vapor atomic absorption spectroscopy. Individuals with obesity, as indicated by a higher BMI, percentage of body fat, and visceral fat area, had elevated serum levels of Ca, Mg, Fe, Al, Sr, Pb, and Hg. Concentrations of Al, Cu, K, Sb, Zn, and Pb significantly affected biochemical and liver function markers in women with overweight or obesity. Elements such as Cu and Al were associated with increased total cholesterol. The correlation analysis between total antioxidant status and Cu, Al, and Ni confirmed associations in both groups. Our findings underscore the importance of addressing excess body weight and obesity in relation to risk elements. The results of the research could be beneficial in identifying potential targets for the treatment or prevention of comorbidities in people with obesity.

High-Hydrostatic-Pressure-Stabilized White Grape Pomace to Improve the Oxidative Stability of Dry-Cured Sausages ("Salchichón").

White grape pomace (winery by-product) stabilized by blanching and high hydrostatic pressure has recently been successful at delaying lipid oxidation in burgers. The aim of this study was to investigate whether it can also delay lipid oxidation in dry-cured sausages, and to compare its effect when added at 0.5 and 3% with those of synthetic additives (sodium nitrite and ascorbic acid) and no additives (Control) in lipid and protein oxidation, the instrumental color, the sensory characteristics, and the volatile compounds. The pomace (68.7 ± 7.4 mmol Trolox g-1) was as effective as the additives at preventing lipid oxidation, resulting in values 3.2-3.8 times lower than the Control sausages. However, the pomace was not effective at decreasing the microbial counts, improving the instrumental and sensory color and the volatile compound profile, and decreasing the off-odor and off-flavor developed in the Control sausages. The lack of a detrimental effect of the pomace at 0.5% on the volatile compounds and the sensory characteristics and its benefits to delay lipid oxidation suggest that it might be useful to improve the oxidative stability. Conversely, at 3%, with a detrimental effect on some sensory characteristics and no benefits over the lower dosage, is not advisable.

Improved triamcinolone acetonide-eluting contact lenses based on cyclodextrins and high hydrostatic pressure assisted complexation.

Contact lenses (CLs) constitute an advantageous platform for the topical release of corticosteroids due to their prolonged contact with the eye. However, the lipophilic nature of corticosteroids hampers CLs' ability to release therapeutic amounts. Two approaches to improve loading and release of triamcinolone acetonide (TA) from poly(2-hydroxyethyl methacrylate)-based hydrogels were investigated: adding 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) to the monomers solution before polymerization (HEMA/i-CD) and an hydrogels' post-treatment with HP-ß-CD (HEMA/p-CD). The effect of HP-ß-CD and sterilization by high hydrostatic pressure (HHP) on the hydrogel properties (water content, oxygen and ion permeability, roughness, transmittance, and stiffness) was evaluated. The HEMA/i-CD hydrogels had stronger affinity for TA, sustaining its release for one day. HHP sterilization promoted the formation of cyclodextrin-TA complexes within the hydrogels, improving their drug-loading capacity ¼60 %. Cytotoxicity and irritability tests confirmed the safety of the therapeutic CLs. TA released from the hydrogels permeated through ocular tissues ex vivo and showed anti-inflammatory activity. Finally, a previously validated mathematical model was used to estimate the ability of the TA-loaded CLs to deliver therapeutic drug concentrations to the posterior part of the eye. Overall, HP-ß-CD-containing CLs are promising candidates for the topical ocular application of TA as an alternative delivery system to intraocular injections.

Epigenetic reprogramming as a key to reverse ageing and increase longevity.

The pursuit for the fountain of youth has long been a fascination amongst scientists and humanity. Ageing is broadly characterized by a cellular decline with increased susceptibility to age-related diseases, being intimately associated with epigenetic modifications. Recently, reprogramming-induced rejuvenation strategies have begun to greatly alter longevity research not only to tackle age-related defects but also to possibly reverse the cellular ageing process. Hence, in this review, we highlight the major epigenetic changes during ageing and the state-of-art of the current emerging epigenetic reprogramming strategies leveraging on transcription factors. Notably, partial reprogramming enables the resetting of the ageing clock without erasing cellular identity. Promising chemical-based rejuvenation strategies harnessing small molecules, including DNA methyltransferase and histone deacetylase inhibitors are also discussed. Moreover, in parallel to longevity interventions, the foundations of epigenetic clocks for accurate ageing assessment and evaluation of reprogramming approaches are briefly presented. Going further, with such scientific breakthroughs, we are witnessing a rise in the longevity biotech industry aiming to extend the health span and ideally achieve human rejuvenation one day. In this context, we overview the main scenarios proposed for the future of the socio-economic and ethical challenges associated with such an emerging field. Ultimately, this review aims to inspire future research on interventions that promote healthy ageing for all.

Pachydysostosis of the fibula in a case of familial adenomatous polyposis.

BACKGROUND: Familial Adenomatous Polyposis (FAP) is a colorectal cancer (CRC) predisposition syndrome caused by germline APC mutations and characterised by an increased risk of CRC and colonic polyps and, in certain forms, of specific prominent extraintestinal manifestations, namely osteomas, soft tissue tumours and dental anomalies. Pachydysostosis of the fibula is a rare clinical entity defined by unilateral bowing of the distal portion of the fibula and elongation of the entire bone, without affectation of the tibia. CLINICAL REPORT: We report a 17-year-old male, who presented with a non-progressive bowing of the right leg detected at 18 months of age caused by a fibula malformation (later characterized as pachydysostosis) and a large exophytic osteoma of the left radius, noticed at the age of 15 years, without gastrointestinal symptoms. There was no relevant family history. Detailed characterisation revealed multiple osteomas, skin lesions and dental abnormalities, raising the hypothesis of FAP. This diagnosis was confirmed by genetic testing [c.4406_4409dup p.(Ala1471Serfs*17) de novo mutation in the APC gene] and endoscopic investigation (multiple adenomas throughout the colon, ileum and stomach). DISCUSSION: This case report draws attention to the phenotypic spectrum of skeletal manifestations of FAP: this patient has a congenital fibula malformation, not previously associated with this syndrome, but which is likely to have been its first manifestation in this patient. This clinical case also illustrates the challenges in the early diagnosis of FAP, especially without family history, and highlights the importance of a multidisciplinary approach and the adequate study of rare skeletal abnormalities.

Structure and properties of Quercus robur acorn starch extracted by pulsed electric field technology.

Pulsed electric field (PEF) technology was used to extract starch from Q. robur flours using low-intensity electric fields (0 and 0.1 kV/cm) and study the impact of PEF on the structure and properties of acorn starch concerning commercial starch. PEF technology is an advantageous method for starch extraction than the aqueous steeping from an industrial perspective since reduces extraction time and allows for continuous processing of larger suspension volumes. PEF technology preserved the amylose and amylopectin contents, hydrogen bonds, and diffraction patterns, as well as the starch native properties. Hence, PEF could be used to obtain native starches, but future studies should verify its economic viability. Acorn starches have lower damaged starch content, gelatinization temperatures, enthalpies, improved pseudoplastic behavior, reduced in-vitro digestibility, and lower resistance to deformation compared to commercial corn starch. The higher solubility and swelling power of acorn starches up to 80 °C make them a suitable food additive in fermented yogurt and milk products and thus help to value acorn and acorn starches. Hence, acorns can be used to obtain native starches, a food ingredient with a wide range of food and non-food usage, using PEF.

Genetic profile of syndromic retinitis pigmentosa in Portugal.

PURPOSE: Retinitis pigmentosa (RP) comprises a genetically and clinically heterogeneous group of inherited retinal degenerations, where 20-30% of patients exhibit extra-ocular manifestations (syndromic RP). Understanding the genetic profile of RP has important implications for disease prognosis and genetic counseling. This study aimed to characterize the genetic profile of syndromic RP in Portugal. METHODS: Multicenter, retrospective cohort study. Six Portuguese healthcare providers identified patients with a clinical diagnosis of syndromic RP and available genetic testing results. All patients had been previously subjected to a detailed ophthalmologic examination and clinically oriented genetic testing. Genetic variants were classified according to the American College of Medical Genetics and Genomics; only likely pathogenic or pathogenic variants were considered relevant for disease etiology. RESULTS: One hundred and twenty-two patients (53.3% males) from 100 families were included. Usher syndrome was the most frequent diagnosis (62.0%), followed by Bardet-Biedl (19.0%) and Senior-Løken syndromes (7.0%). Deleterious variants were identified in 86/100 families for a diagnostic yield of 86.0% (87.1% for Usher and 94.7% for Bardet-Biedl). A total of 81 genetic variants were identified in 25 different genes, 22 of which are novel. USH2A and MYO7A were responsible for most type II and type I Usher syndrome cases, respectively. BBS1 variants were the cause of Bardet-Biedl syndrome in 52.6% of families. Best-corrected visual acuity (BCVA) records were available at baseline and last visit for 99 patients (198 eyes), with a median follow-up of 62.0 months. The mean BCVA was 56.5 ETDRS letters at baseline (Snellen equivalent ~ 20/80), declining to 44.9 ETDRS letters (Snellen equivalent ~ 20/125) at the last available follow-up (p < 0.001). CONCLUSION: This is the first multicenter study depicting the genetic profile of syndromic RP in Portugal, thus contributing toward a better understanding of this heterogeneous disease group. Usher and Bardet-Biedl syndromes were found to be the most common types of syndromic RP in this large Portuguese cohort. A high diagnostic yield was obtained, highlighting current genetic testing capabilities in providing a molecular diagnosis to most affected individuals. This has major implications in determining disease-related prognosis and providing targeted genetic counseling for syndromic RP patients in Portugal.

Dexamethasone phosphate and penetratin co-eluting contact lenses: a strategy to enhance ocular drug permeability.

Contact lenses (CLs) have been suggested as drug delivery platforms capable of increasing the drug residence time on the cornea and therefore its bioavailability. However, when targeting the posterior segment of the eye, the drug released from CLs still encounters the barrier effect of the ocular tissues, which considerably reduces the efficacy of administration. This work aims at the development of CLs able to simultaneously deliver an anti-inflammatory drug (dexamethasone sodium phosphate) and a cell-penetrating peptide (penetratin), the latter acting as a drug carrier across the tissues. Hydroxyethyl methacrylate (HEMA)-based hydrogels were functionalized with acrylic acid (AAc) and/or aminopropyl methacrylamide (APMA) to serve as CL materials with increased affinity for the drug and peptide. APMA-functionalized hydrogels sustained the dual release for 8 h, which is compatible with the wearing time of daily CLs. Hydrogels demonstrated suitable light transmittance, swelling capacity and in vitro biocompatibility. The anti-inflammatory activity of the drug was not compromised by the presence of the peptide nor by sterilization. The ocular distribution of the drug after 6 h of CL wearing was evaluated in vivo in rabbits and revealed that the amount of drug in the cornea and aqueous humor significantly increased when the drug was co-delivered with penetratin.

Effect of high cyclic hydrostatic pressure on osteogenesis of mesenchymal stem cells cultured in liquefied micro-compartments.

Bone resident cells are constantly subjected to a range of distinct mechanical loadings, which generates a complex microenvironment. In particular, hydrostatic pressure (HP) has a key impact on modulation of cell function and fate determination. Although HP is a constant mechanical stimulus, its role in regulating the osteogenesis process within a defined 3D microenvironment has not been comprehensively elucidated. Perceiving how environmental factors regulate the differentiation of stem cells is essential for expanding their regenerative potential. Inspired by the mechanical environment of bone, this study attempted to investigate the influence of different ranges of cyclic HP on human adipose-derived mesenchymal stem cells (MSCs) encapsulated within a compartmentalized liquefied microenvironment. Taking advantage of the liquefied environment of microcapsules, MSCs were exposed to cyclic HP of 5 or 50 MPa, 3 times/week at 37 °C. Biological tests using fluorescence staining of F-actin filaments showed a noticeable improvement in cell-cell interactions and cellular network formation of MSCs. These observations were more pronounced in osteogenic (OST) condition, as confirmed by fluorescent staining of vinculin. More interestingly, there was a significant increase in alkaline phosphatase activity of MSCs exposed to 50 MPa magnitude of HP, even in the absence of osteoinductive factors. In addition, a greater staining area of both osteopontin and hydroxyapatite was detected in the 50 MPa/OST group. These findings highlight the benefit of hydrostatic pressure to regulate osteogenesis of MSCs as well as the importance of employing simultaneous biochemical and mechanical stimulation to accelerate the osteogenic potential of MSCs for biomedical purposes.

High Pressure and Pasteurization Effects on Dairy Cream.

Dairy cream, a common ingredient in various dishes and food products, is susceptible to rapid microbial growth due to its high water activity (≈0.97) and pH (≈6.7). Thus, it requires proper processing conditions to ensure food safety and extend shelf life. High-pressure processing (HPP) has emerged as a nonthermal food pasteurization method, offering an alternative to conventional heat-based techniques to obtain tastier, fresh-like, and safe dairy products without undesirable heat-induced alterations. This study assessed the impact of HPP (450 and 600 MPa for 5 and 15 min at 7 °C) and thermal pasteurization (75 °C for 15 s) on the microbiological and physicochemical attributes of dairy cream immediately after processing and throughout refrigerated storage (4 °C). HPP-treated samples remained microbiologically acceptable even on the 51st day of storage, unlike thermally pasteurized samples. Moreover, HPP decreased inoculated Escherichia coli and Listeria innocua counts by more than 6 log units to undetectable levels (1.00 log CFU/mL). pH, color (maximum variation of ΔE* up to 8.43), and fatty acid profiles remained relatively stable under varying processing conditions and during storage. However, viscosity exhibited higher values for HPP-treated samples (0.028 ± 0.003 Pa·s) compared to thermally processed ones (0.016 ± 0.002 Pa·s) by the 28th day of storage. Furthermore, volatile compounds (VOCs) of all treated samples presented a tendency to increase throughout storage, particularly acids and aliphatic hydrocarbons. These findings show HPP's potential to significantly extend the shelf life of highly perishable dairy cream by at least 15 days compared to thermal pasteurization.

Modification of Acorn Starch Structure and Properties by High Hydrostatic Pressure.

Despite being rich in starch, over half of acorn production is undervalued. High hydrostatic pressure was used to modify the properties of Q. pyrenaica (0.1 and 460 MPa for 20 min) and Q. robur (0.1 and 333 MPa for 17.4 min) acorn starches to obtain high-valued ingredients. Pressure significantly altered the span distribution and heterogeneity of the acorn starch granules depending on the species, but their morphology was unaffected. Pressurization increased the amylose/amylopectin ratio and damaged starch contents, but the effect was more prominent in Q. pyrenaica than in Q. robur. However, the polymorphism, relative crystallinity, gelatinization temperatures, and enthalpies were preserved. The pressure effect on the starch properties depended on the property and species. The solubility, swelling power, and acorn gels' resistance towards deformation for both species decreased after pressurization. For Q. pyrenaica starch, the in vitro digestibility increased, but the pseudoplastic behavior decreased after pressurization. No differences were seen for Q. robur. Regarding the commercial starch, acorn starches had lower gelatinization temperatures and enthalpies, lower in vitro digestibility, lower resistance towards deformation, superior pseudoplastic behavior, and overall higher solubility and swelling power until 80 °C. This encourages the usage of acorn starches as a new food ingredient.

The Impact of HPP-Assisted Biocontrol Approach on the Bacterial Communities' Dynamics and Quality Parameters of a Fermented Meat Sausage Model.

Traditional foods are increasingly valued by consumers, whose attention and purchase willingness are highly influenced by other claims such as 'natural', 'sustainable', and 'clean label'. The purpose of the present study was to evaluate the impact of a novel non-thermal food processing method (i.e., HPP-assisted biocontrol combining mild high hydrostatic pressure, listeriophage Listex, and pediocin PA-1 producing Pediococcus acidilactici) on the succession of bacterial communities and quality of a fermented sausage model. A comparative analysis of instrumental color, texture, and lipid peroxidation revealed no significant differences (p > 0.05) in these quality parameters between non- and minimally processed fermented sausages throughout 60-day refrigerated storage (4 °C). The microbiota dynamics of biotreated and untreated fermented sausages were assessed by 16S rRNA next-generation sequencing, and the alpha and beta diversity analyses revealed no dissimilarity in the structure and composition of the bacterial communities over the analyzed period. The innovative multi-hurdle technology proposed herein holds valuable potential for the manufacture of traditional fermented sausages while preserving their unique intrinsic characteristics.

Effects of Sequential Combination of Moderate Pressure and Ultrasound on Subsequent Thermal Pasteurization of Liquid Whole Egg.

As an alternative to commercial whole egg thermal pasteurization (TP), the sequential combination of moderate pressure (MP) and/or ultrasound (US) pre-treatments prior to a shorter TP was evaluated. The use of US alone or in combination with MP or TP resulted in an inactivation that was far from that of commercial TP. Nevertheless, when these three technologies were combined (MP-US-TP, 160 MPa/5 min-50% amplitude/1 min-60 °C/1.75 min), a safety level comparable to that of commercial TP was established. This was likely due to a decrease in the thermal resistance of Salmonella Senftenberg 775/W caused by MP and US pre-treatments. Regarding liquid whole egg (LWE) properties, using raw LWE as a reference, TP and MP treatments each decreased protein solubility (7-12%), which was accompanied by a viscosity increment (41-59%), whereas the US-only and MP-US-TP treatments improved protein solubility (about 4%) and reduced viscosity (about 34%). On average, all treatments lowered the emulsifying properties of LWE by 35-63%, with the MP-US-TP treatment having a more dramatic impact than commercial TP. In addition, the US-only, MP-only, and MP-US-TP treatments had the greatest impact on the volatile profile of LWE, lowering the concentration of the total volatile components. In comparison to commercial TP, LWE treated with MP-US-TP exhibited greater protein solubility (19%), lower viscosity (56%), and comparable emulsifying stability, but with a decreased emulsifying capacity (39%) and a lower total volatile compounds content (77%). Considering that a combined treatment (MP-US-TP) is lethally equivalent to commercial TP, but the latter better retained the quality properties of raw LWE, including volatiles, the application of MP followed by US pre-treatments before a shorter TP did not demonstrate significant advantages on quality parameters in comparison to commercial TP.

Impact of Fermentation and Pasteurization on the Physico-Chemical and Phytochemical Composition of Opuntia ficus-indica Juices.

Opuntia ficus-indica fruits are a source of valuable compounds, presenting a high nutritional value and several health benefits. However, due to its low shelf life and increased production, there are considerable post-harvest losses of this cactus fruit. So, ways need to be found to drain the increased production of this fruit that is being wasted. The chemical composition of prickly pear makes it an appealing substrate for fermentation. This study investigates the production of fermented beverages produced from Opuntia ficus-indica cv 'Rossa' and evaluates the effects of different fermentation times (18 and 42 h) and post-fermentation pasteurization by high-pressure (500 MPa for 10 min) and temperature (71.1 °C for 30 s) on the physico-chemical and biological characteristics of the produced beverages. According to the results, the beverage produced from 48 h of fermentation has an alcohol content value of 4.90 ± 0.08% (v/v) and a pH of 3.91 ± 0.03. These values contribute to an extended shelf life and improved organoleptic characteristics compared to the sample fermented for 18 h. Additionally, the longer fermentation resulted in 50% fewer total soluble solids, 90% less turbidity, and lower pH when compared to the sample fermented for 18 h. Moreover, overall, high-pressure processing demonstrates better retention of "fresh-like" characteristics, along with higher levels of phytochemical compounds and antioxidant capacity, similar to those observed in the juice for SO•- and NO•-scavenging abilities.

A chemical study of yoghurt produced under isostatic pressure during storage.

Yoghurt fermented under sub-lethal high pressure (10, 20, 30 and 40 MPa at 43 °C), and afterward placed under refrigeration (4 °C for 23 days) was studied and compared with yoghurt fermented at atmospheric pressure (0.1 MPa). For a deeper analysis, metabolite fingerprinting by nuclear magnetic resonance (NMR), sugars and organic acids assessment by high performance liquid chromatography (HPLC), total fatty acids (TFA) determination and quantification by gas chromatography with a flame ionization detector (GC-FID) were performed. Metabolomic analyses revealed that only 2,3-butanediol, acetoin, diacetyl and formate vary with the increase of pressure and probable relation with pressure influenced diacetyl reductase, acetoin reductase and acetolactate decarboxylase. Yoghurts fermented at 40 MPa had the lowest content in lactose (39.7 % of total sugar reduction) and the less content in TFA (56.1 %). Further research is of interest to understand more about fermentation processes under sub-lethal high pressure.

Combining high pressure and electric fields towards Nannochloropsis oculata eicosapentaenoic acid-rich extracts.

Nannochloropsis oculata is naturally rich in eicosapentaenoic acid (EPA). To turn this microalga into an economically viable source for commercial applications, extraction efficiency must be achieved. Pursuing this goal, emerging technologies such as high hydrostatic pressure (HHP) and moderate electric fields (MEF) were tested, aiming to increase EPA accessibility and subsequent extraction yields. The innovative approach used in this study combined these technologies and associated tailored, less hazardous different solvent mixtures (SM) with distinct polarity indexes. Although the classical Folch SM with chloroform: methanol (PI 4.4) provided the highest yield concerning total lipids (166.4 mglipid/gbiomass), diethyl ether: ethanol (PI 3.6) presented statistically higher values in terms of EPA per biomass, corresponding to 1.3-fold increase. When SM were used in HHP and MEF, neither technology independently improved EPA extraction yields, although the sequential combination of technologies did result in 62% increment in EPA extraction. Overall, the SM and extraction methodologies tested (HHP-200 MPa, 21 °C, 15 min, followed by MEF processing at 40 °C, 15 min) enabled increased EPA extraction yields from wet N. oculata biomass. These findings are of high relevance for the food and pharmaceutical industries, providing viable alternatives to the "classical" extraction methodologies and solvents, with increased yields and lower environmental impact. KEY POINTS: • Et2O: EtOH is a less toxic and more efficient alternative to Folch solvent mixture • HHP or MEF per se was not able to significantly increase EPA extraction yield • Combinations of HHP and MEF technologies increased both lipids and EPA yields.

Comparing Different Packaging Conditions on Quality Stability of High-Pressure Treated Serra da Estrela Cheeses during Cold Storage.

Serra da Estrela cheese with a Protected Designation of Origin (PDO) is a traditional cheese that is wrapped in paper without vacuum. High-pressure processing (HPP), which requires vacuum packaging of the cheese, has been used for its cold pasteurization to overcome safety issues. In this study, two packaging systems were studied: non-vacuum greaseproof paper wrapping package and vacuum packaging in plastic film. Lactococci, lactobacilli, enterococci, and total mesophiles reached ca. 8 log cfu g-1 and 4-6 log cfu g-1 in control (unpasteurized) and HPP-treated cheeses, respectively, with no significant differences between packaging systems. Spoilage microorganisms' viable cell numbers were reduced to <3 log cfu g-1 (quantification limit) in HPP-treated cheeses, independently of the packaging system. Yeasts and molds reached >5 log cfu g-1 in non-vacuum paper-wrapped cheeses. A vacuum-packaging system enabled better control of cheese proteolysis, which was revealed to be closer to that of the original control cheese values at the end of the 10-month storage period. In addition, cheese stored under vacuum film packaging became harder than non-vacuum paper-wrapped cheeses at each time point. Overall, conventional non-vacuum paper wrapping is adequate for short storage periods (<3 months), but for long periods vacuum packaging in plastic film is preferable.

High Hydrostatic Pressure in the Modulation of Enzymatic and Organocatalysis and Life under Pressure: A Review.

The interest in high hydrostatic pressure (HHP) is mostly focused on the inactivation of deleterious enzymes, considering the quality-related issues associated with enzymes in foods. However, more recently, HHP has been increasingly studied for several biotechnological applications, including the possibility of carrying out enzyme-catalyzed reactions under high pressure. This review aims to comprehensively present and discuss the effects of HHP on the kinetic catalytic action of enzymes and the equilibrium of the reaction when enzymatic reactions take place under pressure. Each enzyme can respond differently to high pressure, mainly depending on the pressure range and temperature applied. In some cases, the enzymatic reaction remains significantly active at high pressure and temperature, while at ambient pressure it is already inactivated or possesses minor activity. Furthermore, the effect of temperature and pressure on the enzymatic activity indicated a faster decrease in activity when elevated pressure is applied. For most cases, the product concentration at equilibrium under pressure increased; however, in some cases, hydrolysis was preferred over synthesis when pressure increased. The compiled evidence of the effect of high pressure on enzymatic activity indicates that pressure is an effective reaction parameter and that its application for enzyme catalysis is promising.