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Objective: To determine the sensitivity of electrical impedance spectroscopy (EIS) measurements for the detection of a small concentration of breast cancer cells in suspension, previously labeled with magnetic nanoparticles (MNPs) and separated. Additionally, the relation of electrical impedance to the expression of molecular markers was established. Approach: MCF-7, MDA-MB-231 and SK-BR-3 breast cancer cell lines at different concentrations (50, 500 and 5000 cells/500 μl) were labeled with a magnetic nanoparticle–antibody (MNP-ab) bioconjugate that recognizes the corresponding molecular markers (EpCAM, MUC-1 and HER-2, respectively). Electrical bioimpedance spectra (100 Hz to 1 MHz) were recorded in each case. Main results: At the frequency centered at 100 KHz, EIS displayed a greater sensitivity for magnitude when using 50 cells/500 μl (4.11 ± 0.23, 8.81 ± 1.73 and 17.5 ± 3.61 ohms for MCF-7, MDA-MB-231 and SK-BR-3, respectively). There were no significant differences between the phases of impedance tested at the greatest sensitivity of 1 KHz (−78.05 ± 0.53, −79.23 ± 0.93 and −75.26 ± 0.72 degrees for MCF-7, MDA-MB-231 and SK-BR-3, respectively). Significance: Under the present conditions, EIS was sensitive enough to detect a low concentration of breast cancer cells (50 cells/500 μl) and classify the distinct cells lines with a signature impedance pattern, as long as the MNP concentration was very low (⩽0.125 mg MNPs/50 000 cells). The relative expression of the molecular markers on each cancer cell line was related to the magnitude of the electrical impedance. [ABSTRACT FROM AUTHOR]

In spite of the natural harsh marine environments and continuous global change stressors affecting the Levant basin, the Israeli marine flora in the Eastern Mediterranean Sea is quite diverse, with about 300 recognized species. Such high seaweed biodiversity for a small maritime area is remarkable compared to the ca. 1200 species described for the entire Eastern Mediterranean Sea. Since about the year 1890, the Levant basin has been hosting over 115 seaweeds species that migrated from the Indo-Pacific through the Suez Canal. Indeed, approximately 16% of the marine flora is regarded as invasive or exotic to the Israeli shores, in a process that constantly reshapes seaweed populations and their biodiversity. In spite of significant contributions by Israeli scientists to the general biology and technologies for seaweed cultivation worldwide, Israel has little historical and cultural tradition of commercial seaweed cultivation, or use. At present, only two commercial companies are engaged in land-based seaweed cultivation (Ulva sp. and Gracilaria sp.) with a number of products marketed locally and abroad. Recently, offshore cultivation and biorefinery approaches have been explored, but not yet commercialized. [ABSTRACT FROM AUTHOR]

To foster the development of macroalgal biomass for biorefinery applications, we tested two orthogonal techniques for rapid phenotyping of the green macroalga Ulva based on its glucose, rhamnose, xylose and glucuronic acid contents as derived for reference by acid hydrolysis. Partial Least Squares (PLS) regression analyses, calculation of slopes and correlations across different spectral ranges/frequencies were used to predict the monosaccharide contents using two complementary methods: near infrared reflection spectroscopy (NIRS) and microelectromechanical systems (MEMS) resonating membrane vibrometry. Both methods were found to perform sufficiently well in monosaccharide mixtures and to enable quantitative assessment of different monosaccharide contents with the relative Root Mean Square Error of Prediction (%RMSEP) ranging from 8 to 16% (with similar accuracy when using PLS analyses). The best estimation was found for rhamnose and glucose contents, whereas xylose and uronic acid content predictions were found to be less accurate using PLS analyses. For the two latter components, slopes across different spectral ranges and frequencies at certain signals provided better estimates for their concentrations (e.g. for NIRS slopes: R 2 values in the range 0.55–0.66 and with higher accuracy for MEMS: between 0.75 and 0.90). This result is pivotal for opening new perspective to the construction of simple, multi-functional sensors for biomass downstream processing control in biorefinery and biometric applications. [ABSTRACT FROM AUTHOR]

Wound healing is an incredibly complex biological process that often results in thickened collagen-enriched healed tissue called scar. Cutaneous scars lack many functional structures of the skin such as hair follicles, sweat glands, and papillae. The absence of these structures contributes to a number of the long-term morbidities of wound healing, including loss of function for tissues, increased risk of re-injury, and aesthetic complications. Scar formation is a pervasive factor in our daily lives; however, in the case of serious traumatic injury, scars can create long-lasting complications due to contraction and poor tissue remodeling. Within this report we target the expression of connective tissue growth factor (CTGF), a key mediator of TGFβ pro-fibrotic response in cutaneous wound healing, with controlled local delivery of RNA interference. Through this work we describe both a thorough in vitro analysis of nanolayer coated sutures for the controlled delivery of siRNA and its application to improve scar outcomes in a third-degree burn induced scar model in rats. We demonstrate that the knockdown of CTGF significantly altered the local expression of αSMA, TIMP1, and Col1a1, which are known to play roles in scar formation. The knockdown of CTGF within the healing burn wounds resulted in improved tissue remodeling, reduced scar contraction, and the regeneration of papillary structures within the healing tissue. This work adds support to a number of previous reports that indicate CTGF as a potential therapeutic target for fibrosis. Additionally, we believe that the controlled local delivery of siRNA from ultrathin polymer coatings described within this work is a promising approach in RNA interference that could be applied in developing improved cancer therapies, regenerative medicine, and fundamental scientific research. [ABSTRACT FROM AUTHOR]

Fossil resources-free sustainable development can be achieved through a transition to bioeconomy, an economy based on sustainable biomass-derived food, feed, chemicals, materials, and fuels. However, the transition to bioeconomy requires development of new energy-efficient technologies and processes to manipulate biomass feed stocks and their conversion into useful products, a collective term for which is biorefinery. One of the technological platforms that will enable various pathways of biomass conversion is based on pulsed electric fields applications (PEF). Energy efficiency of PEF treatment is achieved by specific increase of cell membrane permeability, a phenomenon known as membrane electroporation. Here, we review the opportunities that PEF and electroporation provide for the development of sustainable biorefineries. We describe the use of PEF treatment in biomass engineering, drying, deconstruction, extraction of phytochemicals, improvement of fermentations, and biogas production. These applications show the potential of PEF and consequent membrane electroporation to enable the bioeconomy and sustainable development. [ABSTRACT FROM AUTHOR]

Exergy analysis methods place a major emphasis on technological and economical optimization of energy systems but have limited ability to address their environmental impacts. Although several approaches, such as Extended Exergy Accounting and Exergonomics, have been proposed to include pollution costs in the energy system optimization, environmental impacts of energy systems are much more complex, as energy systems affect the entire eco-systems services in the areas they are installed. Therefore, energy systems should minimize the adverse impacts on the surrounding eco-system services. In this paper we introduce a method for analysis and optimization of energy systems using technological, economic, and environmental exergy flows. For environmental exergy flow analysis, we included the eco-exergy component in the system objective function. Eco-exergy provides a Second Law derived measurement of the eco-system ability to do work. The disruption of eco-system services, such as reduction of biodiversity, is reflected in a reduction of eco-exergy. Therefore, we propose to include the minimization of the eco-exergy losses in the sustainable energy systems optimization. To demonstrate the method, we analyzed the eco-exergy losses associated with the reduction of the birds' biomass in the EU (European Union) fields dedicated to renewable energy systems. Specifically, we show that the EU renewable energy installations in 2008 lead to addition, today unaccountable, 5 PJ exergetic losses associated with the reduction in the biodiversity and biomass. [ABSTRACT FROM AUTHOR]

When high-amplitude, short-duration pulsed electric fields are applied to cells and tissues, the permeability of the cell membranes and tissue is increased. This increase in permeability is currently explained by the temporary appearance of aqueous pores within the cell membrane, a phenomenon termed electroporation. During the past four decades, advances in fundamental and experimental electroporation research have allowed for the translation of electroporation-based technologies to the clinic. In this review, we describe the theory and current applications of electroporation in medicine and then discuss current challenges in electroporation research and barriers to a more extensive spread of these clinical applications. [ABSTRACT FROM AUTHOR]

The profile of regional distribution of chemicals in tissue is important for the development of novel locally applied therapies. Here we introduce an analytical approach for the temporal and spatial regional distribution of chemicals in living tissues. The new aspect of this model is in local chemical concentration dependence on the tissue perfusion rate, in addition to diffusion and metabolism rates. Using the Duhamel theorem we performed dimensionless and dimensional analysis for local chemical distribution and report on steady-state and transient solutions for tissues with the first-order kinetic rate of clearance by perfusion and metabolism. The predictions of our model are in good correlation with clinically observed data on percutaneous ethanol ablation of liver. Our results emphasize the importance of mathematical modeling that includes perfusion in the planning of therapies with local chemical distribution in living tissues. [ABSTRACT FROM AUTHOR]

We report an all-in-one platform – ScanDrop – for the rapid and specific capture, detection, and identification of bacteria in drinking water. The ScanDrop platform integrates droplet microfluidics, a portable imaging system, and cloud-based control software and data storage. The cloud-based control software and data storage enables robotic image acquisition, remote image processing, and rapid data sharing. These features form a “cloud” network for water quality monitoring. We have demonstrated the capability of ScanDrop to perform water quality monitoring via the detection of an indicator coliform bacterium, Escherichia coli, in drinking water contaminated with feces. Magnetic beads conjugated with antibodies to E. coli antigen were used to selectively capture and isolate specific bacteria from water samples. The bead-captured bacteria were co-encapsulated in pico-liter droplets with fluorescently-labeled anti-E. coli antibodies, and imaged with an automated custom designed fluorescence microscope. The entire water quality diagnostic process required 8 hours from sample collection to online-accessible results compared with 2–4 days for other currently available standard detection methods. [ABSTRACT FROM AUTHOR]

While single cell heterogeneity is present in all biological systems, most studies cannot address it due to technical limitations. Here we describe a nano-liter droplet microfluidic-based approach for stimulation and monitoring of surfaceand secreted markers of live single immune dendritic cells (DCs) as well as monitoring the live T cell/DC interaction. This nano-liter in vivo simulating microenvironment allows delivering various stimuli reagents to each cell and appropriate gas exchanges which are necessary to ensure functionality and viability of encapsulated cells. Labeling bioassay and microsphere sensors were integrated into nano-liter reaction volume of the droplet to monitor live single cell surface markers and secretion analysis in the time-dependent fashion. Thus live cell stimulation, secretion and surface monitoring can be obtained simultaneously in distinct microenvironment, which previously was possible using complicated and multi-step in vitro and in vivo live-cell microscopy, together with immunological studies of the outcome secretion of cellular function. [ABSTRACT FROM AUTHOR]

We have developed a sensitive, specific, rapid and low cost picoliter microsphere-based platform for bioanalyte detection and quantification. In this method, a biological sample, biosensing microspheres, and fluorescently labeled detection (secondary) antibodies are co-encapsulated to capture the analyte (here: human anti-tetanus immunoglobulin G) on the surface of the microsphere in microfluidic pL-sized droplets. The absorption of the analyte and detecting antibodies on the microsphere concentrate the fluorescent signal in correlation with analyte concentration. Using our platform and commercially available antibodies, we were able to quantify anti-tetanus antibodies in human serum. In comparison to standard bulk immunosorbent assays, the microfluidic droplet platform presented here reduces the reagent volume by four orders of magnitude, while fast reagent mixing reduces the detection time from hours to minutes. We consider this platform to be a major leap forward in the miniaturization of immunosorbent assays and to provide a rapid and low cost tool for global point-of-care. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Tissue ablation is an essential procedure for the treatment of many diseases. In the last decade, a nonthermal tissue ablation using intensive pulsed electric fields, called nonthermal irreversible electroporation (NTIRE), has rapidly emerged. The exact mechanisms responsible for cell death by NTIRE, however, are currently unknown. Nevertheless, the technique's remarkable ability to ablate tissue in the proximity of larger blood vessels, to preserve tissue architecture, short procedure duration, and shortened postoperative recovery period rapidly moved NTIRE from bench to bed side. This work provides an overview on the development of NTIRE, its current state-of-the-art, challenges, and future needs. [ABSTRACT FROM PUBLISHER]

• Solar-thermal seaweed biorefinery with offshore intensively grown biomass was analysed. • Co-production of protein, hydrochar, ethanol, distilled water and electricity was analysed. • The overall First Law efficiency is 32% and can reach 40% with energy recovery from aqueous phase. • Seaweed, electricity and protein prices are key for the profitability of the biorefinery. • The lowest environmental impacts are achieved with HDPV cage, PV electricity, and rail transport. Combing solar energy with biomass processing facilities are emerging systems for efficient use of solar energy for electricity generation, energy storage, and production of renewable materials. In this work, we propose a novel combination of solar thermal energy systems with marine macroalgae biorefinery, which requires energy inputs for biomass cultivation and processing. In this work, we analyze a 10-ton dry weight per hour capacity pilot-scale hybrid solar seaweed biorefinery, located at the Mishor Rotem near Dimona, the current location for solar-thermal projects in Israel, where seaweed biomass supply comes from a hypothetical offshore farm located 15 km offshore. Our energy and mass balance analysis show that the overall First Law efficiency of the hybrid solar seaweed biorefinery system for the co-production of protein, hydrochar, ethanol, distilled water, and electricity is 32% and can exceed 40% with additional waste stream recycling. Our cost-benefit analysis of the proposed solar-seaweed biorefinery shows that the prices of seaweed, electricity, and protein are the key drivers of the profitability of the production process. The environmental impacts of the hybrid solar-seaweed biorefinery with intensified offshore cultivated biomass were quantified under various seaweed cultivation, transportation, and processing strategies. [ABSTRACT FROM AUTHOR]

Standard approaches to evaluate scar formation within histological sections rely on qualitative evaluations and scoring, which limits our understanding of the remodelling process. We have recently developed an image analysis technique for the rapid quantification of fibre alignment at each pixel location. The goal of this study was to evaluate its application for quantitatively mapping scar formation in histological sections of cutaneous burns. To this end, we utilized directional statistics to define maps of fibre density and directional variance from Masson's trichrome-stained sections for quantifying changes in collagen organization during scar remodelling. Significant increases in collagen fibre density are detectable soon after burn injury in a rat model. Decreased fibre directional variance in the scar was also detectable between 3 weeks and 6 months after injury, indicating increasing fibre alignment. This automated analysis of fibre organization can provide objective surrogate endpoints for evaluating cutaneous wound repair and regeneration. [ABSTRACT FROM AUTHOR]

Clinical misdiagnosis between cutaneous squamous cell carcinoma (cSCC) and basal cell carcinoma (BCC) poses treatment challenges and carries risks of recurrence, metastases and increased morbidity and mortality. We aimed to identify discriminant proteins markers for cSCC and BCC using a minimally invasive proteome sampling method called e‐biopsy, employing electroporation for non‐thermal cell permeabilization and machine learning. E‐biopsy facilitated ex vivo proteome extraction from 21 cSCC and 21 BCC pathologically validated human cancers. LC/MS/MS profiling of 126 proteomes was followed by machine learning analysis to identify proteins distinguishing cSCC from BCC. For identified panel validation, we used proteomes sampled by e‐biopsy from unrelated 20 cSCC and 46 BCC human cancers, and differential expression analysis of published transcriptomics. Cornulin, the most commonly chosen discriminative biomarker by machine learning models, was also validated using fluorescent immunohistochemistry. One hundred and ninety‐two proteomes sampled from one hundred eight patients were analysed. Machine learning‐based approaches resulted in a set of 11 potential biomarker proteins that can be used to construct a patient classification model with 95.2% average cross‐validation accuracy, BCC precision of 93.6 ± 14.5%, cSCC precision of 98.4 ± 7.2%, specificity of 97.7 ± 11.8% and sensitivity 92.7 ± 15.3%. Protein–protein interaction analysis revealed a novel interaction network connecting 10 of the 11 resulted proteins. Histological and transcriptomic validation confirmed cornulin as a discriminant marker significantly lower in cSCC than in BCC. E‐biopsy combined with machine learning provides a novel approach to molecular sampling from skin for biomarker detection and differential expression analysis between cSCC and BCC. [ABSTRACT FROM AUTHOR]

Background: The incidence rates of cutaneous squamous cell carcinoma (cSCC) and basal cell carcinoma (BCC) skin cancers are rising, while the current diagnostic process is time‐consuming. We describe the development of a novel approach to high‐throughput sampling of tissue lipids using electroporation‐based biopsy, termed e‐biopsy. We report on the ability of the e‐biopsy technique to harvest large amounts of lipids from human skin samples. Materials and Methods: Here, 168 lipids were reliably identified from 12 patients providing a total of 13 samples. The extracted lipids were profiled with ultra‐performance liquid chromatography and tandem mass spectrometry (UPLC‐MS‐MS) providing cSCC, BCC, and healthy skin lipidomic profiles. Results: Comparative analysis identified 27 differentially expressed lipids (p < 0.05). The general profile trend is low diglycerides in both cSCC and BCC, high phospholipids in BCC, and high lyso‐phospholipids in cSCC compared to healthy skin tissue samples. Conclusion: The results contribute to the growing body of knowledge that can potentially lead to novel insights into these skin cancers and demonstrate the potential of the e‐biopsy technique for the analysis of lipidomic profiles of human skin tissues. [ABSTRACT FROM AUTHOR]

Recent developments in personalized medicine are based on molecular measurement steps that guide personally adjusted medical decisions. A central approach to molecular profiling consists of measuring DNA, RNA, and/or proteins in tissue samples, most notably in and around tumors. This measurement yields molecular biomarkers that are potentially predictive of response and of tumor type. Current methods in cancer therapy mostly use tissue biopsy as the starting point of molecular profiling. Tissue biopsies involve a physical resection of a small tissue sample, leading to localized tissue injury, bleeding, inflammation and stress, as well as to an increased risk of metastasis. Here we developed a technology for harvesting biomolecules from tissues using electroporation. We show that tissue electroporation, achieved using a combination of high-voltage short pulses, 50 pulses 500 V cm−1, 30 µs, 1 Hz, with low-voltage long pulses 50 pulses 50 V cm−1, 10 ms, delivered at 1 Hz, allows for tissue-specific extraction of RNA and proteins. We specifically tested RNA and protein extraction from excised kidney and liver samples and from excised HepG2 tumors in mice. Further in vivo development of extraction methods based on electroporation can drive novel approaches to the molecular profiling of tumors and of tumor environment and to related diagnosis practices. [ABSTRACT FROM AUTHOR]

The use of high-value fatty acids (FA), omega-3 (n-3), and omega-6 (n-6) from seaweed could relieve the pressure from natural wild fish sources and reduce overfishing worldwide. This research is designed to explore how the harvest season (winter, spring, and summer) and culture period influence the biomass production yield, lipid content, and FA composition in the green seaweed Ulva sp. The studied seaweed was grown in plastic sleeves with flowthrough seawater in Mikhmoret (East Mediterranean, Israel) from July 2019 to December 2020. The Ulva species was identified as Ulva lactuca Linnaeus by DNA barcoding using rbcL, ITS, and tufA markers. No detectable genetic variability in U. lactuca samples was found throughout the research period. A quantitative examination of the lipid and FA content in U. lactuca in different harvest seasons revealed that the maximum content of lipids was in the summer, and that of polyunsaturated fatty acids (PUFAs) was in the winter. The PUFA profile included eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, n-3, and n-6, where the n-6/n-3 ratio was comparable with the desired range for a balanced nutritional diet. [ABSTRACT FROM AUTHOR]

The production of animal and terrestrial plant proteins has environmental impacts that could limit their sustainable supply. Proteins from microalgae and macroalgae can support the growing animal-free protein demand sustainably, using non-arable land and seawater. However, current technologies and processes developed for terrestrial plant protein processing have multiple limitations when applied to micro and macroalgae, thus, providing an opportunity for emerging technologies. Here we critically review various methods used for micro and macroalgal cell disruption for protein extraction. The pulsed electric field (PEF) algal biomass process is focused on further and highlighted as a green approach for protein extraction from algae. The potential of high-voltage pulsed electric fields and electroporation technologies for algal protein processing are discussed in detail. Current state of the art, challenges, and perspectives are also discussed. The current review will aid in establishing PEF for processing algal biomass to extract valuable compounds including proteins using a non-thermal and chemical-free approach, thus providing an environmentally friendly solution for protein extraction from algae along with highlighting upscaling and processing challenges. [ABSTRACT FROM AUTHOR]

Offshore macroalgae production could provide an alternative source of biomass for food, materials and energy. However, the offshore environment in general, specifically the Eastern Mediterranean Sea (EMS) offshore, is a high energy and low nutrients environment, thus challenging for macroalgae farming. In this study, we experimentally investigated the impact of season, depth, and pre-cultivation fertilization duration on the growth rates and chemical composition of offshore Ulva biomass, and developed a predictive model tailored to offshore conditions, capable of estimating both biomass growth rate and nitrogen content. Specifically, we measured Ulva biomass growth rate and internal nitrogen in the nitrogen-poor EMS a few kilometers offshore the Israeli coast at various depths and on-shore pre-cultivation fertilization schedules. Based on these data, we constructed a predictive cultivation model of Ulva offshore growth, which allows for the optimization of fertilization requirements for offshore cultivation. This study provides new insights on the effects of seasonality, depth, and pre-cultivation fertilization duration on growth rates and chemical composition of offshore Ulva sp. biomass production. [ABSTRACT FROM AUTHOR]

Basal cell carcinoma (BCC) is the most common type of skin cancer. Due to multiple, potential underlying molecular tumor aberrations, clinical treatment protocols are not well-defined. This study presents multisite molecular heterogeneity profiles of human BCC based on RNA and proteome profiling. Three areas from lesions excised from 9 patients were analyzed. The focus was gene expression profiles based on proteome and RNA measurements of intra-tumor heterogeneity from the same patient and inter-tumor heterogeneity in nodular, infiltrative, and superficial BCC tumor subtypes from different patients. We observed significant overlap in intra- and inter-tumor variability of proteome and RNA expression profiles, showing significant multisite heterogeneity of protein expression in the BCC tumors. Inter-subtype analysis has also identified unique proteins for each BCC subtype. This profiling leads to a deeper understanding of BCC molecular heterogeneity and potentially contributes to developing new sampling tools for personalized diagnostics therapeutic approaches to BCC. [ABSTRACT FROM AUTHOR]

Due to its large, exclusive economic zone, India has considerable potential for implementing large-scale cultivation of macroalgae. However, such cultivation requires the availability of, and access to, sites where technical, legal, governmental, and environmental factors are favorable. This review discusses the challenges that have held back the development of seaweed cultivation in India. The review is based on a literature survey and informal discussions with industry-related personnel. It cites the strong need for clear and definitive policies related to access to and use of coastal waters to enable the Indian seaweed industry to reach its full potential. The main challenges that the expansion of macroalgal cultivation in India face are related to legal and regulatory aspects that can be resolved by focusing the policy issues on providing planning tools toward success. In addition, there is a strong need for an adequate bioeconomy that clearly defines the need for marine macroalgal biomass for food, chemicals, and biofuels. Furthermore, the Indian government needs to allocate sufficient funds for accelerating seaweed R&D in areas of seaweed cultivation, harvesting, processing technologies, and their implementation in the local industry. [ABSTRACT FROM AUTHOR]

Offshore macroalgae production could provide an alternative source of biomass for food, materials and energy. However, the offshore environment in general, specifically the Eastern Mediterranean Sea (EMS) offshore, is a high energy and low nutrients environment, thus challenging for macroalgae farming. In this study, we experimentally investigated the impact of season, depth, and pre-cultivation fertilization duration on the growth rates and chemical composition of offshore Ulva biomass, and developed a predictive model tailored to offshore conditions, capable of estimating both biomass growth rate and nitrogen content. Specifically, we measured Ulva biomass growth rate and internal nitrogen in the nitrogen-poor EMS a few kilometers offshore the Israeli coast at various depths and on-shore pre-cultivation fertilization schedules. Based on these data, we constructed a predictive cultivation model of Ulva offshore growth, which allows for the optimization of fertilization requirements for offshore cultivation. This study provides new insights on the effects of seasonality, depth, and pre-cultivation fertilization duration on growth rates and chemical composition of offshore Ulva sp. biomass production. [ABSTRACT FROM AUTHOR]

Assessment of abiotic impact on seaweed chemical composition is required for their use as crops and feedstock for biorefinery. This study investigated how seasonality and the culture age of Ulva lactuca grown in plastic sleeves in a marine photobioreactor with flow-through seawater in Mikhmoret onshore (East Mediterranean, Israel), affected the composition of the amino acids and monosaccharides. Fresh samples of young (14 days) and old (21 days) U. lactuca were sampled from the photobioreactor cultivation system from January to December 2020. DNA sequences of all studied seaweed samples, which corresponded to the U. lactuca sequence, were identified using the rbcL and ITS markers as well as phylogenetic analysis. In addition, the protein and carbohydrate contents as well as the amino acid and monosaccharide profiles were determined and quantified by thermal acid hydrolysis, silyl-based derivatization, and gas chromatography-mass spectroscopy (GC–MS) analysis. The biomass components of U. lactuca were more influenced by season than by their culture age. The total protein in the dry weight of U. lactuca was significantly higher in winter (170–237 mg g−1) than in summer (64–123 mg g−1), whereas the monosaccharide content was significantly higher in summer (117–182 mg g−1) than in winter (88–98 mg g−1). These findings indicated that seasonality is an important parameter that must be considered to predict the best time to harvest U. lactuca biomass for biorefinery and food processing. Finally, an in-vitro digestibility study indicated that U. lactuca has the potential to compete with plants as a suitable protein source. [ABSTRACT FROM AUTHOR]

Degenerative skin diseases affect one third of individuals over the age of sixty. Current therapies use various physical and chemical methods to rejuvenate skin; but since the therapies affect many tissue components including cells and extracellular matrix, they may also induce significant side effects, such as scarring. Here we report on a new, non-invasive, non-thermal technique to rejuvenate skin with pulsed electric fields. The fields destroy cells while simultaneously completely preserving the extracellular matrix architecture and releasing multiple growth factors locally that induce new cells and tissue growth. We have identified the specific pulsed electric field parameters in rats that lead to prominent proliferation of the epidermis, formation of microvasculature, and secretion of new collagen at treated areas without scarring. Our results suggest that pulsed electric fields can improve skin function and thus can potentially serve as a novel non-invasive skin therapy for multiple degenerative skin diseases. [ABSTRACT FROM AUTHOR]

Irreversible electroporation (IRE) is an emerging, minimally invasive technique for solid tumors ablation, under clinical investigation for cancer therapy. IRE affects only the cell membrane, killing cells while preserving the extracellular matrix structure. Current reports indicate tumors recurrence rate after IRE averaging 31% of the cases, of which 10% are local recurrences. The mechanisms for these recurrences are not known and new explanations for incomplete cell death are needed. Using finite elements method for electric field distribution, we show that presence of vascular structures with blood leads to the redistribution of electric fields leading to the areas with more than 60% reduced electric field strength in proximity to large blood vessels and clustered vessel structures. In an in vivo rat model of liver IRE ablation, we show that cells located in the proximity of larger vessel structures and in proximity of clustered vessel structures appear less affected by IRE ablation than cells in the tissue parenchyma or in the proximity of small, more isolated vessels. These findings suggest a role for 'electric field sinks' in local tumors recurrences after IRE and emphasize the importance of the precise mapping of the targeted organ structure and conductivity for planning of electroporation procedures. [ABSTRACT FROM AUTHOR]

Seagriculture, which can provide offshore grown macroalgae biomass would play a significant role in bioeconomy. Nevertheless, seagriculture development has been hindered by the lack of laboratory photobioreactors that enable fundamental and pilot scale macroalgae research. In this work, a macroalgae photobioreactor (MPBR) was developed and integrated into the building. The MPBR operation was demonstrated for 6 months with cultivation of Cladophora sp., Ulva compressa and Ulva rigida green macroalgae species isolated from 3 sites at the Eastern Mediterranean coast. The growth rate, protein, ash, specific energy density, rhamnose, xylose, arabinose, glucose, galactose and glucuronic acid content of the cultivated species were quantified. The maximum accumulated energy rates were 0.033 Wh L −1 d −1 for Cladophora sp., 0.081 Wh L −1 d −1 for U. compressa and 0.029 Wh L −1 d −1 for U. rigida . This work provides a detailed design of an indoor, urban photobioreactor for cultivation, maintenance and energy balance analysis of macroalgae biomass for biorefinery. [ABSTRACT FROM AUTHOR]

Biomass to fuel programs are under research and development worldwide. The largest biomass programs are underway in industrialized countries. In the coming decades, however, developing countries will be responsible for the major increase in transportation fuel demand. Although the lack of existing large-scale infrastructure and primary resources preclude oil refining in developing countries, this provides an opportunity for the rapid implementation of small-scale distributed biorefineries to serve multiple communities locally. The principles for biorefinery design, however, are still in their infancy. This review sets a precedent in combining thermodynamic, metabolic, and sustainability analyses for biorefinery design. We exemplify this approach through the design and optimization of a marine biorefinery for an average town in rural India. In this combined model, we include sustainability and legislation factors, intensive macro algae Ulva farming, and metabolic modeling of the biological two-step conversion of Ulva feedstock by a yeast ( Saccharomyces cerevisiae), and then by a bacterium ( Escherichia coli), into bioethanol. We hope that the model presented here will be useful in considering practical aspects of biorefinery design. © 2013 Society of Chemical Industry and John Wiley & Sons, Ltd [ABSTRACT FROM AUTHOR]

Marine macroalgal biomass is a promising sustainable feedstock for biorefineries. However, the development of macroalgal biomass for industrial cultivation and processing has been slow. In terrestrial plants, highthroughput phenotyping provides rapid imaging methods to select specimens with required properties, rapidly transforming traditional breeding techniques. To foster the development of macroalgal biomass for biorefinery applications, we developed a near-infrared spectrometrybased method for rapid phenotyping of the macroalga Ulva fasciata based on its glucose, rhamnose, xylose and glucuronic acid contents. Spectral slopes were calculated as indicative of major carbohydrate content change. In addition, different spectral indices were generated to distinguish between low and high contents of glucose, rhamnose, xylose and glucuronic acid in wet and dry biomass. Since glucose is a major monosaccharide in Ulva that is fermentable to bioethanol, as an example of future application, we developed a multivariate data analysis based on partial least squares regression to predict its content in dry and wet biomass samples solely from reflectance data. These methods could provide a useful, high-throughput tool to rapidly select thalli with high carbohydrate content for further propagation and to be used for feedstock development for marine biorefineries. [ABSTRACT FROM AUTHOR]

The design of effective electroporation protocols for molecular delivery applications requires the determination of transport parameters including diffusion coefficient, membrane resealing, and critical electric field strength for electroporation. The use of existing technologies to determine these parameters is time-consuming and labor-intensive, and often results in large inconsistencies in parameter estimation due to variations in the protocols and setups. In this work, we suggest using a set of concentric electrodes to screen a full range of electric field strengths in a single test to determine the electroporation-induced transmembrane transport parameters. Using Calcein as a fluorescent probe, we developed analytical methodology to determine the transport parameters based on the electroporation-induced pattern of fluorescence loss from cells. A monolayer of normal human dermal fibroblast (NHDF) cells were pre-loaded with Calcein and electroporated with an applied voltage of 750 V with 10 and 50 square pulses with 50 μs duration. Using our analytical model, the critical electric field strength for electroporation was found for the 10 and 50 pulses experiments. An inverse correlation between the field strength and the molecular transport time decay constant, and a direct correlation between field strength and the membrane permeability were observed. The results of this work can simplify the development of electroporation-assisted technologies for research and therapies. [ABSTRACT FROM AUTHOR]

Excision tissue biopsy, while central to cancer treatment and precision medicine, presents risks to the patient and does not provide a sufficiently broad and faithful representation of the heterogeneity of solid tumors. Here we introduce e-biopsy—a novel concept for molecular profiling of solid tumors using molecular sampling with electroporation. As e-biopsy provides access to the molecular composition of a solid tumor by permeabilization of the cell membrane, it facilitates tumor diagnostics without tissue resection. Furthermore, thanks to its non tissue destructive characteristics, e-biopsy enables probing the solid tumor multiple times in several distinct locations in the same procedure, thereby enabling the spatial profiling of tumor molecular heterogeneity.We demonstrate e-biopsy in vivo, using the 4T1 breast cancer model in mice to assess its performance, as well as the inferred spatial differential protein expression. In particular, we show that proteomic profiles obtained via e-biopsy in vivo distinguish the tumors from healthy breast tissue and reflect spatial tumor differential protein expression. E-biopsy provides a completely new molecular sampling modality for solid tumors molecular cartography, providing information that potentially enables more rapid and sensitive detection at lesser risk, as well as more precise personalized medicine. [ABSTRACT FROM AUTHOR]

This study proposes a process based on the application of high-voltage pulsed electric field (PEF) in combination with pH shift for protein and solute release from fresh edible seaweed from Gracilaria sp.. The PEF was administered using a custom-made generator within and electroporation cell with sliding electrodes. The applied PEF parameters comprised 1000 V applied voltage, 50 µs pulse duration, 200 pulses delivered at 3 Hz frequency, and an inter-electrode gap ranging from 3.9 to 4.4 mm. The pH shift extraction was performed through a combination of fractions extracted sequentially at pH-shift steps spanning the pH range of 1 to 12. Both PEF and pH played pivotal roles in the fractionation of Gracilaria sp., enhancing protein release compared to water extraction, PEF pre-treatment, or pH shift alone. The combination of PEF and sequential pH washes (pH 1 to 12) resulted in a total solute release of 9.61 ± 0.04% and protein release of 28.02 ± 0.2% with essential amino acid (EAA) content of 63.6 ± 5.3%, and branched amino acids (BCAA) content of 3.47 ± 0.06%. It is noteworthy that PEF significantly increased the EAA and BCAA content of the protein extracts, irrespective of pH shifts. These findings underscore the promising potential of PEF pre-treatment in extracting proteins from edible seaweeds. [ABSTRACT FROM AUTHOR]

Seeding of skin cancer cells following diagnostic or therapeutic surgical procedures can occur and might cause local recurrences. As current preferred therapy for skin malignancy is surgical excision, seeding of tumour cells by manipulating malignant tissue or suturing can be another factor leading to recurrences.To evaluate whether genetic material and malignant cells adhere to standard suture materials.This prospective study included patients who underwent excision of skin lesions. Monofilament and braided sutures were examined. Sutures were passed through the observed tumour or healthy skin margins and were examined for DNA material and cells by cytological analysis, cell culture and characterization, and DNA analysis.Twenty-two patients and 148 sutures were included. DNA quantification showed DNA material on all sutures, with no significant difference between braided and monofilament sutures. Cytological analysis showed that all slides prepared from cell blocks contained normal squamous and atypical cells. Cell culture and characterization showed viable cells adhering to the sutures under direct light microscopy. Cell cultures showed rapid proliferation of epithelial cells from squamous cell carcinoma specimens.Suture materials carry DNA material and cells, including malignant cells of cutaneous origin and may seed them at distant sites. [ABSTRACT FROM AUTHOR]

Neoteric phenomena such as climate change, scarce water availability and excessive fertilizer usage necessitate an augmentation of resource utilisation efficiencies in the agricultural sector. There is a need to reorient the agroecosystems to curb stress on environmental resources while meeting rising socio-economic objectives under changing hydro-climatic conditions. Considering this, optimal land allocation for diversified agriculture is essential. We propose a combinatorial optimisation approach for land allocation considering agronomic, socio-economic, environmental and hydro-climatic objectives using bio-inspired optimization algorithms. The stochastic approach tackles the problem of optimal agricultural land allocation for crops in a multidimensional context by simultaneously addressing the conflicting goals of farm-level risk management as well as district-level contingency planning. The efficiencies and sensitivity of the proposed framework are assessed through a case study of the Dharwad district in Karnataka, India using the data (water and fertilizer consumption and cost, crop type, cultivable land, man and machine hours, etc.) from the year 2019–2020. Results indicate that Multi-objective Genetic Algorithm (MOGA) is more capable of optimising agricultural resources management by suggesting optimal land allocation for diversified crop planning. Although Cuckoo Search (CS) and Particle Search Optimisation (PSO) also produced productive Pareto fronts, they were observed to be less effective than MOGA. The annual increase in profits and crop yield obtained using MOGA are 103% and 97% respectively, while water usage is reduced by 5% compared to the conventional routines in Dharwad. The proposed hydro-agronomic decision support framework (DSF) can be utilised to assist the AI-enabled crop planning process for the sustainable management of agroecosystems. [Display omitted] • Artificial intelligence (AI) framework for multi-scalar diversified agriculture. • Socio-economic, agronomic, hydro-climatic and environmental objectives for improving profits. • Comparison of MOPSO, MOGA and MOCS with traditional approach. • Multi-objective genetic algorithm improves profit and crop yield. [ABSTRACT FROM AUTHOR]

A major concern in tissue biopsies with a needle is missing the most lethal clone of a tumor, leading to a false negative result. This concern is well justified, since needle-based biopsies gather tissue information limited to needle size. In this work, we show that molecular harvesting with electroporation, e-biopsy, could increase the sampled tissue volume in comparison to tissue sampling by a needle alone. Suggested by numerical models of electric fields distribution, the increased sampled volume is achieved by electroporation-driven permeabilization of cellular membranes in the tissue around the sampling needle. We show that proteomic profiles, sampled by e-biopsy from the brain tissue, ex vivo, at 0.5mm distance outside the visible margins of mice brain melanoma metastasis, have protein patterns similar to melanoma tumor center and different from the healthy brain tissue. In addition, we show that e-biopsy probed proteome signature differentiates between melanoma tumor center and healthy brain in mice. This study suggests that e-biopsy could provide a novel tool for a minimally invasive sampling of molecules in tissue in larger volumes than achieved with traditional needle biopsies. [ABSTRACT FROM AUTHOR]

In this study we describe a simple lab-on-a-chip (LOC) biosensor approach utilizing well mixed microfluidic device and a microsphere-based assay capable of performing near real-time diagnostics of clinically relevant analytes such cytokines and antibodies. We were able to overcome the adsorption kinetics reaction rate-limiting mechanism, which is diffusion-controlled in standard immunoassays, by introducing the microsphere-based assay into well-mixed yet simple microfluidic device with turbulent flow profiles in the reaction regions. The integrated microsphere-based LOC device performs dynamic detection of the analyte in minimal amount of biological specimen by continuously sampling micro-liter volumes of sample per minute to detect dynamic changes in target analyte concentration. Furthermore we developed a mathematical model for the well-mixed reaction to describe the near real time detection mechanism observed in the developed LOC method. To demonstrate the specificity and sensitivity of the developed real time monitoring LOC approach, we applied the device for clinically relevant analytes: Tumor Necrosis Factor (TNF)-α cytokine and its clinically used inhibitor, anti-TNF-α antibody. Based on the reported results herein, the developed LOC device provides continuous sensitive and specific near real-time monitoring method for analytes such as cytokines and antibodies, reduces reagent volumes by nearly three orders of magnitude as well as eliminates the washing steps required by standard immunoassays. [ABSTRACT FROM AUTHOR]

Multi-scale macroalgae growth models are required for the efficient design of sustainable, economically viable, and environmentally safe farms. Here, we develop a multi-scale model for Ulva sp. macroalgae growth and nitrogen sequestration in an intensive cultivation farm, regulated by temperature, light, and nutrients. The model incorporates a range of scales by incorporating spatial effects in two steps: light extinction at the reactor scale (1 m) and nutrient absorption at the farm scale (1 km). The model was validated on real data from an experimental reactor installed in the sea. Biomass production rates, chemical compositions, and nitrogen removal were simulated under different seasons, levels of dilution in the environment and water-exchange rate in the reactor. This multi-scale model provides an important tool for environmental authorities and seaweed farmers who desire to upscale to large bioremediation and/or macroalgae biomass production farms, thus promoting the marine sustainable development and the macroalgae-based bioeconomy. Zollmann et al. develop a multi-scale model for Ulva sp. macroalgae growth, biochemical composition, and nitrogen sequestration, regulated by temperature, light and nutrients. Their results demonstrate that this model can be used to design environmentally friendly and economically sustainable seaweed farms. [ABSTRACT FROM AUTHOR]

Extracellular matrix (ECM) provides structural support for cell growth, attachments and proliferation, which greatly impact cell fate. Marine macroalgae species Ulva sp. and Cladophora sp. were selected for their structural variations, porous and fibrous respectively, and evaluated as alternative ECM candidates. Decellularization–recellularization approach was used to fabricate seaweed cellulose-based scaffolds for in-vitro mammalian cell growth. Both scaffolds were confirmed nontoxic to fibroblasts, indicated by high viability for up to 40 days in culture. Each seaweed cellulose structure demonstrated distinct impact on cell behavior and proliferation rates. The Cladophora sp. scaffold promoted elongated cells spreading along its fibers' axis, and a gradual linear cell growth, while the Ulva sp. porous surface, facilitated rapid cell growth in all directions, reaching saturation at week 3. As such, seaweed-cellulose is an environmentally, biocompatible novel biomaterial, with structural variations that hold a great potential for diverse biomedical applications, while promoting aquaculture and ecological agenda. [ABSTRACT FROM AUTHOR]

Controlling the life cycle of the green macroalga Ulva (Chlorophyta) is essential to maintain its efficient aquaculture. A fundamental shift in cultivation occurs by transforming the thallus cells into gametangia and sporangia (sporulation), with the subsequent release of gametes and zoids. Sporulation occurrence depends on algal age and abiotic stimuli and is controlled by sporulation inhibitors. Thus, quantification of sporulation intensity is critical for identifying the biotic and abiotic factors that influence the transition to reproductive growth. Here, we propose to determine the sporulation index by measuring the number of released gametes using flow cytometry, in proportion to the total number of thallus cells present before the occurrence of the sporulation event. The flow cytometric measurements were validated by manually counting the number of released gametes. We observed a variation in the autofluorescence levels of the gametes which were released from the gametangia. High autofluorescence level correlated to phototactically active behaviour of the gametes. As autofluorescence levels varied between different groups of gametes related to their mobility, flow cytometry can also determine the physiological status of the gametes used as feedstock in seaweed cultivation. [ABSTRACT FROM AUTHOR]

Blue economy, the sustainable and effective use of ocean resources for economic growth, is a major challenge to coastal communities. Marine macroalgae are potentially sustainable feedstock for future food, materials, chemicals, and fuels. For seaweed biorefinery, the fractionation of the biomass to co-produce multiple products is crucial in the efficient valorization of the marine biomass. In this work, we developed a protocol for co-production of six different products from the green macroalga Ulva ohnoi using green extraction methods. A total of 90.31 ± 1.94% of the initial biomass was recovered in separated products. The fraction of the recovered products from initial dry weight biomass was 45.42 ± 1.91% salts, 3.67 ± 1.38% starch, 3.81 ± 1.26% lipids, 13.88 ± 0.40% ulvan, 14.83 ± 1.06% proteins, and 8.70 ± 1.87% cellulose. A potential revenue analyses, based on these experimental data and current market prices, suggests that total the revenue fluctuates between US$1.56 and US$3.93 kg−1 of dry biomass and depends on recovered products fraction in the seaweed biomass and products market value. [ABSTRACT FROM AUTHOR]

Intelligent biorefinery design that addresses both the composition of the biomass feedstock as well as fermentation microorganisms could benefit from dedicated tools for computational simulation and computer-assisted optimization. Here we present the BioLego Vn2.0 framework, based on Microsoft Azure Cloud, which supports large-scale simulations of biomass serial fermentation processes by two different organisms. BioLego enables the simultaneous analysis of multiple fermentation scenarios and the comparison of fermentation potential of multiple feedstock compositions. Thanks to the effective use of cloud computing it further allows resource intensive analysis and exploration of media and organism modifications. We use BioLego to obtain biological and validation results, including (1) exploratory search for the optimal utilization of corn biomasses—corn cobs, corn fiber and corn stover—in fermentation biorefineries; (2) analysis of the possible effects of changes in the composition of K. alvarezi biomass on the ethanol production yield in an anaerobic two-step process (S. cerevisiae followed by E. coli); (3) analysis of the impact, on the estimated ethanol production yield, of knocking out single organism reactions either in one or in both organisms in an anaerobic two-step fermentation process of Ulva sp. into ethanol (S. cerevisiae followed by E. coli); and (4) comparison of several experimentally measured ethanol fermentation rates with the predictions of BioLego. [ABSTRACT FROM AUTHOR]

Offshore cultivation of marine macroalgae is a potential sustainable resource for fuel, food, and chemicals. Offshore, the high productivity of macroalgae cultivation depends on external nitrogen supply. The current work examines the idea of supplying nitrogen for Ulva sp. cultivation in the oligotrophic Eastern Mediterranean Sea (EMS) by artificial upwelling of nutrient-rich deep seawater (DSW). Growth rates, protein, and starch contents of Ulva sp. were measured for time varying fertilizations with nitrate concentrations corresponding to nutrient concentrations of DSW at increasing depths of the EMS. A maximal relative growth rate of 7.4% was measured for fertilizing ten times per week with 5.8 μM, which corresponds to the artificial upwelling from the depth of 700 m at EMS. Protein and starch contents ranged between 1–6 and 8–15% of dry weight. Finally, yields and energetic costs of DSW pumping were modeled for an example case of 10-ha offshore farm. The model predicts a high productivity but low energetic efficiency, which can be improved by coupling the biomass production with offshore power sources such as ocean thermal energy conversion. [ABSTRACT FROM AUTHOR]

Subcritical water hydrolysis and carbonization of the biomass are an emerging green technology for seaweed biomass processing. In this work, a novel approach for co-generation of two energy streams from seaweed biomass (fermentable sugars and solid hydrochar) with subcritical water from a green macroalgae Ulva sp. was developed. It was found that for the released of glucose, xylose, rhamnose, fructose, and galactose, the process temperature is the most significant parameter, followed by salinity, solid load, and treatment time. For the formation of fermentation inhibitor 5-hydroxymethylfurfural (5-HMF), temperature also was the most important parameter, followed by residence time, salinity, and solid load. The optimum parameters for maximal release of total sugars under minimum formation of 5-HMF were 170 °C (800 kPa abs.), 5% solid loading, 40 min residence time, and 100% salinity. The hydrochar yield was 19.4% and hydrochar high heating value was 20.2 ± 1.31 MJ kg−1. These results provide new detailed information on the subcritical hydrolysis and carbonization of Ulva sp. biomass and show co-production of fermentable monosaccharides and hydrochar. [ABSTRACT FROM AUTHOR]

Enhancing water diffusivity shortens meat processing time and saves energy and costs. One of the processes that can enhance water diffusivity in tissues is high-voltage, short-pulsed electric fields (PEF). However, for industrial PEF process development, there is a need in adaptable laboratory instruments. Here we report on a laboratory PEF generator, based on insulated-gate monopolar transistor switching, coupled with sliding positive electrode for the enhancement of water diffusivity in chicken breast muscle. The system generates rectangular monopolar pulses with a voltage amplitude up to 1000 V, current up to 160 A, pulse duration of 5 to 100 μs, and a frequency of pulse repetition of 1–16 Hz. The energy conversion efficiency of the developed PEF generator is 88%. We found that applying 120 pulses at 1000 V (~ 500 V mm−1), and a pulse duration of 50 μs at 1 Hz, on the chicken breast muscle, increased the effective diffusivity of water by 13–24% and reduced convective air drying time by 6.4–15.3%. These results provide new information on the design of laboratory equipment to improve and optimize meat pre-processing on a small scale. Flexible, small-scale PEF equipment is a necessary step for the industrial development of new processes which could reduce equipment size and process energy consumption in the meat industry. [ABSTRACT FROM AUTHOR]

Marine biorefineries, based on macroalgal (seaweed) feedstocks, could provide sustainable alternative sources of food, energy, and materials. Green macroalgae, with their unique chemical composition, can contribute to marine biorefinery systems associated with a wide range of potential products. This review discusses the challenge of developing industrially relevant and environmentally-friendly green seaweed biorefineries. First, we review potential products from green seaweeds and their co-production, the key element in an integrated biorefinery. Second, we discuss large-scale cultivation, hydrothermal treatments, fermentation, anaerobic digestion, and emerging green solvents, pulsed electric field, microwave, and ultrasound processing technologies. Finally, we analyse the main polysaccharides in green seaweeds: sulfated polysaccharides, starch, and cellulose, as products of a cascading biorefinery, with emphasis on applications and technological challenges. We provide a comprehensive state-of-the-art review of green seaweed as feedstock for the biorefinery, analysing opportunities and challenges in the field. [ABSTRACT FROM AUTHOR]

Increasing biomass production yields is a critical challenge for macroalgae biorefineries. The continuous tumbling and mixing of free‐floating algae through water or airflow has been shown to increase the productivity of algae in land‐based cultivation systems. This approach has not been tested thoroughly in offshore cultivation. We report, here, a field feasibility study on the increase in green macroalga Ulva sp. growth rates in offshore cages, achieved by the combined effect of tumbling and mixing of the algae using influxes of water and air. The experimental system was tested in a shallow coastal area in central Israel, in the eastern Mediterranean Sea. A maximum daily growth rate of 19.2%, areal productivity of 33.72 g dry weight (DW) day−1 m−2, and volumetric yields of 37.78 g DW day−1 m−3, together with 38.47 ± 0.01% ash and 5.28% protein content on a dry matter basis were achieved in the cages with intensified cultivation in the first week of May 2017. Our study shows that cultivation with tumbling and mixing of biomass with air, and water exchange with the environment is a feasible method to increase Ulva sp. biomass productivity offshore. © 2019 Society of Chemical Industry and John Wiley & Sons, Ltd [ABSTRACT FROM AUTHOR]

Abstract Visual-Near-Infra-Red (VIS/NIR) spectroscopy has led the revolution in high-throughput phenotyping methods used to determine chemical and structural elements of organic materials. In the current state of the art, spectrophotometers used for imaging techniques are either very expensive or too large to be used as a field-operable device. In this study we developed a Sparse NIR Optimization method (SNIRO) that selects a pre-determined number of wavelengths that enable quantification of analytes in a given sample using linear regression. We compared the computed complexity time and the accuracy of SNIRO to Marten's test, to forward selection test and to LASSO all applied to the determination of protein content in corn flour and meat and octane number in diesel using publicly available datasets. In addition, for the first time, we determined the glucose content in the green seaweed Ulva sp., an important feedstock for marine biorefinery. The SNIRO approach can be used as a first step in designing a spectrophotometer that can scan a small number of specific spectral regions, thus decreasing, potentially, production costs and scanner size and enabling the development of field-operable devices for content analysis of complex organic materials. Graphical abstract Image 1 Highlights • A Sparse NIR Optimization method (SNIRO) for selecting a given number of significant wavelengths from spectra was developed. • The computed complexity time and the accuracy of SNIRO was compared to Marten's test, to forward selection test and to LASSO. • SNIRO was used to determine protein content in corn flour and meat, and octane number in diesel using public NIR datasets. • SNIRO was used to determine the glucose content in the green seaweed Ulva sp. [ABSTRACT FROM AUTHOR]

Highlights • Method for selective allergens extraction with pulsed electric fields. • Protein extraction with the pulsed electric field from Ulva sp. was optimized. • Allergy assessment of proteins extracted from the seaweed Ulva sp. • Proteomic analysis of Ulva sp. Abstract Extraction of protein from macroalgae, currently defined as "novel food", is challenging and limited information about the health impacts of these proteins is available. Here, we report on a non-thermal, chemical-free green macroalgae Ulva sp. protein extraction by osmotic shock combined with pulsed electric fields (PEF) followed by hydraulic pressure. The extracted proteins were identified and annotated to allergens using sequence similarity. The allergenicity potential of PEF extracted proteins was compared to osmotic shock extracts and complete Ulva sp. proteome, extracted with the thermochemical method. The PEF extracts contained 'superoxide dismutase' (SOD), a known food allergen, osmotic shock extract contained 'troponin C', and thermochemical extract contained two additional potential food allergens 'aldolase A' and 'thioredoxin h'. This study shows an importance and the need for deep investigation of algal proteins and protein extraction technology health impacts prior to large-scale release to the market of "novel food" derived proteins. [ABSTRACT FROM AUTHOR]

Abstract With rising global population and decreasing available land and fresh water resources, the oceans provide an attractive domain for sourcing nutrients. The marine macroalgae Ulva sp. and Gracilaria sp. are candidate raw biomass. Ulva sp. has high growth rate and Gracilaria sp. has high protein content and their seasonal growth is complementary, allowing almost year-round high yield protein production. In this study, we aimed at developing an effective process, to yield a high macroalgae protein content concentrate suitable for food application, and studying the digestibility, amino acid composition and antioxidant properties of the obtained algal protein concentrates (APCs). We developed a new protein extraction protocol, and compared it to several published protocols. The developed protocol is food-grade, and yielded APC from Ulva and form Gracilaria , containing 70 and 86% protein respectively. The amino acid compositions of the APCs suggest their possible use as sources of essential amino acids. Simulated gastro-intestinal digestion showed that APCs proteolysis of at least 89% can be reached. We found that the APCs exhibit antioxidant activity, which is similar to that of known protein isolates in the hydrogen atom transfer mechanism, but 10 to 20 times higher in the single electron transfer mechanism. These results suggest that polyphenolic compounds might be still present in the APCs and contribute to their antioxidant activity. Our results suggest that the protein concentrates extracted from Ulva sp. and Gracilaria sp. seem to be promising sustainable sources for human nutrition thanks to their essential amino acids content, digestibility and antioxidant properties. Graphical abstract Image 1 Highlights • Macroalgae – a new and renewable source for extracted nutrients. • Food grade extraction yielded high purity protein powders. • Extracted proteins contain good levels of essential amino acids. • Protein concentrates exhibit high antioxidant activity. • Extracted proteins were ∼90% digestible under simulated gastro-intestinal conditions. [ABSTRACT FROM AUTHOR]