Alternative Crops for the European Tobacco Industry: A Systematic Review.

Tobacco (Nicotiana tabacum L.) is a major industrial crop that has being cultivated for centuries for the manufacturing of cigarettes, cigars, and other smoking products. Due to its negative effects on both human health and the environment, the European Union has adopted strict policies that aspire to reduce the consumption of tobacco. Herbal cigarettes are alternative smoking products that are often advertised as healthier than conventional tobacco cigarettes and are especially popular in Asian markets. Even though the available literature suggests that they are equally detrimental to human health, the introduction of tobacco-alternative crops (TACs) to the European tobacco industry could smoothen the abandonment of tobacco, and eventually smoking products altogether, in the EU. The aim of the present systematic review was to compile a list of possible TACs that could be incorporated in the European smoking industry, and highlight their strengths and weaknesses. The most dominant crops in the literature (and in the existing market products) were calendula (Calendula officinalis L.), mullein (Verbascum thapsus L.), ginseng (Panax ginseng C.A.Mey.), tea (Camellia sinensis (L.) Kuntze), chamomile (Matricaria chamomilla L.), and mentha (Mentha spp.). Even though these crops are promising, further research is required for their incorporation in the European tobacco industry.

Nuclear magnetic resonance analysis of extra virgin olive oil: classification through secoiridoids.

BACKGROUND: Extra virgin olive oil (EVOO), a natural product with a multidisciplinary role, has been and is continuing to be studied from several points of view. Among them, its chemical analysis is of major importance and several methods have been used. Nuclear magnetic resonance (NMR) spectroscopy has inherent advantages, among them monitoring the chemical constituents without the need for a separation technique and without, for instance, possible carry-over effects. Additionally, several magnetic resonance spectroscopic techniques can provide a novel powered insight into the nature and properties of a sample under study. Moreover, -omics procedure can reveal new information and can lead to the classification of populations under study. The main objective of the present work was the possible classification of the EVOO samples based on their aldehyde content using a proposed unreferenced 1 H-NMR spectroscopic quantification method combined with a metabolomic approach. Moreover, the study of the impact of such elevated aldehyde content on several spectra regions of importance in the proton NMR spectra led to the proposal of a possible new isomer indicator. RESULTS: Univariate analysis of 12 EVOO samples showed that oleacein, oleocanthal, elenolic acid, hydroxytyrosol/hydroxytyrosol derivatives and tyrosol/tyrosol derivatives strongly differentiated two classes of EVOO: OEH (for high aldehyde EVOO content) and OE (for non-high aldehyde content). Moreover, we pointed out the 'impact' of such elevated secoiridoid and derivatives content, through their moieties' units, on a range of several resonances of the 1 H-NMR spectrum. The metabolomic approach demonstrated the classification of EVOO samples based on their secoiridoid and derivatives content. Multivariate analysis showed a strong influence on the discrimination of the EVOO classes based on the protons resonating at the aldehyde region of the 1 H-NMR spectrum; the aldehyde protons corresponding to 5S,4R-ligstrodial and 5S,4R-oleuropeindial, oleacein, oleocanthal, elenolic acid, p-HPEA-EA, 3,4-DHPEA-EA, 5S,4R- and 5S,4S-ligstrodial and the proton corresponding to a new compound were reported for the first time. This isomer compound, reported for the first time, could serve as a possible indicator for EVOO classification. CONCLUSIONS: An unreferenced quantification method was proposed and EVOO samples were classified into two classes: OEH and OE, according to their aldehyde content, gaining thus probably higher nutrient and possible pharmacological value. Moreover, we point out the 'impact' of such elevated aldehyde content on several spectral regions of the 1 H spectrum. Finally, a new compound was detected in the OEH samples and is reported for the first time. This compound could possibly be an indicator. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A 68-Year-Old Patient With Dyspnea and Hypoxemia After Total Hip Arthroplasty.

CASE PRESENTATION: A 68-year-old patient with obesity (BMI, 4 7 kg/m2) was transferred to the ED of our hospital because of dyspnea and pronounced hypoxemia. The patient underwent total right hip arthroplasty in an outside hospital because of osteoarthritis; there was no history of trauma. After 48 h, she experienced dyspnea with severe hypoxemia. The next day she was transferred to our hospital. Her history was notable for arterial hypertension and depression, but not heart failure. Her medications included candesartan (16 mg once daily) and sertraline (100 mg once daily). Perioperatively, she received enoxaparin 4.000 International Units subcutaneously once daily. There was no family history of respiratory diseases. The patient currently smokes (50 pack-years) with no recent increase in her habit and denied vaping, alcohol consumption, illicit drug use, and any home or occupational exposures. Prior to surgery, the family of the patient reported that she maintained modest mobility despite her osteoarthritis and was able to fulfill her daily activities. Interestingly, she reported a similar event of severe dyspnea and hypoxemia after total knee arthroplasty 3 years earlier; however, no further details were available.

Molecular and Genetic Biomarkers in Idiopathic Pulmonary Fibrosis: Where Are We Now?

Idiopathic pulmonary fibrosis (IPF) represents a chronic progressive fibrotic interstitial lung disease of unknown cause with an ominous prognosis. It remains an unprecedent clinical challenge due to its delayed diagnosis and unpredictable clinical course. The need for accurate diagnostic, prognostic and predisposition biomarkers in everyday clinical practice becomes more necessary than ever to ensure prompt diagnoses and early treatment. The identification of such blood biomarkers may also unravel novel drug targets against IPF development and progression. So far, the role of diverse blood biomarkers, implicated in various pathogenetic pathways, such as in fibrogenesis (S100A4), extracellular matrix remodelling (YKL-40, MMP-7, ICAM-1, LOXL2, periostin), chemotaxis (CCL-18, IL-8), epithelial cell injury (KL-6, SP-A, SP-D), autophagy and unfolded protein response has been investigated in IPF with various results. Moreover, the recent progress in genetics in IPF allows for a better understanding of the underlying disease mechanisms. So far, the causative mutations in pulmonary fibrosis include mutations in telomere-related genes and in surfactant-related genes, markers that could act as predisposition biomarkers in IPF. The aim of this review is to provide a comprehensive overview from the bench to bedside of current knowledge and recent insights on biomarkers in IPF, and to suggest future directions for research. Large-scale studies are still needed to confirm the exact role of these biomarkers.

Effect of Arbuscular Mycorrhizal Fungi on Nitrogen and Phosphorus Uptake Efficiency and Crop Productivity of Two-Rowed Barley under Different Crop Production Systems.

Arbuscular Mycorrhizal Fungi (AMF) constitute a ubiquitous group of soil microorganisms, affecting plant and soil microorganism growth. Various crop management practices can have a significant impact on the AM association. This study investigated the AMF inoculation contribution on growth and productivity of two-rowed barley crop by identifying the underlying mechanisms both in conventional and organic cropping systems. A two-year field trial was set up as a split-plot design with 2 main plots [AMF inoculation: with (AMF+) and without (AMF-)] and five sub-plots (fertilization regimes: untreated, 100% recommended dose of fertilizer in organic and inorganic form, and 60% recommended dose of fertilizer in organic and inorganic form) in three replications. According to the results, AMF+ plants presented higher plant height and leaf area index (LAI), resulting in increased biomass and, as a result, higher seed yield. With regard to the quality traits, including the nitrogen and phosphorus uptake and their utilization indices, the AMF inoculated plants showed higher values. Furthermore, the level of fertilization, particularly in an inorganic form, adversely affected AMF root colonization. Consequently, it was concluded that substitution of inorganic inputs by organic, as well as inputs reduction, when combined with AMF inoculation, can produce excellent results, thus making barley crop cultivation sustainable in Mediterranean climates.

On the Path towards a "Greener" EU: A Mini Review on Flax (Linum usitatissimum L.) as a Case Study.

Due to the pressures imposed by climate change, the European Union (EU) has been forced to design several initiatives (the Common Agricultural Policy, the European Green Deal, Farm to Fork) to tackle the climate crisis and ensure food security. Through these initiatives, the EU aspires to mitigate the adverse effects of the climate crisis and achieve collective prosperity for humans, animals, and the environment. The adoption or promotion of crops that would facilitate the attaining of these objectives is naturally of high importance. Flax (Linum usitatissimum L.) is a multipurpose crop with many applications in the industrial, health, and agri-food sectors. This crop is mainly grown for its fibers or its seed and has recently gained increasing attention. The literature suggests that flax can be grown in several parts of the EU, and potentially has a relatively low environmental impact. The aim of the present review is to: (i) briefly present the uses, needs, and utility of this crop and, (ii) assess its potential within the EU by taking into account the sustainability goals the EU has set via its current policies.

Herbicide Use in the Era of Farm to Fork: Strengths, Weaknesses, and Future Implications.

Climate change mitigation is a major concern of the European Union (EU). In 2019, the EU presented the European Green Deal (EGD), a new environmental strategy that aimed to neutralize climate change by 2050. Within its policy areas, the EGD included the Farm to Fork (F2F) Strategy that aims to reduce pesticide use by 50%, by 2030. This reduction was proposed due to the supposed negative effects of pesticides on the environment and its biota. Among the different pesticide groups (herbicides, fungicides, insecticides, etc.) though, herbicides are perhaps the hardest to reduce. This review aimed to shed light to any factors that might hinder the reduction of herbicide use; thus, the implementation of the Farm to Fork Strategy underlines some of its weaknesses and highlights key points of a viable herbicide reduction-related policy framework. The literature suggests that integrated weed management (IWM) consists perhaps the most suitable approach for the reduction of herbicides in the EU. Even though it is too soon to conclusively assess F2F, its success is not impossible.

Development of Enriched Oil with Polyphenols Extracted from Olive Mill Wastewater.

The extraction of olive oil produces significant residual olive-mill wastewater (OMW). The composition of OMW varies according to the type of olive, the fruit's ripeness level, and the extraction method (traditional one-pressing system or continuous systems based on centrifugation of the olive pulp). In olive-producing countries, OMW is important for the environment and the economy and is also a low-cost source of polyphenolic compounds with high antioxidant properties. Olive oil's properties, such as its anti-atherogenic, anti-inflammatory, anti-aging, and immunological modulator effects, have been attributed to its polyphenols. In this study, the cloud point extraction (CPE) method was used to recover polyphenolic compounds from OMW. The total micellar phase of the three recoveries was 24.2% and the total polyphenols (after sonication) was 504 mg GAE/Kg. Furthermore, the addition of polyphenols recovered from OMW enhanced the olive oil and extended its shelf life without changing its organoleptic properties. There was a 42.2% change in polyphenols after 0.5% enrichment of micellar dispersions. Thus, it is suggested that the CPE method could lead to better waste management in the olive oil industry and improve the nutritional quality of food products.

The Role of Alternative Crops in an Upcoming Global Food Crisis: A Concise Review.

Achieving Food Security (FS) is perhaps our most challenging aspiration. Despite our best efforts, millions of people around the globe are malnourished or live with hunger. The state of the geo-political scene, as well as the COVID-19 pandemic, have recently brought forth fears of a Global Food Crisis (GFC). Here, we present the factors that threaten FS and could trigger a GFC, examine the potential of alternative crops (ACs) as a measure against an upcoming GFC, and highlight the key aspects of the ACs introduction process in new regions. ACs could enhance FS, yet their success is premised on the adoption of sustainable practices and the implementation of food strategies that aim to promote healthy consumer behaviours.

The Influence of Fertilization and Plant Density on the Dry Matter Yield and Quality of Black Mustard [Brassica nigra (L.) Koch]: An Alternative Forage Crop.

Black mustard [Brassica nigra (L.) Koch] is mainly cultivated as a seed crop, and there is a lack of information on biomass quality and its potential for animal feeding. A 2-year field experiment was set up in a split-plot design with 2 main plots (plant densities: 46 and 76 plants m-2), 4 sub-plots (fertilization levels: control, compost, urea with and without urease and nitrification inhibitors) and 3 replications for each treatment. The highest dry matter yield (17.55-18.34 tn ha-1) was observed in high-density plots fertilized with urea fertilizer coated with double (nitrification and urease) inhibitors. In terms of the qualitive parameters of total above-ground biomass, the highest crude protein (CP) content was achieved in plots with low density and urea with double inhibitors. Moreover, the highest neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents of above-ground biomass were found under compost and urea with double inhibitors. The high ADF, NDF and relatively high CP content characterized that black mustard aerial biomass can meet the requirements of lactating animals, and therefore the production of black mustard biomass as a forage crop could be of great importance. As a conclusion, black mustard cultivated at plant densities higher than 46 plants m-2 and under inorganic fertilization, especially with urea coated with double inhibitors, could be successfully used as a novel forage crop in ruminants' diets.

Reintroducing Flax (Linum usitatissimum L.) to the Mediterranean Basin: The Importance of Nitrogen Fertilization.

An increasing interest has been reported regarding the reintroduction of flax in the Mediterranean region. The aim of this present study was to evaluate the effects of nitrogen (N) fertilization on the performance of flax cv. Everest, under Mediterranean climate conditions. A two-year study was carried out in 2018-2019, in Western Greece. The experiment was set-up in a randomized complete block design with four replications and six treatments of different N fertilization rates (0, 20, 30, 40, 50, and 60 kg N ha-1). Measurements included plant biomass, the leaf area index (LAI), the yield, and the Growth Degree Days (GDDs) required for full seed maturity. The N uptake of flax was also evaluated utilizing the Nitrogen Harvesting (NHI) and Nitrogen Utilization Efficiency (NUtE) indices. Although the highest fertilization rate (60N) increased the yield by 35.4% (2018) and 23.1% (2019), a GDDs and N indices assessment revealed that it noted the lowest efficiency and may lead to significant yield losses, as it significantly prolonged the crop cycle. On the contrary, even though fertilization rates of 20 and 30 kg N ha-1 increased the yield only by 7% and 15% (on average), they were more efficient, and prolonged the crop cycle less (compared to 60N).

Effect of Rhizophagus irregularis on Growth and Quality of Cannabis sativa Seedlings.

Rhizophagus irregularis is an arbuscular mycorrhiza fungus that can enhance plant nutrition and reduce transplant shock on seedlings. The present study aims to evaluate the effects of this fungus on the quality of cannabis (Cannabis sativa L.) seedlings.A greenhouse float system experiment was conducted in a completely randomized design with three treatments. The treatments included the application of 40, 80 and 120 fungus spores per L of nutrient solution (AMF1, AMF2 and AMF3, respectively). The evaluation was performed based on the agronomic characteristics of the seedlings (root and stem length and weight, stem diameter), N and P content, survival rate, and the Dickson's quality index (DQI). Results indicated that root length and stem dry weight were significantly increased (by 34.14% and 21.4%, respectively) in the AMF3 treatment. The biomass of the seedlings' roots, the fresh weight and the N content were not affected by the AMF. On the contrary, survival rate, P content and DQI were significantly increased in AMF3 (by 5%, 24.3% and 12.4% respectively). Overall, our findings suggest that the application of high doses of Rhizophagus irregularis (AMF3) on float system-produced cannabis seedlings results in a considerable increment of their quality.

RNA Whole-Mount In Situ Hybridization Proximity Ligation Assay (rISH-PLA), an Assay for Detecting RNA-Protein Complexes in Intact Cells.

Techniques for studying RNA-protein interactions have lagged behind those for DNA-protein interactions as a consequence of the complexities associated with working with RNA. This unit describes a method for the adaptation of the In Situ Hybridization-Proximity Ligation Assay (ISH-PLA) to the study of RNA regulation (rISH-PLA). The rISH-PLA assay allows the identification of a given RNA-protein complex at subcellular and single-cell resolution, thus avoiding the lack of spatial resolution and sensitivity associated with assaying heterogeneous cell populations from which conventional RNA-protein interaction detection techniques suffer. This technique will be particularly usefully for studying the activity of RNA binding proteins (RBPs) in complex mixtures of cells, for example tissue sections or whole embryos. © 2017 by John Wiley & Sons, Inc.

RNA Whole-Mount In situ Hybridisation Proximity Ligation Assay (rISH-PLA), an Assay for Detecting RNA-Protein Complexes in Intact Cells.

Techniques for studying RNA-protein interactions have lagged behind those for DNA-protein complexes as a consequence of the complexities associated with working with RNA. Here we present a method for the modification of the existing In Situ Hybridisation-Proximity Ligation Assay (ISH-PLA) protocol to adapt it to the study of RNA regulation (rISH-PLA). As proof of principle we used the well-characterised interaction of the Xenopus laevis Staufen RNA binding protein with Vg1 mRNA, the complex of which co-localises to the vegetal pole of Xenopus oocytes. The applicability of both the Stau1 antibody and the Locked Nucleic Acid probe (LNA) recognising Vg1 mRNA were independently validated by whole-mount Immunohistochemistry and whole-mount in situ hybridisation assays respectively prior to combining them in the rISH-PLA assay. The rISH-PLA assay allows the identification of a given RNA-protein complex at subcellular and single cell resolution, thus avoiding the lack of spatial resolution and sensitivity associated with assaying heterogenous cell populations from which conventional RNA-protein interaction detection techniques suffer. This technique will be particularly usefully for studying the activity of RNA binding proteins (RBPs) in complex mixtures of cells, for example tissue sections or whole embryos.

Kappa-casein based electrochemical and surface plasmon resonance biosensors for the assessment of the clotting activity of rennet.

We report for the first time the development of kappa-casein (κ-CN)-based electrochemical and surface plasmon resonance (SPR) biosensors for the assessment of the clotting activity of rennet. Electrochemical biosensors were developed over gold electrodes modified with a self-assembled monolayer of dithiobis-N-succinimidyl propionate, while SPR measurements were performed on regenerated carboxymethylated dextran gold surfaces. In both types of biosensor, κ-CN molecules were immobilized onto modified gold surfaces through covalent bonding. In electrochemical biosensors, interactions between the immobilized κ-CN molecules and chymosin (the active component of rennet) were studied by performing cyclic voltammetry, differential pulsed voltammetry, and electrochemical impedance spectroscopy (EIS) measurements, using hexacyanoferrate(II)/(III) couple as a redox probe. κ-CN is cleaved by rennet at the Phe105-Met106 bond, producing a soluble glycomacropeptide, which is released to the electrolyte, and the positively charged insoluble para-κ-casein molecule, which remains attached to the surface of the electrode. This induced reduction of the net negative charge of the sensing surface, along with the partial degradation of the sensing layer, results in an increase of the flux of the redox probe, which exists in the solution, and consequently, to signal variations, which are associated with the increased electrocatalysis of the hexacyanoferrate(II)/(III) couple on the gold surface. SPR experiments were performed in the absence of the redox probe and the observed SPR angle alterations were solely attributed to the cleavage of the immobilized κ-CN molecules. Various experimental variables were investigated and under the selected conditions the proposed biosensors were successfully tried to real samples. The ratios of the clotting power units in various commercial solid or liquid samples, as they are calculated by the EIS-based data, were almost identical to those obtained with a reference method. In addition, EIS measurements showed an excellent reproducibility, lower than 5%.

Impedimetric biosensor for the assessment of the clotting activity of rennet.

Cheese production is relied upon the action of rennet (a mixture of chymosin and pepsin) onto casein micelles of milk. For the first time, the monitoring of this interaction with electrochemical impedance spectroscopy (EIS) was used to develop a faradic impedimetric biosensor for the assessment of the clotting activity of rennet, using hexacyanoferrate(II)/(III) couple as a redox probe. Gold electrodes were modified with self-assembled monolayers of different thiols (thioctic acid, dithiobis-N-succinimidyl propionate, and cysteamine), and (artificial) casein micelles were immobilized on the modified gold surfaces. The proposed method is based on the measurement of charge-transfer resistance (R(ct)) changes attributed to the degradation of the negatively charged immobilized casein micelles by rennet to neutral biostructures. This action results in the increase of the flux of the redox probe, which exists in the bulk solution, to the surface of the electrode and, consequently, in the decrease of R(ct). Experimental parameters such as the micelle loading, the reaction time, the concentration of rennet, and the working pH, were optimized. Besides EIS measurements, cyclic voltammetry, FT-IR, and atomic force microscopy (AFM) experiments were also performed before and after the interaction of the immobilized micelles with rennet. Finally, the proposed biosensors were successfully tried for various commercial samples.

Decrease of heat shock protein levels and cell populations by wine phenolic extracts.

The effect of red and white wine total extracts and phenolic fractions on heat shock protein (Hsp) levels in tumor cells and on tumor and endothelial cell populations in vitro has been investigated. Total extracts of red wines decreased Hsp70 and Hsp27 levels and the numbers of tumor and endothelial cells. Several red and white wine fractions significantly decreased Hsp27 levels, and some of them had also an effect on Hsp70 levels. A red wine fraction rich in polymeric flavanols and a white wine one rich in phenolic acids, flavonols, and tyrosol strongly lowered Hsp27 levels. Some red and white wine fractions strongly reduced tumor cell numbers, whereas most of them decreased endothelial cell numbers to variable extents. The present results indicate that wine phenolics decrease Hsp levels in tumor cells and tumor and endothelial cell populations. These properties may be important in the potent anticarcinogenic action of wine phenolics.