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In the scope of vegetarian diets, cobalamin (CBL or vitamin B12) remains a particular concern, as it is mostly absent from natural, vegetarian foods. As such, vegetarians must rely on supplementation or enriched food products, to fulfil their recommended daily intake of the vitamin. The potential of microalgae is of extreme importance in this context, given the fact that they may absorb exogenous CBL and display significant levels of the vitamin. Therefore, it is crucial to establish fast and reliable B12 determination methods, to accurately characterize these products or ingredients and conclude about their role as CBL sources. To that extent, this work presents the application of the AOAC's final action method 2014.02 on the determination of total CBL in selected, commercial, microalgae and cyanobacteria powder samples. Four variants of Chlorella vulgaris, namely organic, smooth, white and honey, as well as Spirulina (Arthrospira platensis) were investigated. High-performance liquid chromatography coupled with ultraviolet (UV), diode-array detection (HPLC-DAD) was implemented in the analysis of microalgal extracts. After slight adaptation and optimisation of the extraction and analysis procedures, namely to allow lower sample amounts, it was determined that the different commercial Chlorella products, organic, smooth, white and honey have total CBL concentrations of 1063 ± 158 μg (100 g)-1, 482 ± 55 μg (100 g)-1, 681 ± 87 μg (100 g)-1 and 660 ± 68 μg (100 g)-1, respectively. Analysis of Spirulina did not yield any measurable levels of CBL in any of the purified extracts. Despite the obtained results displaying the same order of magnitude previously determined in similar microalgal products, these CBL values are amongst the highest concentrations found. Therefore, even considering the known limitations in bioaccessibility and bioavailability of CBL in microalgae, it is possible to conclude that these results stand for the inclusion of these products in vegetarian foods or as food supplements, as good potential sources of CBL, which is generally lacking in vegetarian diets. [ABSTRACT FROM AUTHOR]

Investigations of population structuring in wild species are fundamental to complete the bigger picture defining their ecological and biological roles in the marine realm, to estimate their recovery capacity triggered by human disturbance and implement more efficient management strategies for fishery resources. The Blackspot Seabream ( Pagellus bogaraveo , Brünnich 1768) is a commercially valuable deep-water fish highly exploited over past decades. Considering its exploitation status, deepening the knowledge of intraspecific variability, genetic diversity, and differentiation using high-performing molecular markers is considered an important step for a more effective stock assessment and fishery management. With one of the largest efforts conceived of and completed by countries overlooking the Atlantic and Mediterranean coasts in recent years, a total of 320 individuals were collected from different fishing grounds in the Mediterranean Sea and Atlantic Ocean and analysed using 29 microsatellite loci. We applied multiple statistical approaches to investigate the species' connectivity and population structure across most of its described distribution area. Considering the incomplete knowledge regarding the migratory behaviour of adults, here we suggest the importance of egg and larval dispersal in sustaining the observed genetic connectivity on such a large geographical scale.

Background: Opioid receptors are naloxone-sensitive (MOP [mu: μ], DOP [delta: δ], and KOP [kappa: κ]) and naloxone-insensitive Nociceptin/Orphanin FQ (N/OFQ) peptide receptor (NOP). Clinically, most opioid analgesics target MOP. Angiogenesis is the formation of new blood vessels and involves endothelial cell activation, proliferation, and migration. The effect of opioids on this process is controversial with no data for NOP receptor ligands., Methods: We used patient-derived human umbilical vein endothelial cells (HUVECs) treated with media from the Michigan Cancer Foundation-7 (MCF-7) breast cancer cells or vascular endothelial growth factor (VEGF; 10 ng ml -1 ) and fibroblast growth factor (FGF; 10 ng ml -1 ) as angiogenic stimuli. We have measured (i) NOP/MOP messenger RNA, (ii) receptor protein using N/OFQ ATTO594 and Dermorphin ATTO488 as fluorescent probes for NOP and MOP, and (iii) NOP/MOP function in a wound healing assay (crude measure of migration that occurs during angiogenesis)., Results: HUVEC lines from 32 patients were used. Using all 32 lines, mRNA for NOP but not MOP was detected. This was unaffected by media from MCF-7 cells or VEGF/FGF. There was no binding of either N/OFQ ATTO594 (NOP) or Dermorphin ATTO488 (MOP) in the absence or presence of angiogenic stimuli (six lines tested). In the absence of MOP mRNA, this was expected. Whilst MCF-7 conditioned medium (not VEGF/FGF) reduced wound healing per se (14 lines tested), there was no effect of N/OFQ (NOP ligand) or morphine (MOP ligand)., Conclusions: Media from MCF-7 breast cancer cells or VEGF/FGF as angiogenic stimuli did not influence NOP translation into receptor protein. MOP was absent. In the absence of constitutive or inducible MOP/NOP, there was no effect on wound healing as a measure of angiogenesis., (© 2022 The Author(s).)

We investigate how in the context of Corporate Venture Capital (CVC), the investment decisions affect the likelihood of their subsequent exit strategies. We use OLS and probit regression as well as Weibull distribution of residual values, given its reliability and validity for studying lifetime analysis. Based on a sample of 8722 VC-backed ventures with the first investment dates between 1999 and 2018 in United States (US) and United Kingdom (UK), the results show that the presence of CVCs positively affects the funding amounts and the duration of the investment. CVC funds are more generous and more patient than Independent Venture Capital (IVC) funds regarding their investments in ventures. Moreover, the findings provide evidence that the exit strategies are directly influenced by the funding amounts and the duration of the investment which are influenced, in turn, by the fund type. Greater funding increases the likelihood of IPO exit which is reduced by longer investment duration. Our results are robust to alternative estimation methods, namely two-stage treatment-effects regressions. These results help the various stakeholders (VC funds, investors, ventures) make crucial decisions regarding investment amounts and duration, and exit., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Background: Genome-wide association studies have identified many genetic loci that are robustly associated with coronary artery disease (CAD). However, the underlying biological mechanisms are still unknown for most of these loci, hindering the progress to medical translation. Evidence suggests that the genetic influence on CAD susceptibility may act partly through vascular smooth muscle cells (VSMCs)., Methods: We undertook genotyping, RNA sequencing, and cell behavior assays on a large bank of VSMCs (n>1499). Expression quantitative trait locus and splicing quantitative trait locus analyses were performed to identify genes with an expression that was influenced by CAD-associated variants. To identify candidate causal genes for CAD, we ascertained colocalizations of VSMC expression quantitative trait locus signals with CAD association signals by performing causal variants identification in associated regions analysis and the summary data-based mendelian randomization test. Druggability analysis was then performed on the candidate causal genes. CAD risk variants were tested for associations with VSMC proliferation, migration, and apoptosis. Collective effects of multiple CAD-associated variants on VSMC behavior were estimated by polygenic scores., Results: Approximately 60% of the known CAD-associated variants showed statistically significant expression quantitative trait locus or splicing quantitative trait locus effects in VSMCs. Colocalization analyses identified 84 genes with expression quantitative trait locus signals that significantly colocalized with CAD association signals, identifying them as candidate causal genes. Druggability analysis indicated that 38 of the candidate causal genes were druggable, and 13 had evidence of drug-gene interactions. Of the CAD-associated variants tested, 139 showed suggestive associations with VSMC proliferation, migration, or apoptosis. A polygenic score model explained up to 5.94% of variation in several VSMC behavior parameters, consistent with polygenic influences on VSMC behavior., Conclusions: This comprehensive analysis shows that a large percentage of CAD loci can modulate gene expression in VSMCs and influence VSMC behavior. Several candidate causal genes identified are likely to be druggable and thus represent potential therapeutic targets.

Objectives: Several new cancer therapies targeting signaling pathways involved in the growth and progression of cancer cells were developed as personalized medicine. Our study aimed to identify epidermal growth factor receptor (EGFR) mutations for TKI treatment in non-small-cell lung cancer (NSCLC) Tunisian patients. Methods: Analysis of the TKI sensitivity mutations in exons 18 to 21 of the EGFR gene and exon 15 of the B-raf gene was performed in 79 formalin fixed-paraffin embedded (FFPE) NSCLC samples using pyrosequencing. Results: EGFR mutations were detected in 34 cases among 79 (43%), with the predominance of the L861Q in exon 21 found in 35.3% of the cases (12 out of 34). Deletions in exon 19 were found in 8 cases (23.5%), and only one young male patient had the T790M mutation. Three patients harbored composite EGFR mutations (p.E746_A750del/p.L861R, p.E746_S752>V/p.S768I, and p.G719A/p.L861Q). Furthermore, the EGFR mutated status was significantly more frequent in female patients (p = 0.019), in non-smoker patients (p = 0.008), and in patients with metastasis (p = 0.044). Moreover, the B-raf V600E was identified in 5 EGFR negative patients among 39 analyzed samples (13.15%). Conclusion: The p.L861Q localized in exon 21 of the EGFR gene was the most common mutation identified in our patients (35.3%), whereas the “classic” EGFR mutations such as Del19 and p.L858R were found in 23.5% and 11.7% of the cases, respectively. Interestingly, most of p.L861X mutation-carrying patients showed good response to TKI treatment. Altogether, our findings suggest a particular distribution of the EGFR-TKIs sensitivity mutations in Tunisian NSCLC patients.

Research Background: Microalgae represent an emergent sustainable source of bioactive compounds such as antioxidants, vitamins, minerals and polyunsaturated fatty acids that can ameliorate the nutritional characteristics of foods. The biochemical composition of microalgae could be modulated by varying the culture conditions to enhance the accumulation of biomolecules of interest. The aim of this work is to optimise the nutri-functional properties of two microalgae that can be used in food production., Experimental Approach: Nannochloropsis gaditana L2 and Chlorella sp. SM1 were screened for growth, biochemical composition and radical scavenging activity employing four different growth media (algal, BG-11, f/2 and Conway) with different nutrient composition. The feasibility of using Chlorella sp. SM1 cultivated in BG-11 medium, in an under-investigated Mediterranean dairy product ricotta cheese and its effect on the sensory attributes was investigated. Additionally, Arthrospira platensis was used as reference in sensory analysis., Results and Conclusions: Nitrate- and phosphate-rich media (BG-11 and algal) enhanced the biomass productivity. However, the highest lipid production (23.10 and 11.86 mg/(L·day) by strains SM1 and L2 respectively) and carbohydrate content (34.79 and 44.84% by SM1 and L2 respectively) were obtained with the nitrate-deficient f/2 medium. Regardless of the used medium, the lipid profile of Chlorella sp. SM1 and N. gaditana L2 remained adequate for different applications with >50% C16-18 as the main fatty acids. Significant increase in oleic acid (C18:1) content was recorded in response to nitrogen deficiency, being the highest in SM1 in f/2 medium (34%). Nitrogen deficiency was also found to enhance phenolic compound (expressed as gallic acid equivalents, 48.8 and 35.1 mg/g in SM1 and L2 respectively) and carotenoid contents (2.2 and 2.0 mg/g in SM1 and L2 respectively). Due to its interesting antioxidant potential, Chlorella sp. SM1 was used at different mass fractions (0.2, 1 and 1.5%) to enrich the ricotta cheese. The sample with 0.2% Chlorella sp. SM1 was found to give the most appreciated product., Novelty and Scientific Contribution: This study presents the production of an innovative ricotta cheese using Chlorella sp. as a functional ingredient, without altering the manufacturing procedure, while maintaining acceptable sensorial characteristics. The biochemical composition of the used strains varied depending on the culture medium composition, which enabled the accumulation of phytonutrients of interest., Competing Interests: CONFLICT OF INTEREST The authors confirm that they have no conflicts of interest with respect to the work described in this manuscript.

Objectives: The aim of our study was to assess the knowledge of parturients about breastfeeding and to analyze the factors influencing this level of knowledge., Methods: It was a prospective, descriptive and analytical, cross-sectional study carried out over a period of six months including 500 parturients who gave birth at the maternity unit of the National Center for Maternity and Neonatalogy of Tunis during the period from July to December 2020. Sociodemographic, obstetrical and breastfeeding related data were collected through individual interviews carried out before leaving the postpartum service using a questionnaire written in Tunisian dialectal Arabic., Results: The average age of the mothers was 31.5 years. Housewives accounted for 73.3 % of cases. Forty-four percent of the women surveyed were first-time mothers. The main source of information was relatives (74.8 %). The prevalence of breastfeeding was 93.8 %. Knowledge about breasfeeding was insufficient in 76.4 % of cases. The level of knowledge was insufficient in particular regarding signs of effective breastfeeding, signs of awakening of the newborn, lactogenesis, and diet of breastfeeding women. Unemployment of the parturient age over 30 and primiparity negatively influenced the knowledge of parturients., Conclusion: The level of knowledge of parturients was heterogeneous but generally insufficient. Health education programs on MA targeting subjects most at risk of insufficient knowledge should be put in place in order to improve the knowledge base and hopefully improve the breastfeeding rates., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

The aim of this work was to develop functional couscous in a traditional Tunisian manner (hand rolling), enriched in algae biomass (6% w/w ). Four Chlorella vulgaris ( C. vulgaris ) biomasses and one mixture of C. vulgaris and two macroalgae biomasses ( Ulva rigida and Fucus vesiculosus ) were used. The C. vulgaris strain was subjected to random mutagenesis and different culture conditions (Allmicroalgae), resulting in different pigmentations and biochemical compositions. Couscous samples were characterized in terms of nutritional properties, oscillatory rheology properties and digestibility. All biomasses provided a significant supplementation of nutrients and excellent acceptance. The enrichment resulted in lower firmness, higher viscoelastic functions (G' and G″) and a significant improvement in the cooking quality. Major differences between couscous samples with different microalgae were observed in protein and mineral contents, fully meeting Regulation (EC) No. 1924/2006 requirements for health claims made on foodstuffs. The amount of digested proteins was also higher in algae-containing samples. The fatty acid profile of the enriched couscous varied in a biomass-specific way, with a marked increase in linolenic acid (18:3 ω3) and a decrease in the ω6/ω3 ratio. Sensory analysis revealed that microalgae-containing products could compete with conventional goods with an added advantage, that is, having an ameliorated nutritional value using algae as a "trendy" and sustainable ingredient.