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In-service teachers implementing a science, technology, engineering, and math (STEM) curriculum centered on computational thinking (CT) work with unique content and pedagogical experiences. Understanding how curriculum design and teacher professional development affect curriculum implementation can help researchers understand the critical aspects of supporting the teacher in "just right" ways to learn and teach an embedded CT curriculum. We qualitatively analyzed 22 teachers' discourse through a case study approach. We identified how CT is afforded and constrained through curriculum design and teacher professional development support. Teachers expressed that the supports that were critical to their confidence and perceived ability to teach CT were (a) a program that provides "just right" support, (b) a program that provides options and individualization, (c) an internal personal identity that embraces continual innovation and learning, and (d) an educational system that is encouraging and supportive of the effort and creativity it requires to implement innovative and intensive embedded CT programs. These findings can inform how to support teachers to integrate CT within existing STEM curricula as a core scientific and mathematical practice and to foster student interest in computer science.

Abstract Due to the fast-changing global climate, conventional agricultural systems have to deal with more unpredictable and harsh environmental conditions leading to compromise food production. The application of phytonanotechnology can ensure safer and more sustainable crop production, allowing the target-specific delivery of bioactive molecules with great and partially explored positive effects for agriculture, such as an increase in crop production and plant pathogen reduction. In this study, the effect of free pterostilbene (PTB) and poly(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) loaded with pterostilbene was investigated on Solanum lycopersicum L. metabolism. An untargeted NMR-based metabolomics approach was used to examine primary and secondary metabolism whereas a targeted HPLC–MS/MS-based approach was used to explore the impact on defense response subjected to anti-oxidant effect of PTB, such as free fatty acids, oxylipins and them impact on hormone biosynthesis, in particular salicylic and jasmonic acid. In tomato leaves after treatment with PTB and PLGA NPs loaded with PTB (NPs + PTB), both NPs + PTB and free PTB treatments increased GABA levels in tomato leaves. In addition, a decrease of quercetin-3-glucoside associated with the increase in caffeic acid was observed suggesting a shift in secondary metabolism towards the biosynthesis of phenylpropanoids and other phenolic compounds. An increase of behenic acid (C22:0) and a remodulation of oxylipin metabolism deriving from the linoleic acid (i.e. 9-HpODE, 13-HpODE and 9-oxo-ODE) and linolenic acid (9-HOTrE and 9-oxoOTrE) after treatment with PLGA NPs and PLGA NPs + PTB were also found as a part of mechanisms of plant redox modulation. To the best of our knowledge, this is the first study showing the role of PLGA nanoparticles loaded with pterostilbene in modulating leaf metabolome and physiology in terms of secondary metabolites, fatty acids, oxylipins and hormones. In perspective, PLGA NPs loaded with PTB could be used to reshape the metabolic profile to allow plant to react more quickly to stresses.

The resistance to thyroid hormone syndrome (RTHβ) occurs uncommonly and requires a high level of clinical suspicion and specific investigations to reach a precise diagnosis and to avoid unnecessary and potentially harmful therapies. We report a case of a young male patient referred to our unit for SARS-CoV-2 infection and atrial fibrillation with elevated thyroid hormones and non-suppressed thyroid-stimulating hormone (TSH), for which antithyroid therapy was prescribed. A mood disorder was reported in the medical history. The family history was unknown as the patient was adopted. Thyroid-specific antibodies were undetectable, and thyroid ultrasound revealed a normal thyroid gland without nodules. After the resolution of SARS-CoV-2 infection, the diagnostic workup continued, and the pituitary MRI revealed a small area ascribable to a microadenoma. Due to atrial fibrillation, the execution of the T3 test was contraindicated. The octreotide long-acting release (LAR) test showed an initial reduction of free thyroid hormones levels at first administration, which was consistent with the presence of a TSH-secreting pituitary tumour, although an escape from the response was observed after the following two injections of octreotide LAR. Indeed, the genetic investigation revealed a variant in heterozygosity of the THRβ gene (Pro453Ser), thus leading to an RTHβ diagnosis, and, therefore, medical treatment with triiodothyroacetic acid was initiated. After 2 years from the SARS-CoV-2 infection, the patient continues the follow-up at our outpatient clinic, and no other medical interventions are needed.

Licorice processing waste was pyrolyzed at different temperatures (500 and 700 °C) to obtain biochar (BC500 and BC700) for use as a slow-release fertilizer on Solanum lycopersicum. The materials were characterized through BET analysis, SEM, elemental analysis, pHzc, and pyrolysis temperature effect was evaluated. The biochars were functionalized by the impregnation method to enrich them with nitrogen, phosphorus, and potassium (NPK), and desorption tests were performed in aqueous solution at different pHs (5 and 7). The pseudo-second-order model described well the release of all 3 macronutrients tested, BC500 was found to have slower release kinetics due to smaller pore size, reaching adsorption/desorption equilibrium after 14 days, compared with 10 for BC700, Kdes were lower in all 3 cases and NPK content was higher, initial pH did not change the release kinetics. BC500 was selected as an agricultural soil conditioner by testing at both different dosages of BC (0–25 %) and different NPK ratios (3:1:4 and 4:1:3). The treatment significance was evaluated. The best treatment resulted in BC dosage of 25 % nutrient ratio 4:1:3 which increased, compared to the control, total chlorophyll content (+38 %) and carotenoids (+15 %).

The current study investigates parents’ perceptions of their child’s assessment, focusing on their responses to the Italian version of the Parents’ Experience of Assessment Scale (QUEVA-G). Twenty parents, who voluntarily agreed to be contacted after completing the questionnaire, participated in qualitative interviews to gain deeper insights into their assessment experiences. A thematic analysis was conducted on the interview transcriptions, highlighting three primary domains of parental experience: (1) parental perceptions of the assessment process; (2) effects of the assessment; and (3) parental perceptions of their relationship with their children’s teachers. The findings indicate that the QUEVA-G accurately captures most areas of interest as well as reveals unexplored aspects.

This paper describes the psychometric properties of the Italian version of the Parent Experience of Assessment Scale. Overall, 185 participants took part in the study. Confirmatory factor analysis and structural equation modeling tested the scale structure and its relationship with clients’ satisfaction. Reliability and multivariate analysis of variance measured the factors’ consistency and the differences among different typologies of assessment. Results replicated the original five factors structure of the scale (Parent-Assessor Relationship and Collaboration; New Understanding of the Child; Child-Assessor Relationship; Systemic Awareness; Negative Feelings). Full scale and individual factors’ reliability ranged from high to excellent. Structural equation modeling showed that Parent-Assessor Relationship and Collaboration and New Understanding of the Child factors had the strongest direct effects on parents’ General Satisfaction, measured by the Client Satisfaction Questionnaire. A multivariate analysis of variance showed that the type of assessment, the children’s age and the way the scale was completed impacted on the outcomes of the QUEVA-G. Results suggest that the Italian version of the Parent Experience of Assessment Scale is a valid and reliable tool for assessing parents’ experience of their child’s assessment.

In compensation for the diminishing time spent on science in school, afterschool programs are taking an increasingly larger role in STEM education, with over 69 percent of programs in the U.S. offering some type of STEM programming (Afterschool Alliance, 2015). As the demand for afterschool STEM programs increases, so too does the need for trained educators and staff members. Access to high-quality, accessible, and inexpensive professional development is widely recognized as foundational to implementing high-quality programming that supports and enriches youth (e.g., Miller & Hall, 2007; Vandell, Reisner, & Pierce, 2007). Though afterschool staff and leaders may appreciate how professional development benefits program quality, implementation brings a whole set of challenges (Bradshaw, 2015). Many afterschool educators do not have flexibility in their jobs to attend off-site trainings, or they work multiple jobs and so do not have the time to travel. One study found that, although afterschool staff generally found professional development useful, only 26 percent had regular opportunities to participate (Huang & Dietel, 2011). This article shares promising practices in virtual professional development for afterschool educators. According to the authors, their experience is with STEM professional development, their strategies can be adapted to other disciplines as well.

The yellow pea (Pisum sativum L. ‘Eso’, sin. Lathyrus oleracaeus Lam.(YP)) is an annual herbaceous plant that belongs to the Fabaceae family. Peas, along with other legumes, are an excellent source of proteins and essential amino acids; the yellow variety is known for maintaining a good protein profile even if subjected to industrial processing. However, the presence of antinutrients, such as phytates and oligosaccharides, limits its consumption as a fresh legume to its use as a source of isolated proteins or for animal feed. The aim of the study is to evaluate the changes in the entire phytochemical profile of YP seeds as a function of the harvest time. YPs harvested at about 40, 50, 60, and 70 days from sowing were examined by high-resolution NMR spectroscopy employing 1H-NMR, 1H-1H TOCSY, and 1H-13C HSQC. In total, 40 molecular species were identified and quantified; it was observed that there was a monotonous decrease in amino acids, carbohydrates, and secondary metabolites as a function of time. Antinutrient levels increased, but only in later sampling times. This study identified the optimal harvest time for yellow peas “Eso” in the fortieth day from sowing, adding new information about the best nutritional outcome for humans.

BackgroundHypothermic neonates need to be promptly rewarmed but there is no strong evidence to support a rapid or a slow pace of rewarming. This study aimed to investigate the rewarming rate and its associations with clinical outcomes in hypothermic neonates born in a low-resource setting.MethodsThis retrospective study evaluated the rewarming rate of hypothermic inborn neonates admitted to the Special Care Unit of Tosamaganga Hospital (Tanzania) in 2019–2020. The rewarming rate was calculated as the difference between the first normothermic value (36.5–37.5°C) and the admission temperature, divided by the time elapsed. Neurodevelopmental status at 1 month of age was assessed using the Hammersmith Neonatal Neurological Examination.ResultsMedian rewarming rate was 0.22°C/h (IQR: 0.11–0.41) in 344/382 (90%) hypothermic inborn infants, and was inversely correlated to admission temperature (correlation coefficient −0.36, p

Abstract Background Accurate gestational age (GA) determination allows correct management of high-risk, complicated or post-date pregnancies and prevention or anticipation of prematurity related complications. Ultrasound measurement in the first trimester is the gold standard for GA determination. In low- and middle-income countries elevated costs, lack of skills and poor maternal access to health service limit the availability of prenatal ultrasonography, making it necessary to use alternative methods. This study compared three methods of GA determination: Last Normal Menstrual Period recall (LNMP), New Ballard Score (NBS) and New Ballard Score corrected for Birth Weight (NBS + BW) with the locally available standard (Ultrasound measurement in the third trimester) in a low-resource setting (Tosamaganga Council Designated Hospital, Iringa, Tanzania). Methods All data were retrospectively collected from hospital charts. Comparisons were performed using Bland Altman method. Results The analysis included 70 mother-newborn pairs. Median gestational age was 38 weeks (IQR 37–39) according to US. The mean difference between LNMP vs. US was 2.1 weeks (95% agreement limits − 3.5 to 7.7 weeks); NBS vs. US was 0.2 weeks (95% agreement limits − 3.7 to 4.1 weeks); NBS + BW vs. US was 1.2 weeks (95% agreement limits − 1.8 to 4.2 weeks). Conclusions In our setting, NBS + BW was the least biased method for GA determination as compared with the locally available standard. However, wide agreement bands suggested low accuracy for all three alternative methods. New evidence in the use of second/third trimester ultrasound suggests concentrating efforts and resources in further validating and implementing the use of late pregnancy biometry for gestational age dating in low and middle-income countries.

Abstract Botrytis cinerea, responsible for grey mold diseases, is a pathogen with a broad host range, affecting many important agricultural crops, in pre and post harvesting of fruits and vegetables. Commercial fungicides used to control this pathogen are often subjected to photolysis, volatilization, degradation, leaching, and runoff during application. In this context, the use of a delivery system, based on poly (lactic-co-glycolic acid) nanoparticles (PLGA NPs) represents an innovative approach to develop new pesticide formulations to successfully fight B. cinerea infections. In order to study NPs uptake, B. cinerea conidia and mycelium were treated with PLGA NPs loaded with the high fluorescent probe coumarin 6 (Cu6-PLGA NPs) and analyzed under ApoTome fluorescence microscopy. The observations revealed that 50 nm Cu6-PLGA NPs penetrated into B. cinerea conidia and hyphae, as early as 10 min after administration. Pterostilbene, a natural compound, and fluopyram, a synthetic antifungal, were entrapped in PLGA NPs, added to B. cinerea conidia and mycelium, and their antifungal activity was tested. The results revealed that the compounds loaded in NPs exhibited a higher activity against B. cinerea. These results lay the foundations for the use of PLGA NPs as a new strategy in plant pest management.

Abstract Keratinocytes, the main cell type of the skin, are one of the most exposed cells to environmental factors, providing a first defence barrier for the host and actively participating in immune response. In fact, keratinocytes express pattern recognition receptors that interact with pathogen associated molecular patterns and damage associated molecular patterns, leading to the production of cytokines and chemokines, including interleukin (IL)-6. Herein, we investigated whether mechanical energy transported by low intensity ultrasound (US) could generate a mechanical stress able to induce the release of inflammatory cytokine such IL-6 in the human keratinocyte cell line, HaCaT. The extensive clinical application of US in both diagnosis and therapy suggests the need to better understand the related biological effects. Our results point out that US promotes the overexpression and secretion of IL-6, associated with the activation of nuclear factor-κB (NF-κB). Furthermore, we observed a reduced cell viability dependent on exposure parameters together with alterations in membrane permeability, paving the way for further investigating the molecular mechanisms related to US exposure.

In this study, we investigated the biostimulant effect of fungal culture filtrates obtained from Chaetomium globosum and Minimedusa polyspora on growth performance and metabolomic traits of chicory (Cichorium intybus) plants. For the first time, we showed that M. polyspora culture filtrate exerts a direct plant growth-promoting effect through an increase of biomass, both in shoots and roots, and of the leaf area. Conversely, no significant effect on morphological traits and biomass yield was observed in C. intybus plants treated with C. globosum culture filtrate. Based on 1H-NMR metabolomics data, differential metabolites and their related metabolic pathways were highlighted. The treatment with C. globosum and M. polyspora culture filtrates stimulated a common response in C. intybus roots involving the synthesis of 3-OH-butyrate through the decrease in the synthesis of fatty acids and sterols, as a mechanism balancing the NADPH/NADP+ ratio. The fungal culture filtrates differently triggered the phenylpropanoid pathway in C. intybus plants: C. globosum culture filtrate increased phenylalanine and chicoric acid in the roots, whereas M. polyspora culture filtrate stimulated an increase of 4-OH-benzoate. Chicoric acid, whose biosynthetic pathway in the chicory plant is putative and still not well known, is a very promising natural compound playing an important role in plant defense. On the contrary, benzoic acids serve as precursors for a wide variety of essential compounds playing crucial roles in plant fitness and defense response activation. To the best of our knowledge, this is the first study that shows the biostimulant effect of C. globosum and M. polyspora culture filtrates on C. intybus growth and metabolome, increasing the knowledge on fungal bioresources for the development of biostimulants.

Xanthones are a class of secondary metabolites produced by plant organisms. They are characterized by a wide structural variety and numerous biological activities that make them valuable metabolites for use in the pharmaceutical field. This review shows the current knowledge of the xanthone biosynthetic pathway with a focus on the precursors and the enzymes involved, as well as on the cellular and organ localization of xanthones in plants. Xanthone biosynthesis in plants involves the shikimate and the acetate pathways which originate in plastids and endoplasmic reticulum, respectively. The pathway continues following three alternative routes, two phenylalanine-dependent and one phenylalanine-independent. All three routes lead to the biosynthesis of 2,3′,4,6-tetrahydroxybenzophenone, which is the central intermediate. Unlike plants, the xanthone core in fungi and lichens is wholly derived from polyketide. Although organs and tissues synthesizing and accumulating xanthones are known in plants, no information is yet available on their subcellular and cellular localization in fungi and lichens. This review highlights the studies published to date on xanthone biosynthesis and trafficking in plant organisms, from which it emerges that the mechanisms underlying their synthesis need to be further investigated in order to exploit them for application purposes.

In the present work, nineteen analogues of 1-[(2,2-Diphenyl-1,3-dioxolan-4-yl)methyl]-4-(2-methoxyphenyl)piperazine 5 and N-[2-(2-Methoxyphenoxy)ethyl]-2,2-diphenyl-1,3-dioxolane-4-methanamine 18 were synthesized. The compounds were tested for binding affinity at 5-HT1AR and α1-AR subtypes. They were also tested using functional assays as α1-AR antagonists and the most promising were tested for functional activity at 5-HT1AR, where they were shown to behave as agonists. The results highlight that the replacement of the 1,3-dioxolane ring with a 1,3-oxathiolane or a 1,3-dithiolane moiety leads to an overall reduction in in-vitro affinity at the α1-AR, while affinity, potency and efficacy were strongly enhanced at the 5-HT1A receptor. Overall, the nitrogen-containing aromatic moieties scarcely affect the affinity at the 5-HT1A receptor, while reducing potency and increasing efficacy. The oxidation of the sulphur atom in the 1,3-oxathiolane to give sulfoxides and solfones has a negative effect on affinity and potency at both receptor systems. Regardless of the effect on the other parameters, selectivity toward 5-HT1AR with respect to the α1-AR is often favoured, but never the contrary. The most striking result is the inversion of selectivity. In fact, while the lead 5 is 100-fold selective for α1-AR, the new derivatives, although to differing degrees, are selective for 5-HT1AR.

Abstract Ultrasound (US) induced transient membrane permeabilisation has emerged as a hugely promising tool for the delivery of exogenous vectors through the cytoplasmic membrane, paving the way to the design of novel anticancer strategies by targeting functional nanomaterials to specific biological sites. An essential step towards this end is the detailed recognition of suitably marked nanoparticles in sonoporated cells and the investigation of the potential related biological effects. By taking advantage of Synchrotron Radiation Fourier Transform Infrared micro-spectroscopy (SR-microFTIR) in providing highly sensitive analysis at the single cell level, we studied the internalisation of a nanoprobe within fibroblasts (NIH-3T3) promoted by low-intensity US. To this aim we employed 20 nm gold nanoparticles conjugated with the IR marker 4-aminothiophenol. The significant Surface Enhanced Infrared Absorption provided by the nanoprobes, with an absorbance increase up to two orders of magnitude, allowed us to efficiently recognise their inclusion within cells. Notably, the selective and stable SR-microFTIR detection from single cells that have internalised the nanoprobe exhibited clear changes in both shape and intensity of the spectral profile, highlighting the occurrence of biological effects. Flow cytometry, immunofluorescence and murine cytokinesis-block micronucleus assays confirmed the presence of slight but significant cytotoxic and genotoxic events associated with the US-nanoprobe combined treatments. Our results can provide novel hints towards US and nanomedicine combined strategies for cell spectral imaging as well as drug delivery-based therapies.

Background: The poor quality of care received by mothers and neonates in many limited-resource countries represents a main determinant of newborn mortality. Small and sick hospitalized newborns are the highest-risk population, and they should be one of the prime beneficiaries of quality-of-care interventions. This study aimed to evaluate the impact on neonatal mortality of quality improvement interventions which were implemented at Tosamaganga Council Designated Hospital, Iringa, Tanzania, between 2016 and 2020. Methods: A retrospective comparison between pre- and post-intervention periods was performed using the chi-square test and Fisher’s exact test. Effect sizes were reported as odds ratios with 95% confidence intervals. Results: The analysis included 5742 neonates admitted to the Special Care Unit (2952 in the pre-intervention period and 2790 in the post-intervention period). A decrease in mortality among infants with birth weight between 1500 and 2499 g (overall: odds ratio 0.49, 95% confidence interval 0.27–0.87; inborn: odds ratio 0.50, 95% confidence interval 0.27–0.93) was found. The analysis of cause-specific mortality showed a decrease in mortality for asphyxia (odds ratio 0.33, 95% confidence interval 0.12–0.87) among inborn infants with birth weight between 1500 and 2499 g. Conclusions: A quality improvement intervention was associated with decreased mortality among infants with birth weight between 1500 and 2499 g. Further efforts are needed to improve prognosis in very-low-birth-weight infants.

Multi-responsive nanomaterials based on the self-limited assembly of plasmonic nanoparticles are of great interest due to their widespread employment in sensing applications. We present a thorough investigation of a hybrid nanomaterial based on the protein-mediated aggregation of gold nanoparticles at varying protein concentration, pH and temperature. By combining Small Angle X-ray Scattering with extinction spectroscopy, we are able to frame the morphological features of the formed fractal aggregates in a theoretical model based on patchy interactions. Based on this, we established the main factors that determine the assembly process and their strong correlation with the optical properties of the assemblies. Moreover, the calibration curves that we obtained for each parameter investigated based on the extinction spectra point out to the notable flexibility of this nanomaterial, enabling the selection of different working ranges with high sensitivity. Our study opens for the rational tuning of the morphology and the optical properties of plasmonic assemblies to design colorimetric sensors with improved performances.

Recent political events in the European Union (EU) highlighted a growing dissatisfaction of citizens in several EU regions with the EU institutions’ management of socio-economic and financial challenges. This eventually led to a political legitimization crisis, whose drivers are partially shared among EU regions and partially area-specific. However, the relation between citizens’ identification with the EU project and the regions’ characteristics has not been analysed yet. In this article, we fill in this gap by addressing tree research questions: i) To what extent do EU citizens identify with Europe and the EU project? ii) Do European regions have different patterns and level of identification?, and iii) Are the results driven by specific socio-economic variables? Answering these questions is crucial to inform a more inclusive and resilient design of the EU Cohesion Policy in a crucial period for reforming the EU. To this purpose, we develop a novel probabilistic classification model, IdentEU, which embeds with the concept of individual identification with Europe. We use micro-level data from a survey implemented within the PERCEIVE project. We find that the influencing variables that mostly affect (citizens and) regions’ identification with the European project are trust in the EU institutions; the effectiveness of EU Cohesion Policy and spending; the level of corruption. These issues gain relevance at the light of three main challenges that affected the EU socio-economic development path in the last decade, i.e. the 2008 financial crisis, the globalization process, and Brexit.

Background: Deviations from normothermia affect early mortality and morbidity, but the impact on neurodevelopment of the survivors is unclear. We aimed to investigate the relationship between neonatal temperature at admission and the risk of cerebral palsy (CP) at one month of age in a low-resource setting. Methods: This retrospective study included all inborn neonates admitted to the Special Care Unit of Tosamaganga Hospital (Tanzania) between 1 January 2019 and 31 December 2020. The neurological examination at one month of age was performed using the Hammersmith method. The relationship between the admission temperature and the risk of CP was investigated using logistic regression models, with temperature modeled as the non-linear term. Results: High/moderate risk of CP was found in 40/119 (33.6%) of the neonates at one month of age. A non-linear relationship between the admission temperature and moderate/high risk of CP at one month of age was found. The lowest probability of moderate/high risk of CP was estimated at admission temperatures of between 35 and 36 °C, with increasing probability when departing from such temperatures. Conclusions: In a low-resource setting, we found a U-shaped relationship between the admission temperature and the risk of CP at one month of life. Expanding the analysis of the follow-up data to 12–24 months of age would be desirable in order to confirm and strengthen such findings.

The use of organic substances in integrated pest management can contribute to human- and environment-safe crop production. In the present work, a combination of organic biostimulants (Fullcrhum Alert and BioVeg 500) and an inorganic corroborant (Clinogold, zeolite) was tested for the effects on the plant response to the quarantine pest tomato leaf curl New Delhi virus (ToLCNDV). Biostimulants were applied to healthy and infected greenhouse-grown zucchini plants, and the vegetative parameters and viral titer were evaluated. Although no antiviral effects were observed in terms of both virus replication and symptom expression, these biostimulants were shown to influence plant fitness. A significant increase in biomass and in leaf, flower, and fruit production was induced in both healthy and infected plants. Biostimulants also enhanced the production of metabolites commonly involved in plant response to virus infection, such as carbohydrates, phenylpropanoids and free amino acids. These results encourage new field trials to evaluate the actual productivity of infected plants after treatments and the possible application of organic biostimulants in agriculture.

We investigated the efficacy of different probiotics and bacteria used in dairy products to prevent the development of obesity and ameliorate established obesity in C57BL/6J mice fed a high fat diet (HFD). For the preventive efficacy, Lactobacillus rhamnosus GG, L. acidophilus LA1/K8, a mixture of Bifidobacterium lactis Bi1, B. breve Bbr8 and B. breve BL10 (B. mix), or a mixture of L. bulgaricus Lb2 and S. termophilus Z57 were given to mice before (2 weeks) and together with HFD (12 weeks). B. mix was selected for its anti-obesity activity and assessed in established obesity by feeding mice the HFD for 24 weeks and B. mix for the last 12 weeks. We found that B. mix was able to both prevent and ameliorate established obesity by reducing weight gain, adipose tissue fat accumulation, adipocyte size, and macrophage and CD4+ T cell infiltration, improving lipid profile and regulating leptin and cytokine secretion.

Red beetroot (RB) is a well-known health-promoting food consumed worldwide. RB is commonly used in food processing and manufacturing thanks to the high content of components that can also be employed as natural coloring agents. These bioactive molecules vary their concentration depending on beetroot seasonality, harvest time and climate conditions. The first objective of this study was to evaluate the variation of the RB phytochemical profile related to the root development during three different harvest times, using an 1H-NMR-based metabolomic approach. Changes of carbohydrates and secondary metabolite concentrations were observed from July to September. Secondly, we compared the metabolic profiles of the final processed beet juices in three different production years to observe the effect of climate conditions on the RB’s final product metabotype. A PCA analysis performed on juice extracts showed that production years 2016 and 2017 were characterized by a high content of choline and betaine, while 2018 by a high content of amino acids and dopamine and a low content of inorganic nitrates. This study suggests that the harvest time and roots growth conditions could be used to modulate the RB phytochemical profile, according to the final requirements of use, food or coloring agent source.

Abstract Among different therapeutic applications of Ultrasound (US), transient membrane sonoporation (SP) - a temporary, non-lethal porosity, mechanically induced in cell membranes through US exposure - represents a compelling opportunity towards an efficient and safe drug delivery. Nevertheless, progresses in this field have been limited by an insufficient understanding of the potential cytotoxic effects of US related to the failure of the cellular repair and to the possible activation of inflammatory pathway. In this framework we studied the in vitro effects of very low-intensity US on a human keratinocyte cell line, which represents an ideal model system of skin protective barrier cells which are the first to be involved during medical US treatments. Bioeffects linked to US application at 1 MHz varying the exposure parameters were investigated by fluorescence microscopy and fluorescence activated cell sorting. Our results indicate that keratinocytes undergoing low US doses can uptake drug model molecules with size and efficiency which depend on exposure parameters. According to sub-cavitation SP models, we have identified the range of doses triggering transient membrane SP, actually with negligible biological damage. By increasing US doses we observed a reduced cells viability and an inflammatory gene overexpression enlightening novel healthy relevant strategies.

The merging of the molecular specificity of Raman spectroscopy with the extraordinary optical properties of metallic nanoarchitectures is at the heart of Surface Enhanced Raman Spectroscopy (SERS), which in the last few decades proved its worth as powerful analytical tool with detection limits pushed to the single molecule recognition. Within this frame, SERS-based nanosensors for localized pH measurements have been developed and employed for a wide range of applications. Nevertheless, to improve the performances of such nanosensors, many key issues concerning their assembling, calibration and stability, that could significantly impact on the outcome of the pH measurements, need to be clarified. Here, we report on the detailed characterization of a case study SERS-active pH nanosensor, based on the conjugation of gold nanoparticles with the pH-sensitive molecular probe 4-mercaptobenzoic acid (4MBA). We analyzed and optimized all the aspects of the synthesis procedure and of the operating conditions to preserve the sensor stability and provide the highest responsiveness to pH. Exploiting the dependence of the SERS spectrum on the protonation degree of the carboxylic group at the edge of the 4MBA molecules, we derived a calibration curve for the nanosensor. The extrapolated working point, i.e., the pH value corresponding to the highest sensitivity, falls at pH 5.6, which corresponds to the pKa value of the molecule confined at the nanoparticle surface. A shift of the pKa of 4MBA, observed on the molecules confined at the nanostructured interface respect to the bulk counterpart, unveils the opportunity to assembly a SERS-based pH nanosensor with the ability to select its working point in the sensitivity region of interest, by acting on the nanostructured surface on which the molecular probe is confined. As a proof-of-concept, the nanosensor was successfully employed to measure the extracellular pH of normal and cancer cells, demonstrating the capability to discriminate between them.

The 5-hydroxytryptamine (5-HT1A) receptors represent an attractive target in drug discovery. In particular, 5-HT1A agonists and partial agonists are deeply investigated for their potential role in the treatment of anxiety, depression, ischaemic brain disorder and more recently, of pain. On the other hand, 5-HT1A antagonists have been revealed promising compounds in cognition disorders and, lately, in cancer. Thus, the discovery of 5HT1A ligands is nowadays an appealing research activity in medicinal chemistry. In this work, Comparative Molecular Fields Analysis (CoMFA) and Comparative Molecular Similarity Index Analysis (CoMSIA) were applied on an in-house library of 5-HT1A ligands bearing different chemical scaffolds in order to elucidate their affinity and selectivity for the target. Following this procedure, a number of structural modifications have been drawn for the development of much more effective 5-HT1AR ligands.

The carrot (Daucus carota L.), one of the most important vegetable crops in the world, is recognized as a source of different compounds with healthy properties. Due to their high content of anthocyanins, purple carrots have been used as a natural colorant source to face the increasing demand of consumers for non-synthetic products. However, the root developmental stage can greatly affect the phytochemical composition and, in this regard, the identification of chemical biomarkers for the optimal harvest time would be of paramount interest both from a nutritional point of view and for the agri-food industry. In the present work, the metabolic profiling of purple carrots was monitored over four months using high-resolution 1H NMR spectroscopy. Several metabolites were identified, and their quantitative variations allowed for the investigation of the carrot development processes. The metabolic profile analysis showed an increase in amino acid, NAD, and caffeic acid levels during carrot development. A more tardive harvest in December entailed an increase in levels of luteolin-7-O-glucoside, chlorogenic acid, falcarinol, and γ-aminobutyrate, and a decrease in carotenoids and ω-6 fatty acid. The results showed how the harvest time affects the composition in terms of flavonoids, phenols, and polyacetylenes, therefore, improving the bioactive compound content.

Phenolic compounds, the most widely distributed class of natural products in the plants, show several biological properties including antifungal activity. Phenolics contained in grapes can be classified in two main groups, flavonoids and non-flavonoids compounds. Variability and yield extraction of phenolic and polyphenolic compounds from different matrices of Vitis vinifera depends of cultivar, climate, soil condition and process technology. Unripe grapes, berry skins and seeds, leaves, canes and stems and not-fermented and fermented pomaces represent large reusable and valuable wastes from agricultural and agro-industrial processes. This review summarizes studies that examine the extraction method, chemical characterization, and antifungal activity of phenolic and polyphenolic compounds from edible and non-edible V. vinifera matrices against human fungal pathogens. In the world, around one billion people have fungal diseases related to skin, nail or hair and around 150 million have systemic diseases caused by fungi. Few studies on antifungal activity of plant extracts have been performed. This review provides useful information for the application of V. vinifera phenolics in the field of antifungals for human use.

Polymeric nanoparticle-based carriers are promising agents to deliver drugs to cells. Vitis vinifera phenolic compounds are known for their antifungal activity against Candida albicans. The aim of the present study was to investigate the antifungal activity of pterostilbene or crude extracts from non-fermented grape pomace, entrapped in poly(lactic-co-glycolic) acid nanoparticles (NPs), with diameters of 50 and 150 nm, on Candida biofilm. The fluorescent probe coumarin 6 was used to study the uptake of poly(lactic-co-glycolic)acid (PLGA) NPs in planktonic cells and biofilm. The green fluorescent signal of coumarin 6 was observed in Candida biofilm after 24 and 48 hours. Both pterostilbene and crude pomace extract entrapped in NPs exerted a significantly higher anti-biofilm activity compared to their free forms. The entrapment efficiency of both pterostilbene and crude pomace extract in PLGA NPs was ~90%. At 16 µg/mL, pterostilbene loaded in PLGA NPs reduced biofilm formation of 63% and reduced mature biofilm of 50%. Moreover, at 50 µg/mL, the pomace extract loaded in NPs reduced mature biofilm of 37%. These results strongly suggest that PLGA NPs are promising nanodevices for the delivery of antifungal drugs as the crude grape pomace extract, a by-product of white wine making.

A conditional β-convergence model and a distance-based weight matrix are used to analyse the economic convergence among European NUTS-2 regions over the period 1980-2006. A Spatial Autoregressive Model which identifies two spatial regimes and spatial dependence finds that the convergence process among EU regions is affected by polarization into two clusters defined both on a geographical and economic criterion, which converge at different rates towards different steady states. This result confirms the hypothesis that a methodology which uses spatial econometric techniques is needed to model spatial effects, and that otherwise the estimates are likely to be inefficient or even biased.