Neem oil nanoemulsions: characterisation and antioxidant activity

The aim of the present work is to develop nanoemulsions (NEs), nanosized emulsions, manufactured for improving the delivery of active pharmaceutical ingredients. In particular, nanoemulsions composed of Neem seed oil, contain rich bioactive components, and Tween 20 as nonionic surfactant were prepared. A mean droplet size ranging from 10 to 100nm was obtained by modulating the oil/surfactant ratio. Physicochemical characterisation was carried out evaluating size, f-potential, microviscosity, polarity and turbidity of the external shell and morphology, along with stability in simulated cerebrospinal fluid (CSF), activity of Neem oil alone and in NEs, HEp-2 cell interaction and cytotoxicity studies. This study confirms the formation of NEs by Tween 20 and Neem oil at different weight ratios with small and homogenous dimensions. The antioxidant activity of Neem oil alone and in NEs was comparable, whereas its cytotoxicity was strongly reduced when loaded in NEs after interaction with HEp-2 cells

Single-Walled Carbon Nanotubes as Enhancing Substrates for PNA-Based Amperometric Genosensors

A new amperometric sandwich-format genosensor has been implemented on single-walled carbon nanotubes screen printed electrodes (SWCNT-SPEs) and compared in terms of performance with analogous genoassays developed using the same methodology on non-nanostructured glassy carbon platforms (GC-SPE). The working principle of the genosensors is based on the covalent immobilization of Peptide Nucleic Acid (PNA) capture probes (CP) on the electrode surface, carried out through the carboxylic functions present on SWCNT-SPEs (carboxylated SWCNT) or electrochemically induced on GC-SPEs. The sequence of the CP was complementary to a 20-mer portion of the target DNA; a second biotin-tagged PNA signalling probe (SP), with sequence complementary to a different contiguous portion of the target DNA, was used to obtain a sandwich hybrid with an Alkaline Phosphatase-streptavidin conjugate (ALP-Strp). Comparison of the responses obtained from the SWCNT-SPEs with those produced from the non-nanostructured substrates evidenced the remarkable enhancement effect given by the nanostructured electrode platforms, achieved both in terms of loading capability of PNA probes and amplification of the electron transfer phenomena exploited for the signal transduction, giving rise to more than four-fold higher sensitivity when using SWCNT-SPEs. The nanostructured substrate allowed to reach limit of detection (LOD) of 71 pM and limit of quantitation (LOQ) of 256 pM, while the corresponding values obtained with GC-SPEs were 430 pM and 1.43 nM, respectively

Homogeneous Floquet time crystal protected by gauge invariance

We show that homogeneous lattice gauge theories can realize nonequilibrium quantum phases with long-range spatiotemporal order protected by gauge invariance instead of disorder. We study a kicked $\mathbb{Z}_2$-Higgs gauge theory and find that it breaks the discrete temporal symmetry by a period doubling. In a limit solvable by Jordan-Wigner analysis we extensively study the time-crystal properties for large systems and further find that the spatiotemporal order is robust under the addition of a solvability-breaking perturbation preserving the $\mathbb{Z}_2$ gauge symmetry. The protecting mechanism for the nonequilibrium order relies on the Hilbert space structure of lattice gauge theories, so that our results can be directly extended to other models with discrete gauge symmetries.Comment: 6 pages and 4 figures + 3 pages and 3 figures of Supplementary Informatio

Characterization of the vaginal microbiota in Italian women with endometriosis. Preliminary study

Purpose This cross-sectional study aims to assess the interplay between the vaginal microbiota and endometriosis. Methods 123 consecutive Italian fertile women, aged between 20 and 40 years old, were enrolled during a routine gynecological consultation; 24 were diagnosed with endometriosis and 99 did not complain of any gynecological disease. All women underwent a vaginal swab for the evaluation of the composition and diversity of vaginal microbiota by means of 16 s rDNA metagenomic sequencing. Results Compared to women with no gynecological disease, the vaginal microbiota in women with endometriosis showed a similar abundance of Lactobacillus spp.; however, a statistically significant lower abundance in the genera Pseudomonas (p < 0.01), Bifidobacterium (p < 0.05), Novispirillum (p < 0.0000001) and Sphingomonas (p < 0.0000001), and a statistically significant increase in the abundance of the genera Escherichia (p < 0.00001), Megasphaera (p < 0.00001), and Sneathia (p < 0.0001) were observed. Conclusions There is a complex interplay between vaginal microbiota composition and endometriosis, showing a distinct microbial signature in the bacterial genera usually found in dysbiosis

A Folding-Based Electrochemical Aptasensor for the Single-Step Detection of the SARS-CoV-2 Spike Protein

Efficient and timely testing has taken center stage in the management, control, and monitoring of the current COVID-19 pandemic. Simple, rapid, cost-effective diagnostics are needed that can complement current polymerase chain reaction-based methods and lateral flow immunoassays. Here, we report the development of an electrochemical sensing platform based on single-walled carbon nanotube screen-printed electrodes (SWCNT-SPEs) functionalized with a redox-tagged DNA aptamer that specifically binds to the receptor binding domain of the SARS-CoV-2 spike protein S1 subunit. Single-step, reagentless detection of the S1 protein is achieved through a binding-induced, concentration-dependent folding of the DNA aptamer that reduces the efficiency of the electron transfer process between the redox tag and the electrode surface and causes a suppression of the resulting amperometric signal. This aptasensor is specific for the target S1 protein with a dissociation constant (K-D) value of 43 +/- 4 nM and a limit of detection of 7 nM. We demonstrate that the target S1 protein can be detected both in a buffer solution and in an artificial viral transport medium widely used for the collection of nasopharyngeal swabs, and that no cross-reactivity is observed in the presence of different, non-target viral proteins. We expect that this SWCNT-SPE-based format of electrochemical aptasensor will prove useful for the detection of other protein targets for which nucleic acid aptamer ligands are made available

Multi-Platform Characterization of Cerebrospinal Fluid and Serum Metabolome of Patients Affected by Relapsing-Remitting and Primary Progressive Multiple Sclerosis

Background: Multiple sclerosis (MS) is a chronic immunemediated disease of the central nervous system with a highly variable clinical presentation and disease progression. In this study, we investigate the metabolomics profile of patients affected by relapsing-remitting MS (RRMS)and primary progressive MS (PPMS), in order to find potential biomarkers to distinguish between the two forms. Methods: Cerebrospinal Fluid CSF and blood samples of 34 patients (RRMS n = 22, PPMS n = 12) were collected. Nuclear magnetic resonance (H-1-NMR) and mass spectrometry (coupled with a gas chromatography and liquid chromatography) were used as analytical techniques. Subsequently, a multivariate statistical analysis was performed; the resulting significant variables underwent U-Mann-Whitney test and correction for multiple comparisons. Receiver Operating Characteristic ROC curves were built and the pathways analysis was conducted. Results: The analysis of the serum and the CSF of the two classes, allowed the identification of several altered metabolites (lipids, biogenic amines, and amino acids). The pathways analysis indicated the following pathways were affected: Glutathione metabolism, nitrogen metabolism, glutamine-glutamate metabolism, arginine-ornithine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis etc. Conclusion: The analysis allowed the identification of a set of metabolites able to classify RRMS and PPMS patients, each of whom express different patterns of metabolites in the two biofluids

Analytical methods and experimental quality in studies targeting carbonyls in electronic cigarette aerosols

We provide an extensive review of 14 studies (11 independent and three industry-funded) on emissions generated by Electronic Cigarettes (ECs), specifically focusing on the evaluation of carbonyls present in these emissions and emphasizing a meticulous evaluation of their analytical methods and experimental procedures. Since the presence of carbonyl by-products in EC aerosol is concerning, it is important to evaluate the reliability of emission studies quantifying these compounds by verifying their compliance with the following criteria of experimental quality: authors must 1) supply sufficient information on the devices and experimental procedures to allow for potentially reproducing or replicating the experiments, 2) use of appropriate puffing protocols that approach consumer usage as best as possible, 3) use of appropriate analytical methods and 4) usage of blank samples to avoid false positive detection. Outcomes were classified in terms of the fulfilment of these conditions as reliable in seven studies, partially reliable in five studies, and unreliable in two studies. However, only five studies used blank samples and six studies failed the reproducibility criterion. Carbonyl yields were far below their yields in tobacco smoke in all reproducible studies, even in the partially reliable ones, thus supporting the role of ECs (when properly tested and operated) as harm reduction products. This review highlights the necessity to evaluate the quality of laboratory standards in testing EC emissions to achieve an objective assessment of the risk profile of ECs

179. Correcting the Bleeding Phenotype in Hemophilia Ausing Lentivirally FVIII-Corrected Endothelial Cells Differentiated from Hemophilic Induced Pluripotent Stem Cell (iPSC)

Hemophilia A (HA) is a bleeding disorder caused by factor VIII (FVIII) gene mutations.Somatic cells can be reprogrammed to generate autologous, disease-free iPSCs, then differentiated into cell targetsrelevant for gene and cell therapy. Our aim is to develop a novel HA treatment strategy generating FVIII-corrected patient-specific iPSCs from peripheral blood cells anddifferentiating them into functional endothelial cells (ECs), secreting FVIII after transplantation

Branched-chain amino acids influence the immune properties of microglial cells and their responsiveness to pro-inflammatory signals

AbstractThe branched-chain amino acids (BCAAs) valine, leucine and isoleucine are essential amino acids involved in several important brain functions. Although commonly used as nutritional supplements, excessive intake of BCAAs might favour the establishment of neurotoxic conditions as indicated by the severe neurological symptoms characterising inherited disorders of BCAA catabolism such as maple syrup urine disease (MSUD). Recent evidence indicates that BCAAs induce excitotoxicity through mechanisms that require the presence of astrocytes. In the present study, we evaluated the effects of BCAAs on microglia, the main immune cells of the brain. As an experimental model we used primary microglial cells harvested from mixed glial cultures that had been kept in normal or high BCAA medium (H-BCAA). We show that H-BCAA microglial cells exhibit a peculiar phenotype characterized by a partial skewing toward the M2 state, with enhanced IL-10 expression and phagocytic activity but also increased free radical generation and decreased neuroprotective functions. We suggest that such an intermediate M1/M2 phenotype might result in a less efficient microglial response, which would promote the establishment of a low grade chronic inflammation and increase the likelihood of neurodegeneration. Although based on in vitro evidence, our study adds on to an increasing literature indicating that the increasing use of dietary integrators might deserve consideration for the possible drawbacks. In addition to excitotoxicity, the altered immune profile of microglia might represent a further mechanism by which BCAAs might turn into toxicants and facilitate neurodegeneration

Salt stress alters root meristem definition, vascular differentiation and metabolome in Sorghum bicolor (L.) genotypes

Knowledge on salt tolerance requires further investigation, particularly in plants of agro-food interest. Sorghum is a potentially useful plant because it is a emerging food species that combines high levels of salt tolerance with interesting nutritional characteristics. In sorghum different genotypes respond differently to saline stress and the early events characterizing the salt stress tolerance are not yet fully understood. Moreover, the number of salt resistant genotypes needs to be extended. The genotypes Bianca and Tonkawa are two possible candidates for extending sorghum cultivation to soils characterized by high levels of salinity. The root is the first organ that responds to soil conditions, especially during the initial stages of plant developmental. The research aim was to analyse the root system responses to salt stress (NaCl) of Bianca and Tonkawa genotypes to identify the morpho-functional and metabolic changes that occur during the initial stages of the root system development and to use them as discriminating parameters for assessing the different plant’s susceptibility to the salt. The results showed that salt stress negatively affected many morphological and cyto-histological root parameters, from seed germination to root system establishment. The salt altered the root meristem organization and quiescent centre (QC) definition, but similarly in both genotypes. By contrast, it reduced primary root (PR) length and induced a more extended oxidative stress in the adventitious roots (ARs) and lateral root primordia (LRPs) of Tonkawa in comparison with Bianca. The stele area and the number of protoxylem and phloem elements in the ARs were more reduced in 150 mM NaCl-treated Tonkawa seedlings in comparison with those of Bianca. Moreover, the salt enhanced lignin deposition in protoxylem, early metaxylem and endodermis and changed the root metabolic profiles significantly increasing the levels of leucine, isoleucine, alanine, proline, trigonelline, allantoin and glutamine in Bianca compared to Tonkawa. Altogether, specific morpho-anatomical and metabolic differences between the genotypes were identified as discriminating markers of genotype salt susceptibility