SERS Quantification of Galunisertib Delivery in Colorectal Cancer Cells by Plasmonic-Assisted Diatomite Nanoparticles

AbstractThe small molecule Galunisertib (LY2157299, LY) shows multiple anticancer activities blocking the transforming growth factor‐β1 receptor, responsible for the epithelial‐to‐mesenchymal transition (EMT) by which colorectal cancer (CRC) cells acquire migratory and metastatic capacities. However, frequent dosing of LY can produce highly toxic metabolites. Alternative strategies to reduce drug side effects can rely on nanoscale drug delivery systems that have led to a medical revolution in the treatment of cancer, improving drug efficacy and lowering drug toxicity. Here, a hybrid nanosystem (DNP‐AuNPs‐LY@Gel) made of a porous diatomite nanoparticle decorated with plasmonic gold nanoparticles, in which LY is retained by a gelatin shell, is proposed. The multifunctional capability of the nanosystem is demonstrated by investigating the efficient LY delivery, the enhanced EMT reversion in CRCs and the intracellular quantification of drug release with a sub‐femtogram resolution by surface‐enhanced Raman spectroscopy (SERS). The LY release trigger is the pH sensitivity of the gelatin shell to the CRC acidic microenvironment. The drug release is real‐time monitored at single‐cell level by analyzing the SERS signals of LY in CRC cells. The higher efficiency of LY delivered by the DNP‐AuNPs‐LY@Gel complex paves the way to an alternative strategy for lowering drug dosing and consequent side effects

Does the use of dietary supplements enhance athletes’ sport performances? A systematic review and a meta-analysis

Background: The consumption of dietary supplements has increased in recent years. Despite their widespread use, there is confusion about effects on sport performances. The aim of this study was to investigate association between use of supplements and enhance of athletes’ sports performance. Methods: A review and a meta-analysis of studies conducted on Dietary Supplements and Sports between 2003 and 2013 were performed. Enhancement on sport performances was considered as outcome. The following aspects related to enhancement were considered: ergogenic effect (EE), time to exhaustion (TTE), muscular endurance (ME), post-exercise recovery (PER) and body mass (BM). With respect to meta-analysis, data on level of post Exercise Glucose (GpE [mg/dL]) and level of post exercise Lactate (LpE [mmol/L]) were considered as  indicators of TTE, PER and EE. Similarly, Change in Body Mass (CBM) [kg] was used as indicator of BM. Results: The most investigated dietary supplements were: Creatine, Carbohydrates, Beta-alanine, Proteins. The qualitative analysis evaluating the effect of supplements on sports listed by the International Olympic Committee has achieved interesting results: supplements didn’t show statistically significant effects when compared to placebo in more than 48% of papers. For the quantitative analysis, 15 studies were considered. The meta-analysis showed that there was no significant effect of Beta-alanine, Creatine and Carbohydrates on LpE and GpE. Furthermore, a non-significant increase in BM was observed in athletes undergoing Creatine compared to placebo. Conclusion: Considering the increasing attention to this topic, it would be interesting to investigate the existing awareness about effectiveness and possible risks of supplements

Microfluidic-Assisted Production of Gastro-Resistant Active-Targeted Diatomite Nanoparticles for the Local Release of Galunisertib in Metastatic Colorectal Cancer Cells

The oral route is highly desirable for colorectal cancer (CRC) treatment because it allows concentrating the drug in the colon and achieving a localized effect. However, orally administered drugs are often metabolized in the liver, resulting in reduced efficacy and the need for higher doses. Nanoparticle-based drug delivery systems can be engineered to prevent the diffusion of the drug in the stomach, addressing the release at the target site, and enhancing the efficacy of the delivered drug. Here, an orally administrable galunisertib delivery system is developed with gelatin-covered diatomite nanoparticles targeting the ligand 1-cell adhesion molecule (L1-CAM) on metastatic cells, and further encapsulated in an enteric matrix by microfluidics. The gastro-resistant polymer protects the nanoparticles from the action of the digestive enzymes and allows for a sustained release of galunisertib at the intestinal pH. The efficacy of antibody-antigen interactions to drive the internalization of nanoparticles in the targeted cells is investigated in CRC cells expressing abnormal (SW620) or basal levels (Caco-2, HT29-MTX) of L1-CAM. The combination of local drug release and active targeting enhances the effect of the delivered galunisertib, which inhibits the migration of the SW620 cells with greater efficiency compared to the free drug.Peer reviewe

Microfluidic-Assisted Production of Gastro-Resistant Active-Targeted Diatomite Nanoparticles for the Local Release of Galunisertib in Metastatic Colorectal Cancer Cells (Adv. Healthcare Mater. 6/2023)

Nanoparticle Drug DeliveryThe encapsulation of nanoparticles in hydroxypropyl methyl cellulose is key to treat colorectal cancer through oral administration. In article 2202672 by Ilaria Rea, Hélder A. Santos, and co-workers, gelatin-coated diatomite nanoparticles loaded with galunisertib are functionalized with an anti-L1-CAM antibody that targets metastatic colon cancer cells. These nanoparticles are then encapsulated in a gastro-resistant matrix using microfluidics. When the nanoparticles interact with the targeted cells, the gastro-resistant coating dissolves and galunisertib is released

Toward Multi-Parametric Porous Silicon Transducers Based on Covalent Grafting of Graphene Oxide for Biosensing Applications

Graphene oxide (GO) is a two-dimensional material with peculiar photoluminescence emission and good dispersion in water, that make it an useful platform for the development of label-free optical biosensors. In this study, a GO-porous silicon (PSi) hybrid device is realized using a covalent chemical approach in order to obtain a stable support for biosensing applications. Protein A, used as bioprobe for biosensing purposes, is covalently linked to the GO, using the functional groups on its surface, by carbodiimide chemistry. Protein A bioconjugation to GO-PSi hybrid device is investigated by atomic force microscopy (AFM), scanning electron microscopy (SEM), water contact angle (WCA) measurements, Fourier transform infrared (FTIR) spectroscopy, steady-state photoluminescence (PL), and fluorescence confocal microscopy. PSi reflectance and GO photoluminescence changes can thus be simultaneously exploited for monitoring biomolecule interactions as in a multi-parametric hybrid biosensing device

PNA-based graphene oxide/porous silicon hybrid biosensor: towards a label-free optical assay for Brugada Syndrome

Peptide nucleic acid (PNA) is a synthetic DNA mimic that outperforms the properties of traditional oligonucleotides (ONs). On account of its outstanding features, such as remarkable binding affinity towards complementary DNA or RNA as well as high thermal and chemical stability, PNA has been proposed as a valuable alternative to the ON probe in gene-sensor design. In this study, a hybrid transducer made-up of graphene oxide (GO) nano-sheets covalently grafted onto a porous silicon (PSi) matrix has been investigated for the early detection of a genetic cardiac disorder, the Brugada syndrome (BS). A functionalization strategy towards the realization of a potential PNA-based device is described. A peptide nucleic acid (PNA), able to detect the SCN5A associated with the BS has been properly synthesized and used as a bioprobe for the realization of a proof-of-concept label-free optical PNA-biosensor. PSi reflectance and GO photoluminescence (PL) signals were simultaneously exploited for the monitoring of the device functionalization and response

Pro-active monitoring and social interventions at community level mitigate the impact of coronavirus (COVID-19) epidemic on older adults' mortality in Italy: A retrospective cohort analysis

BackgroundThe COVID-19 epidemic in Italy has severely affected people aged more than 80, especially socially isolated. Aim of this paper is to assess whether a social and health program reduced mortality associated to the epidemic.MethodsAn observational retrospective cohort analysis of deaths recorded among > 80 years in three Italian cities has been carried out to compare death rate of the general population and "Long Live the Elderly!" (LLE) program. Parametric and non-parametric tests have been performed to assess differences of means between the two populations. A multivariable analysis to assess the impact of covariates on weekly mortality has been carried out by setting up a linear mixed model.ResultsThe total number of services delivered to the LLE population (including phone calls and home visits) was 34,528, 1 every 20 day per person on average, one every 15 days during March and April. From January to April 2019, the same population received one service every 41 days on average, without differences between January-February and March-April. The January-April 2020 cumulative crude death rate was 34.8% (9,718 deaths out of 279,249 individuals; CI95%: 34.1-35.5) and 28.9% (166 deaths out of 5,727 individuals; CI95%:24.7-33.7) for the general population and the LLE sample respectively. The general population weekly death rate increased after the 11th calendar week that was not the case among the LLE program participants (p<0.001). The Standardized Mortality Ratio was 0.83; (CI95%: 0.71-0.97). Mortality adjusted for age, gender, COVID-19 weekly incidence and prevalence of people living in nursing homes was lower in the LLE program than in the general population (p<0.001).ConclusionsLLE program is likely to limit mortality associated with COVID-19. Further studies are needed to establish whether it is due to the impact of social care that allows a better clients' adherence to the recommendations of physical distancing or to an improved surveillance of older adults that prevents negative outcomes associated with COVID-19

Cyclic Peptoids as Mycotoxin Mimics: An Exploration of Their Structural and Biological Properties

Cyclic peptoids have recently emerged as important examples of peptidomimetics for their interesting complexing properties and innate ability to permeate biological barriers. In the present contribution, experimental and theoretical data evidence the intricate conformational and stereochemical properties of five novel hexameric peptoids decorated with N-isopropyl, N-isobutyl, and N-benzyl substituents. Complexation studies by NMR, in the presence of sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate (NaTFPB), theoretical calculations, and single-crystal X-ray analyses indicate that the conformationally stable host/guest metal adducts display architectural ordering comparable to that of the enniatins and beauvericin mycotoxins. Similarly to the natural depsipeptides, the synthetic oligolactam analogues show a correlation between ion transport abilities in artificial liposomes and cytotoxic activity on human cancer cell lines. The reported results demonstrate that the versatile cyclic peptoid scaffold, for its remarkable conformational and complexing properties, can morphologically mimic related natural products and elicit powerful biological activities

Development of Surface Chemical Strategies for Synthesizing Redox-Responsive Diatomite Nanoparticles as a Green Platform for On-Demand Intracellular Release of an Antisense Peptide Nucleic Acid Anticancer Agent

Redox-responsive silica drug delivery systems are synthesized by aeco-friendly diatomite source to achieve on-demand release of peptide nucleic acid (PNA) in tumor reducing microenvironment, aiming to inhibit the immune check-point programmed cell death 1 receptor/programmed cell death receptor ligand 1 (PD-1/PD-L1) in cancer cells. The nanoparticles (NPs) are coated with polyethylene glycol chains as gatekeepers to improve their physicochemical properties and control drug release through the cleavable disulfide bonds (S-S) in a reductive environment. This study describes different chemical conditions to achieve the highest NPs' surface functionalization yield, exploring both multistep and one-pot chemical functionalization strategies. The best formulation is used for covalent PNA conjugation via the S-S bond reaching a loading degree of 306 +/- 25 mu g (PNA) mg(DNPs)(-1). These systems are used for in vitro studies to evaluate the kinetic release, biocompatibility, cellular uptake, and activity on different cancer cells expressing high levels of PD-L1. The obtained results prove the safety of the NPs up to 200 mu g mL(-1) and their advantage for controlling and enhancing the PNA intracellular release as well as antitumor activity. Moreover, the downregulation of PD-L1 observed only with MDA-MB-231 cancer cells paves the way for targeted immunotherapy.Peer reviewe

Rare Variants in Autophagy and Non-Autophagy Genes in Late-Onset Pompe Disease: Suggestions of Their Disease-Modifying Role in Two Italian Families

Pompe disease is an autosomal recessive disorder caused by a deficiency in the enzyme acid alpha-glucosidase. The late-onset form of Pompe disease (LOPD) is characterized by a slowly progressing proximal muscle weakness, often involving respiratory muscles. In LOPD, the levels of GAA enzyme activity and the severity of the clinical pictures may be highly variable among individuals, even in those who harbour the same combination of GAA mutations. The result is an unpredictable genotype–phenotype correlation. The purpose of this study was to identify the genetic factors responsible for the progression, severity and drug response in LOPD. We report here on a detailed clinical, morphological and genetic study, including a whole exome sequencing (WES) analysis of 11 adult LOPD siblings belonging to two Italian families carrying compound heterozygous GAA mutations. We disclosed a heterogeneous pattern of myopathic impairment, associated, among others, with cardiac defects, intracranial vessels abnormality, osteoporosis, vitamin D deficiency, obesity and adverse response to enzyme replacement therapy (ERT). We identified deleterious variants in the genes involved in autophagy, immunity and bone metabolism, which contributed to the severity of the clinical symptoms observed in the LOPD patients. This study emphasizes the multisystem nature of LOPD and highlights the polygenic nature of the complex phenotype disclosed in these patients