Nano-immunotherapy: Overcoming tumour immune evasion.

Immunotherapy is emerging as a groundbreaking cancer treatment, offering the unprecedented opportunity to effectively treat and in several cases, even cure previously untreatable malignancies. Anti-tumour immunotherapies designed to amplify T cell responses against defined tumour antigens have long been considered effective approaches for cancer treatment. Despite a clear rationale behind such immunotherapies, extensive past efforts were unsuccessful in mediating clinically relevant anti-tumour activity in humans. This is mainly because tumours adopt specific mechanisms to circumvent the host´s immunity. Emerging data suggest that the full potential of cancer immunotherapy will be only achieved by combining immunotherapies designed to generate or amplify anti-tumour T cell responses with strategies able to impair key tumour immune-evasion mechanisms. However, many approaches aimed to re-shape the tumour immune microenvironment (TIME) are commonly associated with severe systemic toxicity, require frequent administration, and only show modest efficacy in clinical settings. The use of nanodelivery systems is revealing as a valid means to overcome these limitations by improving the targeting efficiency, minimising systemic exposure of immunomodulatory agents, and enabling the development of novel combinatorial immunotherapies. In this review, we examine the emerging field of therapeutic modulation of TIME by the use of nanoparticle-based immunomodulators and potential future directions for TIME-targeting nanotherapies.

Pseudocrossidium replicatum (Taylor) R.H. Zander is a fully desiccation-tolerant moss that expresses an inducible molecular mechanism in response to severe abiotic stress.

KEY MESSAGE: The moss Pseudocrossidium replicatum is a desiccation-tolerant species that uses an inducible system to withstand severe abiotic stress in both protonemal and gametophore tissues. Desiccation tolerance (DT) is the ability of cells to recover from an air-dried state. Here, the moss Pseudocrossidium replicatum was identified as a fully desiccation-tolerant (FDT) species. Its gametophores rapidly lost more than 90% of their water content when exposed to a low-humidity atmosphere [23% relative humidity (RH)], but abscisic acid (ABA) pretreatment diminished the final water loss after equilibrium was reached. P. replicatum gametophores maintained good maximum photosystem II (PSII) efficiency (Fv/Fm) for up to two hours during slow dehydration; however, ABA pretreatment induced a faster decrease in the Fv/Fm. ABA also induced a faster recovery of the Fv/Fm after rehydration. Protein synthesis inhibitor treatment before dehydration hampered the recovery of the Fv/Fm when the gametophores were rehydrated after desiccation, suggesting the presence of an inducible protective mechanism that is activated in response to abiotic stress. This observation was also supported by accumulation of soluble sugars in gametophores exposed to ABA or NaCl. Exogenous ABA treatment delayed the germination of P. replicatum spores and induced morphological changes in protonemal cells that resembled brachycytes. Transcriptome analyses revealed the presence of an inducible molecular mechanism in P. replicatum protonemata that was activated in response to dehydration. This study is the first RNA-Seq study of the protonemal tissues of an FDT moss. Our results suggest that P. replicatum is an FDT moss equipped with an inducible molecular response that prepares this species for severe abiotic stress and that ABA plays an important role in this response.

Angiotensin-Converting Enzyme (ACE) genetic variation and longevity in Peruvian older people: a cross-sectional study.

Background: Some studies have suggested that the insertion(I)/deletion(D) polymorphism of the Angiotensin-Converting Enzyme (ACE) gene may be associated with human longevity, especially in centenarians. However, this association is still controversial. Besides, there have been no studies in Peruvians.Aim: To describe the age distribution of the ACE polymorphism in a convenience sample of Peruvian older people.Subjects and methods: This was a cross-sectional study in 104 Geriatric Day Hospital patients in Lima, Perú. The ACE polymorphism was determined in all patients. For the purpose of association with age, the sample was divided into four categories: young (< 65), youngest-old (65-74), middle-old (75-84) and oldest-old (85 or more).Results: The distribution of genotype frequencies was consistent with a population in Hardy-Weinberg equilibrium (p = 0.62). The number (%) of D/D, I/D and I/I genotypes in the young was 2 (14.3%), 3 (21.4%) and 9 (64.3%), respectively; in youngest-old: 4 (11.4%), 15 (42.9%) and 16 (45.7%); in middle-old: 6 (12.2%), 20 (40.8%) and 23 (46.9%); and in oldest-old: 0 (0.0%), 4 (66.7%) and 2 (33.3%). A chi-square analysis showed no significant differences in genotype distribution between age groups (p = 0.647).Conclusion: No significant age differences were found in the distribution of the ACE polymorphism in this sample. Further studies with greater statistical power are recommended.

Effects of an antioxidant beverage on biomarkers of oxidative stress in Alzheimer's patients.

PURPOSE: The purpose of the study was to test whether daily consumption of a beverage with high antioxidant power, combining extracts of green tea and apple over a period of 8 months, would affect blood and urinary concentrations of biomarkers of oxidative stress in Alzheimer's patients. METHODS: The study included 100 subjects, 48 of them were Alzheimer's patients, aged 76.5 ± 3.5 years, and 52 were control subjects, aged 79 ± 4 years, without dementia. Three blood and urine samples were taken from each participant, the first (T i) before starting the antioxidant or placebo beverage intake, the second (T m) 4 months after the antioxidant or placebo beverage intake and the third (T f) 8 months after the antioxidant or placebo beverage intake, and concentrations of biomarkers of oxidative stress were measured on serum, lysed erythrocytes or urine by UV-Vis spectrophotometry or by competitive in vitro enzyme-linked immunosorbent assay, according to the parameter analyzed. RESULTS: The administration of the antioxidant beverage to the Alzheimer's patients prevented the decrease in total antioxidant status in the moderate phase of the disease (T i = 1.40 ± 0.10 mmol/L vs T f = 1.20 ± 0.08 mmol/L), increased values of glutathione peroxidase and superoxide dismutase in initial (165 and 24 % respectively) and moderate phase (75 and 85 % respectively), and prevented the increase in protein carbonyls in moderate phase (T i = 0.17 ± 0.07 nmol/mg protein vs T f = 0.21 ± 0.06 nmol/mg protein), with a significant decrease in protein carbonyls since the fourth month of the intake in initial phase (T m = 0.21 ± 0.06 nmol/mg protein vs T f = 0.11 ± 0.05 nmol/mg protein). CONCLUSION: Our results suggest that antioxidant beverage could be used as a natural complementary therapy for alleviate or decrease the oxidative stress effects in the stages of Alzheimer's disease.

Inhibition of iNOS as a novel effective targeted therapy against triple-negative breast cancer.

INTRODUCTION: Triple-negative breast cancer (TNBC) is an aggressive form of breast cancer with no effective targeted therapy. Inducible nitric oxide synthase (iNOS) is associated with poor survival in patients with breast cancer by increasing tumor aggressiveness. This work aimed to investigate the potential of iNOS inhibitors as a targeted therapy for TNBC. We hypothesized that inhibition of endogenous iNOS would decrease TNBC aggressiveness by reducing tumor initiation and metastasis through modulation of epithelial-mesenchymal transition (EMT)-inducing factors. METHODS: iNOS protein levels were determined in 83 human TNBC tissues and correlated with clinical outcome. Proliferation, mammosphere-forming efficiency, migration, and EMT transcription factors were assessed in vitro after iNOS inhibition. Endogenous iNOS targeting was evaluated as a potential therapy in TNBC mouse models. RESULTS: High endogenous iNOS expression was associated with worse prognosis in patients with TNBC by gene expression as well as immunohistochemical analysis. Selective iNOS (1400 W) and pan-NOS (L-NMMA and L-NAME) inhibitors diminished cell proliferation, cancer stem cell self-renewal, and cell migration in vitro, together with inhibition of EMT transcription factors (Snail, Slug, Twist1, and Zeb1). Impairment of hypoxia-inducible factor 1α, endoplasmic reticulum stress (IRE1α/XBP1), and the crosstalk between activating transcription factor 3/activating transcription factor 4 and transforming growth factor ß was observed. iNOS inhibition significantly reduced tumor growth, the number of lung metastases, tumor initiation, and self-renewal. CONCLUSIONS: Considering the effectiveness of L-NMMA in decreasing tumor growth and enhancing survival rate in TNBC, we propose a targeted therapeutic clinical trial by re-purposing the pan-NOS inhibitor L-NMMA, which has been extensively investigated for cardiogenic shock as an anti-cancer therapeutic.

[Efficacy of disinfection treatments using essential oils and ultrasound on tomato fruits inoculated with Escherichia coli and impact on antioxidant activity]. / Eficiencia de la desinfección con aceites esenciales y ultrasonido sobre Escherichia coli inoculada en frutos de tomate y el impacto sobre la actividad antioxidante.

Fresh produce often harbors a great number of microorganisms; hence, its growing demand may constitute a risk for consumers. The aim of this study was to evaluate the efficacy of several disinfection procedures against enterotoxigenic Escherichia coli (ETEC) inoculated on tomato fruits and the conservation of the antioxidant properties of these disinfected fruits. Fruits were immersed for 5 or 10min in oregano or thyme essential oil dispersions (5, 10ppm), with or without ultrasound treatment. Antioxidant activity of disinfected fruits was determined as the ability to scavenge 2,2-diphenyl-1-pricrylhydrazyl (DPPH) radicals and was reported as percentage of inhibition (%I). The most efficient disinfectant treatments showing significant differences (p≤.05) between the reductions log10 CFU/g (S) of ETEC were those using 10ppm oregano for 10min, with S=3.05 in individual treatments and S=4.03 in mixed treatments. The highest %I was obtained with individual sonication treatments (69.52 and 72.48), while in combined treatments the %I values increased with thyme oil 5ppm and ultrasound for 5min (51.27%) and 10min (53.31%).

A new antioxidant beverage produced with green tea and apple.

Green tea and apple are natural products with health benefits. These healthy properties are linked closely to the antioxidant compounds, mainly phenolic compounds. These antioxidant compounds have a potential for preventing and treating cancer, cardiovascular, inflammatory and neurodegenerative diseases in humans. The aim of the present work was to design a new beverage with high antioxidant power combining extracts of green tea and apple, studying the antioxidant composition and activity, organoleptic properties (colour) and stability status during storage at different temperatures. The majority compounds identified in the beverage were flavan-3-ols, being the (-)-epigallocatechin-3-gallate which had the highest concentration. After storage, floridzine was the compound with lower decrease of concentration. The new designed beverage had a good colour, and high antioxidant activity and stability at room temperature, so that the beverage needs no refrigeration, showing potential for the development of new healthy functional beverages.

Microencapsulation of Tecoma stans Extracts: Bioactive Properties Preservation and Physical Characterization Analysis.

Bioactive compounds from medicinal plants have applications in the development of functional foods. However, since they are unstable, encapsulation is used as a conservation alternative. This work aimed to assess the bioactive properties (antioxidant and hypoglycemic) of different extracts, including the infusion, as well as their spray-dried microencapsulates from Tecoma stans leaves. A factorial design was proposed to determine the best extraction conditions, based on ABTS and DPPH inhibition. Maltodextrin (MD), arabic gum (AG), and a 1:1 blend (MD:AG) were used as encapsulating agents. Moreover, characterization through physicochemical properties, gas chromatography/mass spectrometry (GC-MS) and scanning electron microscopy (SEM) of the best two powders based on the bioactive properties were analyzed. The results showed that the combination of stirring, water, and 5 min provided the highest inhibition to ABTS and DPPH (35.64 ± 1.25 mg Trolox/g d.s. and 2.77 ± 0.01 g Trolox/g d.s., respectively). Spray drying decreased the antioxidant activity of the extract while preserving it in the infusion. The encapsulated infusion with MD:AG had the highest hypoglycemic activity as it presented the lowest glycemic index (GI = 47). According to the results, the microencapsulates could potentially be added in foods to enhance nutritional quality and prevent/treat ailments.

Advances in Lipid Nanoparticles for mRNA-Based Cancer Immunotherapy.

Over the past decade, messenger RNA (mRNA) has emerged as potent and flexible platform for the development of novel effective cancer immunotherapies. Advances in non-viral gene delivery technologies, especially the tremendous progress in lipid nanoparticles' manufacturing, have made possible the implementation of mRNA-based antitumor treatments. Several mRNA-based immunotherapies have demonstrated antitumor effect in preclinical and clinical studies, and marked successes have been achieved most notably by its implementation in therapeutic vaccines, cytokines therapies, checkpoint blockade and chimeric antigen receptor (CAR) cell therapy. In this review, we summarize recent advances in the development of lipid nanoparticles for mRNA-based immunotherapies and their applications in cancer treatment. Finally, we also highlight the variety of immunotherapeutic approaches through mRNA delivery and discuss the main factors affecting transfection efficiency and tropism of mRNA-loaded lipid nanoparticles in vivo.

Lipid-Based Vectors for Therapeutic mRNA-Based Anti-Cancer Vaccines.

Cancer vaccines have been widely explored as a key tool for effective cancer immunotherapy. Despite a convincing rationale behind cancer vaccines, extensive past efforts were unsuccessful in mediating significantly relevant anti-tumor activity in clinical studies. One of the major reasons for such poor outcome, among others, is the low immunogenicity of more traditional vaccines, such as peptide-, protein- and DNA- based vaccines. Recently, mRNA emerged as a promising alternative to traditional vaccine strategies due to its high immunogenicity, suitability for large-scale and low-cost production, and superior safety profile. However, the clinical application of mRNA-based anti-cancer vaccines has been limited by their instability and inefficient in vivo delivery. Recent technological advances have now largely overcome these issues and lipid-based vectors have demonstrated encouraging results as mRNA vaccine platforms against several types of cancers. This review intends to provide a detailed overview of lipid-based vectors for the development of therapeutic mRNA-based anti-tumor vaccines.

Codelivery of mRNA with α-Galactosylceramide Using a New Lipopolyplex Formulation Induces a Strong Antitumor Response upon Intravenous Administration.

Recently, the use of mRNA-based vaccines for cancer immunotherapy has gained growing attention. Several studies have shown that mRNA delivered in a vectorized format can generate a robust and efficient immune response. In this work, a new lipopolyplex vector (multi-LP), incorporating the immune adjuvant α-galactosylceramide (α-GalCer) and a multivalent cationic lipid, was proposed for the in vivo delivery of mRNA into antigen-presenting cells. We demonstrate that dendritic cells (DCs) can be targeted in vivo by intravenous administration of a α-GalCer-/mRNA-loaded multi-LP vector, without the need for its functionalization with cell-specific antibodies or ligands. The multi-LP nanoparticles loaded with a reporter mRNA efficiently led to high expression of the enhanced green fluorescence protein in DCs both in vitro and in vivo, exhibiting an intrinsic selectivity for DCs. Finally, the TRP2-mRNA/α-GalCer-based multi-LP vaccine induced a significant therapeutic effect against a highly malignant B16-F10 melanoma tumor. This study provides the first evidence that a combination of antigen-mRNA and α-GalCer can be used as an effective antitumor vaccine, inducing strong innate and adaptive immune responses.

Lipopolyplex potentiates anti-tumor immunity of mRNA-based vaccination.

mRNA-based vaccines have the benefit of triggering robust anti-cancer immunity without the potential danger of genome integration from DNA vaccines or the limitation of antigen selection from peptide vaccines. Yet, a conventional mRNA vaccine comprising of condensed mRNA molecules in a positively charged protein core structure is not effectively internalized by the antigen-presenting cells. It cannot offer sufficient protection for mRNA molecules from degradation by plasma and tissue enzymes either. Here, we have developed a lipopolyplex mRNA vaccine that consists of a poly-(ß-amino ester) polymer mRNA core encapsulated into a 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine/1,2-dioleoyl-sn-glycero-3-phosphatidyl-ethanolamine/1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000 (EDOPC/DOPE/DSPE-PEG) lipid shell. This core-shell structured mRNA vaccine enters dendritic cells through macropinocytosis. It displayed intrinsic adjuvant activity by potently stimulating interferon-ß and interleukin-12 expression in dendritic cells through Toll-like receptor 7/8 signaling. Dendritic cells treated with the mRNA vaccine displayed enhanced antigen presentation capability. Mice bearing lung metastatic B16-OVA tumors expressing the ovalbumin antigen were treated with the lipopolyplex mRNA, and over 90% reduction of tumor nodules was observed. Collectively, this core-shell structure offers a promising platform for mRNA vaccine development.

Label-Free Isothermal Amplification Assay for Specific and Highly Sensitive Colorimetric miRNA Detection.

We describe a new method for the detection of miRNA in biological samples. This technology is based on the isothermal nicking enzyme amplification reaction and subsequent hybridization of the amplification product with gold nanoparticles and magnetic microparticles (barcode system) to achieve naked-eye colorimetric detection. This platform was used to detect a specific miRNA (miRNA-10b) associated with breast cancer, and attomolar sensitivity was demonstrated. The assay was validated in cell culture lysates from breast cancer cells and in serum from a mouse model of breast cancer.

Porous silicon microparticle potentiates anti-tumor immunity by enhancing cross-presentation and inducing type I interferon response.

Micro- and nanometer-size particles have become popular candidates for cancer vaccine adjuvants. However, the mechanism by which such particles enhance immune responses remains unclear. Here, we report a porous silicon microparticle (PSM)-based cancer vaccine that greatly enhances cross-presentation and activates type I interferon (IFN-I) response in dendritic cells (DCs). PSM-loaded antigen exhibited prolonged early endosome localization and enhanced cross-presentation through both proteasome- and lysosome-dependent pathways. Phagocytosis of PSM by DCs induced IFN-I responses through a TRIF- and MAVS-dependent pathway. DCs primed with PSM-loaded HER2 antigen produced robust CD8 T cell-dependent anti-tumor immunity in mice bearing HER2+ mammary gland tumors. Importantly, this vaccination activated the tumor immune microenvironment with elevated levels of intra-tumor IFN-I and MHCII expression, abundant CD11c+ DC infiltration, and tumor-specific cytotoxic T cell responses. These findings highlight the potential of PSM as an immune adjuvant to potentiate DC-based cancer immunotherapy.

Eficiencia de la desingección con aceites esenciales y ultrasonido sobre Escherichia coli inoculada en frutos de tomate y el impacto sobre la actividad antioxidante / Efficacy of disinfection treatments using essential oils and ultrasound on tomato fruits inoculated with escherichia coli and impact on antioxidant activity

La creciente demanda de frutos frescos puede constituir un riesgo para la salud de los consumidores, teniendo en cuenta la gran variedad de microorganismos que estos suelen albergar. El objetivo del presente estudio fue evaluar la eficacia de varios procedimientos de desinfección sobre Escherichia coli enterotoxigénica (enterotoxigenic E. coli [ETEC]) inoculada en tomate y la conservación de las propiedades antioxidantes de los frutos desinfectados. Los frutos fueron sumergidos durante 5 o 10min en dispersiones de aceites esenciales de orégano o tomillo (5 o 10ppm), combinados o no con la aplicación de ultrasonido. La actividad antioxidante se determinó por la neutralización del radical 2,2-difenil-1-pricrilhidrazil (DPPH) y se reportó como porcentaje de inhibición (%I). Los tratamientos de desinfección más eficaces para una significativa reducción log10 UFG/g (S) de ETEC fueron con 10ppm de aceite de orégano durante 10min, con reducciones S=3,05 en tratamientos individuales y S=4,03 en mixtos. Los %I más altos se lograron con tratamientos individuales con sonicación (69,52 y 72,48) y en tratamientos combinados con aceite de tomillo 5ppm y ultrasonido durante 5 y 10min, con valores de 51,27 y 53,31%, respectivamente

Fresh produce often harbors a great number of microorganisms; hence, its growing demand may constitute a risk for consumers. The aim of this study was to evaluate the efficacy of several disinfection procedures against enterotoxigenic Escherichia coli (ETEC) inoculated on tomato fruits and the conservation of the antioxidant properties of these disinfected fruits. Fruits were immersed for 5 or 10min in oregano or thyme essential oil dispersions (5, 10ppm), with or without ultrasound treatment. Antioxidant activity of disinfected fruits was determined as the ability to scavenge 2,2-diphenyl-1-pricrylhydrazyl (DPPH) radicals and was reported as percentage of inhibition (%I). The most efficient disinfectant treatments showing significant differences (p≤.05) between the reductions log10 CFU/g (S) of ETEC were those using 10ppm oregano for 10min, with S=3.05 in individual treatments and S=4.03 in mixed treatments. The highest %I was obtained with individual sonication treatments (69.52 and 72.48), while in combined treatments the %I values increased with thyme oil 5ppm and ultrasound for 5min (51.27%) and 10min (53.31%)