Bioactivity analysis of calcium silicate-based sealers and repair cements on the phenotype and cytokine secretion profile of CD14+ monocytes: An ex vivo study.

AIM: This study evaluated the immune bioactivity of testing media (TM) obtained from different calcium silicate-based sealers and cements on monocyte morphology, activation, differentiation and cytokine secretion. METHODS: Blood-derived CD14+ monocytes were isolated and cultured for 5 days with 25% TM from the following calcium silicate-based materials: TotalFill BC RRM Fast-Set Putty, Biodentine, TotalFill BC Sealer and BioRoot-Root-Canal-Sealer (RCS). A resin-based endodontic cement was used as a control. The expression of surface markers such as CD86, HLA-DR, CD16, CD309 and CD209, and cytokine secretion were analysed by flow cytometry. Data were analysed using the one-way repeated measures analysis of variance (anova) multiple comparison test and a Holm-Sidak multiple comparison post-hoc test (p < .05). RESULTS: This comparative analysis revealed that monocytes co-cultured with calcium silicate-based materials showed a spindle-shaped morphology compared with the round shape observed in the control. Regarding activation markers, BioRoot-RCS and Biodentine significantly increased CD86 expression compared with the control sample, whereas no significant differences (p > .05) were observed in HLA-DR expression. In addition, no differences were observed among the differentiation markers. When the inflammatory cytokines were analysed, BioRoot-RCS increased the secretion of IL-1ß, IL-6, IL-10 and TNF-α, whereas BioRoot-RCS and Biodentine significantly decreased IL-8 production (p < .05). CONCLUSIONS: These data showed that the calcium silicate-based materials tested changed the morphology of CD14+ monocytes; however, only BioRoot-RCS and Biodentine significantly upregulated CD86. In addition, BioRoot-RCS was the sealer with the highest immunomodulatory properties for cytokine production which means that it can contribute with the in vivo healing process and regeneration of periapical lesions.

Analysis of Tumor-Infiltrating T-Cell Transcriptomes Reveal a Unique Genetic Signature across Different Types of Cancer.

CD8+ and CD4+ T-cells play a key role in cellular immune responses against cancer by cytotoxic responses and effector lineages differentiation, respectively. These subsets have been found in different types of cancer; however, it is unclear whether tumor-infiltrating T-cell subsets exhibit similar transcriptome profiling across different types of cancer in comparison with healthy tissue-resident T-cells. Thus, we analyzed the single cell transcriptome of five tumor-infiltrating CD4-T, CD8-T and Treg cells obtained from different types of cancer to identify specific pathways for each subset in malignant environments. An in silico analysis was performed from single-cell RNA-sequencing data available in public repositories (Gene Expression Omnibus) including breast cancer, melanoma, colorectal cancer, lung cancer and head and neck cancer. After dimensionality reduction, clustering and selection of the different subpopulations from malignant and nonmalignant datasets, common genes across different types of cancer were identified and compared to nonmalignant genes for each T-cell subset to identify specific pathways. Exclusive pathways in CD4+ cells, CD8+ cells and Tregs, and common pathways for the tumor-infiltrating T-cell subsets were identified. Finally, the identified pathways were compared with RNAseq and proteomic data obtained from T-cell subsets cultured under malignant environments and we observed that cytokine signaling, especially Th2-type cytokine, was the top overrepresented pathway in Tregs from malignant samples.

Undiagnosed sleep disorder breathing as a risk factor for critical COVID-19 and pulmonary consequences at the midterm follow-up.

INTRODUCTION: Patients with severe COVID-19 develops an acute respiratory distress syndrome (ARDS), requiring admission to the intensive care unit. COVID-19 also reports an increased prevalence of comorbidities, similar to patients with Sleep disorder breathing (SDB). OBJECTIVES: To evaluate the association between undiagnosed SDB and the risk of ARDS and pulmonary abnormalities in a cohort of patients' survivors of COVID-19 between 3 and 6 months after diagnosis. METHODS: Prospective cohort study of patients who developed ARDS during hospitalization due to COVID-19 compared with a control group of patients who had COVID-19 with mild to moderate symptoms. All patients were evaluated between the 12th and 24th week after SARS-CoV-2 infection. The evaluation includes persistent symptoms, lung diffusing capacity of carbon monoxide (DLCO), chest CT scan and home sleep apnea test. SDB was diagnosed by the respiratory disturbance index ≥5 ev/h. The association between SDB and ARDS, the hazards of lung impairment and the hazard ratios (HR) were analyzed. RESULTS: A total of 60 patients were included (ARDS: 34 patients, Control: 26 patients). The mean follow-up was 16 weeks (range 12-24). ARDS reported a high prevalence of SDB (79% vs. 38% in control group). A total of 35% reported DLCO impairment, and 67.6% abnormal chest CT. SDB was independently associated to ARDS, OR 6.72 (CI, 1.56-28.93), p < 0.01, and abnormal Chest CT, HR 17.2 (CI, 1.68-177.4, p = 0.01). Besides, ARDS, days in mechanical ventilation, male gender were also associated with an increased risk of abnormal chest CT. CONCLUSION: Undiagnosed SDB is prevalent and independently associated with ARDS. In addition, undiagnosed SDB increased the hazard of abnormal Chest CT in the midterm. STUDY REGISTER: ISRCTN16865246.

Immunomodulation of T Helper Cells by Tumor Microenvironment in Oral Cancer Is Associated With CCR8 Expression and Rapid Membrane Vitamin D Signaling Pathway.

The immune system plays a key role in the protective response against oral cancer; however, the tumor microenvironment (TME) impairs this anti-cancer response by modulating T helper (Th) responses and promoting an anti-inflammatory environment. Regulatory T cells (Tregs) and Th2 effector cells (Teff) are associated with poor prognosis in oral squamous cell carcinoma (OSCC). However, the main immunomodulatory mechanisms associated with the enrichment of these subsets in OSCC remain unknown. We characterized Th-like lineages in Tregs and Teff and evaluated immunomodulatory changes induced by the TME in OSCC. Our phenotypic data revealed a higher distribution of tumour-infiltrating CCR8+ and Th2-like Treg in OSCC compared with non-malignant samples, whereas the percentages of Th1 cells were reduced in cancer. We then analyzed the direct effect of the TME by exposing T cell subsets to cancer secretomes and observed the OSCC secretome induced CCR8 expression and reduced cytokine production from both subsets. Transcriptomic analysis showed that the co-culture with OSCC secretome induced several gene changes associated with the vitamin D (VitD) signaling pathway in T cells. In addition, proteomic analysis identified the presence of several proteins associated with prostaglandin E2 (PGE2) production by rapid membrane VitD signaling and a reduced presence of the VitD binding protein. Thus, we analyzed the effect of VitD and PGE2 and observed that VitD promotes a regulatory Th2-like response with CCR8 expression whilst PGE2 also modulated CCR8 but inhibited cytokine production in combination with VitD. Finally, we evaluated the presence of CCR8 ligand in OSCC and observed increased chemokine CCL18, which was also able to upregulate CCR8 in activated Th cells. Overall, our data showed the immunomodulatory changes induced by the TME involving CCR8 expression and regulatory Th2 phenotypes, which are associated with PGE2 mediated VitD signaling pathway and CCL18 expression in OSCC.

Platinum-vanadium oxide nanotube hybrids.

The present contribution reports on the features of platinum-based systems supported on vanadium oxide nanotubes. The synthesis of nanotubes was carried out using a commercial vanadium pentoxide via hydrothermal route. The nanostructured hybrid materials were prepared by wet impregnation using two different platinum precursors. The formation of platinum nanoparticles was evaluated by applying distinct reduction procedures. All nanostructured samples were essentially analysed by X-ray diffraction and transmission electron microscopy. After reduction, transmission electron microscopy also made it possible to estimate particle size distribution and mean diameter calculations. It could be seen that all reduction procedures did not affect the nanostructure of the supports and that the formation of metallic nanoparticles is quite efficient with an indistinct distribution along the nanotubes. Nevertheless, the reduction procedure determined the diameter, dispersion and shape of the metallic particles. It could be concluded that the use of H2PtCl6 is more suitable and that the use of hydrogen as reducing agent leads to a nanomaterial with unagglomerated round-shaped metallic particles with mean size of 6-7 nm.

Tuning surface basic properties of nanocrystalline MgO by controlling the preparation conditions.

Different nanocrystalline magnesias were synthesized by precipitation and hydrothermal treatments of aqueous salt solutions in an attempt to tune their surface basicity. CO(2) was chosen as an acidic molecule to probe the basic sites by both temperature-programmed desorption and infrared spectroscopy. All samples were shown to be crystalline, and except that obtained by nitrate decomposition, they all possessed high surface areas. The oxides presented different basic site distributions, evidencing the significant role of the preparation conditions on tuning the surface basicity: while medium-strength basic centers are dominant in the samples prepared by precipitation aging or hydrothermal treatment, the one obtained by precipitation features a roughly equal concentration of medium-strength and strong centers. Infrared spectra revealed that hydrogen carbonate and monodentate and bidentate carbonates were formed in distinct proportion on all oxides. However, the bidentate complexes were shown to have different thermal stabilities; the more stable species are thought to be formed on acid-base pair centers associated with an anionic vacancy. Distinct morphological and structural characteristics were also observed by high-resolution transmission electron microscopy. It was consistently found that the high-surface area samples are formed by aggregates of nanoparticles (2-5 nm) randomly oriented and with a high concentration of structural defects. These findings allowed us to conclude that the surface heterogeneity promoted during synthesis increases the concentration of basic sites and plays an important role in tuning the basicity of the solids.