Global and cell type-specific immunological hallmarks of severe dengue progression identified via a systems immunology approach.

Severe dengue (SD) is a major cause of morbidity and mortality. To define dengue virus (DENV) target cells and immunological hallmarks of SD progression in children's blood, we integrated two single-cell approaches capturing cellular and viral elements: virus-inclusive single-cell RNA sequencing (viscRNA-Seq 2) and targeted proteomics with secretome analysis and functional assays. Beyond myeloid cells, in natural infection, B cells harbor replicating DENV capable of infecting permissive cells. Alterations in cell type abundance, gene and protein expression and secretion as well as cell-cell communications point towards increased immune cell migration and inflammation in SD progressors. Concurrently, antigen-presenting cells from SD progressors demonstrate intact uptake yet impaired interferon response and antigen processing and presentation signatures, which are partly modulated by DENV. Increased activation, regulation and exhaustion of effector responses and expansion of HLA-DR-expressing adaptive-like NK cells also characterize SD progressors. These findings reveal DENV target cells in human blood and provide insight into SD pathogenesis beyond antibody-mediated enhancement.

TiO2 NPs Induce DNA Damage in Lymphocytes from Healthy Individuals and Patients with Respiratory Diseases-An Ex Vivo/In Vitro Study.

Little is known of the effects of nanoparticles in human systems, let alone in diseased individuals and nanotechnology has preceded nanotoxicology. Therefore, the effects of titanium dioxide (TiO2) nanoparticles in peripheral blood lymphocytes from patients with respiratory diseases [lung cancer, chronic obstructive pulmonary disease (COPD) and asthma] were compared with those in healthy Individuals, to determine differences in sensitivity to nanochemical insult. The Comet assay was performed according to recommended guidelines. The micronucleus assay and ras oncoprotein detection were conducted according to published standard methods. The results showed statistically significant concentration-dependent genotoxic effects of TiO2 NPs in both respiratory patient and control groups in the Comet assay. The TiO2 NPs caused DNA damage in a concentration dependent manner in both groups (respiratory and healthy controls) with the exception of the lowest TiO2 concentration (10 µg/ml) which did not induce significant damage in healthy controls (n.s). When OTM data were used to compare the whole patient group and the control group, the patient group had more DNA damage (p > 0.001) with the exception of 10 µg/ml of TiO2 that caused less significant damage to patient lymphocytes (p < 0.05). Similarly, there was an increase in the pattern of cytogenetic damage measured in the MN assay without statistical significance except when compared to the negative control of healthy individuals. Furthermore, when modulation of ras p21 expression was investigated, regardless of TiO2 treatment, only lung cancer and COPD patients expressed measurable ras p21 levels. Results were achieved in the absence of cytotoxicity.

Acaricidal properties of two extracts from Guiera senegalensis J.F. Gmel. (Combrataceae) against Hyalomma anatolicum (Acari: Ixodidae).

Laboratory test were carried out on eggs, larvae, nymphs and adults of Hyalomma anatolicum to determine the acaricidal activities of petroleum ether (PE) and crude ethanolic extracts (EE) from the leaves of Guiera senegalensis J.F. Gmel. (Combrataceae) using immersion method. Stock solutions, of 300 mg/ml (30%) of the each two extract, were prepared. Six two-fold serial dilutions each with three replicates were used. Both extracts, at the highest concentration 150 mg/ml (15%), induced 100% failure of hatching of the treated eggs. The concentrations of PE and EE that induced 50% inhibition of the hatchability (IC50) were 1.71 and 0.508%, respectively. In the larval immersion test (LIT), EE at 15% concentration caused complete mortality while the same concentration of PE resulted in 96% mortality. The mortalities increased with concentrations. There was a correlation between the mortalities and increased concentrations, the values of the linear correlation coefficient (r) for PE and EE were 0.93 and 0.79, and The LC50 and LC99 were 2.08 and 14.09, and 0.787 and 11.054, respectively. At the concentrations of 3.75%, 7.5% and 15%, PE inhibited the molting of the nymphs by 40, 55 and 65%, respectively, while EE induced 46.49, 64.3 and 71.4% inhibition, respectively. The effectiveness of the treatment against unfed adult females was assessed by measuring the feeding performance and egg production using adult immersion test (AIT). Although, there was no mortality in unfed adults, PE and EE inhibited feeding and egg-laying of the survived females by 35-100% and 6.16-100%, respectively. Our results indicated that G. senegalensis is a promising biocontrol candidate as an acaricidal agent against H. anatolicum.

Effect of nanoparticles on human cells from healthy individuals and patients with respiratory diseases.

The effects of titanium dioxide (TiO2) nanoparticles in peripheral blood lymphocytes from patients with respiratory diseases (lung cancer, chronic obstructive pulmonary disease (COPD) and asthma) were compared with those in healthy individuals, to determine differences in sensitivity to nanochemical insult. The observations made show statistically significant concentration-dependent genotoxic effects of TiO2 in both respiratory patient and control groups in the Comet assay. An increase in the pattern of cytogenetic damage measured in the MN assay without statistical significance except when compared to the negative control of healthy individuals was also observed.

Genotoxicity and cytotoxicity of zinc oxide and titanium dioxide in HEp-2 cells.

AIMS: The rapidly growing industrial and medical use of nanomaterials, especially zinc oxide and titanium dioxide, has led to growing concerns about their toxicity. Accordingly, the intrinsic genotoxic and cytotoxic potential of these nanoparticles have been evaluated. MATERIALS & METHODS: Using a HEp-2 cell line, cytotoxicity was tested along with mitochondrial activity and neutral red uptake assays. The genotoxic potential was determined using the Comet and the cytokinesis-blocked micronucleus assays. In addition, tyrosine phosphorylation events were investigated. RESULTS & CONCLUSION: We found concentration- and time-dependent cytotoxicity and an increase in DNA and cytogenetic damage with increasing nanoparticle concentrations. Mainly for zinc oxide, genotoxicity was clearly associated with an increase in tyrosine phosphorylation. Our results suggest that both types of nanoparticles can be genotoxic over a range of concentrations without being cytotoxic.