In Vitro Reduction of Arsenic Bioavailability Using Dietary Strategies.

The main route of human exposure to inorganic arsenic (As) is through the consumption of food and water. Continued exposure to inorganic As [As(III) and As(V)] may cause a variety of diseases, including various types of cancer. The removal of As from these sources is complex, especially for food. One way to decrease As exposure could be by reducing intestinal absorption of it. The aim of this study is to seek dietary strategies (pure compounds, extracts, or supplements) that are capable of reducing the amount of As that is absorbed and reaches systemic circulation. Standard solutions of As(III) and As(V) and bioaccessible fractions of food samples with or without the dietary strategies to be tested were added to colon-derived human cells (NCM460 and HT-29MTX) to determine the apparent permeability (Papp) of As. Results show that transport across the intestinal monolayers is substantial, and the passage of As(III) (Papp = 4.2 × 10-5 cm/s) is greater than that of As(V) (Papp = 2.4 × 10-5 cm/s). Some of the treatments used (iron species, cysteine, grape extract) significantly reduce the transport of both inorganic As standards across the intestinal monolayer, thus decreasing absorption of them. In food samples, the effect of the dietary compounds on inorganic As bioavailability was also observed, especially in the cases of curcumin and cysteine. Compounds that proved effective in these in vitro assays could be the basis for intervention strategies aimed at reducing As toxicity in chronically exposed populations or regular consumers of food products with high As contents.

PRPF8 defects cause missplicing in myeloid malignancies.

Mutations of spliceosome components are common in myeloid neoplasms. One of the affected genes, PRPF8, encodes the most evolutionarily conserved spliceosomal protein. We identified either recurrent somatic PRPF8 mutations or hemizygous deletions in 15/447 and 24/450 cases, respectively. Fifty percent of PRPF8 mutant and del(17p) cases were found in AML and conveyed poor prognosis. PRPF8 defects correlated with increased myeloblasts and ring sideroblasts in cases without SF3B1 mutations. Knockdown of PRPF8 in K562 and CD34+ primary bone marrow cells increased proliferative capacity. Whole-RNA deep sequencing of primary cells from patients with PRPF8 abnormalities demonstrated consistent missplicing defects. In yeast models, homologous mutations introduced into Prp8 abrogated a block experimentally produced in the second step of the RNA splicing process, suggesting that the mutants have defects in proof-reading functions. In sum, the exploration of clinical and functional consequences suggests that PRPF8 is a novel leukemogenic gene in myeloid neoplasms with a distinct phenotype likely manifested through aberrant splicing.

Novel somatic mutations in large granular lymphocytic leukemia affecting the STAT-pathway and T-cell activation.

T-cell large granular lymphocytic (T-LGL) leukemia is a clonal disease characterized by the expansion of mature CD3+CD8+ cytotoxic T cells. It is often associated with autoimmune disorders and immune-mediated cytopenias. Our recent findings suggest that up to 40% of T-LGL patients harbor mutations in the STAT3 gene, whereas STAT5 mutations are present in 2% of patients. In order to identify putative disease-causing genetic alterations in the remaining T-LGL patients, we performed exome sequencing from three STAT mutation-negative patients and validated the findings in 113 large granular lymphocytic (LGL) leukemia patients. On average, 11 CD8+ LGL leukemia cell-specific high-confidence nonsynonymous somatic mutations were discovered in each patient. Interestingly, all patients had at least one mutation that affects either directly the STAT3-pathway (such as PTPRT) or T-cell activation (BCL11B, SLIT2 and NRP1). In all three patients, the STAT3 pathway was activated when studied by RNA expression or pSTAT3 analysis. Screening of the remaining 113 LGL leukemia patients did not reveal additional patients with same mutations. These novel mutations are potentially biologically relevant and represent rare genetic triggers for T-LGL leukemia, and are associated with similar disease phenotype as observed in patients with mutations in the STAT3 gene.

Factors affecting the bioaccessibility of fluoride from seafood products.

Fluoride is considered important for health because of its beneficial effect on the prevention of dental caries and on bone development in the child population. However, excessive intake has negative effects. The main pathway for exposure is oral, through consumption of drinking water, and some food products. Therefore its bioaccessibility (quantity of the element solubilized during the digestive process) is a parameter to be considered when estimating the risk/benefit associated with this element. The aim of the present study was to evaluate the influence of the digestion phase, gastrointestinal digestion factors (pH, pepsin and bile salt concentrations) and the presence of cations on the bioaccessibility of fluoride from seafood products. The results show that the solubilization of fluoride takes place entirely during the gastric phase. Its bioaccessibility is strongly influenced by conditions that favor the formation of insoluble complexes of fluoride with other elements present in the matrix. The factors that are most influential in reducing its bioaccessibility are the increase in pH in the gastric phase, the presence of cations, especially in the intestinal phase, and a low concentration of bile salts.

CBL mutation-related patterns of phosphorylation and sensitivity to tyrosine kinase inhibitors.

Recurrent homozygous CBL-inactivating mutations in myeloid malignancies decrease ubiquitin ligase activity that inactivates SRC family kinases (SFK) and receptor tyrosine kinases (RTK). However, the most important SFK and RTK affected by these mutations, and hence, the most important therapeutic targets, have not been clearly characterized. We compared SFK and RTK pathway activity and inhibitors in acute myeloid leukemia cell lines containing homozygous R420Q mutation (GDM-1), heterozygous deletion (MOLM13) and wild-type (WT) CBL (THP1, U937). As expected with CBL loss, GDM-1 displayed high KIT expression and granulocyte-macrophage colony-stimulating factor (GM-CSF) hypersensitivity. Ectopic expression of WT CBL decreased GDM-1 proliferation but not cell lines with WT CBL. GDM-1, but not the other cell lines, was highly sensitive to growth inhibition by dasatinib (dual SFK and RTK inhibitor, LD50 50 nM); there was less or no selective inhibition of GDM-1 growth by sunitinib (RTK inhibitor), imatinib (ABL, KIT inhibitor), or PP2 (SFK inhibitor). Phosphoprotein analysis identified phosphorylation targets uniquely inhibited by dasatinib treatment of GDM-1, including a number of proteins in the KIT and GM-CSF receptor pathways (for example, KIT Tyr721, STAT3 Tyr705). In conclusion, the promiscuous effects of CBL loss on SFK and RTK signaling appear to be best targeted by dual SFK and RTK inhibition.

Heavy metal, total arsenic, and inorganic arsenic contents of algae food products.

The total arsenic, inorganic arsenic, lead, cadmium, and mercury contents of 18 algae food products currently on sale in Spain were determined. The suitability of the analytical methodologies for this type of matrix was confirmed by evaluating their analytical characteristics. The concentration ranges found for each contaminant, expressed in milligrams per kilogram of dry weight, were as follows: total arsenic, 2.3-141; inorganic arsenic, 0.15-88; lead, < 0.05-1.33; cadmium, 0.03-1.9; and mercury, 0.004-0.04. There is currently no legislation in Spain regarding contaminants in algae food products, but some of the samples analyzed revealed Cd and inorganic As levels higher than those permitted by legislation in other countries. Given the high concentrations of inorganic As found in Hizikia fusiforme, a daily consumption of 1.7 g of the product would reach the Provisional Tolerable Weekly Intake recommended by the WHO for an average body weight of 68 kg. A more comprehensive study of the contents and toxicological implications of the inorganic As present in the algae food products currently sold in Spain may be necessary, which might then be the basis for the introduction of specific sales restrictions.

[Clinical significance of a new technique of microscopic visualization in extrapulmonary tuberculosis]. / Interés clínico de una nueva técnica de visualización microscópica en tuberculosis extrapulmonar.

BACKGROUND: The aim of this work was the clinical investigation of a new method of microscopy to improve the sensitivity in the diagnosis of tuberculosis in comparison with the actual conventional technique, without detriment of specificity. METHODS: A comparative study between Smithwick fluorescence technique and a new technique named big drop fluorescence has been made with a total of 185 clinical samples of extrapulmonary tuberculosis. All samples were carried in parallel with both technique, the Smithwick usually utilized in the majority of laboratories, and a new technique described recently by us, named big drop fluorescence, that use a sample with more concentration and different times of stain, with the same reagents. RESULTS: A sensitivity of 12.5% and 56.2% respectively was obtained with Smithwick and big drop fluorescence techniques. A specificity of 97.0% with big drop fluorescence and 98.2% with Smithwick was obtained. This improvement of sensibility was very important and the lost of specificity was so little that a statistic difference without both techniques was not observed. CONCLUSIONS: The new technique named big drop fluorescence (BDF) shows greater sensitivity in the microscopical diagnosis of extrapulmonary tuberculosis.