Spatial distribution of trace metals and associated transport proteins during bacterial infection.

Innate immune systems alter the concentrations of trace elements in host niches in response to invading pathogens during infection. This work reports the interplay between d-block metal ions and their associated biomolecules using hyphenated elemental techniques to spatially quantify both elemental distributions and the abundance of specific transport proteins. Here, lung tissues were collected for analyses from naïve and Streptococcus pneumoniae-infected mice fed on a zinc-restricted or zinc-supplemented diet. Spatiotemporal distributions of manganese (55Mn), iron (56Fe), copper (63Cu), and zinc (66Zn) were determined by quantitative laser ablation-inductively coupled plasma-mass spectrometry. The murine transport proteins ZIP8 and ZIP14, which are associated with zinc transport, were also imaged by incorporation of immunohistochemistry techniques into the analytical workflow. Collectively, this work demonstrates the potential of a single instrumental platform suitable for multiplex analyses of tissues and labelled antibodies to investigate complex elemental interactions at the host-pathogen interface. Further, these methods have the potential for broad application to investigations of biological pathways where concomitant measurement of elements and biomolecules is crucial to understand the basis of disease and aid in development of new therapeutic approaches.

Dietary zinc and the control of Streptococcus pneumoniae infection.

Human zinc deficiency increases susceptibility to bacterial infection. Although zinc supplementation therapies can reduce the impact of disease, the molecular basis for protection remains unclear. Streptococcus pneumoniae is a major cause of bacterial pneumonia, which is prevalent in regions of zinc deficiency. We report that dietary zinc levels dictate the outcome of S. pneumoniae infection in a murine model. Dietary zinc restriction impacts murine tissue zinc levels with distribution post-infection altered, and S. pneumoniae virulence and infection enhanced. Although the activation and infiltration of murine phagocytic cells was not affected by zinc restriction, their efficacy of bacterial control was compromised. S. pneumoniae was shown to be highly sensitive to zinc intoxication, with this process impaired in zinc restricted mice and isolated phagocytic cells. Collectively, these data show how dietary zinc deficiency increases sensitivity to S. pneumoniae infection while revealing a role for zinc as a component of host antimicrobial defences.

Protective effect of selenium supplementation following oxidative stress mediated by glucose on retinal pigment epithelium.

There are many conditions that affect the retina. However, diabetic retinopathy (RD) as a complication of Diabetes Mellitus continues to be the leading cause of blindness in working people globally. Diabetic retinopathy is an ocular complication of diabetes that is caused by the deterioration of the blood vessels that supply the retina, which has the consequence that the vision deteriorates irreversibly. The retina, and specifically the retinal pigment epithelium (RPE) is the only neural tissue that is exposed directly and frequently to light, which favors the oxidation of lipids that become extremely toxic to the cells of the retina. The RPE is a natural barrier playing an important role in the absorption of light and reduction of light scatter within the eye. In addition, the retina is the tissue that proportionally consumes more oxygen, which generates a high production of reactive oxygen species (ROS). The retina is particularly sensitive to hyperglycemia and oxidative stress. The eye tissues are enriched in certain antioxidants in the form of metabolic enzymes or small molecules. Since selenium is essential for regulating the activity of the enzymes involved in protection against oxidative stress, providing selenium to the ocular tissues could be useful for the treatment of different ocular pathologies. Thus, the aim of this study is to investigate the potential efficacy of selenium in human RPE against glucose-induced oxidative stress and its implications for GPx activity. Chromatographic techniques based on HPLC-ICP-MS will be applied in combination with isotope pattern deconvolution (IPD) to study the effects of selenium supplementation and hyperglycemia in an in vitro model of RPE cells.

Selenium levels and Glutathione peroxidase activity in the plasma of patients with type II diabetes mellitus.

Selenium, an essential trace element, is involved in the complex system of defense against oxidative stress through selenium-dependent glutathione peroxidases (GPx) and other selenoproteins. Because of its antioxidant properties, selenium or its selenospecies at appropriate levels could hinder oxidative stress and so development of diabetes. In this vein, quantitative speciation of selenium in human plasma samples from healthy and diabetic patients (controlled and non-controlled) was carried out by affinity chromatography (AF) coupled on-line to inductively coupled plasma mass spectrometry (ICP-MS) and isotope dilution analysis (IDA). Similarly, it is well known that patients with diabetes who exhibit poor control of blood glucose show a decreased total antioxidant activity. Thus, we evaluated the enzymatic activity of GPx in diabetic and healthy individuals, using the Paglia and Valentine enzymatic method, observing a significant difference (p<0.05) between the three groups of assayed patients (healthy (n=24): 0.61±0.11U/ml, controlled diabetic (n=38): 0.40±0.12U/ml and non-controlled diabetic patients (n=40): 0.32±0.09U/ml). Our results show that hyperglycemia induces oxidative stress in diabetic patients compared with healthy controls. What is more, glycation of GPx experiments demonstrated that it is the degree of glycation of the selenoenzyme (another species of the Se protein) what actually modulates its eventual activity against ROS in type II diabetes mellitus patients.