Explore the vast range of titles available.

BackgroundThe fate of gold nanoparticles, 2, 40 and 100 nm, administered intratracheally to adult female mice was examined. The nanoparticles were traced by autometallography (AMG) at both ultrastructural and light microscopic levels. Also, the gold content was quantified by inductively coupled plasma mass spectrometry (ICP-MS) and neutron activation analysis (NAA). The liver is the major site of deposition of circulating gold nanoparticles. Therefore the degree of translocation was determined by the hepatic deposition of gold. Mice were instilled with 5 intratracheal doses of gold nanoparticles distributed over a period of 3 weeks and were killed 24 h after the last dose. One group of mice were given a single intratracheal dose and were killed after 1 h.ResultsThe instilled nanoparticles were found in lung macrophages already 1 h after a single instillation. In mice instilled treated repeatedly during 3 weeks, the load was substantial. Ultrastructurally, AMG silver enhanced gold nanoparticles were found in lysosome-/endosome-like organelles of the macrophages and analysis with AMG, ICP-MS and NAA of the liver revealed an almost total lack of translocation of nanoparticles. In mice given repeated instillations of 2 nm gold nanoparticles, 1.4‰ (by ICP-MS) to 1.9‰ (by NAA) of the instilled gold was detected in the liver. With the 40 nm gold, no gold was detected in the liver (detection level 2 ng, 0.1‰) except for one mouse in which 3‰ of the instilled gold was found in the liver. No gold was detected in any liver of mice instilled with 100 nm gold (detection level 2 ng, 0.1‰) except in a single animal with 0.39‰ of the dose in the liver.ConclusionWe found that that: (1) inert gold nanoparticles, administered intratracheally are phagocytosed by lung macrophages; (2) only a tiny fraction of the gold particles is translocated into systemic circulation. (3) The translocation rate was greatest with the 2 nm gold particles.

Traumatic brain injury (TBI) is one of the leading causes of disability and death among young people. Although much is already known about secondary brain damage the full range of brain tissue responses to TBI remains to be elucidated. A population of neurons located in cerebral areas associated with higher cognitive functions harbours a vesicular zinc pool co-localized with glutamate. This zinc enriched pool of synaptic vesicles has been hypothesized to take part in the injurious signalling cascade that follows pathological conditions such as seizures, ischemia and traumatic brain injury. Pathological release of excess zinc ions from pre-synaptic vesicles has been suggested to mediate cell damage/death to postsynaptic neurons. In order to substantiate the influence of vesicular zinc ions on TBI, we designed a study in which damage and zinc movements were analysed in several different ways. Twenty-four hours after TBI ZnT3-KO mice (mice without vesicular zinc) were compared to littermate Wild Type (WT) mice (mice with vesicular zinc) with regard to histopathology. Furthermore, in order to evaluate a possible neuro-protective dimension of chemical blocking of vesicular zinc, we treated lesioned mice with either DEDTC or selenite. Our study revealed that chemical blocking of vesicular zinc ions, either by chelation with DEDTC or accumulation in zinc-selenium nanocrystals, worsened the effects on the aftermath of TBI in the WT mice by increasing the number of necrotic and apoptotic cells within the first 24 hours after TBI, when compared to those of chemically untreated WT mice. ZnT3-KO mice revealed more damage after TBI compared to WT controls. Following treatment with DEDTC or selenium an increase in the number of both dead and apoptotic cells were seen in the controls within the first 24 hours after TBI while the degree of damage in the ZnT3-KO mice remained largely unchanged. Further analyses revealed that the damage development in the two mouse strains was almost identical after either zinc chelation or zinc complexion therapy.

Ion transporters of the Slc30A- (ZnT-) family regulate zinc fluxes into sub-cellular compartments. beta-cells depend on zinc for both insulin crystallization and regulation of cell mass. This study examined: the effect of glucose and zinc chelation on ZnT gene and protein levels and apoptosis in beta-cells and pancreatic islets, the effects of ZnT-3 knock-down on insulin secretion in a beta-cell line and ZnT-3 knock-out on glucose metabolism in mice during streptozotocin-induced beta-cell stress. In INS-1E cells 2 mM glucose down-regulated ZnT-3 and up-regulated ZnT-5 expression relative to 5 mM. 16 mM glucose increased ZnT-3 and decreased ZnT-8 expression. Zinc chelation by DEDTC lowered INS-1E insulin content and insulin expression. Furthermore, zinc depletion increased ZnT-3- and decreased ZnT-8 gene expression whereas the amount of ZnT-3 protein in the cells was decreased. Zinc depletion and high glucose induced apoptosis and necrosis in INS-1E cells. The most responsive zinc transporter, ZnT-3, was investigated further; by immunohistochemistry and western blotting ZnT-3 was demonstrated in INS-1E cells. 44% knock-down of ZnT-3 by siRNA transfection in INS-1E cells decreased insulin expression and secretion. Streptozotocin-treated mice had higher glucose levels after ZnT-3 knock-out, particularly in overt diabetic animals. Zinc transporting proteins in beta-cells respond to variations in glucose and zinc levels. ZnT-3, which is pivotal in the development of cellular changes as also seen in type 2 diabetes (e.g. amyloidosis in Alzheimer's disease) but not previously described in beta-cells, is present in this cell type, up-regulated by glucose in a concentration dependent manner and up-regulated by zinc depletion which by contrast decreased ZnT-3 protein levels. Knock-down of the ZnT-3 gene lowers insulin secretion in vitro and affects in vivo glucose metabolism after streptozotocin treatment.

Introduction Qualitative and quantitative methods provide different and complementary insights into patients’ preferences for treatment. Objective The aim of this study was to use a novel, mixed-methods approach employing qualitative and quantitative approaches to generate preliminary insights into patient preferences for the treatment of a rare disease—generalized myasthenia gravis (gMG). Methods We conducted a mixed-methods study to collect exploratory qualitative and quantitative patient preference information and generate informative results within a condensed timeline (about 4 months). Recruitment was facilitated by an international health research firm. Study participants first reviewed a brief document describing six treatment attributes (to facilitate more efficient review of the material during the focus groups) and were then provided a link to complete an online quantitative survey with a single risk threshold task. They then participated in online focus groups, during which they discussed qualitative questions about their experience with gMG treatment and completed up to three quantitative threshold tasks, the first of which repeated the threshold task from the online survey. Results The study elicited both quantitative data on 18 participants’ risk tolerance and qualitative data on their treatment experience, additional treatment attributes of importance, the reasoning behind their preferences, and the trade-offs they were willing to make. Most participants ( n  = 15) chose the same hypothetical treatment in the first threshold task in the online survey and the focus groups. Focus group discussions provided insights into participants’ choices in the threshold tasks, confirmed that all the attributes were relevant, and helped clarify what was important about the attributes. Conclusions Patient preference information can be collected using a variety of approaches, both qualitative and quantitative, tailored to fit the research needs of a study. The novel mixed-methods approach employed in this study efficiently captured patient preference data that were informative for exploratory research, internal decision making, and future research.

Ion transporters of the Slc30A- (ZnT-) family regulate zinc fluxes into sub-cellular compartments. [beta]-cells depend on zinc for both insulin crystallization and regulation of cell mass. This study examined: the effect of glucose and zinc chelation on ZnT gene and protein levels and apoptosis in [beta]-cells and pancreatic islets, the effects of ZnT-3 knock-down on insulin secretion in a [beta]-cell line and ZnT-3 knock-out on glucose metabolism in mice during streptozotocin-induced [beta]-cell stress. In INS-1E cells 2 mM glucose down-regulated ZnT-3 and up-regulated ZnT-5 expression relative to 5 mM. 16 mM glucose increased ZnT-3 and decreased ZnT-8 expression. Zinc chelation by DEDTC lowered INS-1E insulin content and insulin expression. Furthermore, zinc depletion increased ZnT-3- and decreased ZnT-8 gene expression whereas the amount of ZnT-3 protein in the cells was decreased. Zinc depletion and high glucose induced apoptosis and necrosis in INS-1E cells. The most responsive zinc transporter, ZnT-3, was investigated further; by immunohistochemistry and western blotting ZnT-3 was demonstrated in INS-1E cells. 44% knock-down of ZnT-3 by siRNA transfection in INS-1E cells decreased insulin expression and secretion. Streptozotocin-treated mice had higher glucose levels after ZnT-3 knock-out, particularly in overt diabetic animals. Zinc transporting proteins in [beta]-cells respond to variations in glucose and zinc levels. ZnT-3, which is pivotal in the development of cellular changes as also seen in type 2 diabetes (e.g. amyloidosis in Alzheimer's disease) but not previously described in [beta]-cells, is present in this cell type, up-regulated by glucose in a concentration dependent manner and up-regulated by zinc depletion which by contrast decreased ZnT-3 protein levels. Knock-down of the ZnT-3 gene lowers insulin secretion in vitro and affects in vivo glucose metabolism after streptozotocin treatment.

Nano-sized clusters of gold atoms, or alternatively silver, mercury, bismuth, or zinc sulphide/selenide molecules, can be autometallographically silver-enhanced by being placed in a developer containing reducing molecules and silver ions, i.e. an autometallographic developer. A specific recipe has been worked out for each autometallographically traceable metal, and in cases where two or more autometallographic catalysts are present in the same section it is feasible to distinguish one from the other by chemical removal of one or the other of the metals. In the present study we present protocols that allow differentiation and control of specificity of the established autometallographically detectable metals. It is recommended to implement a multi-element analysis, e.g. proton-induced X-ray emission on a few samples to secure the histochemical data.[PUBLICATION ABSTRACT]

The present study demonstrates that cultured macrophages are able to liberate gold ions from metallic gold surfaces, a process suggested to be called \"dissolucytosis\", in a way analogous to the release taking place when metallic implants are placed in a body. Using the ultra-sensitive autometallographic (AMG) technique, we demonstrate that murine macrophages grown on a surface of metallic gold liberate gold ions. Ultra-structural AMG reveals that the gold ions are located in an ultra-thin membrane-like structure, \"the dissolution membrane\", intervened between the macrophages and the metal surface. The presence of AMG silver enhanced gold nanoparticles in the dissolution membrane proves that the release of charged gold atoms takes place extracellularly. The dissolution membrane is most likely secreted and chemically controlled by the \"dissolucytes\", here macrophages, and the membrane is essential for the dissolution of metal implants and particles, which cannot be phagocytosed. Our findings support the notion that whenever a metallic gold surface is attacked by dissolucytes, gold ions are liberated and taken up by surrounding cells. As gold ions can suppress the inflammatory process, it is reasonable to expect that when dissolucytosis takes place in the living organism the liberated gold ions will cause local immunosuppression.

Zinc is intimately involved in insulin metabolism, its major known role being the binding of insulin in osmotically stable hexamers in beta-cell granules. To investigate the anatomical distribution of zinc ions necessary for insulin binding we examined the rat pancreas by autometallography (AMG). AMG demonstrates chelatable zinc and is a sensitive marker for zinc in vesicles and also a surrogate marker for recently described zinc pumps regulating intravesicular zinc metabolism. Zinc ions were found in alpha- and beta-cell granules, primarily in the periphery of the granules. Only occasionally was zinc seen in other islet cell types. AMG allows the study of the microscopic and ultrastructural localisation of free zinc ions in the pancreas. The applicability of the method at the ultrastructural level in particular makes AMG a very sensitive tool in future studies on the role of zinc ions in the pancreas.

Bismuth subnitrate was injected into the triceps surae muscle of 3-month-old male Wistar rats. Sections of lumbar spinal cord (L4-L6) and corresponding dorsal root ganglia were developed by autometallography (AMG) to trace possible bismuth in neuronal somata resulting from retrograde axonal transport. At 3 days after treatment bismuth clusters could be traced by AMG in spinal cord motor neurons and in dorsal root ganglion cells ipsilateral to the injection site. Retrograde transport of bismuth could be avoided by ligation or intraneuronal injection of cholchicine into the sciatic nerve. At the ultrastructural level bismuth was found to be located exclusively in lysosome-like organelles in motor and sensory neuronal somata projecting to the injection site. The present study shows that bismuth is transported retrogradely in both sensory and motor axons if their terminals are exposed to bismuth ions.

Traumatic brain injury results in loss of neurons caused as much by the resulting neuroinflammation as by the injury. Gold salts are known to be immunosuppressive, but their use are limited by nephrotoxicity. However, as we have proven that implants of pure metallic gold release gold ions which do not spread in the body, but are taken up by cells near the implant, we hypothesize that metallic gold could reduce local neuroinflammation in a safe way. Bio-liberation, or dissolucytosis, of gold ions from metallic gold surfaces requires the presence of disolycytes i.e. macrophages and the process is limited by their number and activity. We injected 20–45 μm gold particles into the neocortex of mice before generating a cryo-injury. Comparing gold-treated and untreated cryolesions, the release of gold reduced microgliosis and neuronal apoptosis accompanied by a transient astrogliosis and an increased neural stem cell response. We conclude that bio-liberated gold ions possess pronounced anti-inflammatory and neuron-protective capacities in the brain and suggest that metallic gold has clinical potentials. Intra-cerebral application of metallic gold as a pharmaceutical source of gold ions represents a completely new medical concept that bypasses the blood-brain-barrier and allows direct drug delivery to inflamed brain tissue.