Academic Journal List

In vivo phytochelatins (PCs) and their corresponding Hg–PC complexes were characterized using RPLC-ESI-MS/MS in the roots of Brassica chinensis L. under the stress of a mercury cysteine complex (HgCys2) and/or a mercury humic acid complex (Hg–HA). Results indicated that the presence of Cys and/or HA decreased the Hg uptake in both the roots and shoots of B. chinensis but increased the generation of PCs in the roots compared with those where only HgCl2 was in the culture solutions. A series of Hg–PC complexes were synthesized in vitro for predicting the possible Hg–PC formed in vivo in the HgCys2 and/or Hg–HA stressed roots of B. chinensis. The discovery of in vivo oxidized PC2, PC3 and PC4 and their corresponding HgPC2, HgPC3, HgPC4 and Hg2PC4, which were confirmed by their specific isotope distribution, provided definite evidence for understanding the defense and accumulation mechanism of B. chinensis to Hg, in which the induced PCs play an important role not only in Hg detoxification through forming Hg–PC complexes but also for reducing the oxidative stress induced by Hg2+.

Vanadyl(IV) complexes are anti-diabetogenic agents. Intra-peritoneal administration of bis(allixinato)oxidovanadium(IV) [VO(alx)2] lowers high blood glucose levels in animal models of type 1 and type 2 diabetes. We have examined whether oral administration of VO(alx)2 restores impaired activation in signaling cascades related to glucose metabolism and insulin action, and alters gene expression in the skeletal muscles of streptozotocin (STZ)-induced diabetic mice (STZ-diabetic mice). We report here that daily oral administration of VO(alx)2 lowered high blood glucose levels in the STZ-diabetic mice. The oral administration of VO(alx)2 enhanced phosphorylation of Akt and glycogen synthase kinase-3β (GSK3β), located downstream of the insulin receptor cascade in the skeletal muscles. We analyzed gene expression in the muscles of the diabetic mice before and after insulin or VO(alx)2 treatment. Treating the diabetic mice with insulin or VO(alx)2 normalized the gene expression levels of 152 down-regulated and 11 up-regulated genes, and especially the up-regulation of Cyp2E1 and FoxO1 in the muscles of the diabetic mice. The insulin-mimetic effects of VO(alx)2 in the STZ-induced diabetic mice may be due to the enhancement of protein phosphorylation leading to the activation or inactivation of the transcriptional machinery. Our findings suggest that the insulin-mimetic effects of VO(alx)2 in diabetes may be due to changes in the protein phosphorylations and their gene expression levels.

The interaction of thimerosal, an ethylmercury-containing bactericide and fungicide used as preservative in vaccines and other drugs, with free thiols in proteins has been investigated using gradient reversed phase liquid chromatography (LC) with inductively coupled plasma mass spectrometry (ICP-MS) and electrospray mass spectrometry (ESI-MS) detection. As model proteins, β-lactoglobulin A (18.4 kDa) from bovine milk and human serum albumin (66.5 kDa) have been used. Physiological conditions upon an intravenous injection of thimerosal-containing drugs were mimicked. The formation of ethylmercury–protein adducts was proved and the identification of the binding site of ethylmercury, a free thiol residue in the peptide T13 was achieved after tryptic digestion of β-lactoglobulin A.

Selenometabolites transformed from selenomethionine (SeMet) in wheat germ extract (WGE) were identified by complementary use of HPLC-ICP-MS and HPLC-ESI-MS/MS. Three selenium (Se)-containing peaks tentatively named WGE1, WGE2, and WGE3 were detected by HPLC-ICP-MS. WGE1 had [M]+ at m/z 212 on HPLC-ESI-MS analysis, and its fragment ions indicated that WGE1 is selenomethionine methylselenonium (MeSeMet). WGE2 and WGE3 exhibited absorption at 254 nm and molecular ions at m/z 433 and 447, respectively. Their fragment ions revealed that WGE2 and WGE3 are Se-adenosylselenohomocysteine (AdoSeHcy) and Se-adenosylselenomethionine (AdoSeMet), respectively. Their structures were coincident with the absorption of WGE2 and WGE3 at 254 nm. In addition, a trace amount of AdoSeMet was suggested to also exist in rabbit reticulocyte lysate, a mammalian in vitro translation system, because the transformation of AdoSeMet from SeMet was completely inhibited by (2S)-2-amino-4,5-epoxypentanoic acid (AEPA), a potent inhibitor of AdoMet synthetase. These results suggest that SeMet and methionine (Met) share a common metabolic pathway, i.e., SeMet is not only incorporated into proteins in place of Met but also metabolized to AdoSeMet in higher eukaryotes and MeSeMet in plants.

A time-of-flight mass spectrometer (TOFMS), which employs inductively coupled plasma (ICP) and electrospray ionization (ESI) sources simultaneously, has been designed, constructed, and evaluated for comprehensive elemental speciation analysis. The simultaneous operation of both sources with a single mass spectrometer is an improvement over existing techniques. The mass analyzer shares a third-stage vacuum system, extraction region, acceleration region, field-free region, and two-stage reflectron between both sources. Most of the other components, such as first and second-stage vacuum systems, pre-extraction ion optics, microchannel plate detectors, and data acquisition are independent, to provide the greatest degree of flexibility in source operation and signal optimization. A detailed description is given of the design and optimization of the orthogonal acceleration and spontaneous drift geometry, energy discrimination, and the reflectron and preliminary performance data are presented.

The arsenic metallome in African Green Monkey kidney cells is probed by measuring cytotoxicity, cellular arsenic uptake and speciation studies on arsenic-containing chemical warfare agent degradation products (CWDPs) during cell uptake. Inorganic arsenic compounds and methylated species also were studied during cell uptake as a means of providing cytotoxicity information relative to the CWDPs. The degradation products used were phenylarsine oxide (PAO), phenylarsonic acid (PAA), diphenylarsinic acid (DPAA), triphenylarsine (TPA) and triphenylarsine oxide (TPAO). These are the warfare agent’s primary degradation products. The parent warfare agents (red agents) are diphenylarsine chloride (DA or referred to as Clark I) and diphenylarsine cyanide (DC or Clark II), sternutator agents, sneezing gases able to cause bronchial irritation. Cytotoxicity levels and cellular uptake were compared to those of inorganic species: sodium arsenite, NaAsO2 [As(III)], sodium arsenateNa2HAsO4 [As(V)] and methylated arsenicals such as dimethylarsinic acid (DMA) and methylarsonic acid (MMA). The arsenic uptake was demonstrated in an African Green Monkey kidney cell line, CV-1. Quantification of lactate dehydrogenase activityreleased from damaged/dying cells was then measured via an LDH assay. The purpose of this study is to initially investigate toxicity to cells when exposed to different arsenic containing compounds over different concentrations and time ranges from 3 h to 24 h. Furthermore, exposed cells were then analyzed for different arsenic species by high performance liquid chromatography (HPLC) with inductively coupled plasma mass spectrometry to isolate and speciate arsenic fractions followed by nanoLC electrospray ionization mass spectrometry to analyze the molecular level changes of the arsenic based degradation products in the kidney cells. Metabolic changes to the arsenic species were found, and interestingly, at the lowest uptake levels, cytotoxicities were generally higher for the chemical warfare agent degradation products than the inorganic arsenic species.

This study demonstrates the application of quantitative elemental bio-imaging for the determination of the distribution Cu, Mn, Fe and Zn in Parkinsonism mouse model brains. Elevated concentrations of these metals within the substantia nigra (SN) are suspected to play a role on the development of Parkinson’s disease. Elemental bio-imaging employs laser ablation inductively coupled mass spectrometry (LA-ICP-MS) to construct images of trace element distribution. Quantitative data was produced by ablating the standard tissue sections and recording the mean signal intensity calibrated against multi level matrix matched tissue standards. The concentrations of Fe within the substantia nigra of the lesioned animals increased significantly when compared against control animals. Furthermore, the data was compared against solution nebulisation ICP-MS in which the whole substantia nigra was excised. The trends were the same for both methods; however the elemental bio-imaging method returned significantly higher concentrations. This was caused by dilution from inclusion of surrounding tissue of the SN during the excision procedure.

In this review, we discuss genetic disorders involving altered copper metabolism, particularly in relation to Menkes disease (MD), occipital horn syndrome (OHS), and Wilson’s disease (WD). The responsible genes for MD and WD are ATP7A and ATP7B, respectively. Both proteins encoded by these genes are responsible for transporting copper from the cytosol to the Golgi apparatus. However, the pathology of MD is completely different from that of WD, that is, MD is characterized by a copper deficiency while WD is caused by a toxic excess of copper. The reason for this difference is related to the particular cell types in which the ATP7A and ATP7B proteins are expressed. ATP7A is expressed in almost all cell types except hepatocytes, whereas ATP7B is mainly expressed in hepatocytes. MD and OHS are X-linked recessive disorders characterized by copper deficiency. Typical features of MD, such as neurological disturbances, connective tissue disorders, and hair abnormalities, can be explained by the abnormally low activity of copper-dependent enzymes. The current standard-of-care treatment for MD is parenteral administrations of copper–histidine. When the treatment is initiated in newborn babies prior to two months of age, the neurological degeneration may be prevented, but delayed treatment is considerably less effective. Moreover, copper–histidine treatment does not improve symptoms of the connective tissue disorders. As such, systems for mass screening of neonates for MD should be implemented. At the same time, novel treatments targeting connective tissue disorders need to be developed. OHS is a milder form of MD and is characterized by connective tissue abnormalities. Although formal trials have not been conducted for OHS, OHS patients are typically treated in a similar manner to those with MD. WD is an autosomal recessive disorder characterized by the toxic effects of chronic exposure to high levels of copper. The hepatic and nervous systems are typically most severely affected. Numerous other symptoms can also be observed, however, making an early diagnosis difficult. Chelating agents and zinc are effective for the treatment of WD, but they are ineffective for the patients with fulminant hepatic failure. Some patients with neurological diseases show poor response to chelating agents; here again, early diagnosis and treatment are critical. Screening of newborn babies or infants for WD can help lead to timely diagnosis and treatment. Patients with WD may have a risk of hepatocellular carcinoma despite receiving treatment. An understanding of the relation between WD and hepatocellular carcinoma will provide clues to help prevent hepatocellular carcinoma in patients with WD.

In this review, the authors present a brief overview of metals and their possible roles as determinants in the pathogenesis of diabetes and complications. Of course, due to the complexity of diabetes and its far-reaching complications, it would be difficult to cover every metal that has been implicated in diabetes. Therefore, this review has two main objectives, the first of which is to educate the reader with regards to the types of diabetes and complications, especially in relation to hyperglycemia and anti-oxidant properties. Following an overview of the more cited metals in diabetes, the second objective of this review is to offer some opinions about current needs in the area of metal analysis. Specifically, the challenges for scientists to perform quantitative determinations on biological samples in near-real time with subcellular-level spatial resolution.

The success of cisplatin in clinic has stimulated great interest in the development and application of metal-based drugs for therapeutic and diagnostic purposes. However, the treatment efficiency of metallodrugs suffers from side-effects and drug resistance. To overcome these challenges, targets of these metal-based drugs should be identified in order to understand the molecular mechanisms of actions of these compounds and to the intrinsic or acquired drug resistance by cancer cells and infectious microbes. This review summaries some of the recent developments in the identification of binding proteins and their target sites of platinum-, ruthenium-, gold-, arsenic- and bismuth-containing agents by proteomics and metalloproteomics, which may provide a rational basis for the design of new metal-based drugs.