The relevance of arsenic speciation analysis in health & medicine.

Long term exposure to arsenic through consumption of contaminated groundwater has been a global issue since the last five decades; while from an alternate standpoint, arsenic compounds have emerged as unparallel chemotherapeutic drugs. This review highlights the contribution from arsenic speciation studies that have played a pivotal role in the progression of our understanding of the biological behaviour of arsenic in humans. We also discuss the limitations of the speciation studies and their association with the interpretation of arsenic metabolism. Chromatographic separation followed by spectroscopic detection as well as the utilization of biotinylated pull-down assays, protein microarray and radiolabelled arsenic have been instrumental in identifying hundreds of metabolic arsenic conjugates, while, computational modelling has predicted thousands of them. However, these species exhibit a variegated pattern, which supports more than one hypothesis for the metabolic pathway of arsenic. Thus, the arsenic species are yet to be integrated into a coherent mechanistic pathway depicting its chemicobiological fate. Novel biorelevant arsenic species have been identified due to significant evolution in experimental methodologies. However, these methods are specific for the identification of only a group of arsenicals sharing similar physiochemical properties; and may not be applicable to other constituents of the vast spectrum of arsenic species. Consequently, the identity of arsenic binding partners in vivo and the sequence of events in arsenic metabolism are still elusive. This resonates the need for additional focus on the extraction and characterization of both low and high molecular weight arsenicals in a combinative manner.

Potential of electrical impedance spectroscopy to differentiate between healthy and osteopenic bone.

Osteoporosis involves loss of structural stability of bone due to an increase in bone porosity. Dual energy X-ray absorptometry is used to evaluate bone in terms of quantity. However, it does not give an evaluation of the patient's bone quality. For this, present study has been carried out to assess the structural deterioration of bone using electrical impedance spectroscopy. METHODS: Electrical Impedance Spectroscopy has been applied to evaluate the structural and compositional changes of cortical bone in the frequency range of 50 Hz to 5 MHz for the ovariectomized rat model. Initially, bone resorption in the ovariectomized group has been confirmed by estimating tartaric resistant acid phosphatase levels; morphometric parameters; bone matrix components, hydroxyapatite crystallite size and bone micro architecture. The mid diaphyseal regions from the femora and tibiae of sixty days post ovariectomy and control rats were used for the measurement of dielectric parameters. A dispersion model based analysis has been developed by a complex least square fitting of the dielectric data. FINDINGS: Increased tartaric resistant acid phosphatase levels, altered bone matrix components, hydroxyapatite crystallite size and disturbed microarchitecture in the ovariectomized group give us the confirmation of increased bone resorption following estrogen deficiency. These changes were shown to be reflected by single dispersion model based fitted parameters which shows the considerable change in all the parameters of ovariectomized group compared to the control. INTERPRETATION: It has been demonstrated that the parameters of the dispersion model can reflect the bone structural and compositional changes.

Metallothionein does not sequester arsenic(III) ions in condition of acute arsenic toxicity.

The major cause of toxicity of trivalent arsenicals is due to their interaction with the sulfhydryl groups in proteins. Because of its high content, Metallothionein (MT) provides one of the most favorable conditions for the binding of As(III) ions to it. MT has long been anticipated for providing resistance in case of arsenic (As) toxicity with similar mechanism as in case of cadmium toxicity. The present study investigates whether the sequestration of As ions by MT is one of the mechanisms in providing protection against acute arsenic toxicity. A rat model study on the metal stoichiometric analysis of MT1 isoform isolated from the liver of arsenic treated, untreated and zinc treated animals has been carried out using the combination of particle induced X-ray emission (PIXE) and electrospray ionisation mass spectrometry (ESI-MS). The results revealed the absence of arsenic bound MT1 in the samples isolated from arsenic treated animals. Although, both Cu and Zn ions were present in MT1 samples isolated from all the treatment groups. Moreover, only partially metallated MT1 with varying number of Zn ions were observed in all the groups. These results suggest that the role of MT during acute arsenic toxicity is different from its already established role in case of cadmium toxicity.

Conformational behavior of polyalanine peptides with and without protecting groups of varying chain lengths: population of PP-II structure!

Oculopharyngeal muscular dystrophy (OPMD), a polyalanine myopathy, occurs due to expansion of homo-polyalanine stretch in normal polyadenylating binding protein nuclear 1 (PABPN1) protein from Ala10 to Ala11-17. Therefore, the conformational behavior of polyalanine peptides with n = 10-17, with and without terminal protecting groups, have been investigated with different starting geometries in water by molecular dynamics simulation studies. Alanine peptides are shown to give rise to unordered structure irrespective of starting geometry and not more than two residues at a stretch have the same/similar set of φ, ψ values. However, the final structure with terminal protecting groups look like ß-strand. Unprotected poly-Ala peptides adopt twisted ß-hairpin/multi hairpin like structure with increasing chain length. The number of residues having φ, ψ values in collagen region is found to be less in peptides with unprotected termini as compared to peptides with protected termini of same chain length. The results have been supported by recent synchrotron radiation circular dichroism spectroscopy of polyproline II and unordered secondary structures. Opening of the helical structure in poly-Ala peptides with protecting groups has been shown to take place from C-terminal and in peptides without protecting groups opening of helix starts from both terminals. Further, opening of helix takes more time in poly-Ala peptides without terminal protecting groups. The deviations in amide bond planarity have been discussed and compared with available experimental and computational results.

Assessment of abstract reasoning abilities in alcohol-dependent subjects: an fMRI study.

INTRODUCTION: Chronic alcohol abuse has been traditionally associated with impaired cognitive abilities. The deficits are most evident in higher order cognitive functions, such as abstract reasoning, problem solving and visuospatial processing. The present study sought to increase current understanding of the neuropsychological basis of poor abstract reasoning abilities in alcohol-dependent subjects using functional magnetic resonance imaging (fMRI). METHODS: An abstract reasoning task-based fMRI study was carried out on alcohol-dependent subjects (n = 18) and healthy controls (n = 18) to examine neural activation pattern. The study was carried out using a 3-T whole-body magnetic resonance scanner. Preprocessing and post processing was performed using SPM 8 software. RESULTS: Behavioral data indicated that alcohol-dependent subjects took more time than controls for performing the task but there was no significant difference in their response accuracy. Analysis of the fMRI data indicated that for solving abstract reasoning-based problems, alcohol-dependent subjects showed enhanced right frontoparietal neural activation involving inferior frontal gyrus, post central gyrus, superior parietal lobule, and occipito-temporal gyrus. CONCLUSIONS: The extensive activation observed in alcohol dependents as compared to controls suggests that alcohol dependents recruit additional brain areas to meet the behavioral demands for equivalent task performance. The results are consistent with previous fMRI studies suggesting decreased neural efficiency of relevant brain networks or compensatory mechanisms for the execution of task for showing an equivalent performance.

Metal stoichiometry of isolated and arsenic substituted metallothionein: PIXE and ESI-MS study.

The stoichiometric analysis of the metal induced Metallothionein (MT) is pertinent for understanding the metal-MT interactions. Despite innumerable publications on MT, the literature addressing these aspects is limited. To bridge this gap, PIXE and ESI-MS analysis of the commercial rabbit liver MT1 (an isoform of MT), zinc induced isolated rat liver MT1, apo and Arsenic substituted rabbit liver MT1 have been carried out. These techniques in combination provide information about number and the signature of all the metal ions bound to MT. By using ESI-MS in the rabbit MT1, ions of Zn n MT1 (n = 0, 1, 4, 5, 6, 7) whereas, in rat MT1, the Zn1MT1 and Zn5MT1 ions are observed. PIXE analysis shows that some copper along with zinc is also present in the rabbit as well as rat MT1 which could not be assessed with ESI-MS. During As metallation reaction with rabbit MT1, with increase in arsenic concentration, the amount of arsenic bound to MT1 also increases, though not proportionally. The presence of both Zn and Cu in MT1 on Zn supplementation can be related to the role of MT in Zn and Cu homeostasis. Further, the presence of partially metallated MT1 suggests that MT1 may donate fractional amount of metal from it's fully metallated form to other proteins where Zn acts as a cofactor.

Structural and immunogenic effects of multiple hapten loading on carrier protein.

Haptens are low-molecular-weight compounds that are usually nonimmunogenic in nature. These compounds are, in general, conjugated with carrier proteins to elicit an immune response for antibody production. In this work, we report the effect of multiple hapten loading on carrier protein after conjugation by monitoring the structural and immunogenic properties of the protein. Biochemical conjugation of carboxylated hapten (atrazine derivative) to bovine serum albumin via epsilon-amino groups of lysine residues was monitored by the intrinsic fluorescence intensity of tryptophan residues of protein. A significant blue shift of emission maxima confirmed the conformational changes with increasing molar ratio of hapten:protein. Circular dichroism spectroscopy suggested a decreasing trend for alpha-helical and increased formation of beta-sheet structures in hapten-loaded protein. A further insight was sought by using molecular modeling methods for understanding of structural changes in the native protein post-hapten conjugation. A sequential approach for hapten loading on the carrier confirmed that initial binding could affect the possible binding sites for subsequent incorporation of hapten molecules. These changes play a major role in the immunogenic response of hapten-carrier conjugate. The approach taken to develop this model is promising, and can be generalized for studies with other protein-hapten combinations.