Surface capping and size-dependent toxicity of gold nanoparticles on different trophic levels.

In the present study, the toxicity of gold nanoparticles (Au NPs) was evaluated on various trophic organisms. Bacteria, algae, cell line, and mice were used as models representing different trophic levels. Two different sizes (CIT30 and CIT40) and surface-capped (CIT30-polyvinyl pyrrolidone (PVP)-capped) Au NPs were selected. CIT30 Au NP aggregated more rapidly than CIT40 Au NP, while an additional capping of PVP (CIT30-PVP capped Au NP) was found to enhance its stability in sterile lake water medium. Interestingly, all the forms of NPs evaluated were stable in the cell culture medium during the exposure period. Size- and dose-dependent cytotoxicities were observed in both bacteria and algae, with a strong dependence on reactive oxygen species (ROS) generation and lactate dehydrogenase (LDH) release. CIT30-PVP capped Au NP showed a significant decrease in toxicity compared to CIT30 Au NP in bacteria and algae. In the SiHa cell line, dose- and exposure-dependent decline in cell viability were noted for all three types of Au NPs. In mice, the induction of DNA damage was size and dose dependent, and surface functionalization with PVP reduced the toxic effects of CIT30 Au NP. The exposure to CIT30, CIT40, and CIT30-PVP capped Au NPs caused an alteration of the oxidative stress-related endpoints in mice hepatocytes. The toxic effects of the gold nanoparticles were found to vary in diverse test systems, accentuating the importance of size and surface functionalization at different trophic levels.

Genotoxicity evaluation of 4-carboxyl- 2,6-dinitrophenylazohydroxynaphthalenes in mice.

A short-term in vivo genotoxicity evaluation of 4-carboxyl-2,6-dinitrophenylazohydronaphthalenes (AZ-01 to AZ-04) has been carried out in mice. Aqueous colloidal solutions of the dyes were administered to mice on each day for 5 successive days using gastric gavages. Two end point assessments of the genotoxicity potentials of the dyes were assessed using comet assay and chromosomal aberration studies using the mice bone marrow cells. The dyes were well tolerated at the doses investigated, as there were no deaths or any adverse pharmacotoxic events. Dose-dependent DNA damage (in terms of percentage of tail DNA and Olive tail moment) occurred with AZ-01 and AZ-02, although the effects were significant only with the highest doses. AZ-03 gave similar patterns with those of AZ-01 and AZ-02, while replacement with butanone in AZ-04 altered the observed pattern. Minimal chromosomal damages were obtained for the four dyes, with AZ-01 and AZ-02 giving nonsignificant damages, while the highest dose of AZ-03 produced significant aberrations in terms of breaks. Some minor isochromatid breaks and gaps were also noticed in the dye-treated mice. Mitotic indices in all cases were not significantly different from concomitantly administered vehicle control showing lack of cytotoxicity of the monoazo dyes at these doses. The monoazo dyes show the potential of being utilized as colorants, pending further required tests.

Dihydropyrimidinase-like 3 regulates the inflammatory response of activated microglia.

Microglia, the resident immune cells of the CNS, are known to respond to injuries, infection and inflammation in the CNS by producing proinflammatory cytokines and phagocytosing cell debris and pathogens. In this study, we investigated the expression pattern and role of dihydropyrimidinase-like 3 (Dpysl3), a member of collapsin response mediator protein family, on the inflammatory reaction of microglia. Microarray analysis comparing the global gene expression profile of ameboid and ramified microglia has shown that Dpysl3 is mainly expressed in ameboid microglia in the 5-day postnatal rat brain. Immunohistochemical analysis revealed that Dpysl3 was intensely expressed in ameboid microglial cells in the rat brain till postnatal 7th day and then gradually diminished in ramified microglia of 2 weeks postnatal rat brain. Further, in vitro analysis confirmed that Dpysl3 expression was induced in activated BV-2 microglia treated with lipopolysaccharide (LPS). It is well documented that microglial activation by LPS increased the expression of inducible nitric oxide synthase (iNOS) and proinflammatory cytokines through the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activity in BV-2 microglia. However, siRNA-mediated knockdown of Dpysl3 prevented the LPS-induced expression of iNOS and cytokines including interleukin-1 beta, and tumor necrosis factor-alpha as well as nuclear translocation of NF-κB in microglia. Remarkably, knockdown of Dpysl3 inhibited the migration of activated microglia coupled with deranged actin filament configuration (as revealed by F-actin cytoskeleton expression) in lamellipodia projecting from the cells. Knockdown of Dpysl3 also inhibited the phagocytic ability of activated microglia. These findings suggest that knockdown of Dpysl3 can inhibit activation, migration and phagocytic capability of microglia and consequently reduce neuroinflammation.

Determination of aluminium induced metabolic changes in mice liver: a Fourier transform infrared spectroscopy study.

In this study, we made a new approach to evaluate aluminium induced metabolic changes in liver tissue of mice using Fourier transform infrared spectroscopy analysis taking one step further in correlation with strong biochemical evidence. This finding reveals the alterations on the major biochemical constituents, such as lipids, proteins, nucleic acids and glycogen of the liver tissues of mice. The peak area value of amide A significantly decrease from 288.278±3.121 to 189.872±2.012 between control and aluminium treated liver tissue respectively. Amide I and amide II peak area value also decrease from 40.749±2.052 to 21.170±1.311 and 13.167±1.441 to 8.953±0.548 in aluminium treated liver tissue respectively. This result suggests an alteration in the protein profile. The absence of olefinicCH stretching band and CO stretching of triglycerides in aluminium treated liver suggests an altered lipid levels due to aluminium exposure. Significant shift in the peak position of glycogen may be the interruption of aluminium in the calcium metabolism and the reduced level of calcium. The overall findings exhibit that the liver metabolic program is altered through increasing the structural modification in proteins, triglycerides and quantitative alteration in proteins, lipids, and glycogen. All the above mentioned modifications were protected in desferrioxamine treated mice. Histopathological results also revealed impairment of aluminium induced alterations in liver tissue. The results of the FTIR study were found to be in agreement with biochemical studies and which demonstrate FTIR can be used successfully to indicate the molecular level changes.

A six-hour extrapolated sampling strategy for monitoring mycophenolic acid in renal transplant patients in the Indian subcontinent.

BACKGROUND: Therapeutic drug monitoring for mycophenolic acid (MPA) is increasingly being advocated. The present therapeutic range relates to the 12-hour area under the serum concentration time profile (AUC).However, this is a cumbersome, tedious, cost restricting procedure. Is it possible to reduce this sampling period? AIM: To compare the AUC from a reduced sampling strategy with the full 12-hour profile for MPA. SETTINGS AND DESIGN: Clinical Pharmacology Unit of a tertiary care hospital in South India. Retrospective, paired data. MATERIALS AND METHODS: Thirty-four 12-hour profiles from post-renal transplant patients on Cellcept were evaluated. Profiles were grouped according to steroid and immunosuppressant co-medication and the time after transplant. MPA was estimated by high performance liquid chromatography with UV detection. From the 12-hour profiles the AUC up to only six hours was calculated by the trapezoidal rule and a correction factor applied. These two AUCs were then compared. STATISTICAL ANALYSIS: Linear regression, intra-class correlations (ICC) and a two-tailed paired t-test were applied to the data. RESULTS: Comparing the 12-hour AUC with the paired 6-hour extrapolated AUC, the ICC and linear regression(r2) were very good for all three groups. No statistical difference was found by a two-tailed paired t-test. No bias was seen with a Bland Altman plot or by calculation. CONCLUSION: For patients on Cellcept with prednisolone +/- cyclosporine the 6-hour corrected is an accurate measure of the full 12-hour AUC.

Quantitative evaluation of left ventricle performance from two dimensional echo images.

OBJECTIVES: We sought to quantify the left ventricle systolic dysfunction by a geometric index from two-dimensional (2D) echocardiography by implementing an automated fuzzy logic edge detection algorithm for the segmentation. BACKGROUND: The coronary injuries have repercussions on the left ventricle producing changes on wall contractility, the shape of the cavity, and as a whole changes on the ventricular function. METHODS: 2D echocardiogram and M-mode recordings were performed over the control group and those with the dysfunctions. From 2D recordings, individual frames were extracted for at least five cardiac cycles and then segmentation of left ventricle was done by automated fuzzy systems. In each frame, the volumes are measured and a geometric index, eccentricity ratio (ER), was derived. The endocardial fractional shortening (FS), midwall fractional shortening (mFS), and the relative wall thickness (RWT) were also measured in each case. RESULTS: Depressed value of endocardial FS (20.39 +/- 5.43 vs 34.28 +/- 9.36, P = 0.0046), mFS (33 +/- 8.3 vs 52.5 +/- 11.7, P = 0.0047), and the RWT (0.337 +/- 0.096 vs 0.525 +/- 0.119, P = 0.0002) was observed with dysfunction. ER measured at end-diastole (2.86 +/- 0.703 vs 4.14 +/- 0.38) and end-systole (3.14 +/- 0.79 vs 5.48 +/- 0.74) was found to be decreased in the dysfunction group and more significant at the end-systole (P = 0.00017 vs 6.6E-06). CONCLUSION: This work concludes that the regional and global left ventricle systolic dysfunction can be assessed by the ER measured at end-diastole and end-systole from 2D echocardiogram and may contribute to the high rate of cardiovascular disorders.

Detection of left ventricle systolic dysfunction from shape deformity.

Coronary artery disease producing ischemic cardiomyopathy is the most frequent cause of left ventricular systolic dysfunction. Non-ischemic cardiomyopathies can also produce systolic dysfunction; they may be inherited as genetic disorders or occur sporadically. These coronary injuries have repercussions on the left ventricle producing changes on wall contractility, the shape of the cavity and also changes on ventricular function. This study is focused on the 2D echocardiograms of the left ventricle. Apical two chamber and four chamber view recordings were performed on normal and systolic dysfunction subjects. Individual frames were extracted for at least five cardiac cycles. After pre-processing these images, segmentation of the left ventricle was performed by Fuzzy systems. Then the volumes were measured by single and biplane methods along with the perimeter, short axis length and long axis length in each frame, from which the two indices Sphericity Index (SI) and Normalized Eccentricity Index (NEI) was determined. It was found that the diastolic phase is short in the case of systolic dysfunction, and its volume variation is not uniform as in the normal case. Also, in the case of systolic dysfunction, the span of either the long or short axis length variation is less than 0.5 cm. This depicts that akinesis is in the corresponding direction; the value of SI is less than 2 for systolic dysfunction. A sharp peak is seen at each systole point in the NEI plot and also its variation is smooth in subjects having LVEF > 45%, which is not the case for dysfunction.

Left ventricular systolic dysfunction identification by motion analysis.

This paper presents a new technique for identification of regional dysfunctions in the left ventricle from 2-D echocardiography. It uses a novel left ventricular border tracking algorithm based on Fuzzy inference system. In this paper we show how the regional dysfunction present in the left ventricle can be identified by tracking the movement of centre of mass of left ventricle in a 2D space. The path pattern of that point traced over the cardiac cycles shows variation between the two groups. The main advantage of this proposed approach is the smaller date handling in regional dysfunction identifications unlike other existing methods. The method is illustrated on the real 2D echocardiograph dataset that includes patients having dysfunctions in the left ventricular wall. The diagnostic potential of this method is explained in detail.

A prospective evaluation of leflunomide therapy for cytomegalovirus disease in renal transplant recipients.

AIM: A preliminary observation suggests leflunomide is effective in the treatment of cytomegalovirus (CMV) disease in renal transplant recipients. A prospective evaluation was conducted in renal transplant recipients to study the efficacy of leflunomide in the treatment of CMV disease. PATIENTS AND METHODS: With prior approval and informed consent for therapy and follow-up, 17 consecutive consenting renal transplant recipients with proven CMV disease were treated with leflunomide. CMV disease was defined as a clinical syndrome of fever and/or symptoms of organ involvement, leukopenia, and a positive nested CMV quantitative PCR test at 0.001 microg/5 microL template input, with or without histologic evidence of tissue invasion. Leflunomide metabolite concentrations (A77 1726) were monitored. RESULTS: Of the 17 patients, 14 patients were treated for 6 months for CMV disease the first time; the remaining 3 received leflunomide treatment for relapse after ganciclovir treatment, for a year. Seven patients had fever with viremia and no organ involvement, nine had viremia with involvement of gastrointestinal tract, and one had fever with CMV inclusions in the allograft, with no demonstrable viremia. The three patients with relapse treated with leflunomide responded. Overall, 15 patients (88%) clinically responded to leflunomide therapy and with viral clearance from blood and healing of involved organs. The cost of therapy with intravenous ganciclovir (Cymevene, Roche) for 2 weeks was US 721 dollars while that of leflunomide (Cleft, Cipla Ltd) for 6 months was US 64 dollars. CONCLUSION: Leflunomide treatment for CMV disease in renal transplant recipients is effective, simple, and economical.