Investigation of Antioxidant Content and Capacity in Yellow European Plums

<p>Phenolic content and antioxidant capacity of five Yellow European plums (<i>Prunus domestica</i>) were studied using heat reflux extraction. Fresh plums were extracted at 50°C and 70°C, while freeze dried plums were extracted at 50°C, 60°C, and 70°C. Quantification of phenolic compounds such as ascorbic acid, neochlorogenic acid, and chlorogenic acid, was performed using high performance liquid chromatography. Antioxidant activity was determined by evaluating the scavenging ability of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric (Fe³⁺) free radicals. Total phenolic content and ferric reducing antioxidant potential were highest for freeze dried samples extracted at 60°C whereas extraction at 70°C resulted in the lowest yield. Neochlorogenic acid was the predominant phenolic compound in each plum genotype followed by ascorbic acid and chlorogenic acid. This study demonstrates that there is an adequate amount of health promoting phytochemicals within European plums, hence extraction of these compounds have potential for use towards functional food, nutraceutical, and pharmaceutical industries.</p

Downregulation of the SL biosynthetic pathway in AR isolates.

<p>(A) Changes in the composition of major SL classes among the sequential isolates of <i>C. albicans</i> were assessed as described in methods. Green and black colour of gene in the pathway represents upregulation and no change, respectively. (B) Total SLs are represented as % of the total PGL + SE + SL mass spectral signal after normalization to internal standards and were determined as described in methods. (C) The fold change in expression levels of various SL biosynthetic pathway genes (relative amplification to ACT1) between TW1 (most susceptible to FLC) and TW17 (most resistant to FLC) were determined by RT-PCR. The gel picture is a representative of the RT-PCR analysis performed in replicates. Values in the histogram are means ± SD (n = 3 for all <i>Candida</i> strains). Asterisks “*” represents p<0.05. Lipid data taken from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039812#pone.0039812.s004" target="_blank">Sheet S1</a>, worksheet 4.</p

FLC exposure alters CW integrity in the sequential isolates of C. albicans.

<p>(A) Sequential isolates were tested with CW perturbing agents like Triton X-100 (upto 0.04%), 0.02% SDS, 16 µgml⁻¹ Congo Red. (B) Susceptibility to Calcofluor White (upto 50 µgml⁻¹) was tested. These spot tests were performed as described in methods. (C) Passive diffusion rate of PI was monitored by spectrofluorimeter. Briefly, the cells were pre-incubated with 3 µM PI for 45 min and then centrifuged. The supernatant was taken and the emission spectrum of PI was recorded for each strain. (D) The cells with PI were subjected to flow cytometry analysis to measure the amount of PI accumulated in each strain. The data is represented as the mean fluorescent intensity (n = 2).</p

Upregulation of the sterol biosynthetic pathway in AR isolates.

<p>(A) Changes in the composition of major sterols and its intermediates among the sequential isolates of <i>C. albicans</i> were analyzed as described in methods. Green and black colour of gene in the pathway (A) represents upregulation and no change, respectively. (B) Total SEs are represented as % of the total PGL+ SE + SL mass spectral signal after normalization to internal standards and were determined as described in methods. (C) The fold change in gene expression levels of various <i>ERG</i> genes (relative amplification to <i>ACT1</i>) between TW1 (most susceptible to FLC) and TW17 (most resistant to FLC) were determined by RT-PCR. The gel picture is a representative of the RT-PCR analysis performed in replicates. Values in the histogram are means ± SD (n = 3 for all <i>Candida</i> strains). Asterisks “*” represents p<0.05. Lipid data taken from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039812#pone.0039812.s004" target="_blank">Sheet S1</a>, worksheet 3 and 4.</p

Measurement of mitochondrial membrane dysfunction among TW1 and TW17 isolates.

<p>For the flow cytometry analysis, cells were stained with 100 nM MitoTracker Red CMXRos (which fluoresces upon oxidation in respiring mitochondria). (A) Quadrant plots, (B) histograms and (C) bar graph of flow cytometry analysis between TW1 and TW17 are depicted (n = 4). (D) The red fluorescence of MitoTracker Red CMXRos was also visualized by confocal microscopy.</p

FLC exposure leads to gradual development of a partially compromised CW and affects PG biosynthesis in several other clinical isolates of <i>C. albicans</i>.

<p>A. Several pairs of isogenic clinical isolates of <i>C. albicans</i> were tested with 0.02% SDS, a CW perturbing agent. The spot tests were performed as described in methods. B. PL analysis of various AS/AR clinically matched isogenic isolates by HP-TLC. Briefly, cells were grown overnight (to the exponential growth phase) in YEPD medium. Lipids were extracted as described earlier <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039812#pone.0039812-Bligh1" target="_blank">[18]</a> and separated by thin-layer chromatography using chloroform: methanol: acetic acid (65:28:8) as the solvent system. HP-TLC analysis of each match pair and the corresponding values of each lipid class as mol % are also indicated.</p

Accumulation of odd chain-FA containing PGLs in the FLC resistant isolates.

<p>Total amount of odd chain FA-containing PGL was calculated by adding the normalized amounts of each odd chain FA containing PGL molecular species (namely 31-C, 33-C, 35-C and 37-C containing PGLs). The data is represented as % of total PGL + SL + SE mass spectral signal after normalization to internal standards. Values are mean of 3 independent analyses (n = 3). Asterisks “*” represents p < 0.05. Lipid data taken from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039812#pone.0039812.s004" target="_blank">Sheet S1</a> (worksheet 3), <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039812#pone.0039812.s006" target="_blank">Table S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039812#pone.0039812.s008" target="_blank">Table S3</a>.</p

Carbon Dot Based, Naphthalimide Coupled FRET Pair for Highly Selective Ratiometric Detection of Thioredoxin Reductase and Cancer Screening

The fluorescence resonance energy transfer (FRET) mechanism has been established between carbon dots (CDs) and naphthalimide to monitor the activity of thioredoxin reductase (TrxR), which is often overexpressed in many cancer cells. The naphthalimide moiety was covalently attached to the surface of CDs through a disulfide linkage. In normal cell conditions (when devoid of high concentrations of TrxR), the CDs act as an energy donor and naphthalimide acts as an acceptor, which establishes the FRET pair as interpreted from the emission at λ_em = 565 nm, when excited at λ_ex = 360 nm. However, contrary to this, the elevated levels of TrxR cause the breakage of disulfide bonds and consequently abolishes the FRET pair through the release of the naphthalimide moiety from the surface of CDs. This process was studied by monitoring of fluorescence intensity at λ_em = 565 and 440 nm, when excited at the same wavelength (λ_ex = 360 nm). The TrxR based ratiometric quenching and enhancement of fluorescence intensity offers an interesting opportunity to monitor the enzyme activities and has many advantages over conventional monitoring of fluorescence intensity at a single wavelength to avoid interference of external factors. Fluorescence images of cancer cells in response to the nanosensor were visualized under a confocal microscope. Cytotoxicity study of nanosensor retards the growth of HeLa and MCF-7 cell lines in the presence of visible light. Therefore, the nanosensor also acts as a theranostic agent to diagnose as well as killing of cancer cells

Calcium-induced conformational changes of Thrombospondin-1 signature domain: implications for vascular disease

<p><b>Context:</b> Thrombospondin1 (TSP1) participates in numerous signaling pathways critical for vascular physiology and disease. The conserved signature domain of thrombospondin 1 (TSP1-Sig1) comprises three epidermal growth factor (EGF), 13 calcium-binding type 3 thrombospondin (T3) repeats, and one lectin-like module arranged in a stalk-wire-globe topology. TSP1 is known to be present in both calcium-replete (Holo-) and calcium-depleted (Apo-) state, each with distinct downstream signaling effects.</p> <p><b>Objective:</b> To prepare a homology model of TSP1-Sig1 and investigate the effect of calcium on its dynamic structure and interactions.</p> <p><b>Methods:</b> A homology model of Holo-TSP1-Sig1 was prepared with TSP2 as template in Swissmodel workspace. The Apo-form of the model was obtained by omitting the bound calcium ions from the homology model. Molecular dynamics (MD) simulation studies (100 ns) were performed on the Holo- and Apo- forms of TSP1 using Gromacs4.6.5.</p> <p><b>Results and discussion:</b> After simulation, Holo-TSP1-Sig1 showed significant reorientation at the interface of the EGF1-2 and EGF2-3 modules. The T3 wire is predicted to show the maximum mobility and deviation from the initial model. In Apo-TSP1-Sig1 model, the T3 repeats unfolded and formed coils with predicted increase in flexibility. Apo-TSP1-Sig1model also predicted the exposure of the binding sites for neutrophil elastase, integrin and fibroblast growth factor 2. We present a structural model and hypothesis for the role of TSP1-Sig1 interactions in the development of vascular disorders.</p> <p><b>Conclusion:</b> The simulated model of the fully calcium-loaded and calcium-depleted TSP1-Sig1 may enable the development of its interactions as a novel therapeutic target for the treatment of vascular diseases.</p

Details of <i>T</i>. <i>indotineae</i> infections outside India.

Details of T. indotineae infections outside India.</p