Efficient Sample Tracking With OpenLabFramework

The advance of new technologies in biomedical research has led to a dramatic growth in experimental throughput. Projects therefore steadily grow in size and involve a larger number of researchers. Spreadsheets traditionally used are thus no longer suitable for keeping track of the vast amounts of samples created and need to be replaced with state-of-the-art laboratory information management systems. Such systems have been developed in large numbers, but they are often limited to specific research domains and types of data. One domain so far neglected is the management of libraries of vector clones and genetically engineered cell lines. OpenLabFramework is a newly developed web-application for sample tracking, particularly laid out to fill this gap, but with an open architecture allowing it to be extended for other biological materials and functional data. Its sample tracking mechanism is fully customizable and aids productivity further through support for mobile devices and barcoded labels

Development of a cell-based bioassay for phospholipase A2-triggered liposomal drug release.

The feasibility of exploiting secretory phospholipase A2 (sPLA2) enzymes, which are overexpressed in tumors, to activate drug release from liposomes precisely at the tumor site has been demonstrated before. Although the efficacy of the developed formulations was evaluated using in vitro and in vivo models, the pattern of sPLA2-assisted drug release is unknown due to the lack of a suitable bio-relevant model. We report here on the development of a novel bioluminescence living-cell-based luciferase assay for the monitoring of sPLA2-triggered release of luciferin from liposomes. To this end, we engineered breast cancer cells to produce both luciferase and sPLA2 enzymes, where the latter is secreted to the extracellular medium. We report on setting up a robust and reproducible bioassay for testing sPLA2-sensitive, luciferin remote-loaded liposomal formulations, using 1,2-distearoyl-sn-glycero-3-phosphatidylcholine/1,2-distearoyl-sn-glycero-3-phosphatidylglycerol (DSPC/DSPG) 7:3 and DSPC/DSPG/cholesterol 4:3:3 as initial test systems. Upon their addition to the cells, the liposomes were degraded almost instantaneously by sPLA2 releasing the encapsulated luciferin, which provided readout from the luciferase-expressing cells. Cholesterol enhanced the integrity of the formulation without affecting its susceptibility to sPLA2. PEGylation of the liposomes only moderately broadened the release profile of luciferin. The provided bioassay represents a useful tool for monitoring active drug release in situ in real time as well as for testing and optimizing of sPLA2-sensitive lipid formulations. In addition, the bioassay will pave the way for future in-depth in vitro and in vivo studies

The Effect of Water-Soluble Polysaccharide from Jackfruit (Artocarpus heterophyllus Lam.) on Human Colon Carcinoma Cells Cultured In Vitro

Various phytochemical studies have revealed that jackfruit (Artocarpus heterophyllus Lam.) is rich in bioactive compounds, including carotenoids, flavonoids, volatile acids, tannins, and lectins. The aim of the study was to analyze the biological activity of water-soluble polysaccharide (WSP) isolated from jackfruit and to assess its immunomodulatory, cytotoxic, and anti-oxidative effects on human colon carcinoma cells in vitro. The neutral red (NR) uptake assay revealed no toxic influence of the polymer on the viability of tumor cells (HT29 and SW620). After 24 h and 48 h of incubation, the cellular viability was not lower than 94%. The metabolic activity of the cells (MTT) at the compound concentration of 250 µg/mL was higher than 92% in comparison to the control. WSP (250 µg/mL) exerted no significant effect on the morphology of the cells was determined by May-Grünwald-Giemsa staining. WSP changed nitric oxide (NOx) production by the tumor cells depending on the time of incubation and prior 2-h stimulation of the cells with E. coli 0111:B4 LPS. It significantly stimulated IL-1β production by the tumor cells. The IL-6 level increased but that of IL-10 decreased by a WSP concentration-dependent manner. No such effect was detected in SW620. The WSP had antioxidant properties. In conclusion, water-soluble polysaccharide isolated from A. heterophyllus exhibits significant biological activity towards many types of both normal and cancerous cells. Therefore, it may be considered as a useful agent in the protection of human health or in functional and dietary nutrition

Testing of L2 and L2P2 cells.

<p>Luminescence profiles generated upon the addition of 141 μM luciferin to L2 and L2P2 cells.</p

Testing of luciferin-loaded liposomes.

<p>Luminescence profiles generated upon the addition of luciferin-loaded liposomes to L2P2 and L2 cells. Two formulations were tested (100 nm diameter liposomes), DSPC/DSPG 7:3 (PCPG, 33 μM luciferin and 833 μM lipid) and DSPC/DSPG/cholesterol 4:3:3 (PCPGch, 33 μM luciferin and 265 μM lipid).</p

Testing of the effect of PEGylation.

<p>Luminescence profiles generated upon the addition of PEGylated luciferin-loaded liposomes to L2P2 and L2 cells. The lipopolymer DPPE-PEG 2000 (5 mol%) was added via post insertion to the loaded liposomes. Two formulations were tested (100 nm diameter liposomes), DSPC/DSPG 7:3 with (PCPG, 33 μM luciferin and 833 μM lipid) and DSPC/DSPG/cholesterol 4:3:3 (PCPGch, 107 μM luciferin and 833 μM lipid).</p

Analyses of stable cell lines.

<p>(A) Determination of relative <i>PLA2G2A</i> mRNA levels in L2P2 and control cells, normalized to GAPDH and ACTB levels. Values are referred to Colo 205 cells with high endogenous <i>PLA2G2A</i> mRNA levels [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125508#pone.0125508.ref035" target="_blank">35</a>]. (B) Relative luciferase activity of L2 and L2P2 cells normalized to cell numbers as determined by a cell viability assay as substitute readout. Error bars represent standard error of the mean (SEM).</p

Testing of the bioassay using free luciferin.

<p>Dependence of the maximum of the luminescence profile generated upon the addition of luciferin to L2 cells on (A) number of seeded cells (0–50,000 L2 cells, 10 mM luciferin), (B) luciferin concentration (0–5 mM luciferin, 10,000 L2 cells), and (C) number of days after the seeding of 2,500 L2 cells (1–10 days, 1 mM luciferin). Lines are inserted only as a guide to the eye. Error bars represent the standard deviation (SD).</p

Testing of lysed luciferin-loaded liposomes.

<p>Luminescence profiles generated upon the addition of luciferin-loaded liposomes lysed with 0.5 wt.% Triton X-100 to L2 cells. Two formulations were tested (100 nm diameter liposomes), DSPC/DSPG 7:3 (PCPG, 117 μM luciferin and 880 μM lipid) and DSPC/DSPG/cholesterol 4:3:3 (PCPGch, 74 μM luciferin and 1220 μM lipid).</p

Krakowskie Studia Międzynarodowe nr 1, 2011 (Mniejszości etniczno-religijne a modernizacja w krajach Azji i Afryki)

Kolejny numer „Krakowskich Studiów Międzynarodowych”, poświęcony krajom Afryki i Azji, pokazuje relacje między wspólnotami etniczno-religijnymi a państwem oraz charakteryzuje zmiany społeczne i kulturowe, jakie następują w obrębie tych wspólnot pod wpływem przemian modernizacyjnych. Samego słowa „modernizacja” nie należy rozumieć w odniesieniu do teorii modernizacji, oznaczającej upodobnianie się społeczeństw azjatycko-afrykańskich do zachodnich, lecz jako zmianę społeczno-kulturowego status quo w ogóle. To, że społeczeństwa Azji i Afryki są wieloetniczne i wieloreligijne jest powszechnie znane po dziesiątkach lat badań; równie dobrze znany jest fakt, że w państwach Azji i Afryki zachodzą obecnie poważne przemiany