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CalcuSyn Version 2.0 is the analyzer of combined drug effects, able automatically to quantify phenomena such as synergism and inhibition. The use of combined drug treatments is becoming common in the treatment of cancer and AIDS. CalcuSyn performs multiple drug dose-effect calculations using the Median Effect methods described by T-C Chou and P. Talalay (Trends Pharmacol, Sci. 4, 450-454). CalcuSyn can easily be integrated with other software. Data can be entered via the keyboard or file import either into the grid or directly into the analysis engine. When the grid is used, data can be processed through wizards which make the software easily accessible to new users. Data can be processed both for individual drugs, and for constant-ratio or non-constant-ratio combinations of drugs. CalcuSyn automatically graphs the data and produces reports giving summary statistics on all drugs plus detailed analysis of drug interactions including the Combination Index and EDx (for any value of x). Estimates of accuracy of EDx and CI can be calculated with Monte Carlo simulations or by a highly accurate algebraic estimation algorithm. The plots drawn by CalcuSyn include dose-effect, median-effect, isobolograms and CI-effect. All data and results are easily accessible by mouse-click on the contents tree. CalcuSyn Version 2.0 has Undo and Redo tools. There are flexible arrangements for printing of results and graphs and their export to spreadsheets, wordprocessors, and graphics packages. A comprehensive manual is supplied with the software and there is a detailed Help file. These give a thorough account of the theoretical basis of the analysis methods including the statistical treatment of results. CalcuSyn is suitable for the study of drug mixtures.Comments & References: Suitable for academic labs and pharmaceutical companies. A screenshot and a downloadable demo are available here. This program can be ordered online. Includes user manual as pdf. Orders outside the United Kingdom, please click here.
Calcusyn Software Download
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The human pancreatic adenocarcinoma cell lines, AsPC-1, Capan-2, MIA PaCa-2 and Panc-1, were exposed to troxacitabine or gemcitabine alone or in combination, for 72 h, and the effects on cell growth were determined by electronic particle counting. Synergistic efficacy was determined by the isobologram and combination-index methods of Chou and Talalay. Mechanistic studies addressed incorporation of troxacitabine into DNA and intracellular levels of troxacitabine and gemcitabine metabolites. For in vivo studies, we evaluated the effect of both drugs, alone and in combination, on the growth of established human pancreatic (AsPC-1) tumors implanted subcutaneously in nude mice. Statistical analysis was calculated by a one-way ANOVA with Dunnett as a post-test and the two-tailed unpaired t test using GraphPad prism software.
The four human pancreatic cancer cell lines (AsPC-1, Capan-2, MIA PaCa-2, and Panc-1) were seeded in 12-well plates, allowed to attach for 24 h and exposed to graded concentrations of troxacitabine and gemcitabine, either alone or in combination for 72 h (Fig. 1). Since these growth inhibitory studies demonstrated that gemcitabine was more potent than troxacitabine in three of the four cell lines (summarized in Table 1), combination studies were done using a drug ratio of 1:100 (gemcitabine: troxacitabine) over the range of drug concentrations tested. Combination treatments yielded significantly greater growth inhibition than either agent alone in all cell lines tested (Fig. 1). The isobologram and combination-index methods developed by Chou and Talalay [21] were used to confirm and quantify the synergism observed with gemcitabine and troxacitabine. Isobolograms were constructed for Fa values of 0.50, 0.75, and 0.90, representing 50%, 75% and 90% growth inhibition, respectively (Fig. 2). The CI values at Fa of 0.5 were calculated using CalcuSyn software and are summarized in Table 1. These results indicated that gemcitabine and troxacitabine were synergistic with CI values of 0.41, 0.71, 0.37, and 0.52, respectively, for Panc-1, MIA PaCa-2, AsPC-1 and Capan-2 cells. Both methods indicated synergism across a broad range of concentrations in all four pancreatic tumor cell lines.
Isobolograms of in vitro drug combinations. Isobologram analysis of the combination of gemcitabine and troxacitabine in Panc-1, MIA PaCa-2, AsPc-1 and Capan-2 cells. The individual doses of gemcitabine and troxacitabine to achieve 90% (straight line) growth inhibition (Fa = 0.90), 75% (hyphenated line) growth inhibition (Fa = 0.75), and 50% (dotted line) growth inhibition (Fa = 0.50) were plotted on the x- and y-axes. Combination index (CI) values calculated using Calcusyn software is represented by points above (indicate antagonism between drugs) or below the lines (indicate synergy). (X symbol) ED50, (plus sign) ED75 and (open dotted circle ) ED90.
Thank you for downloading CalcuSyn from our software libraryThe version of CalcuSyn you are about to download is 2.0. The contents of the download are original and were not modified in any way. The software is licensed as trial. Please bear in mind that the use of the software might be restricted in terms of time or functionality. The download was scanned for viruses by our system. We also recommend you to check the files before installation. CalcuSyn antivirus reportThis download is virus-free.This file was last analysed by Free Download Manager Lib 68 days ago.
The polydatin and resveratrol pharmacological interaction was evaluated in vitro on growing and differentiated Caco-2 cell lines by median drug effect analysis calculating a combination index with CalcuSyn software. We have selected a synergistic combination and we have evaluated its effect on the biological and molecular mechanisms of cell death.
On the basis of these results, we have evaluated if the ISBn could be synergistic in inducing cell growth inhibition of Caco-2 cells. Specifically, we evaluated the growth inhibition induced by different concentrations of pol/res combination at 24 h on Caco-2 cell line. We used Calcusyn[28], a dedicated software, to examine the synergism of our treatments. With this software, synergistic conditions occur when the combination index (CI) is
Effect of ISBn alone or in combination on mithocondrial superoxide anions, nitric oxide, and enzyme scavengers activity in Caco-2 cells. (A) After 24 h Caco-2 treatment with polydatin and resveratrol alone or in combination (240 and 100 μM) at 37C, the mitochondrial superoxide anion production was analysed by HE (20 ng mL-1) staining. Dye accumulation was analysed by FACScan flow cytometer (FACScan, Becton Dickinson) by the CellQuest software, and the intensities of the bands were expressed as percent control. For each sample, 2 104 events were acquired. (B) Nitric oxide was measured in medium. Whole cell homogenates were used for evaluated the Catalase (C) and Mn-SOD activity (D). The bars represent means SEM of three independent experiments. Asterisks indicate significant difference between the Caco-2-treated samples compared with control value **P P
Confocal analysis of ISBn treated growing Caco-2 cells stained for iNOS, hsp27, and vimentin. Panel A: Subcellular co-localization of iNOS (red) and vimentin filaments (green) in Caco-2 cells without ISBn (ctr) and with Pol, Res, and Pol-Res combination treatment. The cells were incubated with anti-vimentin, anti-HSP27 and anti-iNOS antibodies. Images were obtained by confocal microscopy. Panel B: Subcellular co-localization of HSP-27 (red) and vimentin filaments (green) in Caco-2 cells without ISBn (ctr) and with Pol, Res, and Pol-Res combination treatment The cells were incubated with anti-vimentin, anti-HSP27 and anti-iNOS antibodies. Images were obtained by confocal microscopy. Panel C: The expression levels of the proteins are reported as intensities quantified with Image J software. Panel D: Caspase-3 activity on growing and differentiated Caco-2 cells without ISBn (ctr) and with Pol, Res, and Pol-Res combination treatment .Caspase-3 activity was determined by incubating whole-cell extracts with 40 μM caspase-3 substrate and measuring production of hydrolysed 7-amido-4-methyl-coumarin (AMC) groups using a multi-label plate reader. The results are representative of four separate experiments; summary data shown are means SEM. 2ff7e9595c
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