Нормативно-правові аспекти дослідження витрат торговельних підприємств

У статті досліджено міжнародні та національні нормативно-правові акти, що розкривають суть та методологічні аспекти формування витрат підприємств у бухгалтерському і податковому обліку. (In the article are investigated standard-legal sources that open methodological aspects of formation of costs of the enterprises in the accounting and tax account.

IC₅₀ of Gemcitabine, Thymoquinone and Betulinic acid at 48 hrs.

<p><i>IC₅₀</i><i>value was calculated on</i><i>Human pancreas adenocarcinoma</i><i>cell lines (A) MIA PaCa-2 (B) PANC-1 and (C) Normal cell line FR2: For individual treatment of Gemcitabine, Thymoquinone and Betulinic acid for 48 hrs (**, significant as compaired to control at p value<0.01, n = 3) in-between the three cell lines.</i></p

Effect of Betulinic acid, Thymoquinone and Gemcitabine alone and (in combination) on growth of human pancreatic tumor cell lines.

<p><i>(A) MIA PaCa-2 and (B) PANC-1 cells were exposed to graded concentrations of Betulinic acid, Thymoquinone or Gemcitabine either alone or in combination at a fixed dose ratio of Betulinic acid vs. Gemcitabine and Thymoquinone vs. Gemcitabine for 48 hrs.(Gemcitabine</i><i>having exposure for 24 hrs only in case of combination), Gemcitabine (squares),</i><i>Betulinic acid and Thymoquinone</i><i>(circle) Gemcitabine+ betulinic acid and Gemcitabine +Thymoquinone (triangle).(Mono therapy (BA, GCB, TQ) Versus Combinations(CI_(GCB:BA), CI_(GCB:TQ))(**, significant difference at p value <0.01, n = 3).</i></p

CI values of Dietary molecules (BA and TQ) in combination with Drug (GCB).

<p>CI*(combination index) obtained from the Fa value which denotes the fraction affected (e.g., Fa of 0.5 is equivalent to a 50% reduction in cell growth). The CI value less than 1 shows synergism, equal to 1 show a additivism while greater than 1 shows antagonism.(++)strong synergism,(+)synergism,(-) antagonism. Cells were exposed to a fixed ratio dose concentration of BA or TQ either alone or along with combination at a dose ratio of GCB verses BA and GCB verses TQ for 48 hrs as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107154#pone-0107154-g002" target="_blank">Figure 2</a>.</p><p>CI values of Dietary molecules (BA and TQ) in combination with Drug (GCB).</p

Synergistic Combination of Gemcitabine and Dietary Molecule Induces Apoptosis in Pancreatic Cancer Cells and Down Regulates PKM2 Expression

<div><p>Gemcitabine, an effective agent in treatment of cancer of pancreas, has undergone failures in many instances after multiple cycles of therapy due to emergence of drug resistance. Combination of dietary compounds with clinically validated drugs has emerged as an effective therapeutic approach to treat pancreatic tumors, refractory to gemcitabine therapy. In order to optimize a possible synergistic combination of Gemcitabine (GCB) with dietary molecules, Betuilnic acid (BA) and Thymoquinone (TQ), stand-alone IC₅₀ dose of GCB, BA and TQ was calculated for pancreatic cancer cell lines. Fixed IC₅₀ dose ratio of the dietary molecules in combination with reduced IC₅₀ dose of GCB was tested on GCB resistant PANC-1 and sensitive MIA PaCa-2 cells for synergism, additive response and antagonism, using calcusyn. Combination index (CI) revealed that pre-treatment of BA and TQ along with GCB synergistically inhibited the cancer cell proliferation in <i>in-vitro</i> experiments. Pyruvate kinase (PK) M2 isoform, a promising target involved in cancer cell metabolism, showed down-regulation in presence of TQ or BA in combination with GCB. GCB with BA acted preferentially on tumor mitochondria and triggered mitochondrial permeability transition. Pre-exposure of the cell lines, MIA PaCa-2 and PANC-1, to TQ in combination with GCB induced apoptosis. Thus, the effectiveness of BA or TQ in combination with GCB to inhibit cell proliferation, induce apoptosis and down-regulate the expression of PKM2, reflects promise in pancreatic cancer treatment.</p></div

miR-101 induces senescence and not apoptosis in MCF7 cells.

<p>(a) Original Flow-cytometry dot-blot of PI Staining Vs Annexin-V Staining, (b) Percentage of Annexin-V positive (apoptotic) and negative cells, (c) β-gal positively stained (senescent) cells, (d) Fold change in number of senescent cells, under control (mock-pEP-Null-transfected, mock-DMSO treated & mock negative with scrambled primers) and experimental conditions (pEP-miR-101 transfection, Etoposide 1 µM treatment & anti-miR-101 transfection). Time dependent increase in senescence in miR-101 transfected cells is also shown (d).</p

UBE2N and SMARCA4 as novel targets of miR-101.

<p>(a) Bio-informatics revealing the presence of miR-101 binding site at 3′UTR of UBE2N and SMARCA4, (b) activity of Luc gene showing a significant decrease in Luciferase activity when miR-101 was over-expressed in cells expressing Luc with 3′UTR of either (i & ii) UBE2N or (iii & iv) SMARCA4, in MCF7 and Hela cells. (c & d) Real-Time PCR showing a change in expression using Sybergreen and UBE2N and SMARCA4 specific primers, (c) under miR-101 and anti-miR-101 transfected conditions, showing decrease & increase, respectively, and (d) after 12 hrs exposure to 1uM, 5 uM and 10 uM etoposide, in MCF7 cells, confirming UBE2N and SMARCA4 as targets of miR-101, which is induced to 10 fold with 1 uM etoposide. At the higher concentrations of etoposide miR-101 expression is reduced leading to increased expression of UBE2N and SMARCA4, again confirming the relationship between miR-101 and the two novel targets.</p

Specific activity of Pyruvate Kinase (PK)-M2 in both of human pancreatic tumor cell lines.

<p><i>(A) MIA PaCa-2 cells treated with Betulinic acid, Thymoquinone and Gemcitabine alone and in combination, CI_(GCB:BA) and</i><i>CI_(GCB:TQ) at 48 hrs. A decrease in the activity in treated cells followed with</i><i>more</i><i>decrease</i><i>in combination(s), CI_(GCB:BA)</i><i>and CI_(GCB:TQ),</i><i>as compared with the control was observed;</i><i>(B)</i><i>In PANC-1 cell line vice versa was observed. Control versus monotherapy (BA, TQ, GCB) and Control versus combinations (CI_(GCB:BA) & CI_(GCB:TQ)), (***, significant difference at p value<0.001, n = 3)</i></p

Mean IC₅₀ Value of drug (GCB) and dietary molecules (BA and TQ) on three cell lines (MIA PaCa-2; PANC-1; FR2) at 48 hrs.

<p>IC₅₀ concentration of the drug (GCB) and Dietary molecules (BA and TQ) in <b>µ</b>M concentration causing 50% growth inhibition of two human pancreatic cancer cell lines(MIA PaCa-2 and PANC-1) and non cancer cell line(FR2) after 48 hrs of treatment.</p><p>Mean IC₅₀ Value of drug (GCB) and dietary molecules (BA and TQ) on three cell lines (MIA PaCa-2; PANC-1; FR2) at 48 hrs.</p

Combination Index (CI) at ED₅₀, ED₇₅, ED₉₀ values of Dietary with drug combination on two pancreas cancer cell lines.

<p>The CI Values at a Fa value of 0.5, 0.75, 0.90 for islobologram (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107154#pone-0107154-g003" target="_blank">Figure 3</a>) were calculated with the CalcusynVersion 2.1 software<b>.</b></p><p>Combination Index (CI) at ED₅₀, ED₇₅, ED₉₀ values of Dietary with drug combination on two pancreas cancer cell lines.</p