TERBToronto Escort Review Board
TERBTobacco Evaluation and Review Board (Vermont)
TERBTime En-Route Bulletin
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With these a and m parameters, the Loewe Additivity response surface (LA) is calculated for each BE and terb. The comparison of the surface obtained with experimental data vs the zero response surface allows the direct visualization of spheres which, if they fall into the surface are indicative of additivism, over the surface indicate synergism and below the surface show antagonism.
The filtered methanolic Baccharis extracts (that have showed synergistic or additive effects with terb in CombiTool) were submitted to a HPLC/UV/ESI-MS Balker micrOTOF Q II/Agilent Technologies LC 1200 Series (Balker Daltonics, MA, USA) chromatograph, equipped with an electrospray ionization source operated in positive mode (ESI+) with Nitrogen as nebulizing gas (1.0 bar) and drying gas (4 L min (-1), 200 [degrees] C); capillary 4500V and end plate offset at 500V; the temperature of the heated capillary gas was set to 200 [degrees] C.
Two-drug combination of terb with the detected flavonoids and ent-clerodanes of Baccharis spp.
rubrum was incubated in 96-well plates (Greiner Labortechnik) for 72 h with serial dilutions of terb (100-1.0 x [10.sup.-4] [micro] g/ml) in addition to a sub-inhibitory concentration (31.25 [micro] g/ml) of each Baccharis component (1-7).
Analysis of the combination effect The nature of the interaction (synergism, additivism, or antagonism) between each natural product (NP, namely 1-7) and terb, as a function of [MIC.sub.50], was assessed by the combination index method (Cl) (Chou 2006) which is represented by the following equation:
CI={([MIC.sub.50], terb, comb.)/([MIC.sub.50] terb)} + {(CNP, comb)/[MIC.sub.50] NP}
where '[MIC.sub.50] terb, comb', is the [MIC.sub.50] value for terb in each two-drug combination and 'C NP, comb' is the fixed concentration of each individual NP in the combination with terb.
The dose reduction index (DRI), which is a measure of how many-fold the dose may be reduced as compared with the doses of each drug alone (Chou 2006), is calculated as follows: DRI = '[MIC.sub.50] terb alone'/'[MIC.sub.50] terb, comb' with each combination partner.
The same procedure was carried out along the columns (1-12) with terb. So, each well contained a unique combination (100 [micro]l) of the two substances (one ent-clerodane and terb).
rubrum growth inhibition of forty four BE and of terb, each one acting alone, was determined with the CLSI microbroth dilution method (CLSI 2008).
Six combinations for each of the forty four BE with terb (total 264 combinations) were prepared and tested against T.
So, in the ratio 250 [micro]g/ml BE-1 /10 MIC terb only 5 of the 44 BE produced a significant enhancement (p < 0.05) of antifungal activity.