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Supplemental Table 1: Descriptive statistics are shown for the time to zero analysis of kill kinetics ; n = 6 replicates per treatment.

Time to Zero Analysis – Descriptive Statistics*
Treatment / Mean / SD / Median / IQR / Min / Max
Dihydrosphingosine / 0.5 / 0 / 0.5 / 0 / 0.5 / 0.5
Lauric Acid / 0.5 / 0 / 0.5 / 0 / 0.5 / 0.5
Phytosphingosine / 1 / 0 / 1 / 0 / 1 / 1
Sapienic Acid / 0.1 / 0 / 0.1 / 0 / 0.1 / 0.1
Sphingosine / 0.5 / 0 / 0.5 / 0 / 0.5 / 0.5
* Descriptive statistics are shown for time to zero analysis of kill kinetics.

Supplemental Table 2: Pairwise comparisons of lipid treatments for time to zero analysis of kill kinetics against P. gingivalis.

Time to Zero Pairwise Comparisons (hours)
Treatment 1 / Median / Treatment 2 / Median / P-values*
Dihydrosphingosine / 0.5 / Lauric Acid / 0.5 / 1.0000
Dihydrosphingosine / 0.5 / Phytosphingosine / 1.0 / 0.0022**
Dihydrosphingosine / 0.5 / Sapienic Acid / 0.1 / 0.0022**
Dihydrosphingosine / 0.5 / Sphingosine / 0.5 / 1.0000
Lauric Acid / 0.5 / Phytosphingosine / 1.0 / 0.0022**
Lauric Acid / 0.5 / Sapienic Acid / 0.1 / 0.0022**
Lauric Acid / 0.5 / Sphingosine / 0.5 / 1.0000
Phytosphingosine / 1.0 / Sapienic Acid / 0.1 / 0.0022**
Phytosphingosine / 1.0 / Sphingosine / 0.5 / 0.0022**
Sapienic Acid / 0.1 / Sphingosine / 0.5 / 0.0022**
*Bonferonni adjusted α = 0.005 for a 5% significance level with ten comparisons.
** Indicates significance after adjustment for multiple comparisons.

Supplemental Table 3: Descriptive statistics are shown for the area under the curve (AUC) analysis of kill kinetics ; n = 6 replicates per treatment.

Area Under the Curve (AUC) Analysis of Kill Kinetics
Treatment / Mean / SD / Median / IQR / Min / Max
Control / 574.05 / 1.19 / 573.97 / 1.68 / 572.65 / 575.72
Dihydrosphingosine / 3.22 / 0.03 / 3.23 / 0.03 / 3.16 / 3.25
Lauric Acid / 1.73 / 0.03 / 1.73 / 0.04 / 1.70 / 1.76
Phytosphingosine / 5.34 / 0.30 / 5.25 / 0.51 / 5.06 / 5.76
Sapienic Acid / 0.00 / 0.00 / 0.00 / 0.00 / 0.00 / 0.00
Sphingosine / 1.28 / 0.09 / 1.27 / 0.05 / 1.19 / 1.45
* AUC represents trapezoidal area calculated over the time interval 0.1 hours to 24 hours.

Supplemental Table 4: Trapezoidal areas under the curve for each lipid treatment against P. gingivalis as a summary measure of viability over the time course. Significance probabilities of these comparisons are shown.

Trapezoidal AUC for each treatment against P. gingivalis
Significance Probabilities (P-Values)**
Treatment 1 / Median / Treatment 2 / Median / P-values*
Control / 574.0 / Dihydrosphingosine / 3.2 / 0.0022
Control / 574.0 / Lauric acid / 1.7 / 0.0022
Control / 574.0 / Phytosphingosine / 5.3 / 0.0022
Control / 574.0 / Sapienic acid / 0.0 / 0.0022
Control / 574.0 / Sphingosine / 1.3 / 0.0022
Dihydrosphingosine / 3.23 / Lauric Acid / 1.73 / 0.0022
Dihydrosphingosine / 3.23 / Phytosphingosine / 5.25 / 0.0022
Dihydrosphingosine / 3.23 / Sapienic Acid / 0.00 / 0.0022
Dihydrosphingosine / 3.23 / Sphingosine / 1.27 / 0.0022
Lauric Acid / 1.73 / Phytosphingosine / 5.25 / 0.0022
Lauric Acid / 1.73 / Sapienic Acid / 0.00 / 0.0022
Lauric Acid / 1.73 / Sphingosine / 1.27 / 0.0022
Phytosphingosine / 5.25 / Sapienic Acid / 0.00 / 0.0022
Phytosphingosine / 5.25 / Sphingosine / 1.27 / 0.0022
Sapienic Acid / 0.00 / Sphingosine / 1.27 / 0.0022
**Bonferonni adjusted α = 0.0033 for a 5% significance level with fifteen comparisons. All P-values obtained are below this threshold and therefore are significant after adjustment for multiple comparisons.
** Significance probability associated with the exact nonparametric Kruskal-Wallis test of the null hypothesis that the distribution of trapezoidal area is the same for all treatment groups designated.

P. gingivalis susceptibility to oral mucosal lipidsCarol L. Fischer

2/24/2011

Supplemental Table 5: Literature review of P. gingivalis upregulated and downregulated proteins in stress responses associated with Heat stress 51-58, O2 oxidative stress 51, 53, 59, H2O2 oxidative stress 51, 59, 60, pH stress 53, 54, heme limitation 61, EtOH stress 54, and response to contact with epithelial cells 62. Upregulation is indicated by ^, while downregulation is indicated by ˅.

Gene / Acc # / Protein/Function / Heat / Oxidative (O2) / Oxidative (H2O2) / pH / Heme limitation / EtOH / Epithelial contact / Sapienic acid
hprA / Glycerate dehydrogenase / ^
ISPg1 / PG0184 / Transposase / ^ / ˅ / ^
infB / PG0255 / Translation initiation factor IF-2 / ^ / ˅ / ^
rgpA
prtT / PG2024
PGN_
1970 / Arg-specific cysteine proteinase / ^ ˅ / ^ ˅ / ^ / ^
rgpB / PG0506 / Arg-specific cysteine proteinase / ^ / ^ ˅ / ^ / ^ / ^
PGN_
1466 / ^ / ^ ˅ / ^ / ^
kgp / Kgp; Lys-gingipain (cysteine protease) / ˅ / ^
groEL / PG0520 / HSP60 / ^ / ^ / ^ / ^ / ^ / ^
pyrG / PG0525 / CTP synthase / ^ / ˅ / ^
ompF / PG0695 / Immunoreactive antigen P32, porin / ^ / ˅ / ^
HYP2 / PG0774 / Hypothetical protein / ^ / ˅ / ^
epsC / PG1135 / Glycosyltransferase / ^ / ˅ / ^
rpsA / PG1297 / Ribosomal protein S1 / ^ / ^ / ^
sodB / PG1545 / Superoxide dismutase Fe-Mn / ^ / ^ ˅ / ^ / ^
humY / PG1551 / HmuY protein / ^ / ˅ / ^
secDF / PG1762 / Protein-export membrane protein / ^ / ^ ˅ / ^
HYP1 / PG1795 / 43-64 kDa LPS-modified surface protein (P27) / ^ / ^ ˅ / ^
cysS / PG1878 / Cysteinyl-tRNA synthetase / ^ / ˅ / ^
htpG / PG0045 / ^ / ^ / ^
dnaK / PG1208 / Heat shock protein 70 / ^ / ^ / ^ / ^
abfD / PG0692 / 4-hydroxy-CoA dehydratase / ˅ / ˅
fimA / FimA, structural subunit of fimbriae / ˅ / ^
HSP90 homologue / ^
aphC / PG0618 / Alkyl hydroperoxide reductase C / ^ / ˅ / ^
aphF / PG0619 / Alkyl hydroperoxide reductase F / ^ / ^
dps / PG0090 / Dps family protein (DNA-binding protein from starved cells) / ^ / ˅
batA / PG1582 / BatA protein / ^
ftn / PG1286 / Ferritin / ^ / ˅ / ^
tpx / PG1729 / Thiol peroxidase / ^ / ^
Hemagglutinin-like protein / ^
PG0548 / Pyruvate ferredoxin/flavodoxin oxidoreductse family protein / ^
sucD / PG0687 / Succinate-semialdehyde dehydrogenase, AFD / ^
4hbD / PG0689 / NAD-dependent 4-hydroxybutyrate dehydrogenase / ^
PG0690 / 4-hydroxybutyrate-CoA transferase / ^
PG0691 / NifU-like protein / ^
abfD / PG0692 / 4-hydroxybutyryl-CoA dehydratase / ^
PG1067 / Hypothetical protein / ^
PG1068 / Conserved hypothetical protein / ^
acdA / PG1076 / Acyl-CoA dehydrogenase, short-chain specific / ^
ackA / PG1081 / Acetate Kinase / ^
pta / PG1082 / Phosphotransacetylase / ^
gdh / PG1232 / Glutamate dehydrogenase, NAD-specific / ^ / ^ / ^
frdB / PG1614 / Fumarate reductase, iron-sulfur protein (FrdB) / ˅
frdA / PG1615 / Fumarate reductase, flavoprotein subunit (FrdA) / ˅
PG0026 / Hypothetical protein (homology to Arg proteases) / ^ ˅
pepO / PG0159 / Endopeptidase PepO / ^
PG0350 / Internalin-related protein / ^
PG1374 / Immunoreactive 47-kDa protein (IrpI) / ^
PG2130 / FimX / ^
PG2131 / PgmA / ^
fimA / PG2132 / Fimbrillin FimA / ^ ˅
PG2133 / Lipoprotein / ^
PG2134 / FimC / ^
PG2135 / FimD / ^
PG2136 / FimE / ^
PG1638 / Thioredoxin family protein / ^
PG1639 / Hypothetical protein / ^
PG1640 / DinF, membrane-spanning MATE efflux pump / ^
PG1641 / PtpA, protein tyrosine phosphatase / ^
PG1642 / Cation-translocating ATPase (ZntA) / ^
PG0616 / Thioredoxin/HBP35 (heme-binding protein) / ^
PG0618 / Alkyl hydroperoxide reductase / ^
PG0619 / Alkyl hydroperoxide reductase subunit F / ^
PG0644 / HtrE (Tla); TonB-linked receptor / ^
PG0645 / HtrD; no known function / ^
PG0646 / HtrC; ABC heme transport system ATP binding protein / ^
PG0647 / HtrB; ABC heme transport system permease / ^
PG0648 / HtrA; ABC heme transport system solute binding protein / ^
PG1043 / FeoB2 / ^
PG1551 / HmuY / ^
hmuR / PG1552 / HmuR / ^
PG1553 / HmuS / ^
PG1554 / Hypothetical protein / ^
PG1555 / TolQ / ^
Pg1556 / HmuV / ^
PG1019 / Hypothetical protein / ^
PG1020 / Conserved hypothetical protein; possible outer membrane receptor protein / ^
PG1858 / Flavodoxin A / ^
PG1874 / Conserved hypothetical protein / ^
PG1875 / Hemolysin A / ^
19 and 50 kDa proteins / ˅
92 and 80 kDa proteins / ^
31 and 26 kDa proteins (putative function: heme uptake/storage) / ^
60 and 68 kDa proteins / ^
htpG / PG0045 / Heat shock protein 90 / ^
htrA / PG0593 / Heat-induced serine protease / ^
slyD / PG1315 / FKBP-type peptidylprolyl isomerase / ^
grpE / PG1775 / Heat shock protein / ^
dnaJ / PG1776 / Heat shock protein 40 / ^
trxB / PG1134 / Thioredoxin reductase / ^
fabG / PG1239 / Acyl carrier protein / ^
fabF / PG1764 / 3-Oxoacyl synthase / ^ / ^
fabH / 3-oxoacyl-[acyl carrier protein] synthase 3 / ^
acpP / PG1765 / Acyl carrier protein / ^
rprY / PG1089 / Transcriptional regulator / ^
tetR / PG1240 / Transcriptional regulator / ^
ompH1 / PG0192 / Cationic outer membrane protein / ^
ompH1 / PG0193 / Cationic outer membrane protein / ^
PG0435 / Capsule biosynthesis / ^
PG2167 / Immunoreactive 53-kDa antigen / ^
PG0419 / Conserved hypothetical / ^
PG0686 / Conserved hypothetical / ^
PG0434 / Hypothetical / ^
PG0654 / Hypothetical / ^
PG1316 / Hypothetical / ^
PG1317 / Hypothetical / ^
PG1635 / Hypothetical / ^
groES / PG0521 / ^
ragA / RagA (Pgm1); 110 kDa / ˅
ragB / RagB (Pgm4); 47-55 kDa / ˅
75 kDa protein (Pgm2) / ˅
Fimbrial proteins / ˅
Arg-X / ^ / ^
serC / phosphoserine aminotransferase / ^
gapA / Glyceraldehyde 3-phosphate dehydrogenase, type 1 / ^ / ^ / ^

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