Journal: Pharmaceutics

Article Title: Species-Specific Susceptibility of Planktonic and Biofilm Forming Candida Strains to Cyclodextrin-Encapsulated Essential Oils

doi: 10.3390/pharmaceutics18040508

Figure Lengend Snippet: Species-level susceptibility thresholds (MIC 90 and EC 10 ) across Candida species for native EO and RAMEB-EO formulations. ( A ) MIC 90 and ( B ) EC 10 values for the native EO condition; ( C ) MIC 90 and ( D ) EC 10 values for the corresponding RAMEB-EO (REO) condition, shown across the four species groups: Ca (red: C. albicans ), Ct (blue: C. tropicalis ), Ck (yellow: C. krusei ), and Cd (green: C. dubliniensis ). Boxplots summarize the distribution within each species (median line with interquartile range), with mean-based overlays shown as indicated in the legend (mean marker; mean ± 1 SE; mean ± 95% CI). Horizontal brackets denote statistically significant between-species differences for the given endpoint/formulation (multiple-comparison-adjusted pairwise contrasts); significance levels are indicated as ** ( p < 0.001) on the plot. Error bar overlays represent mean ± SE and mean ± 95% CI as indicated.

Article Snippet: The oils were subsequently encapsulated in randomly methylated β-cyclodextrin (RAMEB) by CycloLab Cyclodextrin Research & Development Laboratory (Budapest, Hungary) to produce the RAMEB–essential oil inclusion complexes used in the biological experiments.

Techniques: Marker, Formulation, Comparison

Journal: Pharmaceutics

Article Title: Species-Specific Susceptibility of Planktonic and Biofilm Forming Candida Strains to Cyclodextrin-Encapsulated Essential Oils

doi: 10.3390/pharmaceutics18040508

Figure Lengend Snippet: The MIC 90 -EC 10 phase plasticity landscape highlights species- and formulation-dependent separation between inhibitory and sub-inhibitory regimes. ( A ) Two-dimensional susceptibility map plotting MIC 90 ( y -axis) against EC 10 ( x -axis) for all strain–treatment observations. Points are colored by treatment (see legend) and grouped by species (Ca, Ct, Ck, and Cd). Gray dashed guides/boxes indicate the within-species spread of MIC 90 -EC 10 coordinates, emphasizing the differences in susceptibility dispersion (plasticity) among species. ( B ) Distribution and geometry of the phase plasticity window, defined as the separation between EC 10 and MIC 90 . The boxplot summarizes the overall distribution of MIC 90 -EC 10 separation (Δs = EC 10 − MIC 90 ; top), and the scatter shows MIC 90 vs. EC 10 colored by formulation class (RC: antifungal reference controls vs. native EO vs. REO: RAMEB-EO), illustrating how formulation shifts points within the MIC-EC plane. Marginal boxplots (right/top) summarize axis-wise distributions. A larger EC 10 -MIC 90 separation indicates a broader sub-inhibitory window (greater phase plasticity), whereas tighter coupling suggests a narrower transition from partial effect to near-complete inhibition.

Article Snippet: The oils were subsequently encapsulated in randomly methylated β-cyclodextrin (RAMEB) by CycloLab Cyclodextrin Research & Development Laboratory (Budapest, Hungary) to produce the RAMEB–essential oil inclusion complexes used in the biological experiments.

Techniques: Formulation, Dispersion, Inhibition

Journal: Pharmaceutics

Article Title: Species-Specific Susceptibility of Planktonic and Biofilm Forming Candida Strains to Cyclodextrin-Encapsulated Essential Oils

doi: 10.3390/pharmaceutics18040508

Figure Lengend Snippet: Species-stratified survival (CFU) responses to essential oils and RAMEB–EO complexes. Estimated marginal means (EMMeans ± SE) of CFU (species-wise Z-transformed) from linear mixed models are shown for each treatment (L, B, P, T and their RAMEB-complexed counterparts RL, RB, RP, and RT). Rows correspond to Candida species (red: Ca; blue: Ct; yellow: Ck; and green: Cd). The horizontal dashed line indicates the within-species reference level (Z = 0). Negative EMMeans reflect reduced CFUs (greater survival suppression) relative to the species baseline, whereas positive values indicate comparatively higher CFU values. This visualization emphasizes species dependence and formulation-dependent shifts (EO vs. RAMEB–EO) in survival outcomes across the treatment panel (non-significant data). Error bars represent ± standard error (SE) of the estimated marginal means.

Article Snippet: The oils were subsequently encapsulated in randomly methylated β-cyclodextrin (RAMEB) by CycloLab Cyclodextrin Research & Development Laboratory (Budapest, Hungary) to produce the RAMEB–essential oil inclusion complexes used in the biological experiments.

Techniques: Transformation Assay, Formulation

Journal: Pharmaceutics

Article Title: Species-Specific Susceptibility of Planktonic and Biofilm Forming Candida Strains to Cyclodextrin-Encapsulated Essential Oils

doi: 10.3390/pharmaceutics18040508

Figure Lengend Snippet: Treatment-induced oxidative/nitrosative stress signatures across Candida species. ( A ) Global hierarchical clustering (Euclidean distance) of treatment signatures based on the standardized mechanistic marker panel RNS, ROS, CAT1 , GPX1 , and SOD1 (species-wise Z-transformed). ( B – E ) Species-stratified heatmaps showing mean Z-scores for each marker by treatment in ( B ) C. albicans (Ca), ( C ) C. tropicalis (Ct), ( D ) C. krusei (Ck), and ( E ) C. dubliniensis (Cd). The rows denote treatments (AM and FL: antifungal reference controls; MN: oxidative-stress control; L/B/P/T: essential oils; and RL/RB/RP/RT: corresponding RAMEB inclusion complexes). The columns indicate mechanistic readouts; the dashed vertical line separates stress markers (RNS/ROS) from antioxidant gene responses ( CAT1 / GPX1 / SOD1 ). The color scale represents mean Z-transformed deviation within species (red = higher-than-species mean; blue = lower-than-species mean), enabling the direct comparison of mechanistic exposure signatures across treatments while controlling baseline species differences (see for treatment-associated changes in antioxidant gene programs). The shaded grey region identifies the specific genes analyzed in this study.

Article Snippet: The oils were subsequently encapsulated in randomly methylated β-cyclodextrin (RAMEB) by CycloLab Cyclodextrin Research & Development Laboratory (Budapest, Hungary) to produce the RAMEB–essential oil inclusion complexes used in the biological experiments.

Techniques: Marker, Transformation Assay, Control, Comparison

Journal: Pharmaceutics

Article Title: Species-Specific Susceptibility of Planktonic and Biofilm Forming Candida Strains to Cyclodextrin-Encapsulated Essential Oils

doi: 10.3390/pharmaceutics18040508

Figure Lengend Snippet: Planktonic metabolism and viability reveal treatment efficacy and species dependence. Panels show estimated marginal means (EMMeans ± SE) from linear mixed models (REML; Satterthwaite degree of freedom) for planktonic functional endpoints across the experimental treatment set (L, B, P, T, RL, RB, RP, and RT), with species indicated by color (red: Ca; blue: Ct; yellow: Ck; and green: Cd). The y -axis is the species-wise Z-transformed response (negative values indicate a shift below the species mean; positive values indicate a shift above the species mean). The x -axis lists treatments, grouped by parent essential oil (EO) and its corresponding RAMEB inclusion complex (REO). ( A – E ) Endpoint-specific EMMeans profiles (one endpoint per panel) illustrate the consistent treatment-driven modulation of planktonic physiology, while differences in the separation and ordering of species-colored points across the treatments indicate species-dependent efficacy patterns (treatment × species effects). The treatment efficiency levels for the experimental parameters (panel ( A ): planktonic metabolic activity/PMT; panel ( B ): planktonic viability/PVA; panel ( C ): biofilm attached cellular metabolic activity/BMT; panel ( D ): biofilm attached cellular viability/BVA; and panel ( E ): biofilm biomass/BB) decline along the horizontal axis, as indicated by the arrow. The significance levels are indicated as ** ( p < 0.001) on the plot. Treatment efficacy for each experimental parameter was inferred from the direction and magnitude of the standardized response associated with each endpoint.

Article Snippet: The oils were subsequently encapsulated in randomly methylated β-cyclodextrin (RAMEB) by CycloLab Cyclodextrin Research & Development Laboratory (Budapest, Hungary) to produce the RAMEB–essential oil inclusion complexes used in the biological experiments.

Techniques: Functional Assay, Transformation Assay, Activity Assay

Journal: Pharmaceutics

Article Title: Species-Specific Susceptibility of Planktonic and Biofilm Forming Candida Strains to Cyclodextrin-Encapsulated Essential Oils

doi: 10.3390/pharmaceutics18040508

Figure Lengend Snippet: Integrated treatment ranking identifies the most effective regimens across endpoints and highlights EO→RAMEB formulation shifts. The main panel shows treatment efficacy ranking based on the composite efficacy score (estimated marginal means, EMMeans ± SE) derived from linear mixed modeling on the standardized (species-wise Z-transformed) integrated endpoint score. Treatments are ordered left-to-right by increasing overall efficacy (arrow indicates rank direction). The dashed horizontal line denotes the neutral reference (0; no shift from the species-wise mean); more positive values indicate higher composite efficacy on the standardized scale, whereas negative values indicate comparatively weaker efficacy. Asterisks denote treatments that differ significantly from the reference level (Bonferroni-adjusted pairwise comparisons; ** p < 0.001). Inset: Slope plot summarizing the direction and magnitude of formulation effects within each EO family, comparing the parent EO to its corresponding RAMEB inclusion complex (R-EO) using the same integrated score. Upward slopes indicate improved efficacy after complexation, while downward slopes indicate reduced efficacy, illustrating that RAMEB reformulation produces family-specific shifts rather than a uniform advantage across all EOs.

Article Snippet: The oils were subsequently encapsulated in randomly methylated β-cyclodextrin (RAMEB) by CycloLab Cyclodextrin Research & Development Laboratory (Budapest, Hungary) to produce the RAMEB–essential oil inclusion complexes used in the biological experiments.

Techniques: Formulation, Derivative Assay, Transformation Assay

Journal: Pharmaceutics

Article Title: Species-Specific Susceptibility of Planktonic and Biofilm Forming Candida Strains to Cyclodextrin-Encapsulated Essential Oils

doi: 10.3390/pharmaceutics18040508

Figure Lengend Snippet: Multivariate mode-of-action landscape separates stress-intensity and antioxidant-program axes across treatments and species. A principal component analysis (PCA) was performed on the aggregated treatment × species mechanistic signatures (RNS, ROS, CAT1 , GPX1 , and SOD1 ; Z-transformed within species). The biplot shows PC1 (86.22%) versus PC2 (13.08%) for each treatment–species signature. The points are colored by treatment type (RC = antifungal reference controls, EO = essential oils, and REO = RAMEB-EO complexes) and shaped by Candida species (Ca, Ct, Ck, and Cd). The dashed crosshairs indicate the origin (0, 0). The gray arrows depict rotated loading vectors: PC1 captures a dominant stress/redox intensity axis (positive direction aligned with ROS and SOD1 , opposed to GPX1 ), whereas PC2 reflects a CAT1 -centered antioxidant-response axis (positive direction aligned with CAT1 ). The centroid labels (treatment codes) indicate the mean position of each treatment across species. The hash marks (#) denote variables with the strongest contributions to the displayed axes (highest absolute loadings). Note: MN is excluded from this panel to prevent the stress-control anchor from compressing the experimental space; full PCA including MN is provided in the .

Article Snippet: The oils were subsequently encapsulated in randomly methylated β-cyclodextrin (RAMEB) by CycloLab Cyclodextrin Research & Development Laboratory (Budapest, Hungary) to produce the RAMEB–essential oil inclusion complexes used in the biological experiments.

Techniques: Transformation Assay, Control