Lovastatin possesses a fungistatic effect against Candida albicans, but does not trigger apoptosis in this opportunistic human pathogen

The quote only partly states this claim.

After 12 h incubation with lovastatin, cell survival rates are 85±5, 84±12, 94±23, and 78±17% for 0, 12.5, 25, and 50 μg/mL, respectively.

“For example, after 12 h incubation with lovastatin, cell survival rates were 85 Æ 5, 84 Æ 12, 94 Æ 23 and 78 Æ 17% (n = 3; P > 5%; in comparison to 0-h cultures) with increasing lovastatin concentrations of 0, 12.5, 25 and 50 mg mL À1 , respectively.”

May combine two claims.

After 24 h incubation with lovastatin, the ratios of pseudomycelia and hyphae are >90% and ~5%, respectively.

“After 24 h incubation with lovastatin, the ratios of pseudomycelia and hyphae were 4 90% and ffi 5%, respectively.”

The quote only partly states this claim.

Incubation with 12.5-50 μg/mL lovastatin does not significantly decrease colony-forming ability at any tested time point.

“The incubation of C. albicans cells in the presence of 12.5-50 mg mL À1 lovastatin did not cause any significant decrease in the colony-forming ability at any time point tested.”

May combine two claims.

At 24 h cultivation, the ratios of annexin-V-positive and TUNEL-positive protoplasts are 12±3% and 8±3%, compared with 0.20±0.05% and 0.10±0.03% at 8 h.

“At 24 h cultivation time, the ratios of protoplasts with externalized phosphatidylserine (annexin-V-positive protoplasts) and fragmented DNA (TUNEL-positive protoplasts) were 12 Æ 3 and 8 Æ 3%, respectively, in comparison to 8-h cultures where they were 0.20 Æ 0.05 and 0.10 Æ 0.03%, respectively”

May combine two claims.

Vitamin E does not significantly change the growth of Candida albicans but decreases both peroxide content and the ratio of annexin-V-positive protoplasts.

“Vitamin E did not cause significant changes in the growth of C. albicans (data not shown); however, it decreased markedly both the peroxide content of the cells and the ratio of the annexin-V-positive protoplasts (Table 1).”

May combine two claims.

Lovastatin has a dose-dependent growth-inhibitory effect on Candida albicans with decreased glucose utilization.

“Lovastatin had a dose-dependent growth-inhibitory effect on C. albicans (Fig. 3a), with decreased glucose utilization rates (Fig. 3b), which was fungistatic rather than fungicidal.”

May combine two claims.

Lovastatin-treated cultures do not reach stationary phase by 24 h and are physiologically equivalent to 8-h control cultures in biomass, glucose consumption, superoxide, and peroxide.

“It is worth mentioning that in these experiments lovastatin-treated cultures did not reach the stationary phase of growth by 24 h of cultivation and seemed to be physiologically equivalent to 8-h control cultures concerning biomass production (1.0 Æ 0.06 vs. 1.1 Æ 0.06 OD 640 units; Fig. 3a), glucose consumption rates (34.2Æ 2.1 vs. 32 Æ 4.4 g L À1 ; Fig. 3b) as well as superoxide (1.1 Æ 0.1 vs. 1”

May combine two claims.

Lovastatin does not induce apoptosis in Candida albicans, with annexin-V-positive and TUNEL-positive ratios remaining low after 8 h exposure.

“Lovastatin did not induce apoptosis in C. albicans.When lovastatin (12.5 mg mL À1 ) was added at 0 h of cultivation, the ratios of annexin-V-positive protoplasts in exposed and untreated control cultures after 8 h exposure were 0.25 Æ 0.05 and 0.20 Æ 0.05%, respectively, while the ratios of TUNEL-positive protoplasts were 0.09 Æ 0.05 and 0.1 Æ 0.03%, respectively.”

May combine two claims.

EPR measurements show 2Azz values of 6.220±0.025 mT and 5.539±0.025 mT, and S values of 0.864 and 0.678, in lovastatin-treated and untreated cultures, respectively.

“In the treated and untreated cultures, the 2A 0 zz values were 6.220 Æ 0.025 mT and 5.539 Æ 0.025 mT (n = 3), while the S values were 0.864 and 0.678, respectively.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

After prolonged 24-h lovastatin exposure, expression of protein prenylation genes declines, which was attributed to the progressing antifungal effect of the statin.

“After prolonged, 24-h exposures to lovastatin, the expression of protein prenylation genes declined, a result that was attributed to the progressing antifungal effect of the statin (Song et al., 2003).”

• cites Song et al., 2003Not checked yet

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Classified as probably not a substantial claim.

Statins trigger apoptosis in many human tumour cell lines by disturbing prenylation-dependent signal transduction pathways.

“In addition, they trigger apoptosis in many human tumour cell lines by disturbing prenylation-dependent signal transduction pathways (Mo & Elson, 1999, 2004;Kusama et al., 2002Kusama et al., , 2003;;Jakobisiak & Golab, 2003;Wu et al., 2004;Cafforio et al., 2005;Shellman et al., 2005).”

• cites Mo & Elson, 1999, 2004Not checked yet

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• cites Kusama et al., 2002Not checked yet

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• cites Jakobisiak & Golab, 2003Not checked yet

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Probably not a substantial claim: may not pass S3 (load-bearing).

Lovastatin treatment slightly induces expression of ERG9 and ERG11 and protein prenylation genes RAM1, RAM2, and CDC43 in mid-logarithmic Candida albicans cultures.

“Importantly, lovastatin treatments induced slightly the expression of ERG9 (squalene synthase) and ERG11 (14 alanosterol demethylase) in the later part of the sterol biosynthetic pathway, together with the protein prenylation genes RAM1, RAM2 and CDC43 in mid-logarithmic growth cultures (Song et al., 2003).”

• cites Song et al., 2003Not checked yet

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May combine two claims.

The increase in order parameter and hyperfine splitting constant indicates decreased mobility of probe molecules, suggesting decreased plasma membrane fluidity in lovastatin-treated cells.

“The increase of order parameter and hyperfine splitting constant indicates a strong interaction, which resulted in decreased mobility in the environment of the probe molecules, suggesting a decreased plasma membrane fluidity in the lovastatintreated cells.”

May combine two claims.

Lovastatin has an antifungal effect on Candida albicans that is synergistic with fluconazole.

“A more recent study by Song et al. (2003) demonstrated the antifungal effect of lovastatin on the opportunistic human pathogenic fungus Candida albicans, which was synergistic with the action of the 14 a-lanosterol demethylase inhibitor fluconazole.”

• cites Song et al., 2003Not checked yet

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Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Lovastatin interferes with Ras signalling and promotes apoptosis-like cell death in Mucor racemosus.

“Lovastatin, a fungal metabolite produced by Aspergillus terreus and Monascus ruber, interferes with Ras-signalling and promotes apoptosis-like cell death in the fungus Mucor racemosus as well (Roze & Linz, 1998).”

• cites Roze & Linz, 1998Not checked yet

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May combine two claims.

The signalling pathways responsible for apoptosis and the morphogenetic switch must diverge or work independently in stress-exposed Candida albicans cells.

“Our observation that lovastatin induced pseudomycelium and some hyphae formation without provoking the onset of programmed cell death further strengthen the view that the signalling pathways responsible for apoptosis and the morphogenetic switch must diverge at some point (Phillips et al., 2003) or work independently in stress-exposed C. albicans cells.”

• cites Phillips et al., 2003Not checked yet

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May combine two claims.

Highest achievable peak serum levels of lovastatin at doses of 4 mg/kg/day or above are 0.1-3.92 μM (about 40 ng/mL-1.6 μg/mL), well below MIC values of 5-30 μg/mL for Candida albicans.

“The highest achievable peak serum levels (0.1-3.92 mM ffi 40 ng mL À1 -1.6 mg mL À1 ) recorded at doses of 4 mg kg À1 day À1 or above in a clinical Phase I study by Thibault et al. (1996) for lovastatin as an anticancer agent are well below the MIC values (5-30 mg mL À1 ) determined for C. albicans in the presence of this statin (this study; Song et al., 2003).”

• cites Thibault et al., 1996Not checked yet

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• cites Song et al., 2003Not checked yet

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Classified as probably not a substantial claim. May combine two claims.

In apoptosis detection, propidium iodide staining was used to filter necrotic cells, and only annexin-V-positive but propidium-iodide-negative protoplasts were regarded as apoptotic.

“The protocol also included propidium iodide staining for filtering necrotic cells, and only annexin-Vpositive but propidium-iodide-negative protoplasts were regarded as 'apoptotic'.”

May combine two claims.

Proapoptotic doses of acetic acid and H2O2 promote the yeast-to-hyphae morphogenetic switch in Candida albicans, but no such transition is observed prior to or during apoptosis triggered by amphotericin B.

“Proapoptotic doses of acetic acid and H 2 O 2 have been shown to promote the yeast !hyphae morphogenetic switch in C. albicans cultures, but no such morphological transition was observed prior to or during apoptotic cell death triggered by amphotericin B (Phillips et al., 2003).”

• cites Phillips et al., 2003Not checked yet

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May combine two claims.

None of the antioxidant enzymes are affected by lovastatin, indicating a lack of oxidative stress in lovastatin-exposed Candida albicans, which is a prerequisite of fungal apoptosis.

“Importantly, none of these antioxidants was affected by this treatment, which indicates the lack of H 2A ' zz (a) (b) oxidative stress in lovastatin-exposed C. albicans cells, a prerequisite of fungal apoptosis.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Statistical significance of changes in physiological parameters was estimated using Student's t-test, with only P<5% regarded as significant.

“Variations between experiments were estimated by standard deviations (SD), and the statistical significance of changes in physiological parameters was estimated using Student's t-test.Only probability levels of P 5% were regarded as statistically significant.”

Classified as probably not a substantial claim.

MIC determination for lovastatin was performed in SDB according to NCCLS guidelines for antifungal susceptibility testing.

“Minimum inhibitory concentration (MIC) determination for lovastatin was performed in SDB medium according to the National Committee for Clinical Laboratory Standards (NCCLS) guidelines for antifungal susceptibility testing (Song et al., 2003).”

• cites Song et al., 2003Not checked yet

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The Candida albicans AG05 isolate was identified using CHROMagar, the ID32C system, germ tube development in sheep serum, and chlamydospore formation on corn meal agar.

“The species was identified using CHROMagar (Oxoid, Basingstoke, UK) and the ID32C system (API bioMérieux, Nürtingen, Germany), and by the development of germ tubes in sheep serum and the ability to form chlamydospores on corn meal agar (Oxoid).”

Probably not a substantial claim: may not pass S3 (load-bearing).

In each phosphatidylserine exposure and TUNEL staining experiment, 5000 protoplasts were analysed using epifluorescence microscopy.

“In each phosphatidylserine exposure and TUNEL staining experiment, 5000 protoplasts were analysed using epifluorescence microscopy (Leiter et al., 2005).”

• cites Leiter et al., 2005Not checked yet

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The importance of ergosterol in yeast-to-hyphae morphological transitions has been demonstrated.

“The importance of ergosterol in yeast !hyphae morphological transitions, for example via the inhibition of chitin synthase, which is activated during budding, has been demonstrated (Pesti et al., 1981;Chiew et al., 1982).”

• cites Pesti et al., 1981Not checked yet

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• cites Chiew et al., 1982Not checked yet

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Probably not a substantial claim: may not pass S3 (load-bearing).

Means and SD values were calculated from 3 to 5 independent experiments.

“Means and SD values calculated from 3 to 5 independent experiments (n) are presented in the paper.”