Inhibition of the Growth of Pathogenic Candida spp. by Salicylhydroxamic Acid

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth each of six Candida spp. was inhibited significantly by ~13 mM SHAM or 2 mM cyanide, albeit to differing extents. In C. dubliniensis, C. krusei and C. tropicalis the rate of O2 uptake was inhibited by 18-36% by 25 mM SHAM, but this had little or no effect on C. glabrata, C. guilliermondii or C. parapsilosis. Although SHAM substantially inhibited the growth of Candida spp., it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of Candida spp. infection.

Estimating Enzyme Kinetic Parameters from Apparent KMs and Vmaxs

The kinetic properties of enzymes are often reported using the apparent KM and Vmax appropriate to the standard Michaelis-Menten enzyme. However, this model is inappropriate to enzymes that have more than one substrate or where the rate expression does not apply for other reasons. Consequently, it is desirable to have a means of estimating the appropriate kinetic parameters from the apparent values of KM and Vmax reported for each substrate. We provide a means of estimating the range within which the parameters should lie and apply the method to data for glutamate dehydrogenase from the nematode parasite of sheep Teladorsagia circumcincta.

Error Estimates for Calculated Glomerular Filtration Rates

Glomerular filtration rate (GFR) is a measure of kidney function. It is usually estimated from serum concentrations of cystatin C or creatinine although there has been considerable debate in the literature about (i) the best equation to use and (ii) the variability in the correlation between the concentrations of creatinine and cystatin C. The equations for GFR can be written in a general form and from these I calculate the error of the GFR estimates associated with analyte measurement error. These show that the error of the GFR estimates is such that it is not possible to distinguish between the equations over much of the concentration range of either analyte. The general forms of the equations are also used to derive an expression for the concentration of cystatin C as a function of the concentration of creatinine. This equation shows that these analyte concentrations are not linearly related. Clinical reports of cystatin C and creatinine concentration are consistent with the expression derived.

Salicylhydroxamic Acid Inhibits the Growth of Candida albicans

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth Candida albicans was inhibited by SHAM (Ki = 9-15 mM) and cyanide (Ki = 2-4 mM), albeit to differing extents. The rate of O2 uptake was inhibited by less than 10% by 25 mM SHAM and by about 90% by 250 μM KCN. Although SHAM substantially inhibited the growth of C. albicans, it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of C. albicans infection.

The Contribution of Growth Rate to the Pathogenicity of Candida spp.

Fungal infections are becoming more common and the range of susceptible individuals has expanded. While Candida albicans remains the most common infective species, other Candida spp. are becoming increasingly significant. In a range of large-scale studies of candidaemia between 1999 and 2006, about 52% of 9717 cases involved C. albicans, about 30% involved either C. glabrata or C. parapsilosis and less than 15% involved C. tropicalis, C. krusei or C. guilliermondii. However, the probability of mortality within 30 days of infection with a particular species was at least 40% for C. tropicalis, C. albicans, C. glabrata and C. krusei and only 22% for C. parapsilopsis. Clinical isolates of Candida spp. grew at rates ranging from 1.65 h-1 to 4.9 h-1. Three species (C. krusei, C. albicans and C. glabrata) had relatively high growth rates (μm > 4 h-1), C. tropicalis and C. dubliniensis grew moderately quickly (Ôëê 3 h-1) and C. parapsilosis and C. guilliermondii grew slowly (< 2 h-1). Based on these data, the log of the odds of mortality within 30 days of diagnosis was linearly related to μm. From this the underlying probability of mortality is 0.13 (95% CI: 0.10-0.17) and it increases by about 0.09 ± 0.02 for each unit increase in μm. Given that the overall crude mortality is about 0.36, the growth of Candida spp. approximately doubles the rate, consistent with the results of larger case-matched studies of candidaemia.

Induction of Alternative Oxidase Activity in Candida albicans by Oxidising Conditions

Candida albicans ATCC 10231 had low endogenous activity of the alternative oxidase compared with that of C. albicans ATCC 10261. In C. albicans ATCC 10231 the endogenous activity declined as the cultures aged. Alternative oxidase activity could be induced in C. albicans ATCC 10231 by treatment with cyanide, but the induction of this activity required the presence of oxygen which could be replaced, at least in part, with high concentrations of potassium ferricyanide. We infer from this that the expression of the gene encoding the alternative oxidase is under the control of a redoxsensitive transcription factor.