António Manuel Leal Ferreira Mendonça da Fonseca

Antifungal activity of ibuprofen alone and in combination with fluconazole against Candida species.

Ibuprofen, a non-steroidal anti-inflammatory drug, exhibited antimicrobial activity against Candida albicans and non-albicans strains. At 10 mg/ml, ibuprofen showed a rapid cidal activity against exponential growth phase C. albicans, accompanied by rapid and extensive leakage of intracellular K+, permeation to propidium iodide, lysis of spheroplasts and severe membrane ultrastructural alterations. These results indicate that the killing of Candida cells is due to direct damage to the cytoplasmic membrane. At 5 mg/ml, ibuprofen inhibited growth; however, it did not kill the yeasts and did not directly affect the cytoplasmic membrane. Evaluation of yeast metabolic vitality with the fluorescent probe FUN-1 showed that growth inhibition induced by the fungistatic drug concentration was due to metabolic alterations. The combination of ibuprofen with fluconazole resulted in synergic activity with eight of the 12 Candida strains studied, including four of the five fluconazole-resistant strains. The MICs of fluconazole for the fluconazole-resistant strains decreased 2-128-fold when the drug was associated with ibuprofen. When in combination with fluconazole, MICs for ibuprofen decreased by up to 64-fold for all the 12 strains studied. These results point to the practicability of using ibuprofen, alone or in combination with azoles, in the treatment of candidosis, particularly when applied topically, taking advantage of the drugs antifungal and anti-inflammatory properties.

Antifungal activity of local anesthetics against Candida species.

OBJECTIVE: To evaluate the activity of benzydamine, lidocaine, and bupivacaine, three drugs with local anesthetic activity, against Candida albicans and non-albicans strains and to clarify their mechanism of activity. METHODS: The minimal inhibitory concentration (MIC) was determined for 20 Candida strains (18 clinical isolates and two American Type Culture Collection strains). The fungistatic activity was studied with the fluorescent probe FUN-1 and observation under epifluorescence microscopy and flow cytometry. The fungicidal activity of the three drugs was assayed by viability counts. Membrane alterations induced in the yeast cells were evaluated by staining with propidium iodide, by quantitation of intracellular K+ leakage and by transmission electron microscopy of intact yeast cells and prepared spheroplasts. RESULTS: The MIC ranged from 12.5-50.0 microg/mL, 5.0-40.0 mg/mL, and 2.5-10.0 mg/mL for benzydamine, lidocaine, and bupivacaine, respectively. The inhibitory activity of these concentrations could be detected with the fluorescent probe FUN-1 after incubation for 60 minutes. A very fast fungicidal activity was shown by 0.2, 50, and 30 mg/mL of benzydamine, lidocaine, and bupivacaine, respectively. CONCLUSIONS: At lower concentrations, the tested drugs have a fungistatic activity, due to yeast metabolic impairment, while at higher concentrations they are fungicidal, due to direct damage to the cytoplasmic membrane.

Susceptibility to fluconazole of Candida clinical isolates determined by FUN-1 staining with flow cytometry and epifluorescence microscopy.

The susceptibility of clinical Candida isolates to fluconazole was assayed by flow cytometry (FCM) and epifluorescence microscopy (EFM), with FUN-1 staining. In all, 25 clinical isolates of Candida spp. (12 sensitive, 3 dose-dependently sensitive and 10 resistant to fluconazole according to the NCCLS M27-A protocol) were treated with increasing concentrations of fluconazole during 1 or 2 h staining with FUN-1 for 30 min and analysed, respectively, by FCM at 575 nm (FL2) and by EFM. Fluconazole-susceptible strains showed an increased accumulation of FUN-1 in comparison with controls as determined by FCM and a reduced metabolic processing of the probe, confirmed by EFM. Conversely, resistant strains showed decreased FUN-1 staining and were able to process the probe. The fluconazole minimal inhibitory concentrations (MICs) determined by FCM or EFM after FUN-1 staining compared very well with the corresponding values determined by the M27-A protocol, indicating that FUN-1 staining can be used as an alternative to the conventional method. MIC values of resistant strains, with the exception of C. krusei, were lower when treatment with fluconazole followed pre-incubation with 0.1 mM sodium azide, a concentration known to inhibit the activity of efflux pumps. These results show that FUN-1 staining can be used as an alternative and rapid method for the assessment of susceptibility of Candida clinical isolates to fluconazole. Furthermore, the results suggest that resistance of Candida cells to fluconazole, with the exception of C. krusei strains, is likely to be due to the activity of efflux pumps.

Germ tube formation changes surface hydrophobicity of Candida cells

Hydrophobic interaction is generally considered to play an important role in the adherence of microorganisms to eukaryotic cells and also to certain inert surfaces. Using a microbe adhesion assay to hydrocarbons (n-hexadecane), 68 strains of Candida albicans and 30 non-albicans strains were studied. Influence of source of isolate, age of the culture, and percentage of germ tube formation on adhesion were studied. C. albicans blastoconidia were found to be hydrophilic; conversely, blastoconidia of non-albicans strains were slightly more hydrophobic. Germ tube formation was associated with a significant rise in cell surface hydrophobicity.

Como avaliar a percepção de competência e aceitação social de crianças com paralisia cerebral?Estudo inicial para a determinação das propriedades psicométricas da versão portuguesa da Dutch Pictorial Scale of Perceived Competence and Social Acceptance in Children with Cerebral Palsy

The main purpose of this study was to examine the psychometric properties of the Portuguese version of the Dutch Pictorial Scale of Perceived Competence and Social Acceptance in Children with Cerebral Palsy. The sample was made up of 108 children (60 boys and 48 girls) with cerebral palsy, aged between 4 and 9 years, who suffered hemiplegia, diplegia or quadriplegia and had an IQ of 70 or higher. Most of the children (n=98) attended regular schools (integrated in kindergarten or primary school); indeed, only 10 children attended special schools for children with special needs. A sub-sample of 41 children filled out the scale in two different moments, separated one to two weeks maximum. The results of Pearson correlations between the two administrations (ranging 0.80 to 0.98) and the Cronbachs alpha (ranging 0.69 to 0.93) showed good reliability of the Portuguese version of the Dutch Pictorial Scale. Moreover, the inter-correlations between the four subscales (r=0.60 or higher) as well the mean scores associated by children with different characteristics (age, sex, medical condition) to the different subscales supported the internal validity of the instrument. Therefore, this study provides initial evidence about the psychometric properties of the Portuguese version of the Dutch Pictorial Scale, which is why its use is recommended to assess the perceived competence and social acceptance of Portuguese children with cerebral palsy.

How to measure children with cerebral palsy's perception of competence and social acceptance?Preliminary study to determinate the psychometric properties of the portuguese version of the Dutch Pictorial Scale of Perceived Competence and Social Acceptance in Children with Cerebral Palsy

The main purpose of this study was to examine the psychometric properties of the Portuguese version of the Dutch Pictorial Scale of Perceived Competence and Social Acceptance in Children with Cerebral Palsy. The sample was made up of 108 children (60 boys and 48 girls) with cerebral palsy, aged between 4 and 9 years, who suffered hemiplegia, diplegia or quadriplegia and had an IQ of 70 or higher. Most of the children (n=98) attended regular schools (integrated in kindergarten or primary school); indeed, only 10 children attended special schools for children with special needs. A sub-sample of 41 children filled out the scale in two different moments, separated one to two weeks maximum. The results of Pearson correlations between the two administrations (ranging 0.80 to 0.98) and the Cronbachs alpha (ranging 0.69 to 0.93) showed good reliability of the Portuguese version of the Dutch Pictorial Scale. Moreover, the inter-correlations between the four subscales (r=0.60 or higher) as well the mean scores associated by children with different characteristics (age, sex, medical condition) to the different subscales supported the internal validity of the instrument. Therefore, this study provides initial evidence about the psychometric properties of the Portuguese version of the Dutch Pictorial Scale, which is why its use is recommended to assess the perceived competence and social acceptance of Portuguese children with cerebral palsy.

Como avaliar a percepção de competência e aceitação social de crianças com paralisia cerebral?Estudo inicial para a determinação das propriedades psicométricas da versão portuguesa da Dutch Pictorial Scale of Perceived Competence and Social Acceptance in

The main purpose of this study was to examine the psychometric properties of the Portuguese version of the Dutch Pictorial Scale of Perceived Competence and Social Acceptance in Children with Cerebral Palsy. The sample was made up of 108 children (60 boys and 48 girls) with cerebral palsy, aged between 4 and 9 years, who suffered hemiplegia, diplegia or quadriplegia and had an IQ of 70 or higher. Most of the children (n=98) attended regular schools (integrated in kindergarten or primary school); indeed, only 10 children attended special schools for children with special needs. A sub-sample of 41 children filled out the scale in two different moments, separated one to two weeks maximum. The results of Pearson correlations between the two administrations (ranging 0.80 to 0.98) and the Cronbachs alpha (ranging 0.69 to 0.93) showed good reliability of the Portuguese version of the Dutch Pictorial Scale. Moreover, the inter-correlations between the four subscales (r=0.60 or higher) as well the mean scores associated by children with different characteristics (age, sex, medical condition) to the different subscales supported the internal validity of the instrument. Therefore, this study provides initial evidence about the psychometric properties of the Portuguese version of the Dutch Pictorial Scale, which is why its use is recommended to assess the perceived competence and social acceptance of Portuguese children with cerebral palsy.