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 One of the mechanisms of Candida albicans resistance to azole drugs used in antifungal therapy relies on increased expression and presence of point mutations in the ERG11 gene that encodes sterol 14α demethylase (14DM), an enzyme which is the primary target for the azole class of antifungals. The aim of the study was to analyze nucleotide substitutions in the Candida albicans ERG11 gene of azole-susceptible and azole-resistant clinical isolates. The Candida albicans isolates represented a collection of 122 strains selected from 658 strains isolated from different biological materials. Samples were obtained from hospitalized patients. Fluconazole susceptibility was tested in vitro using a microdilution assay. Candida albicans strains used in this study consisted of two groups: 61 of the isolates were susceptible to azoles and the 61 were resistant to azoles. Four overlapping regions of the ERG11 gene of the isolates of Candida albicans strains were amplified and sequenced. The MSSCP (multitemperature single strand conformation polymorphism) method was performed to select Candida albicans samples presenting genetic differences in the ERG11 gene fragments for subsequent sequence analysis. Based on the sequencing results we managed to detect 19 substitutions of nucleotides in the ERG11 gene fragments. Sequencing revealed 4 different alterations: T495A, A530C, G622A and A945C leading to changes in the corresponding amino acid sequence: D116E, K128T, V159I and E266D. The single nucleotide changes in the ERG11 gene did not affect the sensitivity of Candida albicans strains, whereas multiple nucleotide substitutions in the ERG11 gene fragments indicated a possible relation with the increase in resistance to azole drugs.
Background. The inadequate nutritional status of hospitalized patients leading to the malnutrition is one of the crucial clinical problems. Objective. The aim of the study was to assess nutritional status of patients on the basis of the mini nutritional assessment (MNA) questionnaire. Material and Methods. The study group included 120 patients staying on the internal medicine ward. The average age for female and male was 65±9.7 and 63±8.4 years, respectively. To assess malnutrition the anthropometric measurements and the MNA questionnaire was conducted. Results. The average MIS (Malnutrition Indicator Score) value in female and male was 25.0 ± 3.0. Among 29% of women and 18.2% of men the risk of malnutrition occurrence was noted. The incorrect values of BMI (body mass index) were observed more frequently in male compared to female (84.0% vs. 67.0%). Incorrect values of the MAC (mid arm circumference) and CC (calf circumference) were found respectively in 6.6 % and 10.5% of female and in 2.3% of male. 55.3% of women and 75% of men declared taking more than 3 prescription drugs a day. In women were observed a significantly higher deficiency of dairy products, meat, fish, or poultry as well as meals containing legume plants or eggs per week compared to men (21.0% vs. 4.5%). An insufficient daily intake of vegetables was declared by 20.4% of men and 15.8% of women. Conclusions. The occurrence of malnutrition or its risk was observed in about a quarter of studied hospitalized patients. The malnutrition was worsened by a reported loss of appetite, a significant number of drugs being taken, and inadequate nutrition among the patients.
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