The host is the main environment for bacteria, and they also expose to many antibiotics during the treatment of infectious diseases in host body. In this study, it was aimed to investigate possible changes in growth rate and expression levels of three virulence genes (foc/foc, cnf1, and usp) in a uropathogenic E. coli standard strain within the presence of ciprofloxacin, nitrofurantoin, and trimethoprim-sulfamethoxazole. The UPEC C7 strain was grown on tryptic soy broth-TSB (control), TSB + ciprofloxacin, TSB + nitrofurantoin, and TSB + trimethoprim-sulfamethoxazole for determination of both growth rate and gene expression level. Antibiotics were added according to their sub-minimal inhibition concentrations. E-test was used to determine MIC values of antibiotics. Growth changes were measured in absorbance 600 nm during 24-h period. Total RNA isolations were performed after incubation for 24 h at 37 °C. Gene expression levels were determined by quantitative PCR. Tukey's post hoc test was used for statistical analysis. According to absorbance values, it has been shown that only ciprofloxacin and trimethoprim-sulfamethoxazole have lead significant decrease on growth rate. We also detected statistically significant differences in each gene expression levels for all antibiotics via relative quantification analysis. Fold changes in gene expression was found 0.65, 1.42, 0.23 for foc/foc gene; 0.01, 0.01, 2.84 for cnf1 gene; and 0.1, 0.01, 0.01 for usp gene in the presence of ciprofloxacin, nitrofurantoin, and trimethoprim/sulfamethoxazole, respectively. This investigation has shown that antibiotics can play a role as an environmental factor which may determine the pathogenicity of bacteria in vivo.
Magnesium (Mg2+), an essential ion for cells and biological systems, is
involved in a variety of cellular processes, including the formation and breakdown of
microtubules. The results of a previous investigation suggested that as cells grow
the intracellular Mg2+ concentration falls, thereby stimulating formation
of the mitotic spindle. In the present work, we used a Mg2+-deficient
Schizosaccharomyces pombe strain GA2, in which two essential
membrane Mg2+ transporter genes (homologs of ALR1 and
ALR2 in Saccharomyces cerevisae) were deleted,
and its parental strain Sp292, to examine the extent to which low Mg2+
concentrations can affect mitotic spindle formation. The two S.
pombe strains were transformed with a plasmid carrying a GFP-α2-tubulin
construct to fluorescently label microtubules. Using the free
Mg2+-specific fluorescent probe mag-fura-2, we confirmed that
intracellular free Mg2+ levels were lower in GA2 than in the parental
strain. Defects in interphase microtubule organization, a lower percentage of mitotic
spindle formation and a reduced mitotic index were also observed in the GA2 strain.
Although there was interphase microtubule polymerization, the lower level of mitotic
spindle formation in the Mg2+-deficient strain suggested a greater
requirement for Mg2+ in this phenomenon than previously thought.
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