Currently there are several methods to extract bacterial DNA based on
different principles. However, the amount and the quality of the DNA obtained by each
one of those methods is highly variable and microorganism dependent, as illustrated
by coagulase-negative staphylococci (CoNS) which have a thick cell wall that is
difficult to lyse. This study was designed to compare the quality and the amount of
CoNS DNA, extracted by four different techniques: two in-house protocols and two
commercial kits. DNA amount and quality determination was performed through
spectrophotometry. The extracted DNA was also analyzed using agarose gel
electrophoresis and by PCR. 267 isolates of CoNS were used in this study. The column
method and thermal lyses showed better results with regard to DNA quality (mean ratio
of A260/280 = 1.95) and average concentration of DNA (), respectively. All four methods tested provided appropriate DNA
for PCR amplification, but with different yields. DNA quality is important since it
allows the application of a large number of molecular biology techniques, and also
it's storage for a longer period of time. In this sense the extraction method based
on an extraction column presented the best results for CoNS.
The emergence of Extended-Spectrum Beta-Lactamase (ESBL)-producing microorganisms in Brazilian hospitals is a challenge that concerns scientists, clinicians and healthcare institutions due to the serious risk they pose to confined patients. The goal of this study was the detection of ESBL production by clinical strains of Escherichia coli and Klebsiella sp. isolated from pus, urine and blood of patients at Hospital Universitário Santa Maria, Rio Grande Sul, RS, Brazil and the genotyping of the isolates based on bla SHV genes. The ESBL study was carried out using the Combined Disc Method, while Polymerase Chain Reaction (PCR) was used to study the bla SHV genes. Of the 90 tested isolates, 55 (61.1%) were identified as ESBL-producing by the combined disk method. The bla SHV genes were found in 67.8% of these microorganisms. K. pneumoniae predominated in the samples, presenting the highest frequency of positive results from the combined disk and PCR.
Introduction: The present study aimed to assess the antibiotic resistance profiles and detect the presence of the sul2 gene in sulfamethoxazole-susceptible and resistant isolates of Escherichia coli obtained from outpatients and inpatients with urinary tract infections. Methodology: The resistance profiles of 739 strains were assessed and the presence of the sul2 gene in 100 isolates was tested. Results: The antibiotics with the highest resistance rates were ampicillin (57.4%) and trimethoprim-sulfamethoxazole (44.7%). The presence of the gene sul2 was detected in 66.7% of outpatient samples and 67.9% of inpatient samples.Conclusions: Our results demonstrate that E. coli isolates exhibit high resistance to various classes of antibiotics, highlighting the need for developing strategies to help in prescribing antibiotics.
The essential hypertension has been associated with membrane cell damage. The aim of the present study is investigate the relationship between erythrocyte Na+,K+-ATPase and lipoperoxidation in prehypertensive patients compared to normotensive status. The present study involved the prehypertensive patients (systolic: 136 ± 7 mmHg; diastolic: 86.8 ± 6.3 mmHg; n = 8) and healthy men with normal blood pressure (systolic: 110 ± 6.4 mmHg; diastolic: 76.1 ± 4.2 mmHg; n = 8) who were matched for age (35 ± 4 years old). The venous blood samples of antecubital vein (5 mL) were collected into a tube containing sodium heparin as anticoagulant (1000 UI), and erythrocyte ghosts were prepared for quantifying Na+,K+-ATPase activity. The extent of the thiobarbituric acid reactive substances (TBARS) was determined in plasma. The statistical analysis was carried out by Student's t-test and Pearson's correlation coefficient. A P < 0.05 was considered significant. The Na+,K+-ATPase activity was lower in prehypertensive patients compared with normotensive subjects (4.9 versus 8.0 nmol Pi/mg protein/min; P < 0.05). The Na+,K+-ATPase activity correlated negatively with TBARS content (r = −0.6; P < 0.05) and diastolic blood pressure (r = −0.84; P < 0.05). The present study suggests that Na+,K+-ATPase activity reduction and elevation of the TBARS content may underlie the pathophysiological aspects linked to the prehypertensive status.
Resection of the inferior vena cava may be required in the courses of oncological surgeries for the tumors originating from or invading it. Management of the remaining defect depends on the extension of the resection. Partial or complete replacement of the inferior vena cava, with a patch or interposition graft, may be required. Standard techniques for the reconstruction with a prosthetic material or the autologous veins can be associated with the prosthetic graft infection, high cost, long‐standing anticoagulation, technical difficulties, and/or need for extra incisions. The use of the autologous peritoneum represents an easy and inexpensive alternative for the partial and complete inferior vena cava reconstructions.
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