PubMed · 14273375
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P REGNIERS. 1965-01-15. [FEVER FROM UNKNOWN CAUSE].. https://pubmed.ncbi.nlm.nih.gov/14273375/
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Extraction of intact RNA is essential for quantitative gene expression analysis. Isolating high quality RNA from gram-positive bacteria is known to be problematic particularly from organisms that have optimal growth temperatures greater than 45 degrees C. We report a novel extraction protocol for the rapid isolation of fully intact RNA from thermophilic Geobacillus thermoleovorans using a lysing matrix containing a mixture of ceramic and glass beads, triisopropylnaphthalene sulfonic acid (TNS), and p-4-aminosalicyclic acid (PAS). Combining both detergents, TNS and PAS, appeared to increase denaturation of RNases at thermophilic temperatures. Gel electrophoresis revealed that only RNA isolated using the TNS-PAS procedure demonstrated sharp, undegraded 23S, 16S, and 5S ribosomal RNA bands. RNA extracted from geobacilli using commercially available kits was extensively degraded and was not suitable for detecting gene expression. Total RNA yields extracted with the TNS-PAS protocol were greater than eightfold higher than those obtained with available kits. Critically, it was also shown that only RNA isolated with the TNS-PAS-based method was suitable for monitoring thermophile gene expression patterns using RT-PCR analysis.
Arylamine N-acetyltransferase (NAT) enzymes catalyze the addition of an acetyl group from acetyl-CoA to a terminal nitrogen on a suitable substrate such as environmentally relevant compounds and pharmaceuticals. In human, there are two highly polymorphic active allozymes, NAT1 and -2, and one inactive pseudogene, NATP. The expression of these enzymes is tissue-specific such that NAT1 is ubiquitously expressed and NAT2 is confined mainly to liver and colorectal tissues. We hypothesized that these genes would be tissue-specifically transcriptionally regulated, and so we isolated putative proximal control regions for both the NAT genes, which were inserted into luciferase vectors and transiently transfected into human liver and bladder cells. The transfected cells were dosed with 4-aminosalicylic acid, sulfamethazine or solvent and the resulting luciferase activity was measured. We found that both NAT1 and -2 regions were inducible in liver cells by both xenobiotics but only one of the NAT1 regions was inducible again by both xenobiotics in bladder cells. These results suggest that the NAT genes may be tissue-specifically transcriptionally regulated.
The paper presents the results of M. tuberculosis drug resistance analysis. The material was phlegm obtained from 1060 patients treated during 1994-2001 y.y. The tendency of rising of M. tuberculosis drug resistance rate and polyresistance has been established. Unfortunately, a great deal of patients were undercovered with drug resistance M. tuberculosis which can massively grow on cultural medium with peak concentration of streptomycin, kanamycin, rifampicin and PAS(A), as stated above it necessitates to modernize the inspection system directed to determine M. tuberculosis drug resistance.