ABSTRACT
Peptide nucleic acid fluorescence in situ hybridization (PNA FISH) is a molecular diagnostic tool for the rapid detection of pathogens directly from liquid media. The aim of this study was to prospectively evaluate PNA FISH assays in comparison with culture-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) identification, as a reference method, for both blood and cerebrospinal fluid (CSF) cultures, during a 1-year investigation. On the basis of the Gram stain microscopy results, four different PNA FISH commercially available assays were used ('Staphylococcus aureus/CNS', 'Enterococcus faecalis/OE', 'GNR Traffic Light' and 'Yeasts Traffic Light' PNA FISH assays, AdvanDx). The four PNA FISH assays were applied to 956 positive blood cultures (921 for bacteria and 35 for yeasts) and 11 CSF cultures. Among the 921 blood samples positive for bacteria, PNA FISH gave concordant results with MALDI-TOF MS in 908/921 (98.64%) samples, showing an agreement of 99.4% in the case of monomicrobial infections. As regards yeasts, the PNA FISH assay showed a 100% agreement with the result obtained by MALDI-TOF MS. When PNA FISH assays were tested on the 11 CSF cultures, the results agreed with the reference method in all cases (100%). PNA FISH assays provided species identification at least one work-day before the MALDI-TOF MS culture-based identification. PNA FISH assays showed an excellent efficacy in the prompt identification of main pathogens, yielding a significant reduction in reporting time and leading to more appropriate patient management and therapy in cases of sepsis and severe infections.
Subject(s)
Bacteria/isolation & purification , Blood/microbiology , Cerebrospinal Fluid/microbiology , Fungi/isolation & purification , In Situ Hybridization, Fluorescence/methods , Peptide Nucleic Acids , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Adolescent , Adult , Aged , Aged, 80 and over , Bacteremia/diagnosis , Bacteremia/microbiology , Central Nervous System Fungal Infections/diagnosis , Central Nervous System Fungal Infections/microbiology , Child , Child, Preschool , Diagnostic Tests, Routine/methods , Female , Fungemia/diagnosis , Fungemia/microbiology , Humans , Infant , Infant, Newborn , Male , Meningitis, Bacterial/diagnosis , Meningitis, Bacterial/microbiology , Middle Aged , Prospective Studies , Time Factors , Young AdultABSTRACT
We evaluated the role of acetyl-L-carnitine (ALCAR) in protecting primary motoneuron cultures exposed to excitotoxic agents or serum-brain derived neurotrophic factor (BDNF) deprived. To exclude that ALCAR works as a metabolic source, we compared its effects with those of L-carnitine (L-CAR), that seems to have no neurotrophic effect. A concentration of 10 mM ALCAR, but not L-CAR, significantly reduced the toxic effect of 50 microM N-methyl-D-aspartate (NMDA, % viability: NMDA 45.4+/-2.80, NMDA+ALCAR 90.8+/-11.8; P<0.01) and of 5 microM kainate in cultured motoneurons (% viability: kainate 40.66+/-10.73; kainate+ALCAR 63.80+/-13.88; P<0.05). The effect was due to a shift to the right of the dose-response curve for kainate (EC50 for kainate 5.99+/-1.012 microM; kainate+ALCAR 8.62+/-1.13 microM; P<0.05). ALCAR, but not L-CAR, significantly protected against BDNF and serum-deprivation reducing the apoptotic cell death (% viability respect to control: without BDNF/serum 61.8+/-13.3: without BDNF/serum+ALCAR 111.8+/-13.9; P<0.01). Immunocytochemistry showed an increase in choline acethyltransferase and tyrosine kinaseB receptors in motoneurons treated with ALCAR but not with L-CAR. These results suggest that ALCAR treatment improves the motoneurons activity, acting as a neurotrophic factor.
Subject(s)
Acetylcarnitine/pharmacology , Motor Neurons/drug effects , Neuroprotective Agents/pharmacology , Nootropic Agents/pharmacology , Amyotrophic Lateral Sclerosis/drug therapy , Animals , Cells, Cultured , Choline O-Acetyltransferase/metabolism , Fetus/cytology , Motor Neurons/cytology , Neurotoxins/pharmacology , Rats , Rats, Sprague-Dawley , Receptor Protein-Tyrosine Kinases/metabolismABSTRACT
The preliminary heat resistance evaluation of 94 Salmonella strains was carried out in culture medium (Trypticase soy broth, TSB). The heat resistance of three S. typhimurium strains (ATCC 14028, 133 and 1116), a strain each of S. derby B4373, S. potsdam 1133, S. menston 179. S. eppendorf 166, and S. kingston I124 was determined also in pork meat containing curing additives. As expected, the eight Salmonella strains showed greater heat resistance in pork meat than in TSB. At the lowest temperature (58 degrees C), the heat resistance increased 1.5-4 times, and it was most pronounced for the strains being most heat sensitive in TSB. S. potsdam 133 was the most resistant strain in pork meat, with D-values at 58 degrees C, 60 degrees C and 63 degrees C of 4.80, 1.57 and 0.30 min, respectively. The most sensitive strain turned out to be S. kingston 1124, with D-values of 2.79. 0.92 and 0.24 min, at the same temperatures. According to collected data, the heating processes, as applied to cured pork meat, providing an internal temperature of 60 degrees C for 9-10 min or of 63 degrees C for 3-4 min can be expected to provide a > or = 7 D kill of Salmonella belonging to the serotypes studied.
