BackgroundSalicylic acid (SA) is a key defense signal molecule against biotrophic pathogens in plants. Quantification of SA levels in plants is critical for dissecting the SA-mediated immune response. Although HPLC and GC/MS are routinely used to determine SA concentrations, they are expensive and time-consuming. We recently described a rapid method for a bacterial biosensor Acinetobacter sp. ADPWH_lux-based SA quantification, which enables high-throughput analysis. In this study we describe an improved method for fast sample preparation, and present a high-throughput strategy for isolation of SA metabolic mutants.ResultsOn the basis of the previously described biosensor-based method, we simplified the tissue collection and the SA extraction procedure. Leaf discs were collected and boiled in Luria-Bertani (LB), and then the released SA was measured with the biosensor. The time-consuming steps of weighing samples, grinding tissues and centrifugation were avoided. The direct boiling protocol detected similar differences in SA levels among pathogen-infected wild-type, npr1 (nonexpressor of pathogenesis-related genes), and sid2 (SA induction-deficient) plants as did the previously described biosensor-based method and an HPLC-based approach, demonstrating the efficacy of the protocol presented here. We adapted this protocol to a high-throughput format and identified six npr1 suppressors that accumulated lower levels of SA than npr1 upon pathogen infection. Two of the suppressors were found to be allelic to the previously identified eds5 mutant. The other four are more susceptible than npr1 to the bacterial pathogen Pseudomonas syringae pv. maculicola ES4326 and their identity merits further investigation.ConclusionsThe rapid SA extraction method by direct boiling of leaf discs further reduced the cost and time required for the biosensor Acinetobacter sp. ADPWH_lux-based SA estimation, and allowed the screening for npr1 suppressors that accumulated less SA than npr1 after pathogen infection in a high-throughput manner. The highly efficacious SA estimation protocol can be applied in genetic screen for SA metabolic mutants and characterization of enzymes involved in SA metabolism. The mutants isolated in this study may help identify new components in the SA-related signaling pathways.
Disinfection with sodium hypochlorite or chlorhexidine solutions significantly reduces SA and PA biofilms on PVC TTs. Standard home care of reusable pediatric TTs may be improved by use of these readily available solutions.
Background Rib fractures are present in 10% of all trauma patients and 30% of patients with significant chest trauma. Pain from rib fractures results in decreased respiratory effort which can lead to atelectasis and potentially pneumonia and death. Pain control is therefore of utmost importance in preventing the complications of rib fractures by improving respiratory function. Erector spinae plane blocks (ESPB) have been effectively used in elective surgery with subjective and objective improvements in pain. Materials and Methods We sought to evaluate subjective pain and objective evaluation of respiratory effort by way of incentive spirometry levels after administration of an ESPB for patients with rib fractures. Our trauma service applied ESPB over 2 years in patients with rib fractures. Ultrasound guidance was used to administer 50cc of a long-acting local anesthetic at the transverse process underneath the erector spinae muscle group. Evaluation of pain scores and incentive spirometry levels were measured prior to and after the ESPB. Results In total, we obtained data from 45 patients. Mean pre-pain scores were 7.93 with post-pain scores of 4.47 (p < 0.001). Mean pre-block incentive spirometry volumes were 1160 cc with post-block IS of 1495cc (p 0.035). There were no associated complications. Discussion ESPBs are safe and significantly reduce pain scores and increased incentive spirometry volumes after administration. They are easy to perform and can be done by the trauma service, including trainees. ESPB has the potential to reduce pulmonary complications of rib fractures, as well as subjectively improving pain experienced by our trauma patients. Based on our results, we recommend this block as an adjunct to multimodal analgesia for patients with rib fractures.
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