A secreted phosphodiesterase/aIkaline phosphatase, APaseD, was purified from a culture of Bacillus subfilis JH646MS. Its phosphodiesterase activity was reminiscent of an APase isolated and characterized previously. lmmunoassay and N-terminal sequencing showed the two proteins to be identical. Using the first 20 amino acids of the mature protein, a BLAST search of GenBank was used to find an homologous sequence. An exact match was found but in a putative non-coding region. It was hypothesized that there was a base pair deletion in the phoD gene. A DNA fragment internal to the coding region was generated by PCR using template DNA from a strain which produced APaseD. The PCR fragment was cloned and used to interrupt the gene. Western blot analysis of the parent and the mutated strains showed that APaseD was missing in the mutant. Resequencing of the gene revealed a larger ORF encoding a protein similar in size to the 49 kDa APaseD estimated by SDS-PAGE. The promoter was then cloned, sequenced and used in phoD-lacZ promoter fusions which showed that the gene was phosphate-starvation-induced and dependent on PhoP and PhoR for expression.

Objective:The underuse of analgesics, or "oligoanalgesia," is common in emergency departments (EDs). To improve care we must understand our patients' pain experiences as well as our clinical practice patterns. To this end, we examined pain etiology, pain management practices and patient satisfaction in 2 urban EDs. Methods: We conducted a cross-sectional study using structured interviews and chart reviews for patients with pain who presented to either of 2 university-affiliated EDs. We assessed pain etiologies, patient pain experiences, pain management practices, and patient satisfaction with pain management. Results:The 525 study subjects reported high pain intensity levels on presentation, with a median rating of 8 on a 10-point numerical rating scale (NRS). At discharge, pain severity had decreased to a median rating of 4; however, 48% of patients were discharged from the ED in moderate to severe pain (NRS 5-10). Subjects reported spending 57% of their ED stay in moderate to severe pain. Analgesics were administered to only 50% of patients. The mean time to analgesic administration was almost 2 hours. Despite high levels of reported pain at discharge and low rates of analgesic administration, subjects reported high satisfaction with pain management. Conclusions: In the 2 EDs studied, we found high levels of pain severity for our patients, as well as low levels of analgesic use. When used, analgesic administration was often delayed. Despite these findings, patient satisfaction remained high. Despite recent efforts to improve pain management practice; oligoanalgesia remains a problem for our specialty. RÉSUMÉObjectif : La sous-utilisation des analgésiques ou «oligoanalgésie» est courante au département d'urgence (DU). Pour améliorer les soins, nous devons comprendre le niveau de douleur ressentie par nos patients ainsi que nos habitudes de pratique clinique. À cette fin, nous avons examiné l'é-tiologie de la douleur, les pratiques de prise en charge de la douleur et le degré de satisfaction des patients dans deux DU en milieu urbain. Méthodes : Nous avons mené une enquête transversale à l'aide d'entrevues structurées et de revue de dossiers pour des patients en douleur reçus à l'un ou l'autre de deux DU affiliés à une université. Nous avons évalué l'étiologie de la douleur, le niveau de douleur ressentie par les patients, les pratiques de prise en charge de la douleur et la satisfaction des patients face à cette prise en charge.

The tagAB and tagDEF operons, which are adjacent and divergently transcribed, encode genes responsible for cell wall teichoic acid synthesis in Bacillus subtilis. TheBacillus data presented here suggest that PhoP and PhoR are required for direct repression of transcription of the two operons under phosphate starvation conditions but have no regulatory role under phosphate-replete conditions. These data identify for the first time that PhoP∼P has a negative role in Pho regulon gene regulation.

No abstract

The PhoP-PhoR two-component regulatory system controls the phosphate deficiency response in B. subtilis. A number of Pho regulon genes which require PhoP∼P for activation or repression have been identified. The studies reported here were initiated to understand the PhoP-DNA interaction necessary for Pho promoter regulation. The regulatory region of phoD was characterized in detail using oligo-directed mutagenesis, DNase I footprinting, and in vivo transcription assays. These data reveal basic principles of PhoP binding relevant to PhoP’s interaction with other Pho regulon promoters. Our results show that: (i) a dimer of PhoP∼P is able to bind two consensus repeats in a stable fashion; (ii) PhoP binding is highly cooperative within the core promoter region, which is located from −66 to −17 on the coding strand and contains four TT(A/T/C)ACA-like repeats; (iii) specific bases comprising the TT(A/T/C)ACA consensus are essential for transcriptional activation, but the specific base pairs of the intervening sequences separating the consensus repeats are not important for either PhoP binding or promoter activation; (iv) the spacing between two consensus repeats within a putative dimer binding site in the core region is important for both PhoP binding and promoter activation; (v) the exact spacing between two dimer binding sites within the core region is important for promoter activation but less so for PhoP binding affinity, as long as the repeats are on the same face of the helix; and (vi) the 5′ secondary binding region is important for coordinated PhoP binding to the core binding region, making it nearly essential for promoter activation.

No abstract

No abstract

No abstract