Listening – a Possibility for Improved Healthcare

The conversation between a healthcare provider and a patient is one component of obtaining information for diagnosis and treatment. Responsibility rests with healthcare providers to actively lead this conversation so this exchange of information is accurate and complete with nothing added to and nothing left out. The question then becomes, “Do the words of the patient land word for word with their practitioner?”. In the world of “being” or ontology the answer is no. This article will focus on an ontological perceptual constraint known as Already-Always-Listening and introduce the practice of Authentic Listening as access to a transformation in healthcare provider and patient communication

Global Regulatory Functions of the Staphylococcus aureus Endoribonuclease III in Gene Expression

RNA turnover plays an important role in both virulence and adaptation to stress in the Gram-positive human pathogen Staphylococcus aureus. However, the molecular players and mechanisms involved in these processes are poorly understood. Here, we explored the functions of S. aureus endoribonuclease III (RNase III), a member of the ubiquitous family of double-strand-specific endoribonucleases. To define genomic transcripts that are bound and processed by RNase III, we performed deep sequencing on cDNA libraries generated from RNAs that were co-immunoprecipitated with wild-type RNase III or two different cleavage-defective mutant variants in vivo. Several newly identified RNase III targets were validated by independent experimental methods. We identified various classes of structured RNAs as RNase III substrates and demonstrated that this enzyme is involved in the maturation of rRNAs and tRNAs, regulates the turnover of mRNAs and non-coding RNAs, and autoregulates its synthesis by cleaving within the coding region of its own mRNA. Moreover, we identified a positive effect of RNase III on protein synthesis based on novel mechanisms. RNase III–mediated cleavage in the 5′ untranslated region (5′UTR) enhanced the stability and translation of cspA mRNA, which encodes the major cold-shock protein. Furthermore, RNase III cleaved overlapping 5′UTRs of divergently transcribed genes to generate leaderless mRNAs, which constitutes a novel way to co-regulate neighboring genes. In agreement with recent findings, low abundance antisense RNAs covering 44% of the annotated genes were captured by co-immunoprecipitation with RNase III mutant proteins. Thus, in addition to gene regulation, RNase III is associated with RNA quality control of pervasive transcription. Overall, this study illustrates the complexity of post-transcriptional regulation mediated by RNase III

Bacterial Stressors in Minimally Processed Food

Stress responses are of particular importance to microorganisms, because their habitats are subjected to continual changes in temperature, osmotic pressure, and nutrients availability. Stressors (and stress factors), may be of chemical, physical, or biological nature. While stress to microorganisms is frequently caused by the surrounding environment, the growth of microbial cells on its own may also result in induction of some kinds of stress such as starvation and acidity. During production of fresh-cut produce, cumulative mild processing steps are employed, to control the growth of microorganisms. Pathogens on plant surfaces are already stressed and stress may be increased during the multiple mild processing steps, potentially leading to very hardy bacteria geared towards enhanced survival. Cross-protection can occur because the overlapping stress responses enable bacteria exposed to one stress to become resistant to another stress. A number of stresses have been shown to induce cross protection, including heat, cold, acid and osmotic stress. Among other factors, adaptation to heat stress appears to provide bacterial cells with more pronounced cross protection against several other stresses. Understanding how pathogens sense and respond to mild stresses is essential in order to design safe and effective minimal processing regimes

The effect of the form and concentration of nitrogen on geosmin production by an aquatic actinomycete (streptomyces isolate WM1C1).

Actinomycetes are organisms that produce geosmin which is one of the organic compounds responsible for the earthy (musty) odor in water. This study was designed to isolate an actinomycete, confirm it as a geosmin producer, and determine the effects of the form and concentration of nitrogen on the production of geosmin by the isolate. Five actinomycetes were isolated from the muds of Lake Waco; four were identified as Streptorayces and one as a possible Nocardia sp. or Micromonospora sp., and two Streptomyces spp. were confirmed to be geosmin producers by GC-MS. The effects of nitrogen on one Streptomyces sp. were reported as affecting threshold odor number (R.O.N.), biomass production (mg dry wt), and Specific T.O.N. (T.O.N.-mg dry wt ^). The forms of nitrogen used were NO^-N as KNO^, NH^-N as NH^Cl, and organic nitrogen (ORG-N) as asparagine and the concen- tration range of nitrogen was 0.000 mg'L to 0.600 mg * L. T.O.N. values increased linearly as concentrations of NO^-N and NH^-N increased while ORG-N (0.300 mg'L inhibited geosmin production. Biomass production increased linearly as concentrations of NO^-N and ORG-N increased (0.000 mg*L ^ to 0.600 mg'L"'*") but low concentrations of all three nitrogen forms did not promote significantly larger biomass proudction than the nitrogen-free media. Specific T.O.N. increased as concentrations of NH^-N and NO^-N increased with NH^-N producing significantly higher Specific T.O.N. per flask

Growth-dependent activity of the cold shock cspA promoter + 5′ UTR and production of the protein CspA in Staphylococcus aureus Newman

Abstract Background Research involving the cold shock gene cspA of the medically important bacterium Staphylococcus aureus is steadily increasing as the relationships between the activity of this gene at 37 °C and a spectrum of virulence factors (e.g., biofilm formation, capsule production) as well as stress-related genes (e.g., alkaline shock protein, asp-23 and the alternative sigma factor, sigB) are distinguished. Fundamental to each of these discoveries is defining the regulation of cspA and the production of its protein product CspA. Results In this paper, primer extension analysis was used to identify a transcriptional start point at 112 bp upstream of the initiation codon of the cspA coding sequence from S. aureus Newman RNA collected at 37 °C. Based on the location of the putative −10 and −35 sites as well as putative cold shock protein binding sites, a 192 bp sequence containing an 80 bp promoter + a 112 bp 5′ UTR was generated by polymerase chain reaction. The activity of this 192 bp sequence was confirmed in a pLL38 promoter::xylE reporter gene construct. In addition, Western blots were used to confirm the production of CspA at 37 °C and demonstrated that production of the protein was not constitutive but showed growth-dependent production with a significant increase at the 6 h time point. Conclusions The results presented identify another regulatory region for the cold shock gene cspA of S. aureus and show growth-dependent activity of both this cspA regulatory sequence, presented as a 192 bp sequence of promoter + 5′ UTR and the production of the CspA protein at 37 °C. The presence of two active transcription start points, a −112 bp sequence defined in this work and a second previously defined at −514 bp upstream of the cspA initiation codon, suggests the possibility of interactions between these two regions in the regulation of cspA. The growth-dependent production of the cold shock protein CspA supports the availability of this protein to be a modulator of virulence and stress factor genes at 37 °C

CspA Regulates Pigment Production in Staphylococcus aureus through a SigB-Dependent Mechanism

We report that the cold shock protein CspA of Staphylococcus aureus is required for maximal production of pigment. Results from transcriptional studies revealed that loss of CspA resulted in decreased expression of genes needed for the biosynthesis of 4,4′-diaponeurosporene and the alternative sigma factor SigB