Proteomic Approach for Extracting Cytoplasmic Proteins from Streptococcus sanguinis using Mass Spectrometry

Streptococcus sanguinis is a commensal and early colonizer of oral cavity as well as an opportunistic pathogen of infectious endocarditis. Extracting the soluble proteome of this bacterium provides deep insights about the physiological dynamic changes under different growth and stress conditions, thus defining “proteomic signatures” as targets for therapeutic intervention. In this protocol, we describe an experimentally verified approach to extract maximal cytoplasmic proteins from Streptococcus sanguinis SK36 strain. A combination of procedures was adopted that broke the thick cell wall barrier and minimized denaturation of the intracellular proteome, using optimized buffers and a sonication step. Extracted proteome was quantitated using Pierce BCA Protein Quantitation assay and protein bands were macroscopically assessed by Coomassie Blue staining. Finally, a high resolution detection of the extracted proteins was conducted through Synapt G2Si mass spectrometer, followed by label-free relative quantification via Progenesis QI. In conclusion, this pipeline for proteomic extraction and analysis of soluble proteins provides a fundamental tool in deciphering the biological complexity of Streptococcus sanguinis

Who am I and what do I do? A Hermeneutic Phenomenological Study on the Experience of RNs and RPNs Working in the Same Environment.

The aim of this study was to explore the lived experiences of registered nurses (RNs) and registered practical nurses (RPNs) working in intraprofessional dual-scope work environments. The study was conducted using a hermeneutic phenomenological approach. Conversational interviewing was conducted with a purposeful sample of nine nurses who worked in an intraprofessional care area. Two themes emerged from the participant conversations: The Dance and The Fissure. The Dance displays the experiences of empathy, comradery and teamwork lived by the nurses working in the dual-scope environment. The Fissure displays the lived experiences of role ambiguity and challenges nursing in a dual-scope environment. There is importance in giving voice to the stories of the nurses working in the shared care areas, allowing insight into emotions, integrity and wisdom. This research has given light to various issues in the shared care areas for both RNs and RPNs and has provided implications for nursing practice, nursing education, and nursing research

The Impact of Quiet Time on the Sleep Quality of Cardiothoracic Surgery Patients

The purpose of this study was to observe patients\u27 quality of sleep on a cardiothoracic step-down unit before and after institution of an undisturbed quiet time policy from eleven o\u27clock in the evening until five in the morning. It was hypothesized that these patients have improved sleep quality after institution of the Quiet Time protocol. The comparative group, quasi-experimental study utilized the Verran and Snyder-Halpern Sleep Scale and Factors Influencing Sleep Questionnaire to measure sleep disturbance, effectiveness, and supplementation. Results demonstrated a decrease in sleep disturbance with initiation of the Quiet Time, but no significant change in sleep effectiveness and supplementation. Factors most disturbing to sleep included pain and discomfort, bed and ventilation system, procedures performed upon the patient, interruptions, and talking

The effect of DNA structure and restriction enzymes on transformation efficiencies in Neurospora crassa

Addition of the appropriate restriction enzyme to linearized transforming DNA has been shown to increase transformation efficiencies in organisms as diverse as Saccharomyces cerevisiae (Schiestl and Petes 1991 Proc. Nat. Acad. Sci. USA 88:7585-7589) and Dictyostelium discoideum (Kuspa and Loomis 1992 Proc. Nat. Acad. Sci. USA 89:8803- 8807). This process has been described as REMI, for restriction enzyme-mediated integration. We have examined the effect of restriction enzyme addition on transformation efficiencies in Neurospora crassa. The frequency of cotransformation of a qa-2 inl double mutant with two plasmids [one containing the selectable marker qa-2+ (quinate utilization) and the other containing inl+ (inositol)] was also examined, as was the generation of stable versus abortive transformants

Retinal adaptation to spatial correlations

The classical center-surround retinal ganglion cell receptive field is thought to remove the strong spatial correlations in natural scenes, enabling efficient use of limited bandwidth. While early studies with drifting gratings reported robust surrounds (Enroth-Cugell and Robson, 1966), recent measurements with white noise reveal weak surrounds (Chichilnisky and Kalmar, 2002). This might be evidence for dynamical weakening of the retinal surround in response to decreased spatial correlations, which would be predicted by efficient coding theory. Such adaptation is reported in LGN (Lesica et al., 2007), but whether the retina also adapts to correlations is unknown. 

We tested for adaptation by recording simultaneously from ~40 ganglion cells on a multi-electrode array while presenting white and exponentially correlated checkerboards and strips. Measuring from ~200 cells responding to 90 minutes each of white and correlated stimuli, we were able to extract precise spatiotemporal receptive fields (STRFs). We found that a difference-of-Gaussians was not a good fit and the surround was generally displaced from the center. Thus, to assess surround strength we found the center and surround regions and the total weight on the pixels in each region. The relative surround strength was then defined as the ratio of surround weight to center weight. Surprisingly, we found that the majority of recorded cells have a stronger surround under white noise than under correlated noise (p<.05), contrary to naive expectation from theory. The conclusion was robust to different methods of extracting STRFs and persisted with checkerboard and strip stimuli.

To test, without assuming a model, whether the retina decorrelates stimuli, we also measured the pairwise correlations between spike trains of simultaneously recorded neurons under three conditions: white checkerboard, exponentially correlated noise, and scale-free noise. The typical amount of pairwise correlation increased with extent of input correlation, in line with our STRF measurements

Training the Next Generation of Practitioners In Early Intervention and Telepractice: Three University Models

The COVID-19 pandemic continues to shape the provision of family-centered early intervention services for children who are Deaf or Hard-of-Hearing and their families. In programs, schools, and centers, direct in-person contact with families have been significantly curtailed as a means to limit the exposure to and spead of the virus. Emergency remote learning has lead to an increase in telepractice, also referred to as teleintervention, as the designated model of service provision. Most early interventionists, speech-language pathologists, and teachers of the Deaf were not sufficiently trained to suddenly implement emergency remote teaching or telepractice services, but service providers had no option but to forge ahead, often with limited or no prior knowledge and experience with the provision of services using only telecommunications technology. Fortunately, however, some university training programs have integrated telepractice into their curricula and practica experiences for many years, and three of those programs are profiled here