Expression of T4 Lysozyme Gene (gene e) in Streptococcus salivarius subsp. Thermophilus

In present study, we aimed to express T4 Lysozyme gene (gene e) in Streptococcus salivarus subsp. thermophilus to create better probiotics for poultry. The Esherichia coli plasmid, Bluescript II SK +/-harboring gene e named pL1, was converted to a new E. coli-Streptococcus sp. shuttle vector (pL2) by cloning and inserting Streptococcal replication origin of pTRW10 vector into pL1. pL2 plasmid isolated from E. coli was introduced into S. salivarius subsp. thermophilus and Lactococcus lactis cells by electro-transformation. The lysozyme enzymes expressing by these bacteria were found to be active on Micrococcus luteus cells and thereby preventing their growth on assay plates. Thermostability of these enzymes from the recombinant bacteria was also found different from each other. The lysozyme expressed by S. salivarius subsp. thermophilus cells seemed to increase its capacity for thermoresistance and was not denaturated at 70°C for 15 min. In contrast, the enzyme expressed by L.lactis and E. coli cells were easily denaturated when exposed to the same temperature treatment

Median raphe region stimulation alone generates remote, but not recent fear memory traces

The median raphe region (MRR) is believed to control the fear circuitry indirectly, by influencing the encoding and retrieval of fear memories by amygdala, hippocampus and prefrontal cortex. Here we show that in addition to this established role, MRR stimulation may alone elicit the emergence of remote but not recent fear memories. We substituted electric shocks with optic stimulation of MRR in C57BL/6N male mice in an optogenetic conditioning paradigm and found that stimulations produced agitation, but not fear, during the conditioning trial. Contextual fear, reflected by freezing was not present the next day, but appeared after a 7 days incubation. The optogenetic silencing of MRR during electric shocks ameliorated conditioned fear also seven, but not one day after conditioning. The optogenetic stimulation patterns (50Hz theta burst and 20Hz) used in our tests elicited serotonin release in vitro and lead to activation primarily in the periaqueductal gray examined by c-Fos immunohistochemistry. Earlier studies demonstrated that fear can be induced acutely by stimulation of several subcortical centers, which, however, do not generate persistent fear memories. Here we show that the MRR also elicits fear, but this develops slowly over time, likely by plastic changes induced by the area and its connections. These findings assign a specific role to the MRR in fear learning. Particularly, we suggest that this area is responsible for the durable sensitization of fear circuits towards aversive contexts, and by this, it contributes to the persistence of fear memories. This suggests the existence a bottom-up control of fear circuits by the MRR, which complements the top-down control exerted by the medial prefrontal cortex

A qualitative study of cardiovascular disease risk communication in NHS Health Check using different risk calculators: protocol for the RIsk COmmunication in NHS Health Check (RICO) study. BMC family practice, 20(1), 11.

Background NHS Health Check is a national cardiovascular disease (CVD) risk assessment programme for 40–74 year olds in England, in which practitioners should assess and communicate CVD risk, supported by appropriate risk-management advice and goal-setting. This requires effective communication, to equip patients with knowledge and intention to act. Currently, the QRISK®2 10-year CVD risk score is most common way in which CVD risk is estimated. Newer tools, such as JBS3, allow manipulation of risk factors and can demonstrate the impact of positive actions. However, the use, and relative value, of these tools within CVD risk communication is unknown. We will explore practitioner and patient CVD risk perceptions when using QRISK®2 or JBS3, the associated advice or treatment offered by the practitioner, and patients’ responses. Methods RIsk COmmunication in NHS Health Check (RICO) is a qualitative study with quantitative process evaluation. Twelve general practices in the West Midlands of England will be randomised to one of two groups: usual practice, in which practitioners use QRISK®2 to assess and communicate CVD risk; intervention, in which practitioners use JBS3. Twenty Health Checks per practice will be video-recorded (n = 240, 120 per group), with patients stratified by age, gender and ethnicity. Post-Health Check, video-stimulated recall (VSR) interviews will be conducted with 48 patients (n = 24 per group) and all practitioners (n = 12–18), using video excerpts to enhance participant recall/reflection. Patient medical record reviews will detect health-protective actions in the first 12-weeks following a Health Check (e.g., lifestyle referrals, statin prescription). Risk communication, patient response and intentions for health-protective behaviours in each group will be explored through thematic analysis of video-recorded Health Checks (using Protection Motivation Theory as a framework) and VSR interviews. Process evaluation will include between-group comparisons of quantitatively coded Health Check content and post-Health Check patient outcomes. Finally, 10 patients with the most positive intentions or behaviours will be selected for case study analysis (using all data sources). Discussion This study will produce novel insights about the utility of QRISK®2 and JBS3 to promote patient and practitioner understanding and perception of CVD risk and associated implications for patient intentions with respect to health-protective behaviours (and underlying mechanisms). Recommendations for practice will be developed

Electrochemical copolymerization of N-methylpyrrole and 2,2′-bithitiophene; characterization, micro-capacitor study, and equivalent circuit model evaluation

N-methylpyrrole (N-MPy) and 2,2'-bithiophene (BTh) were electrocopolymerized in 0 center dot 2 M acetonitrile-sodium perchlorate solvent-electrolyte couple on a glassy carbon electrode (GCE) by cyclic voltammetry (CV). The resulting homopolymers and copolymers in different initial feed ratios of [N-MPy](0)/[BTh](0) = 1/1, 1/2, 1/5 and 1/10 were characterized by CV, Fourier-transform infrared reflectance attenuated transmittance (FTIR-ATR), scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDX) and electrochemical impedance spectroscopy (EIS). The capacitive behaviours of the modified electrodes were defined via Nyquist, Bode-magnitude, Bode-phase and admittance plots. The equivalent circuit model of R(C(R)(QR)(CR)) was performed to fit theoretical and experimental data. The highest low-frequency capacitance (C (LF)) were obtained as C (LF) = similar to 1 center dot 23 x 10 (-4) mF cm (-2) for P(N-MPy), C (LF) = similar to 2 center dot 09 x 10 (-4) mF cm(-2) for P(BTh) and C (LF) = similar to 5 center dot 54 x 10 (-4) mF cm(-2) for copolymer in the inital feed ratio of [N-MPy](0)/[BTh] (0) = 1/2.Research Foundation of Namik Kemal University, TurkeyNamik Kemal University [NKUBAP.00.10.YL.12.02]Financial support for this work by the Research Foundation of Namik Kemal University, Turkey, (project number: NKUBAP.00.10.YL.12.02), is gratefully acknowledged