Optimization of Propidium Monoazide qPCR (Viability-qPCR) to Quantify the Killing by the Gardnerella-Specific Endolysin PM-477, Directly in Vaginal Samples from Women with Bacterial Vaginosis

Quantification of the number of living cells in biofilm or after eradication treatments of biofilm, is problematic for different reasons. We assessed the performance of pre-treatment of DNA, planktonic cells and ex vivo vaginal biofilms of Gardnerella with propidium monoazide (PMAxx) to prevent qPCR-based amplification of DNA from killed cells (viability-qPCR). Standard PMAxx treatment did not completely inactivate free DNA and did not affect living cells. While culture indicated that killing of planktonic cells by heat or by endolysin was complete, viability-qPCR assessed only log reductions of 1.73 and 0.32, respectively. Therefore, we improved the standard protocol by comparing different (combinations of) parameters, such as concentration of PMAxx, and repetition, duration and incubation conditions of treatment. The optimized PMAxx treatment condition for further experiments consisted of three cycles, each of: 15 min incubation on ice with 50 µM PMAxx, followed by 15 min-long light exposure. This protocol was validated for use in vaginal samples from women with bacterial vaginosis. Up to log2.2 reduction of Gardnerella cells after treatment with PM-477 was documented, despite the complex composition of the samples, which might have hampered the activity of PM-477 as well as the quantification of low loads by viability-qPCR.

LAUGHTER AND EMPATHY

At a biological level, laughter was found to help in dealing with pain and suffering. From a psychological perspective, its effects have been studied both at intrapersonal and interpersonal levels. At an intrapersonal level, laughter contributes to self-regulating emotions (especially lowering the trait anxiety), diminishing the expression of anger, internally or externally, the same time with increasing self-acceptance. At interpersonal level, laughing together builds trust, while being laughed at may be traumatic. In the present study, the objective was to investigate the effects of laughter on empathy, awareness, and acceptance of personal experience. The participants in the study, all women, watched a situational comedy for 1.5 hours, with different life situations which may seem negative, but presented in an amusing manner, and, very important, with a happy end. At the end, they completed the empathy and awareness questionnaires. Statistical analysis, comparing their scores with those in a non-treatment condition, showed that laughing at the situational comedy significantly influenced almost all aspects of empathy, significantly decreasing the personal distress from empathizing with others, but also sensitivity and emotional interest towards them, without significant influence on awareness and acceptance of personal experience.

Effect of treatment condition on perovskite film for perovskite solar cell application

In this study, the Perovskite material CH3NH3PbI3 was prepared using two-step sequential solution deposition technique. The treatment condition for Perovskite film including dipping duration, reaction temperature and annealing temperature was studied. Crystal structure, grain size, and purity of the prepared material were examined using XRD and SEM methods. The results indicate that controlling treatment condition has a significant effect on the crystallinity and purity of Perovskite film. Under suitable condition, the obtained Perovskite material has a tetragonal structure and grain size ranges from 200 to 400 nm. The Perovskite film was then applied as a light-harvesting material in Perovskite solar cell. The device exhibits a power conversion efficiency of 5.18% with JSC of 13.6 mA cm-2, VOC of 0.83 V, and fill factor of 45.9%.

Low-Stress Livestock Handling Protects Cattle in a Five-Predator Habitat

Given the ecological importance of top predators, societies are turning to non-lethal methods for coexistence. Coexistence is challenging when livestock are released within wild predator habitats, even when people supervise or use lethal methods. We report a randomized, controlled design to evaluate low-stress livestock handling (L-SLH), a form of range riding, to deter grizzly (brown) bears, gray wolves, cougars, black bears, and coyotes in Southwestern Alberta. The treatment condition was supervision by two newly hired and trained range riders and an L-SLH practicing range rider. This treatment was compared against a baseline pseudo-control condition of the single experienced range rider working alone. Cattle experienced zero injuries or deaths in either condition. We infer that inexperienced range riders trained and supervised by an experienced rider did not raise or lower the risk to cattle. Also, predators did not shift to the cattle herds protected by fewer range riders. Pending experimental evaluation of other designs, we recommend use of L-SLH.

Efficacy of acupuncture for melasma: a protocol of systematic review and meta-analysis

Review question / Objective: How effective is acupuncture in treating melasma compared to conventional treatment？ Condition being studied: Melasma is a common acquired hyperpigmentation of the skin. The clinical manifestations are light brown or dark brown patches symmetrically distributed on the cheeks, forehead and mandibular with different shades and unclear borders. Melasma particularly affects women during menstruation, especially in thirties and forties Asian women. The incidence of Asian women of childbearing age is as high as 30%. Current treatments for melasma include topical drugs, chemical peeling agents, laser and light treatments, and systemic drugs. Despite the strong demand for treatment, the treatment of melasma is still very challenging, the results are inconsistent, and the recurrence rate is almost constant.

Phase-based treatment versus immediate trauma-focused treatment for post-traumatic stress disorder due to childhood abuse: randomised clinical trial

Background It is unclear whether people with post-traumatic stress disorder (PTSD) and symptoms of complex PTSD due to childhood abuse need a treatment approach different from approaches in the PTSD treatment guidelines. Aims To determine whether a phase-based approach is more effective than an immediate trauma-focused approach in people with childhood-trauma related PTSD (Netherlands Trial Registry no.: NTR5991). Method Adults with PTSD following childhood abuse were randomly assigned to either a phase-based treatment condition (8 sessions of Skills Training in Affect and Interpersonal Regulation (STAIR), followed by 16 sessions of eye-movement desensitisation and reprocessing (EMDR) therapy; n = 57) or an immediately trauma-focused treatment condition (16 sessions of EMDR therapy; n = 64). Participants were assessed for symptoms of PTSD and complex PTSD, and other forms of psychopathology before, during and after treatment and at 3- and 6-month follow-ups. Results Data were analysed with linear mixed models. No significant differences between the two treatments on any variable at post-treatment or follow-up were found. Post-treatment, 68.8% no longer met PTSD diagnostic criteria. Self-reported PTSD symptoms significantly decreased for both STAIR–EMDR therapy (d = 0.93) and EMDR therapy (d = 1.54) from pre- to post-treatment assessment, without significant difference between the two conditions. No differences in drop-out rates between the conditions were found (STAIR–EMDR 22.8% v. EMDR 17.2%). No study-related adverse events occurred. Conclusions This study provides compelling support for the use of EMDR therapy alone for the treatment of PTSD due to childhood abuse as opposed to needing any preparatory intervention.

Estimands in published protocols of randomised trials: urgent improvement needed

Background An estimand is a precise description of the treatment effect to be estimated from a trial (the question) and is distinct from the methods of statistical analysis (how the question is to be answered). The potential use of estimands to improve trial research and reporting has been underpinned by the recent publication of the ICH E9(R1) Addendum on the use of estimands in clinical trials in 2019. We set out to assess how well estimands are described in published trial protocols. Methods We reviewed 50 trial protocols published in October 2020 in Trials and BMJ Open. For each protocol, we determined whether the estimand for the primary outcome was explicitly stated, not stated but inferable (i.e. could be constructed from the information given), or not inferable. Results None of the 50 trials explicitly described the estimand for the primary outcome, and in 74% of trials, it was impossible to infer the estimand from the information included in the protocol. The population attribute of the estimand could not be inferred in 36% of trials, the treatment condition attribute in 20%, the population-level summary measure in 34%, and the handling of intercurrent events in 60% (the strategy for handling non-adherence was not inferable in 32% of protocols, and the strategy for handling mortality was not inferable in 80% of the protocols for which it was applicable). Conversely, the outcome attribute was stated for all trials. In 28% of trials, three or more of the five estimand attributes could not be inferred. Conclusions The description of estimands in published trial protocols is poor, and in most trials, it is impossible to understand exactly what treatment effect is being estimated. Given the utility of estimands to improve clinical research and reporting, this urgently needs to change.

Effects of Oxytocin on Social Comparisons in Intergroup Situations

Oxytocin (OXT) is known to affect various social processes, including social comparisons and intergroup competition. In this study, we examined whether social comparisons in intergroup situations can be modulated by OXT and, if so, how this modulation manifests. Using a double-blind placebo-controlled design, we randomly assigned male participants to either OXT or placebo treatment and then asked them to play a card game with either an in-group or an out-group member. The OXT-treated participants showed a greater social comparison effect in the games with an out-group member than in games with an in-group member. Specifically, the participants in the OXT treatment condition showed a greater acceptance rate for relative gain (downward comparison) and a lower acceptance rate for relative loss (upward comparison) while playing with an out-group member rather than an in-group member. In contrast, no such effect was observed among placebo-treated participants. These findings demonstrate that OXT facilitates intergroup social comparisons with out-group versus in-group members.

Comparative investigations of cryo-treated and untreated inserts on machinability of AISI 1050 by using response surface methodology, ANOVA and Taguchi design

This study aims to focus on the machinability of the AISI 1050 workpieces with cutting inserts, treated under deep cryogenic heat (−146 °C), and with untreated ones, and to investigate the optimization of cutting parameters and heat treatment conditions for surface roughness and cutting force by using Taguchi mixed design and Response Surface Methodology (RSM). The machining experiment was performed on a CNC lathe with machining parameters such as three feed rates, three cutting speeds and a constant depth of cut under dry condition and with heat treatment condition. As is known, Taguchi design L18 (32 21) consists of three factors; cutting parameters with each one of three levels and heat treatment condition with two levels. The results of machining tests were evaluated considering surface roughness, vibrations and cutting force. Furthermore, chip morphology and wear led by cryo-treated and untreated inserts were detected with the aid of a scanning electron microscope. The results demonstrated that cryo-treated (CTI) insert had lower tool wear, vibration, and cutting force than untreated insert (UI) in all conditions. In aspect of chip morphology, untreated inserts had bigger and larger serrations than the treated inserts. In addition, according to Taguchi S/N ratio, optimal cutting parameters and heat treatment conditions were obtained from CHT1, V3, and f1 for the Fc and from CHT1, V1, and f1 for the Ra, respectively. Also, the most significant control factors on surface roughness and cutting force were feed rate depending on ANOVA results and RSM. Validation test results demonstrated that RSM and Taguchi mixed design calculated the cutting force (R2RSM (CTI and UI) = 99.99% and R2Tag. = 99.95%) and surface roughness (R2RSM (CTI) = 99.76%, R2RSM (UI) = 99.59% and R2Tag. = 99.12%). Therefore, RSM and Taguchi mixed design predicts highly well match experimental data with prediction data.