Tools for genetic manipulation of the plant growth-promoting bacterium Azospirillum amazonense

<p>Abstract</p> <p>Background</p> <p><it>Azospirillum amazonense </it>has potential to be used as agricultural inoculant since it promotes plant growth without causing pollution, unlike industrial fertilizers. Owing to this fact, the study of this species has gained interest. However, a detailed understanding of its genetics and physiology is limited by the absence of appropriate genetic tools for the study of this species.</p> <p>Results</p> <p>Conjugation and electrotransformation methods were established utilizing vectors with broad host-replication origins (pVS1 and pBBR1). Two genes of interest - <it>glnK </it>and <it>glnB</it>, encoding PII regulatory proteins - were isolated. Furthermore, <it>glnK</it>-specific <it>A. amazonense </it>mutants were generated utilizing the pK19MOBSACB vector system. Finally, a promoter analysis protocol based on fluorescent protein expression was optimized to aid genetic regulation studies on this bacterium.</p> <p>Conclusion</p> <p>In this work, genetic tools that can support the study of <it>A. amazonense </it>were described. These methods could provide a better understanding of the genetic mechanisms of this species that underlie its plant growth promotion.</p

NRF2 activation in Trp53;p16-deficient mice drives oral squamous cell carcinoma

UNLABELLED: Aberrant activation of the NRF2/NFE2L2 transcription factor commonly occurs in head and neck squamous cell carcinomas (HNSCC). Mouse model studies have shown that NRF2 activation alone does not result in cancer. When combined with classic oncogenes and at the right dose, NRF2 activation promotes tumor initiation and progression. Here we deleted the tumor suppressor genes p16INK4A and p53 (referred to as CP mice), which are commonly lost in human HNSCC, in the presence of a constitutively active NRF2E79Q mutant (CPN mice). NRF2E79Q expression in CPN mice resulted in squamous cell hyperplasia or dysplasia with hyperkeratosis in the esophagus, oropharynx, and forestomach. In addition, CPN mice displayed oral cavity squamous cell carcinoma (OSCC); CP mice bearing wild-type NRF2 expression did not develop oral cavity hyperplasia, dysplasia or OSCC. In both CP and CPN mice, we also observed predominantly abdominal sarcomas and carcinomas. Our data show that in the context of p53 and p16 tumor suppressor loss, NRF2 activation serves oncogenic functions to drive OSCC. CPN mice represent a new model for OSCC that closely reflects the genetics of human HNSCC. SIGNIFICANCE: Human squamous cancers frequently show constitutive NRF2 activation, associated with poorer outcomes and resistance to multiple therapies. Here, we report the first activated NRF2-driven and human-relevant mouse model of squamous cell carcinoma that develops in the background of p16 and p53 loss. The availability of this model will lead to a clearer understanding of how NRF2 contributes to the initiation, progression, and therapeutic response of OSCC

What mental health services should be available after the postnatal period?

IntroductionMothers with severe mental illness may require mental health support through postnatal services. However, little is known about what services are actually provided to support parents after the postnatal period in Europe.AimsTo explore existing services for parents with severe mental illness after the postnatal period across Europe.MethodsMental health specialists from major cities in nine European countries were asked to identify all health and social services available for mothers with psychosis after the postnatal period. They received two case vignettes and completed a data collection sheet for every identified service. Data analysis used semi-quantitative methods to describe the identified services.ResultsA wide range of different services was identified with no systematic coverage of specific target groups or target problems. Likewise, their scope was extremely diverse, ranging from simple telephone advice to multi-professional support for multiple complex problems. Most services targeted parents or families in general but would at least in principle be available for parents with severe mental illness. A much smaller number specialized on targeted help for parents with mental illness.ConclusionsPatchy and heterogeneous service provision may make it difficult to navigate support systems for both patients and professionals. Systematic research is required, e.g. on the use, the costs, and patient experiences in different types of services, so that service provision can be based on some evidence. Given the differences in service provision across European countries, such research might use international comparisons for evaluating the benefits of different types of services for parents with severe illnesses.Disclosure of interestThe authors have not supplied their declaration of competing interest

Characterization of the KATRIN cryogenic pumping section

The KArlsruhe TRItium Neutrino (KATRIN) experiment aims to determine the effective anti-electron neutrino mass with a sensitivity of 0.2 eV/c$^2$ by using the kinematics of tritium $\beta$-decay. It is crucial to have a high signal rate which is achieved by a windowless gaseous tritium source producing 10$^{11}$ $\beta$-electrons per second. These are guided adiabatically to the spectrometer section where their energy is analyzed. In order to maintain a low background rate below 0.01 cps, one essential criteria is to permanently reduce the flow of neutral tritium molecules between the source and the spectrometer section by at least 14 orders of magnitude. A differential pumping section downstream from the source reduces the tritium flow by seven orders of magnitude, while at least another factor of 10$^7$ is achieved by the cryogenic pumping section where tritium molecules are adsorbed on an approximately 3 K cold argon frost layer. In this paper, the results of the cryogenic pumping section commissioning measurements using deuterium are discussed. The cryogenic pumping section surpasses the requirement for the flow reduction of 10$^7$ by more than one order of magnitude. These results verify the predictions of previously published simulations

WELLFOCUS PPT – modified positive psychotherapy to improve well-being in psychosis: study protocol for a pilot randomised controlled trial

BACKGROUND: The promotion of well-being is an important goal of recovery oriented mental health services. No structured, evidence-based intervention exists that aims to increase the well-being in people with severe mental illness such as psychosis. Positive psychotherapy (PPT) is a promising intervention for this goal. Standard PPT was adapted for use with people with psychosis in the UK following the Medical Research Council framework for developing and testing complex interventions, resulting in the WELLFOCUS Model describing the intended impact of WELLFOCUS PPT. This study aims to test the WELLFOCUS Model, by piloting the intervention, trial processes, and evaluation strategy. METHODS/DESIGN: This study is a non-blinded pragmatic pilot RCT comparing WELLFOCUS PPT provided as an 11-session group therapy in addition to treatment as usual to treatment as usual alone. Inclusion criteria are adults (aged 18–65 years) with a main diagnosis of psychosis who use mental health services. A target sample of 80 service users with psychosis are recruited from mental health services across the South London and Maudsley NHS Foundation Trust. Participants are randomised in blocks to the intervention and control group. WELLFOCUS PPT is provided to groups by specifically trained and supervised local therapists and members of the research team. Assessments are conducted before randomisation and after the group intervention. The primary outcome measure is well-being assessed by the Warwick-Edinburgh Mental Well-being Scale. Secondary outcomes include good feelings, symptom relief, connectedness, hope, self-worth, empowerment, and meaning. Process evaluation using data collected during the group intervention, post-intervention individual interviews and focus groups with participants, and interviews with trial therapists will complement quantitative outcome data. DISCUSSION: This study will provide data on the feasibility of the intervention and identify necessary adaptations. It will allow optimisation of trial processes and inform the evaluation strategy, including sample size calculation, for a future definitive RCT. TRIAL REGISTRATION: Current Controlled Trials ISRCTN04199273 – WELLFOCUS study: an intervention to improve well-being in people with psychosis, Date registered: 27 March 2013, first participant randomised on 26 April 2013

Institution-Wide Retreats Foster Organizational Learning and Action at a Comprehensive Cancer Center

Providing safe and informed healthcare for sexual and gender minority (SGM) individuals with cancer is stymied by the lack of sexual orientation and gender identity (SOGI) data reliably available in health records and by insufficient training for staff. Approaches that support institutional learning, especially around sensitive topics, are essential for hospitals seeking to improve practices impacting patient safety and research. We engineered annual institutional retreats to identify and unify stakeholders, promote awareness of gaps and needs, identify initiatives, minimize redundant projects, and coordinate efforts that promote improvements in SGM cancer care, education, and research. The 2022 and 2023 retreats employed a 4-h hybrid format allowing virtual and in-person engagement. Retreat organizers facilitated small-group discussions for brainstorming among participants. We performed descriptive statistics from retreat evaluations. The retreats engaged 104 attendees from distinct departments and roles. Participants expressed robust satisfaction, commending the retreat organization and content quality. Notably, the first retreat yielded leadership endorsement and funding for a Quality Improvement pilot to standardize SOGI data collection and clinical staff training. The second retreat provided a platform for updates on focused efforts across the institution and for receiving direction regarding national best practices for SGM care and research. We report the processes and outcomes of institution-wide retreats, which served as a platform for identifying gaps in organizational healthcare practices and research for SGM individuals with cancer. The strategies described herein may be readily scaled at other cancer hospitals seeking to learn and enact system-wide practice changes that support the needs of SGM patients and families