Heterologous expression of a lycophyte protein enhances angiosperm seedling vigor.

Seedling vigor is a key agronomic trait that determines juvenile plant performance. Angiosperm seeds develop inside fruits and are connected to the mother plant through vascular tissues. Their formation requires plant-specific genes, such as BREVIS RADIX (BRX) in Arabidopsis thaliana roots. BRX family proteins are found throughout the euphyllophytes but also occur in non-vascular bryophytes and non-seed lycophytes. They consist of four conserved domains, including the tandem BRX domains. We found that bryophyte or lycophyte BRX homologs can only partially substitute for Arabidopsis BRX (AtBRX) because they miss key features in the linker between the BRX domains. Intriguingly, however, expression of a BRX homolog from the lycophyte Selaginella moellendorffii (SmBRX) in an A. thaliana wild-type background confers robustly enhanced root growth vigor that persists throughout the life cycle. This effect can be traced to a substantial increase in seed and embryo size, is associated with enhanced vascular tissue proliferation, and can be reproduced with a modified, SmBRX-like variant of AtBRX. Our results thus suggest that BRX variants can boost seedling vigor and shed light on the activity of ancient, non-angiosperm BRX family proteins.

Mixed-methods process evaluation of a residence-based SARS-CoV-2 testing participation pilot on a UK university campus during the COVID-19 pandemic.

BACKGROUND: Regular testing for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is an important strategy for controlling virus outbreaks on university campuses during the COVID-19 pandemic but testing participation rates can be low. The Residence-Based Testing Participation Pilot (RB-TPP) was a novel intervention implemented at two student residences on a large UK university campus over 4 weeks. The aim of the pilot was to increase the frequency of asymptomatic SARS-CoV-2 saliva testing onsite. This process evaluation aimed to determine whether RB-TPP was implemented as planned and identify implementation barriers and facilitators. METHODS: A mixed-methods process evaluation was conducted alongside the RB-TPP. Evaluation participants were students (opting in, or out of RB-TPP) and staff with a role in service provision or student support. Monitoring data were collected from the intervention delivery team and meeting records. Data were collected from students via online survey (n = 152) and seven focus groups (n = 30), and from staff via individual interviews (n = 13). Quantitative data were analysed descriptively and qualitative data thematically. Barriers and facilitators to implementation were mapped to the 'Capability, Opportunity, Motivation-Behaviour' (COM-B) behaviour change framework. RESULTS: Four hundred sixty-four students opted to participate in RB-TPP (98% of students living onsite). RB-TPP was implemented broadly as planned but relaxed social distancing was terminated early due to concerns relating to national escalation of the COVID-19 Delta variant, albeit testing continued. Most students (97.9%) perceived the period of relaxed social distancing within residences positively. The majority engaged in asymptomatic testing (88%); 46% (52% of testers) were fully compliant with pre-determined testing frequency. Implementation was facilitated by convenience and efficiency of testing, and reduction in the negative impacts of isolation through opportunities for students to socialise. Main barriers to implementation were perceived mixed-messages about the rules, ambivalent attitudes, and lack of adherence to COVID-19 protective measures in the minority. CONCLUSIONS: This process evaluation identifies factors that help or hinder the success of university residence-based outbreak prevention and management strategies. RB-TPP led to increased rates of SARS-CoV-2 testing participation among students in university residences. Perceived normalisation of university life significantly enhanced student mental wellbeing. The complexity and challenge generated by multiple lines of communication and rapid adaptions to a changing pandemic context was evident. TRIAL REGISTRATION NUMBER: UKAS 307727-02-01; Pre-results. CLINICALTRIALS: gov Identifier: NCT05045989 ; post-results (first posted, 16/09/21). ETHICAL APPROVAL: Faculty of Medicine & Health Sciences Research Ethics Committee, University of Nottingham (Ref: FMHS 96-0920).

Simultaneous imaging and functional studies reveal a tight correlation between calcium and actin networks.

Tip-growing cells elongate in a highly polarized manner via focused secretion of flexible cell-wall material. Calcium has been implicated as a vital factor in regulating the deposition of cell-wall material. However, deciphering the molecular and mechanistic calcium targets in vivo has remained challenging. Here, we investigated intracellular calcium dynamics in the moss Physcomitrella patens, which provides a system with an abundant source of genetically identical tip-growing cells, excellent cytology, and a large molecular genetic tool kit. To visualize calcium we used a genetically encoded cytosolic FRET probe, revealing a fluctuating tipward gradient with a complex oscillatory profile. Wavelet analysis coupled with a signal-sifting algorithm enabled the quantitative comparison of the calcium behavior in cells where growth was inhibited mechanically, pharmacologically, or genetically. We found that cells with suppressed growth have calcium oscillatory profiles with longer frequencies, suggesting that there is a feedback between the calcium gradient and growth. To investigate the mechanistic basis for this feedback we simultaneously imaged cytosolic calcium and actin, which has been shown to be essential for tip growth. We found that high cytosolic calcium promotes disassembly of a tip-focused actin spot, while low calcium promotes assembly. In support of this, abolishing the calcium gradient resulted in dramatic actin accumulation at the tip. Together these data demonstrate that tipward calcium is quantitatively linked to actin accumulation in vivo and that the moss P. patens provides a powerful system to uncover mechanistic links between calcium, actin, and growth.

Long-Term Growth of Moss in Microfluidic Devices Enables Subcellular Studies in Development.

Key developmental processes that occur on the subcellular and cellular level or occur in occluded tissues are difficult to access, let alone image and analyze. Recently, culturing living samples within polydimethylsiloxane (PDMS) microfluidic devices has facilitated the study of hard-to-reach developmental events. Here, we show that an early diverging land plant, Physcomitrella patens, can be continuously cultured within PDMS microfluidic chambers. Because the PDMS chambers are bonded to a coverslip, it is possible to image P. patens development at high resolution over long time periods. Using PDMS chambers, we report that wild-type protonemal tissue grows at the same rate as previously reported for growth on solid medium. Using long-term imaging, we highlight key developmental events, demonstrate compatibility with high-resolution confocal microscopy, and obtain growth rates for a slow-growing mutant. By coupling the powerful genetic tools available to P. patens with long-term growth and imaging provided by PDMS microfluidic chambers, we demonstrate the capability to study cellular and subcellular developmental events in plants directly and in real time.

Making the case for a 'fifth wave' in public health.

This paper will argue that the UK has seen several phases of public health improvement since the Industrial Revolution, and that each of these can be linked to major shifts in thinking about the nature of society and health itself. The authors are not, however, attempting to delineate firm sequences of events (or imply causality) as this would require a level of analysis of the relationship between economy, society and culture which is beyond the scope of this paper. Rather, it is suggested that each phase of health improvement can be thought of in metaphorical terms as a 'wave'. The first wave is associated with great public works and other developments arising from social responses to the profound disruptions which followed the Industrial Revolution. The second wave saw the emergence of medicine as science. The third wave involved the redesign of our social institutions during the 20th Century and gave birth to the welfare state. The fourth wave has been dominated by efforts to combat disease risk factors and the emergence of systems thinking. Although a trough of public health activity continues from each wave, none exerts the same impact as when it first emerged. This paper will discuss the complex challenges of obesity, inequality and loss of wellbeing, together with the broader problems of exponential growth in population, money creation and energy usage. As exponential growth is unsustainable on a finite planet, inevitable change looms. Taken together, these analyses suggest that a fifth wave of public health development is now needed; one which will need to differ radically from its forerunners. The authors invite others to join them in envisioning its nature and in furthering the debate about future public health.

Unexpected involvement of a second rodent species makes impacts of introduced rats more difficult to detect.

Rodent predators are implicated in declines of seabird populations, and removing introduced rats, often, but not always, results in the expected conservation gains. Here we investigated the relationship between small mammal (Norway rat, wood mouse and pygmy shrew) abundance and Manx shearwater breeding success on the island of Rum, Scotland, and tested whether localised rodenticide treatments (to control introduced Norway rats) increased Manx shearwater breeding success. We found that Manx shearwater breeding success was negatively correlated with late summer indices of abundance for rats and mice, but not shrews. On its own, rat activity was a poor predictor of Manx shearwater breeding success. Rat activity increased during the shearwater breeding season in untreated areas but was supressed in areas treated with rodenticides. Levels of mouse (and shrew) activity increased in areas treated with rodenticides (likely in response to lower levels of rat activity) and Manx shearwater breeding success was unchanged in treated areas (p < 0.1). The results suggest that, unexpectedly, negative effects from wood mice can substitute those of Norway rats and that both species contributed to negative impacts on Manx shearwaters. Impacts were intermittent however, and further research is needed to characterise rodent population trends and assess the long-term risks to this seabird colony. The results have implications for conservation practitioners planning rat control programmes on islands where multiple rodent species are present.

Plant PIEZO homologs modulate vacuole morphology during tip growth.

In animals, PIEZOs are plasma membrane-localized cation channels involved in diverse mechanosensory processes. We investigated PIEZO function in tip-growing cells in the moss Physcomitrium patens and the flowering plant Arabidopsis thaliana PpPIEZO1 and PpPIEZO2 redundantly contribute to the normal growth, size, and cytoplasmic calcium oscillations of caulonemal cells. Both PpPIEZO1 and PpPIEZO2 localized to vacuolar membranes. Loss-of-function, gain-of-function, and overexpression mutants revealed that moss PIEZO homologs promote increased complexity of vacuolar membranes through tubulation, internalization, and/or fission. Arabidopsis PIEZO1 also localized to the tonoplast and is required for vacuole tubulation in the tips of pollen tubes. We propose that in plant cells the tonoplast has more freedom of movement than the plasma membrane, making it a more effective location for mechanosensory proteins.

Enabling well-being in a time of radical change: Integrative public health for the 21st century.

We live in a rapidly changing world; one where existing models for and approaches to health appear to deliver diminishing returns, whilst new public health challenges emerge. This paper outlines an integrative approach to such challenges. Integral theory suggests that key dimensions of human experience, frequently presented in opposition to each other (e.g. subjective-objective; individual-collective), need to be understood as integral to the whole. This is relevant to the public health community because powerful forces within neglected dimensions can undermine or destroy our efforts in other dimensions. This is illustrated in this paper by focusing on the issue of well-being, which illustrates the interconnected ways in which people in affluent societies can suffer from particular problems arising in such society and contribute to broader, global problems. The integral framework is used to show how a more integrative approach to such challenges can transcend some neglected blind-spots within public health. It is argued that public health leaders and practitioners need to apply integrative forms of thinking to their own practice in order to respond more effectively to the complexity of contemporary public health problems.

Correction to: Examining Race/Ethnicity Differences in the Association between the Experience of Workplace Racial Discrimination and Depression or Negative Emotions.

Please note that the name of coauthor Shauna Carlisle was presented incorrectly in this article as published: The correct rendering of her name is Carlisle, S.K. (instead of the incorrect rendering of Carlisle, S.E., which incorrectly notes E. [surname] as her middle initial).

Upper dose thresholds for radiation-induced adaptive response against cancer in high-dose-exposed, cancer-prone, radiation-sensitive Trp53 heterozygous mice.

Trp53 heterozygous mice are radiation-sensitive and cancer-prone. Groups of 7-8-week-old female Trp53 heterozygous mice were exposed to 4 Gy of 60Co gamma radiation at high (0.5 Gy/min) or low (0.5 mGy/min) dose rate. Other groups received 10 or 100 mGy at low dose rate 24 h prior to the 4-Gy dose. Tumor frequency and latency were measured over the animals' life span. Exposure to 10 mGy prior to 4 Gy resulted in a small (approximately 5%) but significant life-span regain and increased latency (approximately 9%) for all malignant tumors taken together, but 100 mGy further reduced life span slightly (approximately 7%). Latency responses were tumor type-specific. The prior 10-mGy exposure resulted in a small (approximately 7%) regain in latency for lymphomas but no change in latency for spinal osteosarcomas. Increasing the adapting dose to 100 mGy eliminated the increase in lymphoma latency and further reduced life span (approximately 8%). A 10-mGy dose prior to 4 Gy at low dose rate had no effects. Adapting exposures had no significant effect on tumor frequency. We conclude that a single low dose induced a small protective response in vivo in Trp53+/- mice, reducing the carcinogenic effects of a subsequent large, high-dose-rate exposure by increasing tumor latency. The upper dose threshold at which low-dose protective effects gave way to detrimental effects was tumor type-specific, as found previously for spontaneous tumors in these same cancer-prone mice (Radiat. Res. 159, 320-327, 2003). However, the upper dose thresholds appear to be lower (below 100 mGy) for radiation-induced tumors than for the same tumors appearing spontaneously.

Low doses of radiation increase the latency of spontaneous lymphomas and spinal osteosarcomas in cancer-prone, radiation-sensitive Trp53 heterozygous mice.

Mice heterozygous for Trp53 are radiation-sensitive and cancer-prone, spontaneously developing a variety of cancer types. Osteosarcomas in the spine lead to paralysis, while lymphomas lead rapidly to death, distinct events that provide objective measures of latency. The effects of a single low-dose (10 or 100 mGy), low-dose-rate (0.5 mGy/min) (60)Co gamma irradiation on lymphoma or spinal osteosarcoma frequency and latency, defined as time of death or of onset of paralysis, respectively, were examined. Compared to spontaneous lymphomas or to spinal osteosarcomas leading to paralysis in unexposed mice, an exposure of 7-8-week-old Trp53(+/-) mice to 10 or 100 mGy had no significant effect on tumor frequency, indicating no effect on tumor initiation. All tumors are therefore assumed to be of spontaneous origin. However, a 10-mGy exposure reduced the risk of both lymphomas and spinal osteosarcomas by significantly increasing tumor latency, indicating that the main in vivo effect of a low-dose exposure is a reduction in the rate at which spontaneously initiated cells progress to malignancy. The effect of this adaptive response persisted for the entire life span of all the animals that developed these tumors. Exposure to 100 mGy delayed lymphoma latency longer than the 10-mGy exposure. However, the 100-mGy dose increased spinal osteosarcoma risk by decreasing overall latency compared to unexposed control mice. That result suggested that this higher dose was in a transition zone between reduced and increased risk, but that the dose at which the transition occurs varies with the tumor type.

Differential expression of the CB2 cannabinoid receptor by rodent macrophages and macrophage-like cells in relation to cell activation.

An in vitro model of multi-step activation, in which cells of macrophage lineage are driven sequentially through inflammatory, primed, and fully activated states, was employed to assess for cannabinoid receptor expression. Murine and rat peritoneal macrophages, murine RAW264.7 and P388D, macrophage-like cells, and neonatal rat brain cortex microglia expressed the cannabinoid receptor type 2 (CB2) differentially in relation to cell activation. The CB2 was undetectable in resident peritoneal macrophages, present at high levels in thioglycolate-elicited inflammatory and interferon gamma (IFNgamma)-primed peritoneal macrophages, and detected at significantly diminished levels in bacterial lipopolysaccharide (LPS)-activated peritoneal macrophages. A comparable pattern of differential expression of the CB2 was noted for murine macrophage-like cells and neonatal rat brain cortex microglia. The cannabinoid receptor type 1 (CB1) was not detected in peritoneal macrophages or murine macrophage-like cells regardless of cell activation state but was present in neonatal rat microglia at low levels. These results indicate that levels of the CB2 in cells of macrophage lineage undergo major modulatory changes in relation to cell activation. Furthermore, since inflammatory and primed macrophages express the highest levels of CB2, the functional activities of macrophages when in these respective states of activation may be the most sensitive to the action of cannabinoids.

Cannabinoid-mediated inhibition of inducible nitric oxide production by rat microglial cells: evidence for CB1 receptor participation.

Activated brain microglial cells release inflammatory mediators such as nitric oxide (NO) that may play important roles in central nervous system antibacterial, antiviral, and antitumor activities. However, excessive release of these factors has been postulated to elicit immune-mediated neurodegenerative inflammatory processes and to cause brain injury. Recent studies using the rat animal model indicate that select cannabinoids may modulate production of these inflammatory factors. Treatment of neonatal rat brain cortical microglial cells with the cannabinoid paired enantiomers CP55940 and CP56667 resulted in a stereoselective differential effect on inducible NO production. The analog CP55940 exerted a dose-dependent inhibition of interferon gamma (IFNy)/bacterial lipopolysaccharide (LPS)-inducible NO production which was significantly greater than that exerted by CP56667. Pretreatment of microglial cells with the CB1 cannabinoid receptor-selective antagonist SR141716A reversed this CP55940-mediated inhibition. MRT-PCR demonstrated the presence of CB1 receptor mRNA within microglial cells consistent with the presence of CB1 receptors. Collectively, these results indicate that the cannabinoid analog CP55940 selectively inhibits inducible NO production by microglial cells and that this inhibition is effected, at least in part, through the CB1 receptor.

Practice and professional development plans (PPDPs): results of a feasibility study.

BACKGROUND: Dissatisfaction with uniprofessional education structures as a means of improving the quality of healthcare has led to proposals to develop ways of integrating professional learning and organisational development. AIMS: Test the feasibility of introducing practice and professional development plans using a centrally sponsored project in Wales. DESIGN: Qualitative observational study. STUDY SAMPLE: All 541 practices in Wales were alerted to the project and invited to apply. A selection process was suggested to Health Authorities but not always efficiently conducted: 23 practices were selected and 18 participated in the process. METHOD: Central funding was made available to health authorities. The project framework was designed by an educational department and conceptualised as the development of personal portfolios linked to one key organisation change in each practice, facilitated by external consultants who would typically hold workshops or other events. An independent researcher using non-participant observation techniques at workshops and practices undertook documentary analysis and fieldwork in four health authorities. RESULTS: Difficulties were encountered with the process of implementing the project: marketing and practice selection inconsistencies delayed the work and it was difficult to recruit practices into the project. The lack of experienced individuals to do the work and practitioner suspicion about perceived 'management' agendas were significant problems. After initial hesitancies most practices appreciated the value of developing wider ownership and commitment to proposed practice changes. Organisations found it difficult to support individual completion of the personal portfolio component of the plans. The ability to develop systems for clinical services was dependent on having already established a culture of effective teamwork in the organisation. CONCLUSIONS: This work supports the view that organisational development has considerable potential for bringing about effective change, and individual contributions could form a valuable component of personal portfolios. We believe that the existing structures in education and management in the health service are not yet able to support these processes. Evidence from the fields of risk management and quality improvement all point to the need to develop effective organisational systems and the results of this feasibility study indicate that alternative models of sustaining organisational development need careful evaluation.

Schistosoma mansoni: morphologic changes induced by maintenance in vitro.

Adults of Schistosoma mansoni were incubated in several culture media at various time intervals ranging from 2 to 96 hr. The morphologic changes induced by the incubation were documented using both scanning and transmission electron microscopy. These included changes in the tegument, esophageal cells, and cecum. Variability was noted among worms within experimental groups and the surface changes on single worms were frequently observed to have patchy distribution. Based on morphologic changes observed, culture media were ranked as adequate, mediocre, or inadequate. RPMI-1640 + 50% fetal calf serum, Eagle's MEM with Earle's salts (no serum), and McCoy's 5A Medium + serum were judged adequate. Basal Eagle's Medium, Triple Eagle Medium, NCTC-135, and Earle's Balanced Salt Solution (all with or without serum) were judged mediocre. Hanks' Basal Salt Solution (with or without serum) was judged as severely inadequate. All media tested gave better results in the presence of serum. These factors point to the necessity for the use of carefully selected culture media, as well as adequate controls and sampling techniques in the interpretation of in vitro experiments with S. mansoni.

Schistosoma mansoni: evaluation of selected preparative procedures for transmission and scanning electron microscopy.

Evaluation of numerous procedures for collection and preparation of Schistosoma mansoni for scanning (SEM) and transmission microscopy (TEM) revealed that some commonly used methods result in introduction of osmotic or mechanical damage. Modification and selection of appropriate procedures allowed elimination of these artifacts. Not eliminated by these procedures, however, were several kinds of spheres and blebs that were either external to the apical plasma membrane of the tegument or budding from it. After careful examination of all procedures, we concluded that these spheres appear to some extent on all schistosomes prepared for either SEM or TEM and may represent either a normal component of the schistosome surface, a constant host contaminant, or an artifact not eliminated by any of the multiple modifications of technique examined.

Effects of parasympathominetics on porcine stillbirth.

To study the effect of 2 parasympathomimetic drugs in reduction of incidence of stillborn pigs, 84 sows and gilts were randomly allotted to 3 treatment groups. Single 1-ml injections of isotonic saline solution, carbachol (2 mg/ml), or neostigmine bromide (5 mg/ml) were subcutaneously administered to pigs of groups A, B, and C, respectively. Injections were given midway through parturition in an attempt to hasten the delivery of the last pigs in the litter, where the corresponding incidence of stillbirth is greatest. The mean time of injection was after the birth of 3.5 pigs. Total stillbirths/litter for carbachol-treated (0.23) and neostigmine-treated (0.21) pies were significantly different (P greater than 0.0001) from those for control pigs (0.88). Before injection, stillbirths/litter for control pigs (0.23) were not significantly different from those for carbachol-treated (0.18) or neostigmine-treated (0.12) pigs. After treatment, stillbirths/litter for carbachol-treated (0.06) and neostigmine-treated (0.09) pigs were both significantly different (P greater than 0.0001) from those for saline solution-treated control pigs (0.65). When injected midway through parturition, carbachol and neostigmine reduced stillbirth rate by reducing stillbirths which occur late in farrowing.

A simple transport system for radiation treatment of specific pathogen-free mice in lifetime studies.

To prevent the introduction of pathogens, specific pathogen-free (SPF) facilities generally have a "once out, never back" policy with respect to animals and materials. In a lifetime study of the long-term effects of ionizing radiation exposure in mice, large numbers of SPF mice needed to be transported from clean-animal barrier labs to a multiuser conventional building for radiation treatment and then back into the animal facility. The conventional building is known to harbor wild mice as well as insects, spiders, and mites, and this situation might potentiate the transfer of wild mouse pathogens to laboratory animals. Introduction of pathogens into the mouse population would jeopardize the entire study, but the radiation treatments were an essential component of the study. These considerations prompted development of a system for transporting individual animals out of and back into the facility without exposure to pathogens. The system consists of reusable transport/treatment vessels and transport protocols designed to minimize the potential for pathogen exposure.