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A focus group was conducted to determine the views of persons with AIDS about how to best conduct the process of establishing advance directives. (Advance directives are instructions regarding future medical care to be implemented should a patient be unable to communicate his/her treatment preferences.) Several hypotheses emerged from this focus group that were then formally tested with a questionnaire. The results of the questionnaire identified (a) predictors of attitudes toward advance directives and (b) predictors of the act of having established a directive. Based upon the findings from the focus group and the questionnaire, recommendations for social workers raising the issue of advance directives can be made. First, time for the client to adjust to seropositive notification should be allowed prior to raising the advance directive issue. Second, the advance directive issue should be raised in the context of empowerment rather than being embedded in other paper work. Third, people who are most likely to be disturbed by the process of establishing advance directives are those who have low health locus of control, low trait optimism, low reliance on active involvement as a coping mechanism, and low reliance on distraction as a coping mechanism. More time should be allotted for people with those characteristics. [ABSTRACT FROM AUTHOR]

Cancer initiates as a consequence of genomic mutations and its subsequent progression relies in part on increased production of ribosomes to maintain high levels of protein synthesis for unchecked cell growth. Recently, cytidine deaminases have been uncovered as sources of mutagenesis in cancer. In an attempt to form a connection between these 2 cancer driving processes, we interrogated the cytidine deaminase family of proteins for potential roles in human ribosome biogenesis. We identified and validated APOBEC3A and APOBEC4 as novel ribosome biogenesis factors through our laboratory's established screening platform for the discovery of regulators of nucleolar function in MCF10A cells. Through siRNA depletion experiments, we highlight APOBEC3A's requirement in making ribosomes and specific role within the processing and maturation steps that form the large subunit 5.8S and 28S ribosomal (r)RNAs. We demonstrate that a subset of APOBEC3A resides within the nucleolus and associates with critical ribosome biogenesis factors. Mechanistic insight was revealed by transient overexpression of both wild-type and a catalytically dead mutated APOBEC3A, which both increase cell growth and protein synthesis. Through an innovative nuclear RNA sequencing methodology, we identify only modest predicted APOBEC3A C-to-U target sites on the pre-rRNA and pre-mRNAs. Our work reveals a potential direct role for APOBEC3A in ribosome biogenesis likely independent of its editing function. More broadly, we found an additional function of APOBEC3A in cancer pathology through its function in ribosome biogenesis, expanding its relevance as a target for cancer therapeutics., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 McCool et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Corn ( Zea mays ) biomass accumulation and nutrient uptake by the six-leaf collar (V6) growth stage are low, and therefore, synchronizing nutrient supply with crop demand could potentially minimize nutrient loss and improve nutrient use efficiency. Knowledge of corn's response to nutrient stress in the early growth stages could inform such nutrient management. Field studies were conducted to assess corn recovery from when no fertilizer application is made until the V6 growth stage, and thereafter, applying fertilizer rates as those in non-stressed conditions. The early season nutrient stress and non-stress conditions received the same amount of nutrients. As the availability of nutrients for plant uptake is largely dependent on soil moisture, corn recovery from the early season nutrient stress was assessed under different soil moisture regimes induced via irrigation scheduling at 50% and 80% field capacity under overhead and subsurface drip irrigation (SSDI) systems. Peanut ( Arachis hypogaea ) was the previous crop under all conditions, and the fields were under cereal rye ( Secale cereale ) cover crop prior to planting corn. At the V6 growth stage, the nutrient concentrations of the early season-stressed crops, except for copper, were above the minimum threshold of sufficiency ranges reported for corn. However, the crops showed poor growth, with biomass accumulation being reduced by over 50% compared to non-stressed crops. Also, the uptake of all nutrients was significantly lower under the early season nutrient stress conditions. The recovery of corn from the early season nutrient stress was low. Compared to non-stress conditions, the early season nutrient stress caused 1.58 Mg ha -1 to 3.4 Mg ha -1 yield reduction. The percent yield reduction under the SSDI system was 37.6-38.2% and that under the overhead irrigation system was 11.7-13%. The high yield reduction from the early season nutrient stress under the SSDI system was because of water stress conditions in the topsoil soil layer. The findings of the study suggest ample nutrient supply in the early season growth stage is critical for corn production, and thus, further studies are recommended to determine the optimum nutrient supply for corn at the initial growth stages., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Amissah, Ankomah, Lee, Perry, Washington, Porter, Virk, Bryant, Vellidis, Harris, Cabrera, Franklin, Diaz-Perez and Sintim.)

While microRNAs and other non-coding RNAs are the next frontier of novel regulators of mammalian ribosome biogenesis (RB), a systematic exploration of microRNA-mediated RB regulation has not yet been undertaken. We carried out a high-content screen in MCF10A cells for changes in nucleolar number using a library of 2603 mature human microRNA mimics. Following a secondary screen for nucleolar rRNA biogenesis inhibition, we identified 72 novel microRNA negative regulators of RB after stringent hit calling. Hits included 27 well-conserved microRNAs present in MirGeneDB, and were enriched for mRNA targets encoding proteins with nucleolar localization or functions in cell cycle regulation. Rigorous selection and validation of a subset of 15 microRNA hits unexpectedly revealed that most of them caused dysregulated pre-rRNA processing, elucidating a novel role for microRNAs in RB regulation. Almost all hits impaired global protein synthesis and upregulated CDKN1A (p21) levels, while causing diverse effects on RNA Polymerase 1 (RNAP1) transcription and TP53 protein levels. We provide evidence that the MIR-28 siblings, hsa-miR-28-5p and hsa-miR-708-5p, potently target the ribosomal protein mRNA RPS28 via tandem primate-specific 3' UTR binding sites, causing a severe pre-18S pre-rRNA processing defect. Our work illuminates novel microRNA attenuators of RB, forging a promising new path for microRNA mimic chemotherapeutics., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Background: This study investigated potential opportunities or challenges for plant-based meat in the Chinese market. A quantitative framework was applied to determine the current level of familiarity and experience with plant-based meat among Chinese consumers, the proportion of consumers who would try or purchase plant-based meat, which demographics within China are most likely to buy plant-based meat and which attitudes are important in driving the purchase intent of plant-based meat., Methods: A pre-registered cross-sectional online survey (N = 1206) was distributed to respondents (matched to China's adult population for gender and age)., Results: Respondents reported a variety of dietary identities, with 43.4% reporting that they were reducing or avoiding meat. The majority of respondents (60.1%) said they had eaten plant-based meat at least once before. Of those who said they had never eaten plant-based meat, 41.9% intended to try it and 31.4% intended to purchase it. The strongest attitudinal predictor of plant-based meat purchase intent was perceived healthiness (β = 0.235, p < 0.001), whereas the strongest demographic predictor of plant-based meat purchase intent was age (β = -0.248, p < 0.001)., Conclusions: The findings of this study suggest that an approach based on increasing opportunities for trial, as well as appealing to specific attitudinal and demographic predictors of plant-based purchase intent, could prove successful in increasing adoption of plant-based and alternative meat products., (© 2022 The Authors. Journal of Human Nutrition and Dietetics published by John Wiley & Sons Ltd on behalf of British Dietetic Association.)

The main components of the essential cellular process of eukaryotic ribosome biogenesis are highly conserved from yeast to humans. Among these, the U3 Associated Proteins (UTPs) are a small subunit processome subcomplex that coordinate the first two steps of ribosome biogenesis in transcription and pre-18S processing. While we have identified the human counterparts of most of the yeast Utps, the homologs of yeast Utp9 and Bud21 (Utp16) have remained elusive. In this study, we find that NOL7 is the likely ortholog of Bud21. Previously described as a tumour suppressor through regulation of antiangiogenic transcripts, we now show that NOL7 is required for early pre-rRNA accumulation and pre-18S rRNA processing in human cells. These roles lead to decreased protein synthesis and induction of the nucleolar stress response upon NOL7 depletion. Beyond Bud21's nonessential role in yeast, we establish human NOL7 as an essential UTP that is necessary to maintain both early pre-rRNA levels and processing.

Our planet is currently facing unprecedented interconnected environmental, societal, and economic dilemmas due to climate change, the outbreak of pandemics and wars, among others. These global challenges pose direct threats to food security and safety and clearly show the urgent need for innovative scientific solutions and technological approaches. Backed by the current alarming situation, many food-related trends have emerged in recent years in response to these global issues. This review looks at two megatrends in agriculture and the food industry; the shift to vegetable diets and the digital transformation in food production and consumption patterns. On one side, several innovative technologies and protein sources have been associated with more sustainable food systems and enhanced nutritional quality and safety. On the other side, many digital advanced technologies (e.g., artificial intelligence, big data, the Internet of Things, blockchain, and 3D printing) have been increasingly applied in smart farms and smart food factories to improve food system outcomes. Increasing adoption of vegetal innovations and harnessing Industry 4.0 technologies along the food supply chain have the potential to enable efficient digital and ecological transitions., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Author(s).)

In eukaryotes, the nucleolus is the site of ribosome biosynthesis, an essential process in all cells. While human ribosome assembly is largely evolutionarily conserved, many of the regulatory details underlying its control and function have not yet been well-defined. The nucleolar protein RSL24D1 was originally identified as a factor important for 60S ribosomal subunit biogenesis. In addition, the PeBoW (BOP1-PES1-WDR12) complex has been well-defined as required for pre-28S rRNA processing and cell proliferation. In this study, we show that RSL24D1 depletion impairs both pre-ribosomal RNA (pre-rRNA) transcription and mature 28S rRNA production, leading to decreased protein synthesis and p53 stabilization in human cells. Surprisingly, each of the PeBoW complex members is also required for pre-rRNA transcription. We demonstrate that RSL24D1 and WDR12 co-immunoprecipitate with the RNA polymerase I subunit, RPA194, and regulate its steady state levels. These results uncover the dual role of RSL24D1 and the PeBoW complex in multiple steps of ribosome biogenesis, and provide evidence implicating large ribosomal subunit biogenesis factors in pre-rRNA transcription control., (Published by Cold Spring Harbor Laboratory Press for the RNA Society.)

Turgor pressure is an essential, but difficult to measure indicator of plant water status. Turgor has been quantified by localized compression of cells or tissues, but a simple method to perform these measurements is lacking. We hypothesized that changes in leaf turgidity can be monitored by uniaxially compressing the leaf lamina and measuring the mechanical stress under a constrained thickness (stress relaxation) and that changes in leaf water content can be monitored by measuring the leaf thickness under constant mechanical stress. Using a simple, custom-built leaf squeeze-flow rheometer, we performed different compression tests on leaves from 13 plant species. The mechanical stress measured during stress relaxation was correlated with leaf bulk turgor pressure (R 2  > 0.95) and thus with balancing pressure (R 2  > 0.94); the leaf thickness measured under constant mechanical stress was correlated with relative water content (R 2  > 0.74). The coefficients of these relationships were related to the leaf bulk osmotic pressure at the turgor-loss point. An idealized average-cell model suggests that, under isothermal conditions, the stationary bulk modulus during compression is largely determined by the bulk osmotic pressure. Our study presents an inexpensive, accessible and automatable method to monitor plant water status noninvasively., (© 2022 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Proteogenomic identification of translated small open reading frames has revealed thousands of previously unannotated, largely uncharacterized microproteins, or polypeptides of less than 100 amino acids, and alternative proteins (alt-proteins) that are co-encoded with canonical proteins and are often larger. The subcellular localizations of microproteins and alt-proteins are generally unknown but can have significant implications for their functions. Proximity biotinylation is an attractive approach to define the protein composition of subcellular compartments in cells and in animals. Here, we developed a high-throughput technology to map unannotated microproteins and alt-proteins to subcellular localizations by proximity biotinylation with TurboID (MicroID). More than 150 microproteins and alt-proteins are associated with subnuclear organelles. One alt-protein, alt-LAMA3, localizes to the nucleolus and functions in pre-rRNA transcription. We applied MicroID in a mouse model, validating expression of a conserved nuclear microprotein, and establishing MicroID for discovery of microproteins and alt-proteins in vivo., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Many unannotated microproteins and alternative proteins (alt-proteins) are coencoded with canonical proteins, but few of their functions are known. Motivated by the hypothesis that alt-proteins undergoing regulated synthesis could play important cellular roles, we developed a chemoproteomic pipeline to identify nascent alt-proteins in human cells. We identified 22 actively translated alt-proteins or N-terminal extensions, one of which is post-transcriptionally upregulated by DNA damage stress. We further defined a nucleolar, cell-cycle-regulated alt-protein that negatively regulates assembly of the pre-60S ribosomal subunit (MINAS-60). Depletion of MINAS-60 increases the amount of cytoplasmic 60S ribosomal subunit, upregulating global protein synthesis and cell proliferation. Mechanistically, MINAS-60 represses the rate of late-stage pre-60S assembly and export to the cytoplasm. Together, these results implicate MINAS-60 as a potential checkpoint inhibitor of pre-60S assembly and demonstrate that chemoproteomics enables hypothesis generation for uncharacterized alt-proteins., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

We conceptualize the journey to ethical veganism in the stages of the transtheoretical model of change, from precontemplation through contemplation, preparation, action, and maintenance. At each stage, we explore the psychological barriers to progressing towards veganism, discuss how they manifest, and explore ways to overcome them. It is hoped that this paper can be used as a guide for animal advocates to identify the stage an individual is at, and understand and overcome the social and psychological barriers they may face to progressing. We argue that, while many people are ignorant of the cruel practices entailed in animal farming, many deliberately avoid thinking about the issue, are unable to appreciate the scale of the issue, and simply tend to favour the status quo. When engaging with the issue of farm animal suffering, meat-eaters are largely driven by cognitive dissonance, which manifests as motivated reasoning aimed at protecting one's image of oneself and one's society. This is facilitated by confirmation bias and complicit media which cater to the preferred views of their meat-eating audience. Even once convinced of veganism, habit and willpower present further barriers to acting on those beliefs. This is all in the context of a speciesist and carnistic culture where meat consumption is normal, farming is noble, and vegans are 'others'. We locate and elucidate each of these biases within the stages of the transtheoretical model and discuss the implications of this model for animal advocates and for further research., (Copyright © 2021. Published by Elsevier Ltd.)

Studies of the regulation of nucleolar function are critical for ascertaining clearer insights into the basic biological underpinnings of ribosome biogenesis (RB), and for future development of therapeutics to treat cancer and ribosomopathies. A number of high-throughput primary assays based on morphological alterations of the nucleolus can indirectly identify hits affecting RB. However, there is a need for a more direct high-throughput assay for a nucleolar function to further evaluate hits. Previous reports have monitored nucleolar rRNA biogenesis using 5-ethynyl uridine (5-EU) in low-throughput. We report a miniaturized, high-throughput 5-EU assay that enables specific calculation of nucleolar rRNA biogenesis inhibition, based on co-staining of the nucleolar protein fibrillarin (FBL). The assay uses two siRNA controls: a negative non-targeting siRNA control and a positive siRNA control targeting RNA Polymerase 1 (RNAP1; POLR1A ), and specifically quantifies median 5-EU signal within nucleoli. Maximum nuclear 5-EU signal can also be used to monitor the effects of putative small-molecule inhibitors of RNAP1, like BMH-21, or other treatment conditions that cause FBL dispersion. We validate the 5-EU assay on 68 predominately nucleolar hits from a high-throughput primary screen, showing that 58/68 hits significantly inhibit nucleolar rRNA biogenesis. Our new method establishes direct quantification of nucleolar function in high-throughput, facilitating closer study of RB in health and disease.

Evidence overwhelmingly supports the view that we need to drastically reduce our consumption of animal products for reasons related to the environment and public health, while moral concerns about the treatment of animals in agriculture are becoming ever more common. As governments increasingly recognize the need to change our food production and alternative protein products become more appealing to consumers, agriculture finds itself in a unique period of transition. How do farmers respond to the changing atmosphere? We present secondary analyses of qualitative and quantitative data to highlight some of the uncertainty and ambivalence about meat production felt throughout the farming community. Survey data from France and Germany reveals that in both countries, those who work in the meat industry have significantly higher rates of meat avoidance than those who do not work in the industry. While non-meat-industry workers are more likely to cite concerns for animals or the environment, meat industry workers more often cite concerns about the healthiness or safety of the products. Concurrently, interviews with people who raise animals for a living suggest that moral concerns among farmers are growing but largely remain hidden; talking about them openly was felt as a form of betrayal. We discuss these findings in the context of the ongoing agricultural transition, observe how tension has manifested as polarization among Dutch farmers, and offer some thoughts about the role of farmers in a new world of alternative proteins., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Alopecia, neurologic defects, and endocrinopathy (ANE) syndrome is a rare ribosomopathy known to be caused by a p.(Leu351Pro) variant in the essential, conserved, nucleolar large ribosomal subunit (60S) assembly factor RBM28. We report the second family of ANE syndrome to date and a female pediatric ANE syndrome patient. The patient presented with alopecia, craniofacial malformations, hypoplastic pituitary, and hair and skin abnormalities. Unlike the previously reported patients with the p.(Leu351Pro) RBM28 variant, this ANE syndrome patient possesses biallelic precursor messenger RNA (pre-mRNA) splicing variants at the 5' splice sites of exon 5 (ΔE5) and exon 8 (ΔE8) of RBM28 (NM&#95;018077.2:c.[541+1&#95;541+2delinsA]; [946G > T]). In silico analyses and minigene splicing experiments in cells indicate that each splice variant specifically causes skipping of its respective mutant exon. Because the ΔE5 variant results in an in-frame 31 amino acid deletion (p.(Asp150&#95;Lys180del)) in RBM28 while the ΔE8 variant leads to a premature stop codon in exon 9, we predicted that the ΔE5 variant would produce partially functional RBM28 but the ΔE8 variant would not produce functional protein. Using a yeast model, we demonstrate that the ΔE5 variant does indeed lead to reduced overall growth and large subunit ribosomal RNA (rRNA) production and pre-rRNA processing. In contrast, the ΔE8 variant is comparably null, implying that the partially functional ΔE5 RBM28 protein enables survival but precludes correct development. This discovery further defines the underlying molecular pathology of ANE syndrome to include genetic variants that cause aberrant splicing in RBM28 pre-mRNA and highlights the centrality of nucleolar processes in human genetic disease., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

Despite growing evidence of the environmental and public health threats posed by today's intensive animal production, consumers in the west remain largely attached to meat. Cultivated meat offers a way to grow meat directly from cells, circumventing these issues as well as the use of animals altogether. The aim of this study was to assess the overall consumer markets and a range of preferences around cultivated meat in the US and the UK relating to nomenclature, genetic modification, health enhancements, and other features. To this end, we recruited large representative samples to participate in an online survey about cultivated meat, and subsequently analyzed segments (a) in the early majority population (guided by the Diffusion of Innovations Model), (b) by generation, and (c) in the general population. Our findings showed a high level of openness (80%) in both the US and UK populations, with 40% somewhat or moderately likely to try and 40% highly likely to try. Younger generations had the greatest openness: 88% of Gen Z, 85% of Millennials, 77% of Gen X, and 72% of Baby Boomers were at least somewhat open to trying cultivated meat. All segments envisioned cultivated meat to be nearly half of their total meat intake. Findings show that consumers prefer the terms 'cultured' and 'cultivated' over 'cell-based' and 'cell-cultured' for use in a social context and on packages, even though they perceive these terms as less descriptive. The most important on-package label was one indicating government assurances, and participants preferred non-GM products over GM products. We also found that US consumers prefer nutritionally superior meat over nutritionally equivalent meat. We discuss implications for product development, messaging, and understanding the likely adoption path of this food innovation.

Cultured meat grown in vitro from animal cells has the potential to address many of the ethical, environmental, and public health issues associated with conventional meat production. However, as well as overcoming technical challenges to producing cultured meat, producers and advocates of the technology must consider a range of social issues, including consumer appeal and acceptance, media coverage, religious status, regulation, and potential economic impacts. Whilst much has been written on the prospects for consumer appeal and acceptance of cultured meat, less consideration has been given to the other aspects of the social world that will interact with this new technology. Here, each of these issues is considered in turn, forming a view of cultured meat as a technology with a diverse set of societal considerations and far-reaching social implications. It is argued that the potential gains from a transition to cultured meat are vast, but that cultural phenomena and institutions must be navigated carefully for this nascent industry to meet its potential., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science.)

We used a widely distributed tree Eucalyptus camaldulensis subsp. camaldulensis to partition intraspecific variation in leaf functional traits to genotypic variation and phenotypic plasticity. We examined if genotypic variation is related to the climate of genotype provenance and whether phenotypic plasticity maintains performance in a changing environment. Ten genotypes from different climates were grown in a common garden under watering treatments reproducing the wettest and driest edges of the subspecies' distribution. We measured functional traits reflecting leaf metabolism and associated with growth (respiration rate, nitrogen and phosphorus concentrations, and leaf mass per area) and performance proxies (aboveground biomass and growth rate) each season over a year. Genotypic variation contributed substantially to the variation in aboveground biomass but much less in growth rate and leaf traits. Phenotypic plasticity was a large source of the variation in leaf traits and performance proxies and was greater among sampling dates than between watering treatments. The variation in leaf traits was weakly correlated to performance proxies, and both were unrelated to the climate of genotype provenance. Intraspecific variation in leaf traits arises similarly among genotypes in response to seasonal environmental variation, instead of long-term water availability or climate of genotype provenance., (© 2020 The Authors New Phytologist © 2020 New Phytologist Trust.)

Ribosome biogenesis is the fine-tuned, essential process that generates mature ribosomal subunits and ultimately enables all protein synthesis within a cell. Novel regulators of ribosome biogenesis continue to be discovered in higher eukaryotes. While many known regulatory factors are proteins or small nucleolar ribonucleoproteins, microRNAs (miRNAs), and long non-coding RNAs (lncRNAs) are emerging as a novel modulatory layer controlling ribosome production. Here, we summarize work uncovering non-coding RNAs (ncRNAs) as novel regulators of ribosome biogenesis and highlight their links to diseases of defective ribosome biogenesis. It is still unclear how many miRNAs or lncRNAs are involved in phenotypic or pathological disease outcomes caused by impaired ribosome production, as in the ribosomopathies, or by increased ribosome production, as in cancer. In time, we hypothesize that many more ncRNA regulators of ribosome biogenesis will be discovered, which will be followed by an effort to establish connections between disease pathologies and the molecular mechanisms of this additional layer of ribosome biogenesis control., (© 2020 The Author(s).)

The significance of shoot surface water uptake (SSWU) has been debated, and it would depend on the range of conditions under which it occurs. We hypothesized that the decline of leaf hydraulic conductance (K leaf ) in response to dehydration may be recovered through SSWU, and that the hydraulic conductance to SSWU (K surf ) declines with dehydration. We quantified effects of leaf dehydration on K surf and effects of SSWU on recovery of K leaf in dehydrated leaves of Avicennia marina. SSWU led to overnight recovery of K leaf , with recovery retracing the same path as loss of K leaf in response to dehydration. SSWU declined with dehydration. By contrast, K surf declined with rehydration time but not with dehydration. Our results showed a role of SSWU in the recovery of leaf hydraulic conductance and revealed that SSWU is sensitive to leaf hydration status. The prevalence of SSWU in vegetation suggests an important role for atmospheric water sources in maintenance of leaf hydraulic function, with implications for plant responses to changing environments., (© 2019 Australian National University New Phytologist © 2019 New Phytologist Trust.)

Clean meat (grown from animal cells rather than rearing animals) has the potential to address many concerns associated with meat production. However, research suggests that the perceived unnaturalness of clean meat could be a barrier to consumer acceptance. This study investigated the efficacy of different messages designed to address consumers' concerns about clean meat naturalness. In an experimental design, participants read one of four messages: clean meat is natural, conventional meat is unnatural, naturalness is not important, or highlighting benefits of clean meat without addressing naturalness. The results indicated that arguing that conventional meat is unnatural resulted in a significant increase in some measures of acceptance compared to other messages. Arguing that clean meat is natural and challenging the appeal to nature were less persuasive, and challenging the appeal to nature resulted in some measures of acceptance being lower than not addressing naturalness. We discuss these results in the context of existing naturalness research and give recommendations for further research., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Greater availability of leaf dark respiration (R dark ) data could facilitate breeding efforts to raise crop yield and improve global carbon cycle modelling. However, the availability of R dark data is limited because it is cumbersome, time consuming, or destructive to measure. We report a non-destructive and high-throughput method of estimating R dark from leaf hyperspectral reflectance data that was derived from leaf R dark measured by a destructive high-throughput oxygen consumption technique. We generated a large dataset of leaf R dark for wheat (1380 samples) from 90 genotypes, multiple growth stages, and growth conditions to generate models for R dark . Leaf R dark (per unit leaf area, fresh mass, dry mass or nitrogen, N) varied 7- to 15-fold among individual plants, whereas traits known to scale with R dark , leaf N, and leaf mass per area (LMA) only varied twofold to fivefold. Our models predicted leaf R dark , N, and LMA with r 2 values of 0.50-0.63, 0.91, and 0.75, respectively, and relative bias of 17-18% for R dark and 7-12% for N and LMA. Our results suggest that hyperspectral model prediction of wheat leaf R dark is largely independent of leaf N and LMA. Potential drivers of hyperspectral signatures of R dark are discussed., (© 2019 John Wiley & Sons Ltd.)

In vitro meat (IVM) grown from animal cells is approaching commercial viability. This technology could enable consumers to circumvent the ethical and environmental issues associated with meat-eating. However, consumer acceptance of IVM is uncertain, and is partly dependent on how the product is framed. This study investigated the effect of different names for IVM on measures of consumer acceptance. Participants (N = 185) were allocated to one of four conditions in an experimental design in which the product name was manipulated to be 'clean meat', 'cultured meat', 'animal free meat', or 'lab grown meat'. Participants gave word associations and measures of their attitudes and behavioural intentions towards the product. The results indicated that those in the 'clean meat' and 'animal free meat' conditions had significantly more positive attitudes towards IVM than those in the 'lab grown meat' condition, and those in the 'clean meat' condition had significantly more positive behavioural intentions towards IVM compared to those in the 'lab grown meat' condition. Mediation analyses indicated that the valence of associations accounted for a significant amount of the observed differences, suggesting that anchoring can explain these differences. We discuss these results in the context of social representations theory and give recommendations for future research., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Pyrrolizidine alkaloids (PAs) are a class of secondary metabolites that function as feeding deterrents in a range of different plant species. In perennial ryegrass (Lolium perenne L.) the only PAs that have been identified are the thesinine-rhamnoside group, which displays significant genetic variation. Homospermidine synthase (HSS) has evolved from deoxyhypusine synthase (DHS) and catalyses the first step in the PA pathway, making it a key candidate for the investigation of genes influencing observed PA trait variation., Results: During PCR amplification and sequence analysis of DHS we identified two putative HSS genes in perennial ryegrass. One of the genes (LpHSS1) was absent in some perennial ryegrass plants. Thesinine-rhamnoside levels were measured using liquid chromatography coupled with mass spectrometry in a diverse association mapping population, consisting of 693 plants free of fungal endophytic symbionts. Association tests that accounted for population structure identified a significant association of absence of the LpHSS1 gene with lower levels of thesinine-rhamnoside PAs. HSS-like gene sequences were identified for other grass species of the Poaceae, including tall fescue, wheat, maize and sorghum., Conclusion: HSS is situated at the crucial first step in the PA pathway making it an important candidate gene for investigation of involvement in PA phenotypic variation. In this study, PA level in perennial ryegrass was strongly associated with the presence or absence of the LpHSS1 gene. A genetic marker, developed for the presence/absence of LpHSS1, may be used for marker-assisted breeding to either lower or increase PAs in breeding populations of perennial or Italian ryegrass to investigate a potential role in the deterrence of herbivore pests. The presence of HSS-like genes in several other Poaceae species suggests that PA biosynthesis may occur in plant family members beyond perennial ryegrass and tall fescue and identifies a potential route for manipulating PA levels.

Polyacrylamides (PAMs), when applied as a soil amendment, purportedly improve soil infiltration, decrease erosion, and reduce offsite agrochemical transport. The effect of PAM on infiltration, erosion, agrochemical transport, and crop yield when applied in furrow to mid-southern US production systems has not been evaluated. The objective of this study was to assess PAM effects on infiltration, erosion, corn ( L.) grain yield, and nitrogen (N) and phosphorus (P) transport when applied at 10 mg L through lay-flat polyethylene tubing. A 2-yr field study was conducted at the Mississippi State Delta Research and Extension Center in Stoneville, MS, on a Dundee silt loam and a Forestdale silty clay loam. The experimental design was a randomized complete block with four replications of each treatment: irrigated plus no PAM (control) and irrigated plus PAM at 10 mg L. Each irrigation event delivered 102 mm of water at 18.9 L m per furrow, and runoff was captured in a holding tank on the lower end of each plot. Pooled over year and soil texture, PAM increased infiltration and corn grain yield by 6% ( ≤ 0.0398). Polyacrylamide effects on the offsite transport of sediment and N and P were inconsistent, varying across year and soil texture. Results indicate that PAM improves infiltration and corn grain yield on silt loam and silty clay loam textured soils; however, further research is required before PAM can be recommended as a best management practice for mitigating erosion and offsite agrochemical transport in mid-southern production systems., (Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.)

Deregulated extracellular signal-regulated kinase (ERK) signaling drives cancer growth. Normally, ERK activity is self-limiting by the rapid inactivation of upstream kinases and delayed induction of dual-specificity MAP kinase phosphatases (MKPs/DUSPs). However, interactions between these feedback mechanisms are unclear. Here we show that, although the MKP DUSP5 both inactivates and anchors ERK in the nucleus, it paradoxically increases and prolongs cytoplasmic ERK activity. The latter effect is caused, at least in part, by the relief of ERK-mediated RAF inhibition. The importance of this spatiotemporal interaction between these distinct feedback mechanisms is illustrated by the fact that expression of oncogenic BRAF V600E , a feedback-insensitive mutant RAF kinase, reprograms DUSP5 into a cell-wide ERK inhibitor that facilitates cell proliferation and transformation. In contrast, DUSP5 deletion causes BRAF V600E -induced ERK hyperactivation and cellular senescence. Thus, feedback interactions within the ERK pathway can regulate cell proliferation and transformation, and suggest oncogene-specific roles for DUSP5 in controlling ERK signaling and cell fate., Competing Interests: The authors declare no conflict of interest.

Objectives: Nuclear factor κB (NF-κB) is a critical activator of inflammatory processes and MTX is one of the most commonly prescribed DMARDs for treatment of RA. We sought to determine whether MTX inhibited NF-κB activity in RA and in lymphocytes and fibroblast-like synoviocytes (FLSs) and to define underlying mechanisms of action., Methods: An NF-κB luciferase reporter plasmid was used to measure NF-κB activation across experimental stimuli. Flow cytometry was used to quantify changes in intracellular protein levels, measure levels of reactive oxygen species and determine apoptosis. Quantitative RT-PCR was used to identify changes in MTX target genes., Results: In T cell lines, MTX (0.1 μM) inhibited activation of NF-κB via depletion of tetrahydrobiopterin (BH4) and increased Jun-N-terminal kinase (JNK)-dependent p53 activity. Inhibitors of BH4 activity or synthesis also inhibited NF-κB activation and, similar to MTX, increased JNK, p53, p21 and JUN activity. Patients with RA expressed increased levels of phosphorylated or active RelA (p65) compared with controls. Levels of phosphorylated RelA were reduced in patients receiving low-dose MTX therapy. In contrast, inhibition of NF-κB activation by MTX was not mediated via BH4 depletion and JNK activation in FLSs, but rather was completely prevented by adenosine receptor antagonists., Conclusion: Our findings support a model whereby distinct pathways are activated by MTX in T cells and FLSs to inhibit NF-κB activation., (© The Author 2014. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Ectopic expression of dual-specificity phosphatase 5 (DUSP5), an inducible mitogen-activated protein (MAP) kinase phosphatase, specifically inactivates and anchors extracellular signal-regulated kinase (ERK)1/2 in the nucleus. However, the role of endogenous DUSP5 in regulating the outcome of Ras/ERK kinase signaling under normal and pathological conditions is unknown. Here we report that mice lacking DUSP5 show a greatly increased sensitivity to mutant Harvey-Ras (HRas(Q61L))-driven papilloma formation in the 7,12-Dimethylbenz[a]anthracene/12-O-tetradecanoylphorbol-13-acetate (DMBA/TPA) model of skin carcinogenesis. Furthermore, mouse embryo fibroblasts (MEFs) from DUSP5(-/-) mice show increased levels of nuclear phospho-ERK immediately after TPA stimulation and fail to accumulate total ERK in the nucleus compared with DUSP5(+/+) cells. Surprisingly, a microarray analysis reveals that only a small number of Ras/ERK-dependent TPA-responsive transcripts are up-regulated on deletion of DUSP5 in MEFs and mouse skin. The most up-regulated gene on DUSP5 loss encodes SerpinB2, an inhibitor of extracellular urokinase plasminogen activator and deletion of DUSP5 acts synergistically with mutant HRas(Q61L) and TPA to activate ERK-dependent SerpinB2 expression at the transcriptional level. SerpinB2 has previously been implicated as a mediator of DMBA/TPA-induced skin carcinogenesis. By analyzing DUSP5(-/-), SerpinB2(-/-) double knockout mice, we demonstrate that deletion of SerpinB2 abrogates the increased sensitivity to papilloma formation seen on DUSP5 deletion. We conclude that DUSP5 performs a key nonredundant role in regulating nuclear ERK activation, localization, and gene expression. Furthermore, our results suggest an in vivo role for DUSP5 as a tumor suppressor by modulating the oncogenic potential of activated Ras in the epidermis.

Patient specific guides (PSGs) are postulated to improve the alignment of components in total knee arthroplasty. Three hundred consecutive total knee arthroplasties performed with either conventional (CON) (n = 185) or Visionaire PSG (n = 115) were evaluated with a CT protocol for coronal limb alignment, coronal and sagittal alignment of individual components and femoral component rotation. There was no statistically significant difference between the two groups in any of the above parameters. In addition, no difference was found in total operative time. PSGs do not offer any benefit over conventional guides in terms improving the coronal alignment of the limb or alignment of individual components., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Pin1 is an emerging oncology target strongly implicated in Ras and ErbB2-mediated tumourigenesis. Pin1 isomerizes bonds linking phospho-serine/threonine moieties to proline enabling it to play a key role in proline-directed kinase signalling. Here we report a novel series of Pin1 inhibitors based on a phenyl imidazole acid core that contains sub-μM inhibitors. Compounds have been identified that block prostate cancer cell growth under conditions where Pin1 is essential., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The peptidyl prolyl cis/trans isomerase Pin1 is a promising molecular target for anti-cancer therapeutics. Here we report the structure-guided evolution of an indole 2-carboxylic acid fragment hit into a series of alpha-benzimidazolyl-substituted amino acids. Examples inhibited Pin1 activity with IC(50) <100nM, but were inactive on cells. Replacement of the benzimidazole ring with a naphthyl group resulted in a 10-50-fold loss in ligand potency, but these examples downregulated biomarkers of Pin1 activity and blocked proliferation of PC3 cells., (Copyright 2009 Elsevier Ltd. All rights reserved.)

The naturally occurring phthalide-containing antibiotics spirolaxine methyl ether, CJ-12,954, CJ-13,013, CJ-13,015, CJ-13,102, CJ-13,103, CJ-13,104 and CJ-13,108, have been reported to exhibit anti-H. pylori activity. However, the exact stereochemistry of spirolaxine methyl ether, CJ-12,954 or CJ-13,013, contributing to this observed activity has not been confirmed. The anti-H. pylori activity of several analogues of spirolaxine methyl ether, CJ-12,954 and CJ-13,013 of defined stereochemistry together with the anti-H. pylori activity of several indole analogues of the simpler phthalide-containing antibiotics CJ-13,102, CJ-13,104, CJ-13,108 and CJ-13,015 is reported herein. A 1:1 mixture of spiroacetals 5b and 6b in which the phthalide substituent exhibited (3R)-stereochemistry was sixty times more active than the corresponding 1:1 mixture of spiroacetals with (3S)-stereochemistry. Notably, the unnatural (2''S)-diastereomer of spirolaxine methyl ether exhibited more potent anti-H. pylori activity than the natural product spirolaxine methyl ether. The 4,6-dimethoxyindoles 9, 10, 11 and 13 were all found to be less active than their parent compounds 1, 2, 3 and 4, respectively. Chain-shortened 4,6-dimethoxyindole analogue 12 of CJ-13,108 3 and 4,6-dimethoxyindole-spiroacetal 13 exhibited weak anti-H. pylori activity thus providing future opportunity for drug discovery programs.

The synthesis of the spiroacetal-containing anti-Helicobacter pylori agents (3S,2''S,5''S,7''S)- (ent-CJ-12,954) and (3S,2''S,5''R,7''S)- (ent-CJ-13,014) has been carried out based on the convergent union of a 1:1 mixture of heterocycle-activated spiroacetal sulfones and with (3S)-phthalide aldehyde . The synthesis of the (3R)-diastereomers (3R,2''S,5''S,7''S)- and (3R,2''S,5''R,7''S)- was also undertaken in a similar manner by union of (3R)-phthalide aldehyde with a 1:1 mixture of spiroacetal sulfones and . Comparison of the (1)H and (13)C NMR data, optical rotations and HPLC retention times of the synthetic compounds (3S,2''S,5''S,7''S)- and (3S,2''S,5''R,7''S)- and the (3R)-diastereomers (3R,2''S,5''S,7''S)- and (3R,2''S,5''R,7''S)-, with the naturally occurring compounds, established that the synthetic isomers and were in fact enantiomeric to the natural products CJ-12,954 and CJ-13,014. The (2S,8S)-stereochemistry in protected dihydroxyketone , the precursor to the mixture of spiroacetal sulfones and was established via union of readily available (S)-acetylene with aldehyde in which the (4S)-stereochemistry was established via asymmetric allylation. Deprotection and cyclization of protected dihydroxyketone afforded an inseparable 1:1 mixture of spiroacetal alcohols and that were converted into a 1:1 inseparable mixture of spiroacetal sulfones and . Phthalide-aldehyde was prepared via intramolecular acylation of bromocarbamate in which the (3S)-stereochemistry was established via asymmetric CBS reduction of ketone .

The first synthesis of the spiroacetal-containing anti-Helicobacter pylori agents ent-CJ-12,954 and ent-CJ-13,014 is reported based on the union of a heterocycle-activated spiroacetal-containing sulfone fragment with a phthalide-containing aldehyde fragment; comparison of the 1H and 13C NMR data, optical rotations and HPLC retention times of the synthetic compounds (3S,2"S,5"S,7"S)-(1a) and (3S,2"S,5"R,7"S)-(2a) and the (3R)-diastereomers (3R,2"S,5"S,7"S)-(1b) and (3R,2"S,5"R,7"S)- (2b) with the naturally occurring compounds established that the synthetic isomers (1a) and (2a) were in fact enantiomeric to the natural products CJ-12,954 and CJ-13,014.

Inflammation may play an essential role in vaso-occlusion in sickle cell disease. Sickle patients have high white counts and elevated levels of serum C-reactive protein (CRP), cytokines, and adhesion molecules. In addition, circulating endothelial cells, leukocytes, and platelets are activated. We examined 4 transgenic mouse models expressing human alpha- and sickle beta-globin genes to determine if they mimic the inflammatory response seen in patients. These mouse models are designated NY-S, Berk-S(Antilles), NY-S/S(Antilles) (NY-S x Berk-S(Antilles)), and Berk-S. The mean white counts were elevated 1.4- to 2.1-fold (P

The rotavirus major inner capsid protein (VP6) has been expressed in Nicotiana benthamiana plants using vectors based on potato virus X (PVX). VP6 was expressed either as a fusion with the PVX coat protein or from an additional subgenomic promoter inserted to enable both VP6 and PVX coat protein to be expressed independently. Both approaches yielded VP6, which retained the ability to form trimers. VP6 expressed from the subgenomic promoter assembled into paracrystalline sheets and tubes. Expression as a fusion protein yielded PVX rods that presented an external "overcoat" of VP6, but unexpectedly, some rotavirus protein also assembled into icosahedral viruslike particles (VLPs). The assembly of viral protein into VLPs suggests that prior display of VP6 on the flexuous PVX rod facilitates the subsequent assembly of VP6 into stable icosahedral particles., (Copyright 2000 Academic Press.)

Patients who underwent a single stage surgical procedure to place two Brånemark implants to stabilise complete mandibular overdentures with bar attachments, required no more postoperative follow-up visits before definitive prosthesis construction than patients who had their implants placed at two separate surgical stages. However, the visits of the single stage group, occurred over a shorter time-scale and more attention was required to tighten or replace their healing abutments.

Recent studies have suggested that giving opioids locally into inflamed tissue may cause analgesia. This antinociceptive effect has been attributed to the interaction of the drug with opioid receptors upregulated by inflammation in the peripheral tissues. We have compared the analgesic effect of intra-articular morphine with that of normal saline and a combination of morphine and its antagonist naloxone after arthroscopy of the temporomandibular joint (TMJ). Twenty-one patients took part in a randomized controlled double-blind trial and received one of these three solutions at the end of operation. The pain scores, time to the first request for analgesia, and the analgesic consumption of the patients in the three groups did not differ significantly at any time during the study period.

Aim: To determine the change in use of a day surgery unit over a 21-year period., Design: Retrospective analysis., Setting: The day surgery unit within a central London dental hospital., Materials: The operating and anaesthetic records of surgical activity in 1973, 1983, 1993 and 1994., Results: A six-fold increase in the total number of dentoalveolar procedures performed was demonstrated during the period studied. However, these procedures represented only 77.6% of surgical activity in 1994 compared with 98.9% in 1973., Conclusions: Day case surgery appears to be advantageous to patients, clinicians, hospital managers and purchasing authorities. The development of more specialist day case services and facilities within "dental centres' is proposed as a means of ensuring that the specialty is able to adapt to the changing patterns of health care in the future.

Recent changes in the classification of psychiatric illnesses have resulted in the term dysmorphophobia being replaced by that of body dysmorphic disorder (BDD). This paper attempts to alert the clinician to the presenting features of the condition and discusses its management, with particular emphasis on the role of surgery and current concepts of pharmacological treatment. A number of case reports are included to illustrate the diversity of this interesting disorder and the difficulties involved in treatment.

Commercially available controls for reducing worker exposure to paint overspray were evaluated in six autobody shops and a spray-painting equipment manufacturer's test facility. Engineering control measures included spray-painting booths, vehicle preparation stations, and spray-painting guns. The controls were evaluated by measuring particulate overspray concentrations in the worker's breathing zone, visualizing the airflow in spray-painting booths and vehicle preparation stations, and measuring airflow volumes and velocities. In addition, respirator usage observations were collected at five of the autobody repair shops, and quantitative fit tests were conducted on existing respirators at three shops. Several conclusions were drawn from this study. Downdraft spray-painting booths provide lower particulate overspray concentrations measured on the worker than crossdraft and semidowndraft spray-painting booths. In the latter two booths, the spray-painting gun can disperse as much as half the paint overspray into the incoming fresh air, increasing worker overspray exposure. Vehicle preparation stations have no walls to contain the overspray and, commonly, a single exhaust fan removes air from the painting area. Airflow patterns suggest that these do not control the paint overspray. Switching from a conventional spray-painting gun to a high-volume low pressure spray-painting gun reduced the particulate overspray concentration by a factor of 2 at a manufacturer's test facility. However, this change did not significantly affect solvent concentrations. Finally, respirator usage in five of the six shops studied was inappropriate. Respirators were poorly maintained and/or did not fit the workers, perhaps due to the absence of a formal respirator program.

Many amphibian populations are declining in a number of geographical locations throughout the world. In most cases, the cause or causes are unknown, but are assumed to result from man-made alterations in the environment. We review existing evidence concerning how environmental xenobiotics could contribute to declines of amphibian populations by impacting growth and development of the young. This paper examines the potential roles of toxicants in: a) affecting the susceptibility of young to disease; b) retarding growth and development of amphibian young; c) affecting the ability of larvae to avoid predation; d) affecting the development of physiological, morphological, or behavioral processes in a manner that subsequently impairs the ability of the young for future reproduction; and e) directly causing mortality of young. These issues are not well studied, and more studies are needed before the roles of environmental xenobiotics in amphibian declines are fully understood.

Careful analysis of electrocautery smoke produced during breast surgery has found organic compounds that are unidentifiable with current analytical techniques. The purpose of this study was to determine the potential mutagenicity of the smoke produced by the electrocautery knife during reduction mammaplasty. Multiple air samples were collected in the operating room during two reduction mammaplasty procedures. Airborne smoke particles were tested for mutagenic potential in both tester strains of Salmonella typhimurium (TA98 and TA100) using the standard Salmonella microsomal test (Ames test). All testing was performed by the Hazard Evaluations and Technical Assistance Branch of the National Institute of Occupational Safety and Health. The smoke produced with the electrocautery knife during reduction mammaplasty was found to be mutagenic to the TA98 strain. The Ames test, an established technique for evaluating the mutagenicity of a substance, was convincingly positive for the smoke collected during the breast surgery. Whether the smoke represents a serious health risk to operating room personnel is not known. Development of techniques to limit electrocautery smoke exposure in the operating room appears to be needed, and surgeons should attempt to minimize their exposure.