Consistently Exploring Nurse Staffing and Neurocritical Care Unit Turnover.

ABSTRACT: BACKGROUND: Staffing models within nursing units have long been a hot topic of discussion. The COVID-19 pandemic exacerbated this discussion by straining the national nursing environment and workforce. Before the pandemic, the neuroscience intensive care unit (NSICU) primarily used an acuity-adjusted staffing model and aimed for a nurse-to-patient ratio of 1:1.5. During and after the pandemic, the NSICU was forced to primarily use a centralized staffing model because of the increased turnover in the hospital at large and a rise in patient census. METHODS : Unit census data in an NSICU were tracked before, during, and after the pandemic alongside utilization of a centralized staffing model in the hospital at large. RESULTS : During this time, the NSICU saw a statistically significant increase in average nurse-to-patient ratio and incidences of both floating and tripled assignments. The NSICU simultaneously saw a 180% increase in nursing turnover. CONCLUSION : Although we cannot prove that a centralized staffing model is directly responsible for higher nursing turnover, its utilization led to greater incidence of poor staffing-reflected in deviation from the nurse-to-patient ratio goal of the unit. Nurse staffing concerns play a large role in nurse satisfaction in the workforce: staffing shortages have been described both as a precursor to and as a consequence of increased nursing turnover.

Signalling between the sexes during pollen tube reception.

Plant reproduction is a complex, highly-coordinated process in which a single, male germ cell grows through the maternal reproductive tissues to reach and fertilise the egg cell. Focussing on Arabidopsis thaliana, we review signalling between male and female partners which is important throughout the pollen tube journey, especially during pollen tube reception at the ovule. Numerous receptor kinases and their coreceptors are implicated in signal perception in both the pollen tube and synergid cells at the ovule entrance, and several specific peptide and carbohydrate ligands for these receptors have recently been identified. Clarifying the interplay between these signals and the downstream responses they instigate presents a challenge for future research and may help to illuminate broader principles of plant cell-cell communication.

The incidence and characteristics of giant cell arteritis in Hispanics and the associated outcomes of ischemic ocular events and stroke.

Giant cell arteritis (GCA) is a large vessel vasculitis whereby up to half of all patients show ocular involvement, highlighting the importance of treating GCA before it leads to irreversible blindness. Most of the research published for GCA and associated adverse ocular events are based on majority Caucasian populations establishing the current belief that Hispanics have a much lower incident of GCA. We sought to investigate the incidence of GCA in Hispanics and characterize associated ocular events, stroke rates, and comorbid diseases. We conducted a retrospective cohort study of GCA-coded medical records in self-identified Hispanics assessed at the University Medical Center Hospital in El Paso, Texas. From 2000 to 2019, there were 68 new cases of GCA in the Hispanics which represents an average incidence of 0.062% with a 95% confidence interval of [0.05, 0.09] of Hispanics over the age of 50 admitted to University Medical Center hospital. Of the subjects diagnosed with GCA, the majority had ocular involvement such as blurry vision (47.06%), ocular pain (26.47%), and blindness (14.71%). We did not observe a history of stroke had a statistically significant difference associated with GCA with ophthalmic disease compared to GCA without ophthalmic disease. Polymyalgia rheumatica (PMR) was identified in only 2.94% of the Hispanic subjects with GCA. Major conclusions in our study advocate Hispanics with GCA are prevalent and unique in its associations with other comorbid diseases. Unlike non-Hispanic White populations, Hispanic subjects with GCA do not show an association with PMR nor an increased association with stroke.

Attention and Memory in Depersonalization-Spectrum Dissociative Disorders: Impact of Selective-Divided Attentional Condition, Stimulus Emotionality, and Stress.

We investigated cognition in depersonalization-spectrum dissociative disorders without comorbid posttraumatic stress disorder to explore evidence for emotionally avoidant information processing. Forty-eight participants with DSM-IV dissociative disorder (DD) (Depersonalization Disorder - 37, Dissociative Disorder NOS -11), 36 participants with Posttraumatic Stress Disorder (PTSD), and 56 healthy controls (HC) were administered the Weschler Adult Intelligence Scale-III (WAIS); the Weschler Memory Scale-III (WMS); and three Stroop tasks: the Standard Stroop, a selective-attention Emotional Stroop using neutral, dissociation, and trauma-related word categories, and a divided-attention Emotional Stroop using comparable words. Participants were also administered a paired-associates explicit and implicit memory test using emotionally neutral and negative words, before and after the Trier Social Stress Test. The DD and HC groups had comparable general intelligence and memory scores, though dissociation severity was inversely related to verbal comprehension and working memory. In the selective-attention condition, DD participants showed greater incidental recall across word categories with comparable interference. However in the divided-attention condition, DD participants significantly favored lesser attentional interference at the expense of remembering words. Across attentional conditions, DD participants had better recall for disorder-related than neutral words. Pre-stress, the DD group demonstrated better explicit memory for neutral versus negative words with reversal after stress, whereas the HC group demonstrated the opposite pattern; implicit memory did not differ. Cognition in the PTSD control group was generally dissimilar to the DD group. The findings in toto provide substantial evidence for emotionally avoidant information processing in DD, vulnerable to the impact of stress, at the level of both attention and memory.

Methods for a composite ecological suitability measure to inform cumulative restoration assessments in Gulf of Mexico estuaries.

Ecosystem management requires a systematic, holistic approach that considers ecological and social outcomes. Effective restoration practices promote a balance of ecological and social goals by addressing ecological integrity, efficiently maximizing benefits while minimizing investment, and encompassing collaborative stakeholder engagement. Socio-ecological assessments can inform adaptive management and be utilized to prioritize restoration activities and monitor restoration effectiveness. In estuarine systems, socio-ecological assessments should evaluate the ability of habitats to support both ecologically and locally important species. The composite measure presented utilizes a combination of ecological and social measures to characterize ecological suitability for individual and multiple Gulf of Mexico estuarine species. The ecological suitability value (ES) for a given spatial unit is based on a suite of biophysical measures of the quality and extent of suitable habitat for each species, the species' trophic importance in a food web context, and the importance of each species in relation to stakeholder values and benefits. ES values for individual spatial units can be aggregated to estimate the distribution of ecological suitability at the estuarine scale. The ES values are calculated using examples for each step in the process. The information provided by ecological suitability characterizations can support restoration prioritization decisions for Gulf of Mexico estuaries and can provide a baseline measure to gauge restoration effectiveness over time to inform cumulative restoration assessments.

Tricolored bats at a southern range edge exhibit partial migration northward in autumn.

BACKGROUND: Animal migration is a widespread global adaptation by which individuals move in response to environmental conditions to reach more favorable conditions. For bats in temperate climates, migration and hibernation are often associated with each other when these bats must migrate to reach suitable overwintering sites. However, differences in movement across the geographical range of a species and the degree to which hibernation drives migratory behavior of bats in subtropical climates, where conditions may remain warm with available prey year-round, remains incomplete. Understanding the migratory strategies of subtropical bats during winter is of increasing importance as they are threatened by stressors such as disease and environmental change. METHODS: We evaluated migration patterns of tricolored bats (Perimyotis subflavus) in Florida, USA, through analysis of stable hydrogen isotope ratios of the fur. We inferred the summer geographic origins of the fur samples and estimated the minimum distance and likely direction traveled by hibernating individuals. We used linear models to examine whether hibernation region, colony size, and an individual's sex affected the distance traveled. RESULTS: We sampled 111 bats hibernating at 40 sites and found that more than half (54.1%) of individuals were residents of the area in which they hibernated. We found that almost half of the sampled bats (43.2%) traveled from southern Florida to overwinter in North Florida. We also documented three individuals that traveled > 100 km from northerly origins, one of which had traveled an estimated minimum distance of 1382 km. We also found that tricolored bats moved farther to reach hibernacula in Northwest Florida and hibernacula with more populous colonies, with no difference in movement between sexes. CONCLUSIONS: Our results indicate a pattern of northward autumnal movements of tricolored bats in the subtropical southeastern portion of their range. We suggest that bats are differentially constrained at the edge of their geographical range, resulting in movement contrary to what is expected. Even though we found that few (2.7%) bats moved into Florida from farther north, those migrants can potentially transfer the fungus that causes the deadly white-nose syndrome, which does not currently occur in the state. Our results support the characterization of tricolored bats as flexible partial migrators, with a rarely exercised capacity for long-distance movements.

A conceptual approach to characterizing ecological suitability: Informing socio-ecological measures for restoration effectiveness.

A combination of ecological and socio-economic outcome indicators is essential for understanding and assessing the effectiveness of the remediation and restoration of degraded ecosystems and revitalizing communities that could benefit from these ecosystem management activities. In this paper, we propose and develop a conceptual approach to characterize ecological suitability that incorporates ecological attributes that support ecosystem structural diversity and functionality, stakeholder values and perceptions, and the benefits derived from ecosystem goods and services. A structured literature review was used to identify existing restoration frameworks and indicators to inform the conceptual foundation for characterizing ecological suitability. The structure of the conceptual approach primarily builds from ecological and social attributes in the International Principles and Standards for the Practice of Ecological Restoration (Gann et al., 2019). We provide a conceptual example of the ecological suitability approach in estuaries. This example is based on habitat suitability and food web characterizations in combination with the provisioning of ecosystem services and desired social benefits to prioritize and evaluate restoration effectiveness. This foundational work sets the stage for developing a composite measure of ecological suitability. The holistic conceptual approach presented complements existing information regarding restoration effectiveness evaluations. Characterizing ecological suitability is a novel way to incorporate ecological and social information and communicate potential restoration outcomes to ecosystem managers and stakeholders.

A. thaliana Hybrids Develop Growth Abnormalities through Integration of Stress, Hormone and Growth Signaling.

Hybrids between Arabidopsis thaliana accessions are important in revealing the consequences of epistatic interactions in plants. F1 hybrids between the A. thaliana accessions displaying either defense or developmental phenotypes have been revealing the roles of the underlying epistatic genes. The interaction of two naturally occurring alleles of the OUTGROWTH-ASSOCIATED KINASE (OAK) gene in Sha and Lag2-2, previously shown to cause a similar phenotype in a different allelic combination in A. thaliana, was required for the hybrid phenotype. Outgrowth formation in the hybrids was associated with reduced levels of salicylic acid, jasmonic acid and abscisic acid in petioles and the application of these hormones mitigated the formation of the outgrowths. Moreover, different abiotic stresses were found to mitigate the outgrowth phenotype. The involvement of stress and hormone signaling in outgrowth formation was supported by a global transcriptome analysis, which additionally revealed that TCP1, a transcription factor known to regulate leaf growth and symmetry, was downregulated in the outgrowth tissue. These results demonstrate that a combination of natural alleles of OAK regulates growth and development through the integration of hormone and stress signals and highlight the importance of natural variation as a resource to discover the function of gene variants that are not present in the most studied accessions of A. thaliana.

Activity-based protein profiling reveals deubiquitinase and aldehyde dehydrogenase targets of a cyanopyrrolidine probe.

Ubiquitin carboxy-terminal hydrolase L1 (UCHL1), a deubiquitinating enzyme (DUB), is a potential drug target in various cancers, and liver and lung fibrosis. However, bona fide functions and substrates of UCHL1 remain poorly understood. Herein, we report the characterization of UCHL1 covalent inhibitor MT16-001 based on a thiazole cyanopyrrolidine scaffold. In combination with chemical proteomics, a closely related activity-based probe (MT16-205) was used to generate a comprehensive quantitative profile for on- and off-targets at endogenous cellular abundance. Both compounds are selective for UCHL1 over other DUBs in intact cells but also engage a range of other targets with good selectivity over the wider proteome, including aldehyde dehydrogenases, redox-sensitive Parkinson's disease related protein PARK7, and glutamine amidotransferase. Taken together, these results underline the importance of robust profiling of activity-based probes as chemical tools and highlight the cyanopyrrolidine warhead as a versatile platform for liganding diverse classes of protein with reactive cysteine residues which can be used for further inhibitor screening, and as a starting point for inhibitor development.

The genetic and epigenetic landscape of the Arabidopsis centromeres.

Centromeres attach chromosomes to spindle microtubules during cell division and, despite this conserved role, show paradoxically rapid evolution and are typified by complex repeats. We used long-read sequencing to generate the Col-CEN Arabidopsis thaliana genome assembly that resolves all five centromeres. The centromeres consist of megabase-scale tandemly repeated satellite arrays, which support CENTROMERE SPECIFIC HISTONE H3 (CENH3) occupancy and are densely DNA methylated, with satellite variants private to each chromosome. CENH3 preferentially occupies satellites that show the least amount of divergence and occur in higher-order repeats. The centromeres are invaded by ATHILA retrotransposons, which disrupt genetic and epigenetic organization. Centromeric crossover recombination is suppressed, yet low levels of meiotic DNA double-strand breaks occur that are regulated by DNA methylation. We propose that Arabidopsis centromeres are evolving through cycles of satellite homogenization and retrotransposon-driven diversification.

Multiplexed Simian Immunodeficiency Virus-Specific Paired RNA-Guided Cas9 Nickases Inactivate Proviral DNA.

Human and simian immunodeficiency virus (HIV and SIV) infections establish lifelong reservoirs of cells harboring an integrated proviral genome. Genome editing CRISPR-associated Cas9 nucleases, combined with SIV-specific guiding RNA (gRNA) molecules, inactivate integrated provirus DNA in vitro and in animal models. We generated RNA-guided Cas9 nucleases (RGNu) and nickases (RGNi) targeting conserved SIV regions with no homology in the human or rhesus macaque genome. Assays in cells cotransfected with SIV provirus and plasmids coding for RGNus identified SIV long terminal repeat (LTR), trans-activation response (TAR) element, and ribosome slip site (RSS) regions as the most effective at virus suppression; RGNi targeting these regions inhibited virus production significantly. Multiplex plasmids that coexpressed these three RGNu (Nu3), or six (three pairs) RGNi (Ni6), were more efficient at virus suppression than any combination of individual RGNu and RGNi plasmids. Both Nu3 and Ni6 plasmids were tested in lymphoid cells chronically infected with SIVmac239, and whole-genome sequencing was used to determine on- and off-target mutations. Treatment with these all-in-one plasmids resulted in similar levels of mutations of viral sequences from the cellular genome; Nu3 induced indels at the 3 SIV-specific sites, whereas for Ni6 indels were present at the LTR and TAR sites. Levels of off-target effects detected by two different algorithms were indistinguishable from background mutations. In summary, we demonstrate that Cas9 nickase in association with gRNA pairs can specifically eliminate parts of the integrated provirus DNA; also, we show that careful design of an all-in-one plasmid coding for 3 gRNAs and Cas9 nuclease inhibits SIV production with undetectable off-target mutations, making these tools a desirable prospect for moving into animal studies. IMPORTANCE Our approach to HIV cure, utilizing the translatable SIV/rhesus macaque model system, aims at provirus inactivation and its removal with the least possible off-target side effects. We developed single molecules that delivered either three truncated SIV-specific gRNAs along with Cas9 nuclease or three pairs of SIV-specific gRNAs (six individual gRNAs) along with Cas9 nickase to enhance efficacy of on-target mutagenesis. Whole-genome sequencing demonstrated effective SIV sequence mutation and inactivation and the absence of demonstrable off-target mutations. These results open the possibility to employ Cas9 variants that introduce single-strand DNA breaks to eliminate integrated proviral DNA.

Novel allele elh of the UBP14 gene affects plant organ size via cell expansion in Arabidopsis thaliana.

Plant organ size control is an essential process of plant growth and development. The regulation of plant organ size involves a complicated network of genetic, molecular interactions, as well as the interplay of environmental factors. Here, we report a temperature-sensitive hypocotyl elongation EMS-generated mutant, hereby referred to as elongated hypocotyl under high-temperature (elh). The elongated hypocotyl phenotype was prominent when the elh seedlings were grown at high temperature, 28°C, but not under the growth temperature of 21°C. We observed significantly larger organ sizes in elh plants, including cotyledons, petals and seeds. In elh plants, the cell sizes in cotyledons and petals were significantly larger than wild type. By measuring the cell density and organ area of cotyledons, petals and mature dissected embryos, we found no differences in total cell numbers in any organ indicating that cell expansion rather than cell proliferation was perturbed in elh. elh plants produced leaves at a slower rate than wild type plants, suggesting that perturbing the balance between cell division and cell expansion is linked to the developmental rate at which leaves are produced.

Investigating the relationship between environmental quality, socio-spatial segregation and the social dimension of sustainability in US urban areas.

This work is intended to assess the relationship between the three dimensions of sustainability (environmental, economic, and social) and residential segregation broadly across US urban areas. Multivariate indices of segregation and sustainability are created using data aggregated to 933 US Census Core-Based Statistical Areas (CBSAs). The relationship between segregation and sustainability (and dimensions, independently) is analyzed across the CBSAs using correlation and spatial analyses. Results show an inverse relationship between segregation and sustainability, where increasing urban segregation corresponds to decreases in sustainability. Social and economic dimensions of sustainability are negatively correlated with segregation while the environmental dimension is positively correlated. Spatial analysis reveals regional associations between the measures with most of the lower sustainability scores and higher segregation scores occuring mainly in the South. The findings demonstrate an inverse relationship between segregation and sustainability as well as between the environmental and social dimensions. These correlations indicate a need to consider inequalities, and the social dimension of sustainability more broadly, when planning urban revitalization efforts. Spatial scale is also imperative to understanding and improving the social outcomes for all members of the community.

Silencing integrated SIV proviral DNA with TAR-specific CRISPR tools.

BACKGROUND: One approach for a functional HIV cure is to prevent transcription from integrated proviral DNA. A critical step in HIV transcription is the Tat protein interaction with the TAR element viral RNA. We tested the strategy of blocking this Tat-TAR interaction in the SIVmac model. METHODS: We designed five CRISPR short guiding RNAs (sgRNAs) targeting the SIVmac TAR element, along with inactive versions of Cas9 (dCas9). These sgRNA constructs were delivered as ribonucleoproteins or plasmid DNA, along with SIV DNA. The constructs were also tested in integrated viral DNA in a cell line chronically infected by SIV. RESULTS: The sgRNAs targeting the coding strand of the TAR element inhibited SIV RNA transcription in association with dCas9-KRAB, but not with dCas9. CONCLUSIONS: Induction of epigenetic modifications may be more effective in inactivating provirus than transcriptional interference and thus may be a better strategy to achieve a functional cure in vivo.

Discovery of a Potent and Selective Covalent Inhibitor and Activity-Based Probe for the Deubiquitylating Enzyme UCHL1, with Antifibrotic Activity.

Ubiquitin carboxy-terminal hydrolase L1 (UCHL1) is a deubiquitylating enzyme that is proposed as a potential therapeutic target in neurodegeneration, cancer, and liver and lung fibrosis. Herein we report the discovery of the most potent and selective UCHL1 probe (IMP-1710) to date based on a covalent inhibitor scaffold and apply this probe to identify and quantify target proteins in intact human cells. IMP-1710 stereoselectively labels the catalytic cysteine of UCHL1 at low nanomolar concentration in cells. We further demonstrate that potent and selective UCHL1 inhibitors block pro-fibrotic responses in a cellular model of idiopathic pulmonary fibrosis, supporting the potential of UCHL1 as a potential therapeutic target in fibrotic diseases.

A malectin domain kinesin functions in pollen and seed development in Arabidopsis.

The kinesin family is greatly expanded in plants compared with animals and, with more than a third up-regulated in expression during cell division, it has been suggested that this expansion facilitated complex plant-specific cytoskeletal rearrangements. The cell cycle-regulated kinesins include two with an N-terminal malectin domain, a protein domain that has been shown to bind polysaccharides and peptides when found extracellularly in receptor-like kinases. Although malectin domain kinesins are evolutionarily deep rooted, their function in plants remains unclear. Here we show that loss of MALECTIN DOMAIN KINESIN 2 (MDKIN2) results in stochastic developmental defects in pollen, embryo, and endosperm. High rates of seed abnormalities and abortion occur in mdkin2 mutants through a partial maternal effect. No additive effect or additional developmental defects were noted in mdkin1 mdkin2 double mutants. MDKIN2 is expressed in regions of cell division throughout the plant. Subcellular localization of MDKIN2 indicates a role in cell division, with a possible secondary function in the nuclei. Our results reveal a non-essential but important role for a malectin domain kinesin during development in plants.

CrRLK1L receptor-like kinases HERK1 and ANJEA are female determinants of pollen tube reception.

Communication between the gametophytes is vital for angiosperm fertilisation. Multiple CrRLK1L-type receptor kinases prevent premature pollen tube burst, while another CrRLK1L protein, FERONIA (FER), is required for pollen tube reception in the female gametophyte. We report here the identification of two additional CrRLK1L homologues, HERCULES RECEPTOR KINASE 1 (HERK1) and ANJEA (ANJ), which act redundantly to promote pollen tube growth arrest at the synergid cells. HERK1 and ANJ localise to the filiform apparatus of the synergid cells in unfertilised ovules, and in herk1 anj mutants, a majority of ovules remain unfertilised due to pollen tube overgrowth, together indicating that HERK1 and ANJ act as female determinants for fertilisation. As in fer mutants, the synergid cell-specific, endomembrane protein NORTIA (NTA) is not relocalised after pollen tube reception; however, unlike fer mutants, reactive oxygen species levels are unaffected in herk1 anj double mutants. Both ANJ and HERK1 associate with FER and its proposed co-receptor LORELEI (LRE) in planta. Together, our data indicate that HERK1 and ANJ act with FER to mediate female-male gametophyte interactions during plant fertilisation.