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When data are sparse and/or predictors multicollinear, current implementation of sparse partial least squares (SPLS) does not give estimates for non-selected predictors nor provide a measure of inference. In response, an approach termed all-possible SPLS is proposed, which fits a SPLS model for all tuning parameter values across a set grid. Noted is the percentage of time a given predictor is chosen, as well as the average non-zero parameter estimate. Using a large number of multicollinear predictors, simulation confirmed variables not associated with the outcome were least likely to be chosen as sparsity increased across the grid of tuning parameters, while the opposite was true for those strongly associated. Lastly, variables with a weak association were chosen more often than those with no association, but less often than those with a strong relationship to the outcome. Similarly, predictors most strongly related to the outcome had the largest average parameter estimate magnitude, followed by those with a weak relationship, followed by those with no relationship. Across two independent studies regarding the relationship between volumetric MRI measures and a cognitive test score, this method confirmed a priori hypotheses about which brain regions would be selected most often and have the largest average parameter estimates. In conclusion, the percentage of time a predictor is chosen is a useful measure for ordering the strength of the relationship between the independent and dependent variables, serving as a form of inference. The average parameter estimates give further insight regarding the direction and strength of association. As a result, all-possible SPLS gives more information than the dichotomous output of traditional SPLS, making it useful when undertaking data exploration and hypothesis generation for a large number of potential predictors.

Objective: To compare: (1) the load and diversity of cultivatable bacterial species isolated from tissue biopsies with cultures from surface swabs, and (2) the ability of each technique to detect methicillin-resistant Staphylococcus aureus (MRSA) in a model of MRSA-infected equine wounds., Study Design: Experimental in vivo study., Animals: Three light-breed adult horses., Methods: Four 2.5 × 2.5 cm full-thickness skin wounds were created on the dorsolateral aspect of each forelimb. Five days later, each wound was inoculated with a pure culture of MRSA (ATCC 43300). One hundred microlitres of 0, 5 × 10 8 , 5 × 10 9 or 5 × 10 10 colony forming units (CFU)/ml was used to inoculate each wound. Surface swabs (Levine technique) and tissue biopsy samples (3 mm punch biopsy) were obtained at 2, 7, 14, and 21 days after inoculation. Quantitative aerobic culture was performed using routine clinical techniques., Results: A similar bacterial profile was identified from the culture of each wound-sampling technique and there was moderate correlation (R = 0.49, P < .001) between the bacterial bioburdens. Agreement was fair (κ = 0.31; 95% CI, 0.129-0.505) between the sampling techniques in identification of MRSA. Methicillin-resistant Staphylococcus aureus was isolated more frequently (P = .016) from cultures of tissue biopsies (79%; 76/96) than from surface swabs (62%; 60/96)., Conclusion: Bacterial load and diversity did not differ between sampling techniques but MRSA was detected more often from the cultures of tissue biopsies., Clinical Significance: Tissue biopsy should be preferred to culture swab in wounds where MRSA is suspected., (© 2022 American College of Veterinary Surgeons.)

Racial and ethnic discrimination persist in science, technology, engineering and mathematics fields, including ecology, evolution and conservation biology (EECB) and related disciplines. Marginalization and oppression as a result of institutional and structural racism continue to create barriers to inclusion for Black people, Indigenous people and people of colour (BIPOC), and remnants of historic racist policies and pseudoscientific theories continue to plague these fields. Many academic EECB departments seek concrete ways to improve the climate and implement anti-racist policies in their teaching, training and research activities. We present a toolkit of evidence-based interventions for academic EECB departments to foster anti-racism in three areas: in the classroom; within research laboratories; and department wide. To spark restorative discussion and action in these areas, we summarize EECB's racist and ethnocentric histories, as well as current systemic problems that marginalize non-white groups. Finally, we present ways that EECB departments can collectively address shortcomings in equity and inclusion by implementing anti-racism, and provide a positive model for other departments and disciplines., (© 2021. Springer Nature Limited.)

Background: Vulvodynia and pudendal neuralgia comprise significant contributors to vulvar-related pain and its impact on daily life., Aim: A retrospective clinical audit was conducted at the Women's Health & Research Institute of Australia, Sydney, to determine the pattern of use and the efficacy of the application of topical amitriptyline 0.5% plus oestriol 0.03% in organogel (AOO), to the vulvar vestibule in reducing the impact of pain on daily life., Materials and Methods: There were 1174 patients who received a script from May 2017 until February 2020: 1054 patients agreed to be contacted and had a valid email address., Results: There were 376 (35.7%) patients who replied. Pain with intercourse was the main indication for use. Treatment was rated effective by 51.2% (95% CI: 35.4-66.8%) of patients less than 30 years of age, 66.7% (95% CI: 57.3-74.9%) of patients 30-50 years of age, and 58.3% (95% CI: 50.9-65.4%) in patients over 50. Stinging at the site of application was the most commonly reported side effect., Conclusion: Topical AOO is an effective and well-tolerated treatment for vulvar pain., (© 2021 The Authors. Australian and New Zealand Journal of Obstetrics and Gynaecology published by John Wiley & Sons Australia, Ltd on behalf of Royal Australian and New Zealand College of Obstetricians and Gynaecologists.)

Many therapeutically important enzymes are present in multiple cellular compartments, where they can carry out markedly different functions; thus, there is a need for pharmacological strategies to selectively manipulate distinct pools of target enzymes. Insulin-degrading enzyme (IDE) is a thiol-sensitive zinc-metallopeptidase that hydrolyzes diverse peptide substrates in both the cytosol and the extracellular space, but current genetic and pharmacological approaches are incapable of selectively inhibiting the protease in specific subcellular compartments. Here, we describe the discovery, characterization, and kinetics-based optimization of potent benzoisothiazolone-based inhibitors that, by virtue of a unique quasi-irreversible mode of inhibition, exclusively inhibit extracellular IDE. The mechanism of inhibition involves nucleophilic attack by a specific active-site thiol of the enzyme on the inhibitors, which bear an isothiazolone ring that undergoes irreversible ring opening with the formation of a disulfide bond. Notably, binding of the inhibitors is reversible under reducing conditions, thus restricting inhibition to IDE present in the extracellular space. The identified inhibitors are highly potent (IC50(app) = 63 nM), nontoxic at concentrations up to 100 μM, and appear to preferentially target a specific cysteine residue within IDE. These novel inhibitors represent powerful new tools for clarifying the physiological and pathophysiological roles of this poorly understood protease, and their unusual mechanism of action should be applicable to other therapeutic targets.

Effective discharge planning can contribute significantly to preventing homelessness. As part of a larger continuum of care, this process can help people reach goals of stable housing, recovery, and increased quality of life in the community. Discharge planning identifies and organizes services a person with mental illness, substance abuse, and other vulnerabilities needs when leaving an institutional or custodial setting and returning to the community. A three-component model is presented, relating discharge planning to institutional assessment and treatment, and to community services. Service challenges, innovative programs (based on a state-by-state analysis), and recommendations for enhancing discharge planning also are discussed.

Cognitive impairments can increase significantly a person's level of risk for becoming or remaining homeless. Five topics are explored that address these impairments in preventing homelessness: (1) the nature and estimated prevalence of cognitive impairments among people who are homeless, especially those with vulnerabilities like mental illness or substance abuse; (2) the multiple origins of these impairments; (3) how these impairments impact services for people at risk for homelessness; (4) good practice approaches to handling cognitive impairments in homeless shelters, supported housing programs, and other service systems for people at risk for homelessness; and, (5) important research and practice issues requiring further action.

Land use has generally been considered a local environmental issue, but it is becoming a force of global importance. Worldwide changes to forests, farmlands, waterways, and air are being driven by the need to provide food, fiber, water, and shelter to more than six billion people. Global croplands, pastures, plantations, and urban areas have expanded in recent decades, accompanied by large increases in energy, water, and fertilizer consumption, along with considerable losses of biodiversity. Such changes in land use have enabled humans to appropriate an increasing share of the planet's resources, but they also potentially undermine the capacity of ecosystems to sustain food production, maintain freshwater and forest resources, regulate climate and air quality, and ameliorate infectious diseases. We face the challenge of managing trade-offs between immediate human needs and maintaining the capacity of the biosphere to provide goods and services in the long term.

Here we show that the VirD2 protein of A. tumefaciens functions as a nuclear localizing protein in plant cells. The nuclear localization signal of VirD2 consists of two regions containing 4-5 basic amino acids (KRPR and RKRER), located within the C-terminal 34 amino acids. These regions conform to the KR/KXR/K motif required for numerous nuclear localized nonplant eukaryotic proteins. Each region independently directs a beta-glucuronidase reporter protein to the nucleus; however, both regions are necessary for maximum efficiency. VirD2 has been shown to be tightly bound to the 5' end of the single-stranded DNA transfer intermediate, T-strand, transferred from Agrobacterium to the plant cell genome. The present results imply that T-strand transport to the plant nucleus is mediated by the tightly attached VirD2 protein via an import pathway common to higher eukaryotes.

We analyzed the extent, reproducibility, and developmental control of genomic rearrangements in the somatic macronucleus of the ciliate Tetrahymena thermophila. To exclude differences caused by genetic polymorphisms, we constructed whole-genome homozygotes, and we compared the homozygous progeny derived from single macronuclear differentiation events. This strategy enabled us to identify a novel form of variable rearrangement and to confirm previous findings that rearranged sequences occur at a high frequency in the Tetrahymena genome. Rearrangements studied here were deletions of both unique and interchromosomally dispersed repetitive DNA sequences involving DNA rejoining of internal, nontelomeric regions of macronuclear DNAs. We showed that although rearrangements of some sequence classes are reproducible among independently developed macronuclei, other specific sequence classes are variably rearranged in macronuclear development. The variable somatic genomes so produced may be the source of phenotypically variant cell lines.

The T-DNA transfer process of Agrobacterium is activated following induction of expression of the Ti plasmid virulence (vir) genes. The virD1 and virD2 gene products are required for the production of nicks at the T-DNA borders and for the generation of free linear single-stranded copies of the T-DNA region, T-strands. T-strands are complexed with proteins in vir-induced bacteria, since T-strands partition to the aqueous/phenol interface in non-Pronasetreated total cell extracts. To determine whether the proteins are tightly associated with T-strands, DNA-protein complexes were purified away from bulk proteins by adsorption to glass beads. A 58-kDa protein was specifically released from vir-induced DNA-protein complexes after treatment with S1 nuclease to digest single-stranded DNA. The 58-kDa protein was identified as VirD2 by using VirD2-specific antibodies. The tight association of VirD2 with T-strands was shown directly by using VirD2-specific antibody to isolate T-strands. The 5' side of the border nick sites on the Ti plasmid was also shown to be tightly associated with protein. The data suggest that after VirD1/VirD2-dependent nicking at the T-DNA borders, the VirD2 protein remains bound to the 5' end of the nick, and the VirD2 protein continues to bind tightly to this 5' end during unwinding (or displacement) of the T-strand from the Ti plasmid T-DNA region. The tight binding of VirD2 to T-strands suggests that this protein has additional functions in T-strand generation and potentially in the later steps of T-DNA transfer.

Transient-expression analysis has shown anaerobic regulation of the alcohol dehydrogenase (Adh1) promoter in a chimeric construct. Chimeric plasmids containing the promoter for the Adh1 gene of maize (Zea mays L.) linked to the coding sequence of the chloramphenicol acetyl transferase (cat) gene were introduced into maize protoplasts using electroporation. Both the introduced Adh1 promoter and the endogenous Adh1 gene promoter are regulated at the RNA level, the O2-tension optimum for induction is the same for both, and both promoters initiate transcription from the same site. The demonstration of regulation of a plant gene promoter in a transient expression system will allow the identification of cis acting sequences responsible for regulation.

Expression of the maize alcohol dehydrogenase 1 (Adh1) gene is transcriptionally regulated under conditions of anaerobic stress. DNA sequences required for the expression of Adh1 have been identified by a functional analysis of in vitro constructed hybrid genes consisting of the Adh1 promoter fused to the chloramphenicol acetyltransferase coding region. A series of 5' deletions, 3' deletions, hybrid promoters, and linker scanning mutants of the Adh-CAT hybrid gene were introduced into maize protoplasts by electroporation and assayed for chloramphenicol acetyltransferase activity after incubation of the protoplasts under different oxygen tensions. The results indicate that a 40-base-pair DNA sequence within the Adh1 promoter is required for anaerobically regulated expression of the hybrid gene. Clustered point mutations in this sequence show that it is composed of two essential regions, each approximately 15 base pairs, separated by a 10-base-pair DNA sequence that does not appear to be important for anaerobic expression. Attachment of this 40-base-pair element to an unrelated promoter shows that this DNA sequence is both necessary and sufficient for induction of gene expression by low oxygen stress.