Visible-light-mediated Diels-Alder reactions under single-chain polymer confinement: investigating the role of the crosslinking moiety on catalyst activity.

Macromolecular scaffolds are rapidly emerging in catalysis owing to the ability to control catalyst placement at precise locations. This spatial proximity allows for enhanced catalyst activity that may not be observed using small molecules. Herein, we describe a triphenylpyrylium (TPT)-based visible-light active single-chain polymer nanoparticle (SCNP) that facilitates the radical cation [4 + 2]-cycloaddition. We find that the catalytic activity is highly dependent on the styrylarene comonomer used, wherein it can act as a redox mediator under confinement, increasing the catalytic turnover (TON) by up to 30 times in comparison to free TPT in solution. Mechanistic studies indicate that TPT excited states are quenched by the acene, with the resultant radical cation formed from naphthalene-based SCNPs able to proceed in oxidizing the dienophile in the elementary step of the reaction, while leading to near quantitative yields of the cycloadduct. The TPT-SCNP demonstrates enhanced photocatalyst efficiency compared to molecular TPT, and is able to be recycled and reused in three iterations of the reaction prior to decreased performance from photobleaching. Our results overall suggest that the confined nature of the SCNP and spatial proximity of acene-based pendants enforces their participation as cocatalytic redox mediators that impart enhanced photoredox catalysis under confinement.

Cross-Cultural Adaptation of the Scale of the Nursing Work Index With Nurses From Seven Countries.

Background and Purpose: The aims of this study are as follows: to adapt and validate the psychometric properties of the Spanish version of the Practice Environment Scale of the Nursing Work Index (PES-NWI) adapted to the nursing home environment across seven Spanish-speaking countries and to cross-culturally adapt the Scale of the Nursing Work Index with nurses from seven countries. Methods: Adaptation process and psychometric validation of the instrument included translation and back-translation, content validity, test-retest reliability, internal consistency, and construct validity. Results: A total of 134 nursing homes belonging to the same religious order were randomly selected from seven Spanish-speaking countries with a sample of 378 nurses. The exploratory factor analysis explained a five-factor structure (56% of the explained variance) with adequate goodness-of-fit indices in the final factor solution. Conclusions: The validation process indicates that the Spanish language version of the PES-NWI with five factors and 31 items, for long-term care facilities for the elderly, is valid and reliable in its current version and can be used to measure the environment of nurses working in clinical practice in Spanish-speaking nursing homes.

Unearthing a Cryptic Biosynthetic Gene Cluster for the Piperazic Acid-Bearing Depsipeptide Diperamycin in the Ant-Dweller Streptomyces sp. CS113.

Piperazic acid is a cyclic nonproteinogenic amino acid that contains a hydrazine N-N bond formed by a piperazate synthase (KtzT-like). This amino acid, found in bioactive natural products synthesized by non-ribosomal peptide synthetases (NRPSs), confers conformational constraint to peptides, an important feature for their biological activities. Genome mining of Streptomyces strains has been revealed as a strategy to identify biosynthetic gene clusters (BGCs) for potentially active compounds. Moreover, the isolation of new strains from underexplored habitats or associated with other organisms has allowed to uncover new BGCs for unknown compounds. The in-house "Carlos Sialer (CS)" strain collection consists of seventy-one Streptomyces strains isolated from the cuticle of leaf-cutting ants of the tribe Attini. Genomes from twelve of these strains have been sequenced and mined using bioinformatics tools, highlighting their potential to encode secondary metabolites. In this work, we have screened in silico those genomes, using KtzT as a hook to identify BGCs encoding piperazic acid-containing compounds. This resulted in uncovering the new BGC dpn in Streptomyces sp. CS113, which encodes the biosynthesis of the hybrid polyketide-depsipeptide diperamycin. Analysis of the diperamycin polyketide synthase (PKS) and NRPS reveals their functional similarity to those from the aurantimycin A biosynthetic pathway. Experimental proof linking the dpn BGC to its encoded compound was achieved by determining the growth conditions for the expression of the cluster and by inactivating the NRPS encoding gene dpnS2 and the piperazate synthase gene dpnZ. The identity of diperamycin was confirmed by High-Resolution Mass Spectrometry (HRMS) and Nuclear Magnetic Resonance (NMR) and by analysis of the domain composition of modules from the DpnP PKS and DpnS NRPS. The identification of the dpn BGC expands the number of BGCs that have been confirmed to encode the relatively scarcely represented BGCs for depsipeptides of the azinothricin family of compounds and will facilitate the generation of new-to-nature analogues by combinatorial biosynthesis.

A yeast-based high-throughput screen identifies inhibitors of trypanosomatid HRG heme transporters with potent leishmanicidal and trypanocidal activity.

OBJECTIVES: New drugs are required to treat neglected diseases caused by trypanosomatid parasites such as Leishmania, Trypanosoma brucei and Trypanosoma cruzi. An Achilles' heel of these parasites is their heme auxotrophy; they have an absolute dependence on scavenging this molecule from the host, and trypanosomatid HRG heme transporters (TrypHRG) play an important role in this process. As these proteins are essential for the parasites and have low similarity with their human orthologue, they have been proposed as attractive therapeutic targets. Here, we have developed two yeast-based assays that allow an inexpensive high-throughput screening of TrypHRG inhibitors within a cellular context. METHODS: We first assessed that Leishmania major, Leishmania donovani and T. brucei HRG proteins were heterologously expressed in the digestive vacuole membrane of a mutant heme auxotrophic yeast strain. Here, TrypHRG imports hemoglobinderived heme into the cytosol, allowing mutant yeast to grow in the presence of low hemoglobin concentrations and promoting the activity of hemeproteins such as catalase, which was used as a reporter of cytosolic heme levels. RESULTS: In the presence of a TrypHRG inhibitor, both catalase activity (test 1) and yeast growth (test 2) were diminished, being easily monitored. The assays were then tested on a pilot scale for HTS purposes using a collection of repurposing drugs and food antioxidants. Some of the TrypHRG inhibitors identified in yeast presented strong trypanocidal and leishmanicidal activity in the submicromolar range, proving the potential of this approach. CONCLUSIONS: Cumulatively, it was shown that the inhibition bioassays developed were robust and applicable to large-scale HTS.

FiCRoN, a deep learning-based algorithm for the automatic determination of intracellular parasite burden from fluorescence microscopy images.

Protozoan parasites are responsible for dramatic, neglected diseases. The automatic determination of intracellular parasite burden from fluorescence microscopy images is a challenging problem. Recent advances in deep learning are transforming this process, however, high-performance algorithms have not been developed. The limitations in image acquisition, especially for intracellular parasites, make this process complex. For this reason, traditional image-processing methods are not easily transferred between different datasets and segmentation-based strategies do not have a high performance. Here, we propose a novel method FiCRoN, based on fully convolutional regression networks (FCRNs), as a promising new tool for estimating intracellular parasite burden. This estimation requires three values, intracellular parasites, infected cells and uninfected cells. FiCRoN solves this problem as multi-task learning: counting by regression at two scales, a smaller one for intracellular parasites and a larger one for host cells. It does not use segmentation or detection, resulting in a higher generalization of counting tasks and, therefore, a decrease in error propagation. Linear regression reveals an excellent correlation coefficient between manual and automatic methods. FiCRoN is an innovative freedom-respecting image analysis software based on deep learning, designed to provide a fast and accurate quantification of parasite burden, also potentially useful as a single-cell counter.

Low Energy Availability and Increased Risk of Relative Energy Deficiency in Sport (RED-S) During a 3767-km Thru-Hike on the Pacific Crest Trail: A Case Study.

Long-distance "thru-hiking" has extraordinary physical demands and has become increasingly popular. This report describes a man (55 y) who thru-hiked the Pacific Crest Trail in 2021 and was at risk of developing the relative energy deficiency in sport (RED-S) syndrome. Hiking distance was 3767 km over 128 d. Eighty-eight days (69%) were full days of hiking, covering 38±8 km/d (mean±SD) in 7.9±1.6 h/d. Exercise energy expenditure above rest (heart rate vs indirect calorimetry regression method) was 2834±1518 kcal/d, total energy expenditure was 5702±1323 kcal/d, and energy intake was 4141 kcal/d. Body mass decreased by 9%, and fat mass (dual-energy X-ray absorptiometry) decreased by 46%. Energy availability (energy intake minus exercise energy expenditure) was 19.3 kcal/d/kg fat-free mass, indicating low energy availability (defined as <30 kcal/d/kg). Dual-energy X-ray absorptiometry-measured spine bone mineral density (BMD) decreased by 8.6%, with little to no decrease in total hip (-1.0%) and femoral neck (-1.5%) BMD. Total cholesterol, low-density lipoprotein cholesterol, and triglycerides increased by 24, 39, and 57%, respectively. Within 8 mo after the hike, BMD and serum lipids nearly or fully returned to baseline. No changes in high-density lipoprotein cholesterol, glycemia, or blood pressure were observed. According to guidelines, these observations are consistent with a moderate risk of RED-S, and a medical evaluation and treatment plan are advisable in order to avoid clinical manifestations (eg, stress fractures, anemia, psychological disturbances). To minimize RED-S risk, it may be prudent for thru-hikers to optimize energy availability by moderating daily hiking distances and/or increasing food intake.

Delftia tsuruhatensis TC1 symbiont suppresses malaria transmission by anopheline mosquitoes.

Malaria control demands the development of a wide range of complementary strategies. We describe the properties of a naturally occurring, non-genetically modified symbiotic bacterium, Delftia tsuruhatensis TC1, which was isolated from mosquitoes incapable of sustaining the development of Plasmodium falciparum parasites. D. tsuruhatensis TC1 inhibits early stages of Plasmodium development and subsequent transmission by the Anopheles mosquito through secretion of a small-molecule inhibitor. We have identified this inhibitor to be the hydrophobic molecule harmane. We also found that, on mosquito contact, harmane penetrates the cuticle, inhibiting Plasmodium development. D. tsuruhatensis TC1 stably populates the mosquito gut, does not impose a fitness cost on the mosquito, and inhibits Plasmodium development for the mosquito's life. Contained field studies in Burkina Faso and modeling showed that D. tsuruhatensis TC1 has the potential to complement mosquito-targeted malaria transmission control.

Mpox Virus: Control of In-Hospital Occupational Transmission Experience from a Tertiary Level Hospital in Milan, Italy.

Mpox has caused a global outbreak since May 2022, particularly affecting people belonging to key populations, but cases among healthcare providers have been reported. The aim of this work is to present the experience of the Infectious Diseases Unit of San Raffaele Scientific Institute, Milan, Italy with respect to infection control and prevention of mpox occupational transmission. Between May-November 2022, 140 individuals were diagnosed with mpox and six required hospitalization. Overall, 12 medical doctors and 22 nurses provided care to people with mpox. A hospital policy aimed at controlling viral transmission was implemented in May 2022. Protective equipment was used for all healthcare providers. One accidental puncture occurred with a scalpel contaminated with blood from a mpox viremic individual (mpox plasma cycle threshold = 36); no mpox related symptoms were observed and mpox testing ruled out transmission. Six months following exposure, neutralizing antibodies were not detectable, ruling out contagion. Overall, we observed no mpox transmission among healthcare workers, despite the number of visits and procedures performed, including bodily-fluids sampling, and even following puncture with contaminated blood. Hospital preparedness for the management of new infectious disease outbreaks, with rapid implementation of policies aimed at controlling infection, is paramount to avoid occupational transmission.

Finding fraud: enforcement, detection, and recoveries after the ACA.

Medicaid Fraud Control Units investigate and prosecute acts of financial fraud and patient abuse within the program. Prior to the expansion of Medicaid under the Affordable Care Act (ACA), federal government MFCU expenditures totaled half a percent of Medicaid expenditures. Following the enrollment of 12 million adults into the Medicaid program under the ACA, expenditures for these units are now less than pre-ACA levels, as a share of program expenses. We use data for states' fraud enforcement efforts in the period 2010-2018 and a difference-in-differences design that exploits states' decision to expand Medicaid under the ACA. States that did expand Medicaid increased their fraud investigations, compared to states that did not expand. Further, civil recoveries and excluded individuals increased after the Medicaid expansion. We find evidence that increases in program scale, in terms of enrollment and utilization, reverted to the mean, facilitating the identification of outlier provider behavior.

Uncovering the Cryptic Gene Cluster ahb for 3-amino-4-hydroxybenzoate Derived Ahbamycins, by Searching SARP Regulator Encoding Genes in the Streptomyces argillaceus Genome.

Genome mining using standard bioinformatics tools has allowed for the uncovering of hidden biosynthesis gene clusters for specialized metabolites in Streptomyces genomes. In this work, we have used an alternative approach consisting in seeking "Streptomyces Antibiotic Regulatory Proteins" (SARP) encoding genes and analyzing their surrounding DNA region to unearth cryptic gene clusters that cannot be identified using standard bioinformatics tools. This strategy has allowed the unveiling of the new ahb cluster in Streptomyces argillaceus, which had not been retrieved before using antiSMASH. The ahb cluster is highly preserved in other Streptomyces strains, which suggests a role for their encoding compounds in specific environmental conditions. By combining overexpression of three regulatory genes and generation of different mutants, we were able to activate the ahb cluster, and to identify and chemically characterize the encoded compounds that we have named ahbamycins (AHBs). These constitute a new family of metabolites derived from 3-amino-4-hydroxybenzoate (3,4-AHBA) known for having antibiotic and antitumor activity. Additionally, by overexpressing three genes of the cluster (ahbH, ahbI, and ahbL2) for the synthesis and activation of 3,4-AHBA, a new hybrid compound, AHB18, was identified which had been produced from a metabolic crosstalk between the AHB and the argimycin P pathways. The identification of this new BGC opens the possibility to generate new compounds by combinatorial biosynthesis.

The Questionnaire of Intention to Help in VAW Cases (QIHVC): Development and preliminary results.

Introduction: Violence against women (VAW) is a worldwide social and health problem of epidemic proportions. This violence is preventable, and bystander programs are one of the possible preventative strategies. The main purpose of this research was to develop a tool that, by applying a contrastive methodology for its application in different forms of violence (forms of gender-based violence, such as intimate partner VAW, sexual harassment at work, and street harassment, and common violence, such as a robbery), would allow measuring the probability of occurrence of bystander response in the face of these types of violence with good evidence of content validity. Method: Firstly (Study 1), an initial version of a measure tool, the Questionnaire of Intention to Help in VAW Cases (QIHVC), was developed; secondly (Study 2), a Delphi (modified) study was carried out to obtain valid, content-based evidence; and finally (Study 3), a pilot study was carried out to evaluate the appropriate functioning of the QIHVC and, if required, to make any necessary adjustments. Results and discussion: The main result is the development of a set of case scenarios and a questionnaire related to its content which constitutes the QIHVC and, in its initial approximation, seems to constitute an adequate and sensible tool to capture the differences between the characterizations of common violence and VAW and in the possible response of bystanders in the face of such violence.

Work-family conflict among hotel housekeepers in the Balearic Islands (Spain).

The massive incorporation of women to the labour market has increased academic and applied interest on work-life issues throughout the years. This article aims to describe the domestic burden and difficulties in work-life balance (WLB) and to understand the intersection of work and family spheres among hotel housekeepers (HHs). A cross-sectional study was conducted through Primary Health Care in the Balearic Islands (Spain); 1,043 HHs were enrolled. 56.7% reported difficulties in WLB. Risk factors for perceiving difficulties in WLB were: living with someone else (regardless of the number of co-habitants), having difficulties making ends meet, being the main person in charge of domestic tasks, having a dependant, having an external locus of control, presenting higher levels of stress at work, working more hours a week and being younger. Protective factors from experiencing work-family conflict (WFC) were job and wage satisfaction. WFC is strongly influenced by individual, economic, labour and domestic factors: these relationships show that labour and domestic spheres are non-separate worlds.

Professional grief among nurses in Spanish public health centers after caring for COVID-19 patients.

AIMS AND OBJECTIVES: The aim of the present study is to investigate the professional grief suffered by nurses in various medical units, after coping with the COVID-19 pandemic for the last 18 months. BACKGROUND: Addressing and acknowledging the reality of professional grief is of fundamental importance to nurses' mental health, as this condition has both professional and personal consequences. DESIGN: A qualitative, content analysis approach was taken. METHODS: Based on 25 interviews with nursing professionals working in different health centers units were performed. The following sampling schemes were used: first, convenience sampling, then nominated sampling, and finally theoretical sampling. RESULTS: From our analysis of the data obtained, three main themes were identified: the impact on nurses of COVID-19 outcomes; the symptoms of professional grief; and cognitive reactions. These core elements interacted with 12 subtopics, including symptoms of grief and the cognitive impact produced. CONCLUSIONS: A large proportion of the nurses consulted in this study have suffered and suffered professional grief and report many related symptoms. In response to the present pandemic and any future occurrence, the question of professional grief needs to be addressed. RELEVANCE TO CLINICAL PRACTICE: To help them cope better with this type of situation, nurses should receive appropriate training. Moreover, healthcare institutions should be made aware of the problem and be encouraged to offer assistance to address the impact produced on nurses by the deaths of their patients. CLINICAL RELEVANCE: This study shows the impact of professional grief on nurses in the context of the COVID-19 pandemic. Nurses are affected personally by the deaths of patients and by alterations to their working conditions. In many cases, this grief remains unresolved and its various symptoms persist.

Workplace bullying and sexual harassment at work among hotel housekeepers in the Balearic Islands (Spain).

Introduction: Hotel housekeepers are close to being a 100% feminized occupational group in Spain. This fact, coupled with some features of the job, places them at high risk of sexual harassment at work and bullying in the workplace. This study aims to explore experiences of sexual harassment at work and workplace bullying among hotel housekeepers in the Balearic Islands. Second, it aims to describe and estimate the prevalence of both phenomena. Methods: This is a mixed-methods study. Ten semi-structured interviews were conducted with key informants, and six focus groups were held with hotel housekeepers. Additionally, a quantitative cross-sectional study (n = 1,043) was undertaken. Results: Most participants in focus groups had been sexually harassed at work. However, they had normalized this kind of situations, not labeling themselves as victims of sexual harassment; and harassment events were seen as unimportant, normal, and unquestioned, as well as being part of their daily work. Hotel housekeepers who were sexually harassed indicated high levels of stress at work and low social support. The prevalence of different workplace bullying behaviors was quite high among hotel housekeepers working in the Balearic Islands. Some were associated with poorer self-rated health, less satisfaction with the job and the salary, lower social support, and higher levels of stress. Despite this, qualitative methods informed us that less severe behaviors were normalized and perceived by hotel housekeepers as intrinsic to their job. Discussion: The results show the high tolerance to less severe expressions of sexual harassment at work and workplace bullying, as well as difficulties in or reluctance to labeling this kind of experiences as such.

Combinatorial biosynthesis yields novel hybrid argimycin P alkaloids with diverse scaffolds in Streptomyces argillaceus.

Coelimycin P1 and argimycins P are two types of polyketide alkaloids produced by Streptomyces coelicolor and Streptomyces argillaceus, respectively. Their biosynthesis pathways share some early steps that render very similar aminated polyketide chains, diverging the pathways afterwards. By expressing the putative isomerase cpkE and/or the putative epoxidase/dehydrogenase cpkD from the coelimycin P1 gene cluster into S. argillaceus wild type and in argimycin mutant strains, five novel hybrid argimycins were generated. Chemical characterization of those compounds revealed that four of them show unprecedented scaffolds (quinolizidine and pyranopyridine) never found before in the argimycin family of compounds. One of these compounds (argimycin DM104) shows improved antibiotic activity. Noticeable, biosynthesis of these quinolizidine argimycins results from a hybrid pathway created by combining enzymes from two different pathways, which utilizes an aminated polyketide chain as precursor instead of lysine as it occurs for other quinolizidines.

Expanding the utility of Marfey's analysis by using HPLC-SPE-NMR to determine the Cß configuration of threonine and isoleucine residues in natural peptides.

The determination of amino acid chirality in natural peptides is typically addressed by Marfey's analysis. This approach relies on the complete hydrolysis of the peptide followed by the reaction of the resulting amino acid pool with Marfey's reagent, a chiral derivatizing agent which turns amino acid enantiomers into diastereomeric pairs which can be resolved by conventional reversed-phase HPLC. However, for certain amino acids possessing a second chiral centre at Cß, the discrimination between the two possible epimers may still be challenging due to the lack of chromatographic resolution. Such is the case of isoleucine and threonine which can also be found in natural nonribosomal peptides as their allo-diastereomers. We describe a new approach based on the extension of Marfey's analysis using HPLC-SPE-NMR to sort out this challenge. Marfey's derivatives of these epimeric amino acids at Cß can be differentiated by their distinct NMR spectra. Thus, simple comparison of the NMR spectra of trapped HPLC peaks with the corresponding spectra of standards enables the unambiguous assignment of the absolute configuration at the second chiral centre in such cases. The general applicability of this approach is showcased for two model cyclic peptides bearing L-Ile and L-Thr.

Personal Traits of the People Who Help: The Case of Bystanders to Violence against Women.

Within the context of emergency situations, the terms witness or bystander are used to refer to individuals involved in oppressive incidents who are neither the victim nor the perpetrator. Among the different types of emergency situations, our study focuses on violence against women (VAW). In keeping with current efforts in the scientific literature on bystander intervention and the evidence currently available, the main focus of this study is to analyze some personal factors that reflect the characteristics or experiences of bystanders and that could have a bearing on their predisposition to help victims of VAW (i.e., empathy, a just world belief system, and expectations of self-efficacy) and later analyze the possible relationship between these personal characteristics and gender or previous experience as a VAW bystander. An opportunity sample of 546 Spanish participants (73.4% women and 26.6% men) between 18 and 56 years of age took part in this study and fill out a sociodemographic data sheet, a questionnaire to evaluate the experience as violence witness designed ad hoc, and the Characteristics of People who Help Questionnaire scale (CPHQ). The results obtained indicate that CPHQ could constitute an adequate measure for the three dimensions analyzed. Female participants are significantly more empathetic than males, but in the case of a just world belief and expectations of self-efficacy the results showed no gender-related differences. Additionally, only a just world belief was clearly influenced by having been a bystander to some form of VAW. In conclusion, this study contributes a proposal for an evaluating instrument featuring three relevant personal characteristics in the development of helping behaviors, presenting some results of interest regarding empathy, a just world belief, and expectations of self-efficacy and their relationship with gender or previous experiences as VAW bystanders. These results obtained suggest an initial path toward future research in the development of interventions with bystander participation in our environment.

Mindfulness-Based Intervention for the Reduction of Compassion Fatigue and Burnout in Nurse Caregivers of Institutionalized Older Persons with Dementia: A Randomized Controlled Trial.

The recent COVID-19 pandemic has severely impacted the mental health of nurses caring for institutionalized older people. Caring in this environment can be complex, with higher levels of burnout and compassion fatigue in staff. It is therefore important to find interventions to increase the well-being of staff. Mindfulness training is known to be effective in treating a variety of physical and mental health conditions. This study sought to conduct a direct evaluation of the effectiveness of a combined online training in two types of mindfulness-based therapies for the reduction of compassion fatigue and burnout in geriatric nurses caring for institutionalized elderly people with dementia. In a randomized controlled trial (n = 39 experimental group, n = 35 control group), we explored whether individuals with high levels of burnout and compassion fatigue would benefit more from an online mindfulness training program. The outcome variable was the ProQoL professional quality of life scale, which was collected at baseline, at six weeks, and at three months after completion of the intervention. Significant differences were found between both groups for the subscales Compassion Fatigue and Burnout (p < 0.05), with a significant improvement in the experimental group (significant effect size). These findings were maintained at three months after the end of the intervention for both compassion fatigue (F1,28 = 18.14, p = 0.003) and burnout (F1,28 = 7.25, p = 0.040). However, there were no differences between groups for the satisfaction subscale. The effect of time and the effects of comparing the two groups after controlling for time were statistically significant for all three subscales of the questionnaire (all p values < 0.001), with effect sizes ranging from small to large (R2 change 0.10-0.47). These data indicate that the experimental condition was more effective, explaining between 10 and 18% more of the variance. A short, online intervention based on mindfulness training appears to be effective for reducing compassion fatigue and burnout in geriatric nurses, with sustained effects over time.