Broad-scale pesticide screening finds anticoagulant rodenticide and legacy pesticides in Australian frogs.

Pesticide contamination poses a significant threat to non-target wildlife, including amphibians, many of which are already highly threatened. This study assessed the extent of pesticide exposure in dead frogs collected during a mass mortality event across eastern New South Wales, Australia between July 2021 and March 2022. Liver tissue from 77 individual frogs of six species were analysed for >600 legacy and contemporary pesticides, including rodenticides. More than a third (36 %) of the liver samples contained at least one of the following pesticides: brodifacoum, dieldrin, DDE, heptachlor/heptachlor epoxide, fipronil sulfone, and 2-methyl-4-chlorophenoxyacetic acid (MCPA). Brodifacoum, a second-generation anticoagulant rodenticide, was found in four of the six frog species analysed: the eastern banjo frog (Limnodynastes dumerilii), cane toad (Rhinella marina), green tree frog (Litoria caerulea) and Peron's tree frog (Litoria peronii). This is the first report of anticoagulant rodenticide detected in wild amphibians, raising concerns about potential impacts on frogs and extending the list of taxa shown to accumulate rodenticides. Dieldrin, a banned legacy pesticide, was also detected in two species: striped marsh frog (Limnodynastes peronii) and green tree frog (Litoria caerulea). The toxicological effects of these pesticides on frogs are difficult to infer due to limited comparable studies; however, due to the low frequency of detection the presence of these pesticides was not considered a major contributing factor to the mass mortality event. Additional research is needed to investigate the effects of pesticide exposure on amphibians, particularly regarding the impacts of second-generation anticoagulant rodenticides. There is also need for continued monitoring and improved conservation management strategies for the mitigation of the potential threat of pesticide exposure and accumulation in amphibian populations.

A systematic review and meta-analysis of English language online patient education materials in breast cancer: Is readability the only story?

BACKGROUND: Online patient education materials (OPEMs) are an increasingly popular resource for women seeking information about breast cancer. The AMA recommends written patient material to be at or below a 6th grade level to meet the general public's health literacy. Metrics such as quality, understandability, and actionability also heavily influence the usability of health information, and thus should be evaluated alongside readability. PURPOSE: A systematic review and meta-analysis was conducted to determine: 1) Average readability scores and reporting methodologies of breast cancer readability studies; and 2) Inclusion frequency of additional health literacy-associated metrics. MATERIALS AND METHODS: A registered systematic review and meta-analysis was conducted in Ovid MEDLINE, Web of Science, Embase.com, CENTRAL via Ovid, and ClinicalTrials.gov in June 2022 in adherence with the PRISMA 2020 statement. Eligible studies performed readability analyses on English-language breast cancer-related OPEMs. Study characteristics, readability data, and reporting of non-readability health literacy metrics were extracted. Meta-analysis estimates were derived from generalized linear mixed modeling. RESULTS: The meta-analysis included 30 studies yielding 4462 OPEMs. Overall, average readability was 11.81 (95% CI [11.14, 12.49]), with a significant difference (p < 0.001) when grouped by OPEM categories. Commercial organizations had the highest average readability at 12.2 [11.3,13.0]; non-profit organizations had one of the lowest at 11.3 [10.6,12.0]. Readability also varied by index, with New Fog, Lexile, and FORCAST having the lowest average scores (9.4 [8.6, 10.3], 10.4 [10.0, 10.8], and 10.7 [10.2, 11.1], respectively). Only 57% of studies calculated average readability with more than two indices. Only 60% of studies assessed other OPEM metrics associated with health literacy. CONCLUSION: Average readability of breast cancer OPEMs is nearly double the AMA's recommended 6th grade level. Readability and other health literacy-associated metrics are inconsistently reported in the current literature. Standardization of future readability studies, with a focus on holistic evaluation of patient materials, may aid shared decision-making and be critical to increased screening rates and breast cancer awareness.

Portable gas chromatography-mass spectrometry method for the in-field screening of organic pollutants in soil and water at pollution incidents.

Environmental pollution incidents generate an emergency response from regulatory agencies to ensure that the impact on the environment is minimised. Knowing what pollutants are present provides important intelligence to assist in determining how to respond to the incident. However, responders are limited in their in-field capabilities to identify the pollutants present. This research has developed an in-field, qualitative analytical approach to detect and identify organic pollutants that are commonly detected by regulatory environmental laboratories. A rapid, in-field extraction method was used for water and soil matrices. A coiled microextraction (CME) device was utilised for the introduction of the extracted samples into a portable gas chromatography-mass spectrometry (GC-MS) for analysis. The total combined extraction and analysis time was approximately 6.5 min per sample. Results demonstrated that the in-field extraction and analysis methods can screen for fifty-nine target organic contaminants, including polyaromatic hydrocarbons, monoaromatic hydrocarbons, phenols, phthalates, organophosphorus pesticides, and organochlorine pesticides. The method was also capable of tentatively identifying unknown compounds using library searches, significantly expanding the scope of the methods for the provision of intelligence at pollution incidents of an unknown nature, although a laboratory-based method was able to provide more information due to the higher sensitivity achievable. The methods were evaluated using authentic casework samples and were found to be fit-for-purpose for providing rapid in-field intelligence at pollution incidents. The fact that the in-field methods target the same compounds as the laboratory-based methods provides the added benefit that the in-field results can assist in sample triaging upon submission to the laboratory for quantitation and confirmatory analysis.

Dual-function antiandrogen/HDACi hybrids based on enzalutamide and entinostat.

The combination of androgen receptor antagonists with histone deacetylase inhibitors (HDACi) has been shown to be more effective than antiandrogens alone in halting growth of prostate cancer cell lines. Here we have designed, synthesized and assessed a series of antiandrogen/HDACi hybrids by combining structural features of enzalutamide with either SAHA or entinostat. The hybrids are demonstrated to maintain bifunctionality using a fluorometric HDAC assay and a bioluminescence resonance energy transfer (BRET) antiandrogen assay. Antiproliferative assays showed that hybrids bearing o-aminoanilide-based HDACi motifs outperformed hydroxamic acid based HDACi's. The hybrids demonstrated selectivity for epithelial cell lines vs. stromal cell lines, suggesting a potentially useful therapeutic window.

Heart failure-the experience of living with end-stage heart failure and accessing care across settings.

BACKGROUND: Heart failure is a complex clinical syndrome affecting an increasing number of the ageing population. Patients and carers require increasing input from specialist palliative care services to both manage symptoms and access support in the last year of life. An integrated clinical service between the local cardiology team at Princess Royal University Hospital and the palliative care team at St. Christopher's Hospice was piloted for patients with end-stage heart failure in Bromley in Kent, UK. This study explored views of patients and carers who participated in the integrated pilot service. METHODS: A qualitative study was conducted in which a convenience sample of patients and carers were invited to participate in focus groups: two bereaved carer groups (n=2, n=2); one patient group (n=4), held between 14th December 2018 and 18th January 2019. Participants were asked to describe their experiences of care received facilitated by a topic guide. Interviews were recorded, transcribed and coded using thematic analysis to identify common themes. RESULTS: Four patients (2:2 M:F) aged between 70 to 87 years and four female carers whom had cared for patients aged between 70 to 96 years who were since deceased, participated in this study. Overall, the service was positively received, and responses were mapped into four key areas; being diagnosed and living with heart failure, referral to palliative care, key helpful components of the care received and finally, unhelpful components of the new service in terms of care. Common themes emerged including understanding of heart failure and its trajectory, communication around palliative care, having a 'broker' for the system, recognition of carer's needs, service responsiveness, and feeling 'in control'. CONCLUSIONS: This qualitative study highlighted important considerations when developing an integrated heart failure and palliative care service. Education about heart failure for patients and carers, but also the integrated multidisciplinary team is crucial to improving detection of deterioration and facilitating communication around Advance Care Planning. The value of the 'expert-carer' should also be promoted and supported in chronic conditions. We recommend a focus on development of integrated services that enable joined-up care or single point of contact for patients and carers.

Breast MRI radiomics for the pretreatment prediction of response to neoadjuvant chemotherapy in node-positive breast cancer patients.

The purpose of this study was to evaluate breast MRI radiomics in predicting, prior to any treatment, the response to neoadjuvant chemotherapy (NAC) in patients with invasive lymph node (LN)-positive breast cancer for two tasks: (1) prediction of pathologic complete response and (2) prediction of post-NAC LN status. Our study included 158 patients, with 19 showing post-NAC complete pathologic response (pathologic TNM stage T0,N0,MX) and 139 showing incomplete response. Forty-two patients were post-NAC LN-negative, and 116 were post-NAC LN-positive. We further analyzed prediction of response by hormone receptor subtype of the primary cancer (77 hormone receptor-positive, 39 HER2-enriched, 38 triple negative, and 4 cancers with unknown receptor status). Only pre-NAC MRIs underwent computer analysis, initialized by an expert breast radiologist indicating index cancers and metastatic axillary sentinel LNs on DCE-MRI images. Forty-nine computer-extracted radiomics features were obtained, both for the primary cancers and for the metastatic sentinel LNs. Since the dataset contained MRIs acquired at 1.5 T and at 3.0 T, we eliminated features affected by magnet strength using the Mann-Whitney U-test with the null-hypothesis that 1.5 T and 3.0 T samples were selected from populations having the same distribution. Bootstrapping and ROC analysis were used to assess performance of individual features in the two classification tasks. Eighteen features appeared unaffected by magnet strength. Pre-NAC tumor features generally appeared uninformative in predicting response to therapy. In contrast, some pre-NAC LN features were able to predict response: two pre-NAC LN features were able to predict pathologic complete response (area under the ROC curve (AUC) up to 0.82 [0.70; 0.88]), and another two were able to predict post-NAC LN-status (AUC up to 0.72 [0.62; 0.77]), respectively. In the analysis by a hormone receptor subtype, several potentially useful features were identified for predicting response to therapy in the hormone receptor-positive and HER2-enriched cancers.

Contributions of TolC Orthologs to Francisella tularensis Schu S4 Multidrug Resistance, Modulation of Host Cell Responses, and Virulence.

Francisella tularensis is a Gram-negative, facultative intracellular pathogen and the causative agent of tularemia. Previous studies with the attenuated live vaccine strain (LVS) identified a role for the outer membrane protein TolC in modulation of host cell responses during infection and virulence in the mouse model of tularemia. TolC is an integral part of efflux pumps that export small molecules and type I secretion systems that export a range of bacterial virulence factors. In this study, we analyzed TolC and its two orthologs, FtlC and SilC, present in the fully virulent F. tularensis Schu S4 strain for their contributions to multidrug efflux, suppression of innate immune responses, and virulence. We found that each TolC ortholog participated in multidrug efflux, with overlapping substrate specificities for TolC and FtlC and a distinct substrate profile for SilC. In contrast to their shared roles in drug efflux, only TolC functioned in the modulation of macrophage apoptotic and proinflammatory responses to Schu S4 infection, consistent with a role in virulence factor delivery to host cells. In agreement with previous results with the LVS, the Schu S4 ΔtolC mutant was highly attenuated for virulence in mice by both the intranasal and intradermal routes of infection. Unexpectedly, FtlC was also critical for Schu S4 virulence, but only by the intradermal route. Our data demonstrate a conserved and critical role for TolC in modulation of host immune responses and Francisella virulence and also highlight strain- and route-dependent differences in the pathogenesis of tularemia.

Bisphenol A Does Not Mimic Estrogen in the Promotion of the In Vitro Response of Murine Dendritic Cells to Toll-Like Receptor Ligands.

Sex hormones affect immune responses and might promote autoimmunity. Endocrine disrupting chemicals such as bisphenol A (BPA) may mimic their immune effects. Conventional dendritic cells (cDCs) are pivotal initiators of immune responses upon activation by danger signals coming from pathogens or distressed tissues through triggering of the Toll-like receptors (TLRs). We generated in vitro murine cDCs in the absence of estrogens and measured the effects of exogenously added estrogen or BPA on their differentiation and activation by the TLR ligands LPS and CpG. Estrogen enhanced the differentiation of GM-CSF-dependent cDCs from bone marrow precursors in vitro, and the selective estrogen receptor modulators (SERMs) tamoxifen and fulvestrant blocked these effects. Moreover, estrogen augmented the upregulation of costimulatory molecules and proinflammatory cytokines (IL-12p70 and TNFα) upon stimulation by TLR9 ligand CpG, while the response to LPS was less estrogen-dependent. These effects are partially explained by an estrogen-dependent regulation of TLR9 expression. BPA did not promote cDC differentiation nor activation upon TLR stimulation. Our results suggest that estrogen promotes immune responses by increasing DC activation, with a preferential effect on TLR9 over TLR4 stimulation, and highlight the influence of estrogens in DC cultures, while BPA does not mimic estrogen in the DC functions that we tested.

TolC-dependent modulation of host cell death by the Francisella tularensis live vaccine strain.

Francisella tularensis is a facultative intracellular, Gram-negative pathogen and the causative agent of tularemia. We previously identified TolC as a virulence factor of the F. tularensis live vaccine strain (LVS) and demonstrated that a ΔtolC mutant exhibits increased cytotoxicity toward host cells and elicits increased proinflammatory responses compared to those of the wild-type (WT) strain. TolC is the outer membrane channel component used by the type I secretion pathway to export toxins and other bacterial virulence factors. Here, we show that the LVS delays activation of the intrinsic apoptotic pathway in a TolC-dependent manner, both during infection of primary macrophages and during organ colonization in mice. The TolC-dependent delay in host cell death is required for F. tularensis to preserve its intracellular replicative niche. We demonstrate that TolC-mediated inhibition of apoptosis is an active process and not due to defects in the structural integrity of the ΔtolC mutant. These findings support a model wherein the immunomodulatory capacity of F. tularensis relies, at least in part, on TolC-secreted effectors. Finally, mice vaccinated with the ΔtolC LVS are protected from lethal challenge and clear challenge doses faster than WT-vaccinated mice, demonstrating that the altered host responses to primary infection with the ΔtolC mutant led to altered adaptive immune responses. Taken together, our data demonstrate that TolC is required for temporal modulation of host cell death during infection by F. tularensis and highlight how shifts in the magnitude and timing of host innate immune responses may lead to dramatic changes in the outcome of infection.

EmrA1 membrane fusion protein of Francisella tularensis LVS is required for resistance to oxidative stress, intramacrophage survival and virulence in mice.

Francisella tularensis is a category A biodefence agent that causes a fatal human disease known as tularaemia. The pathogenicity of F. tularensis depends on its ability to persist inside host immune cells primarily by resisting an attack from host-generated reactive oxygen and nitrogen species (ROS/RNS). Based on the ability of F. tularensis to resist high ROS/RNS levels, we have hypothesized that additional unknown factors act in conjunction with known antioxidant defences to render ROS resistance. By screening a transposon insertion library of F. tularensis LVS in the presence of hydrogen peroxide, we have identified an oxidant-sensitive mutant in putative EmrA1 (FTL_0687) secretion protein. The results demonstrate that the emrA1 mutant is highly sensitive to oxidants and several antimicrobial agents, and exhibits diminished intramacrophage growth that can be restored to wild-type F. tularensis LVS levels by either transcomplementation, inhibition of ROS generation or infection in NADPH oxidase deficient (gp91Phox(-/-)) macrophages. The emrA1 mutant is attenuated for virulence, which is restored by infection in gp91Phox(-/-) mice. Further, EmrA1 contributes to oxidative stress resistance by affecting secretion of Francisella antioxidant enzymes SodB and KatG. This study exposes unique links between transporter activity and the antioxidant defence mechanisms of F. tularensis.

The effect of 17beta-estradiol on the development of modified hemal spines in early-life stage Gambusia holbrooki.

The morphologic development of the gonopodium of male mosquitofish, Gambusia holbrooki, is essential for proper reproductive function and has previously been used as a biomarker for the presence of endocrine-disrupting chemicals (EDCs) in freshwater systems. The development of the gonopodium is accompanied by the modification of the hemal spines on the 14th, 15th, and 16th vertebrae. These form an anchor for the movement of the gonopodium, and their normal development is therefore critical. This development is under hormonal control. It has been hypothesized that the modification of the hemal spines may be used as an end point for understanding the effects of exposure of early-life stage fish to EDCs and to add to the understanding of the differential sensitivities of different life stages. Larval fish were exposed to concentrations (0, 20, 100, and 500 ng/L) of 17beta-estradiol (E(2)) in two experiments using a flow-through system for 8 and 12 weeks. There was a general delay in the development of the three hemal spines when exposed to high concentrations of E(2). However, this delay was not observed for all end points on a particular spine nor across all spines. Fish examined at 8 weeks postpartum showed a more extensive delay at high concentrations than those examined at 12 weeks postpartum, suggesting that later in the life cycle, high levels of exogenous E(2) may be overcome by increasing levels of endogenous androgen. There was a decrease in the number of phenotypic male fish present in the 500-ng/L treatment at 12 weeks postpartum and an apparent decrease in the degree of sexual differentiation at high levels of E(2) exposure. The results suggest that effects of EDC exposure begin very early in a fish's life cycle.

Sexual behavior and impregnation success of adult male mosquitofish following exposure to 17beta-estradiol.

The effects of 17beta-estradiol (E2) on the sexual activity of adult male mosquitofish (Gambusia holbrooki) were assessed. Sexually mature males were exposed to nominal concentrations of 20, 100, and 500 ng/L of E2 and a solvent control (0.00001% ethanol v/v) for 84 days under continuous flow conditions. Following exposure, males exposed to E2 at concentrations of 20 ng/L and above displayed a lower level of sexual activity, measured as the number of approaches and copulation attempts made toward nonexposed females, than the control males. In addition, E2-exposed males were less capable of impregnating females than the control males. Exposure to E2 had no significant effect on spermatozeugmata production or the secondary sexual characteristics, indicating that the reduced impregnation efficiency of the E2-exposed males was most likely due to the decline in sexual activity.

The effect of 17beta-estradiol on the gonopodial development and sexual activity of Gambusia holbrooki.

Many chemicals have recently been demonstrated to possess estrogenic activity and may potentially interfere with normal sexual development. In the present study, we quantified the effects of waterborne exposure to 17beta-estradiol on the gonopodial development and sexual activity of male eastern mosquitofish (Gambusia holbrooki). Juvenile males were exposed during the period of sexual maturation to nominal concentrations of 20, 100, and 500 ng/L of 17beta-estradiol and a solvent control (0.00003% w/v ethanol) for 84 d under continuous-flow conditions. Following exposure, significant differences were found among the test treatments with respect to gonopodial length and degree of gonopodial elongation of the fish. Sexual activity, measured as the number of approaches and copulatory attempts made by the exposed males to nonexposed females, also significantly decreased with increasing concentrations of 17beta-estradiol. Because the degree of gonopodial maturation and frequency of sexual activity are important characteristics for the reproductive success of male mosquitofish, it is suggested that these traits provide sensitive and ecologically relevant endpoints for assessing estrogenic activity under both laboratory--and field-exposure conditions.

Tumor detection by virtual cystoscopy with color mapping of bladder wall thickness.

PURPOSE: We determine the value of color mapping of bladder wall thickness for detection of tumor as a component of virtual cystoscopy. MATERIALS AND METHODS: A total of 31 subjects with hematuria and/or a history of bladder tumor underwent helical computerized tomography of the pelvis after distention of the bladder with air. Three-dimensional (D) models were constructed, and thickness of the wall was color mapped according to a fixed and validated mm. scale. Axial source images and 3-D models were reviewed and graded for the presence of wall thickening. A comparison was made with findings on conventional cystoscopy in 31 patients and pathological specimen in 13. RESULTS: Compared with conventional cystoscopy, the analysis of axial image yielded a sensitivity of 0.80, specificity 0.90, positive predictive value 0.80 and negative predictive value 0.90 for the presence of tumor. Examination of color mapped 3-D renderings resulted in 0.83, 0.36, 0.42 and 0.71, respectively. CONCLUSIONS: Thin axial computerized tomography of the air distended bladder shows promise as a potential screening tool for bladder cancer. The low specificity of color mapped 3-D renderings makes the technique inappropriate for screening. It may valuable for guiding urologists to additional suspicious sites in a patient with a known tumor.