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The influence of ultrasonic activation (UA) on the physicochemical properties of setting time (n = 5), volumetric stability (n = 10), alkalizing activity/calcium ions release (pH/Ca 2+ ) (n = 10) and calcium/phosphate-rich substance formation in simulated body fluid (Hank's balanced salt solution; HBSS) of three cements was evaluated: white MTA-Angelus (MTA), MTA Repair HP (MTAHP) and Biodentine (BIO). It was observed that UA influenced the setting times (min) distinctly, delaying the initial setting time of MTA (40.8/64.6) and accelerating that of BIO (35.6/22.4). The final setting time of MTAHP (154.4/114.6) and BIO (64.4/42.4) were also accelerated (p < 0.05). Volumetric reduction was observed in all groups, with MTA/UA being the most volumetrically stable (0.98%); UA did not influence volumetric stability (p > 0.05). All groups exhibited an alkaline pH, with a disadvantage for MTAHP and MTAHP/UA at 24 h, 72 h and 168 h (p < 0.05). All groups showed Ca 2+ release. The tested cements exhibited a favourable Ca/P ratio on their surface after 28 days., (© 2024 Australian Society of Endodontology Inc.)

Background and Objectives: Soft tissue sarcomas (STS) are a heterogenous group of malignancies of mesenchymal origin. Given recent data linking obesity as well as the pattern of fat distribution with cancer outcomes, we sought to investigate the association of visceral fat area (VFA) and subcutaneous fat area (SFA) with oncologic outcomes in patients with STS undergoing surgery., Methods: We analyzed data from 88 patients with STS diagnosed from 2008 to 2022. Predictor variables included body mass index (BMI), VFA, and SFA. VFA and SFA were obtained from computed tomography of the abdomen and pelvis. Univariable and multivariable Cox regression analysis was used to analyze associations between predictor variables and overall survival and recurrence-free survival., Results: Although BMI was closely correlated with VFA (r = 0.69, p < 0.0001) and SFA (r = 0.80, p < 0.0001), there was no significant association between high BMI, VFA or SFA, and worse oncologic outcomes., Conclusions: Although VFA and SFA are strongly correlated with BMI, we did not observe BMI nor imaging metrics of fat composition to be associated with worse oncologic outcomes. Further research is needed to elucidate any links between body fat content and metabolic or immune factors governing oncologic outcomes in STS., (© 2024 Wiley Periodicals LLC.)

Cultivation of yellow dragon fruit ( Selenicereus megalanthus ) in Peru has recently expanded (Verona-Ruiz et al. 2020). In August 2021, approximately 170 of 1,110 dragon fruit cuttings (15.3%) in the university's nursery (6°26'10'' S; 77°31'25'' W) showed basal rot symptoms. Initial symptoms included small brown spots on the base of stems, expanding towards the top that became soft and watery. All symptomatic plants eventually died, i.e., a severity of 100%. The disease was more prevalent on cuttings during the rooting phase than on well-established cuttings. We collected five symptomatic cuttings from throughout the nursery. Four sections of 1 × 1 cm 2 of tissue adjacent to the diseased area were excised from each cutting, immersed for 1 min in 2% NaClO, rinsed twice with sterile distilled water, placed on potato dextrose agar (PDA) medium (four sections per Petri plate, five plates), and incubated at 25°C for 7 days. Morphologically similar mycelia grew from all sections, and five monosporic isolates were obtained, one per plate. Colonies grew fast, reaching 60 to 64 mm in 7 days, and produced violet-white cottony aerial mycelia with orange sporodochia on PDA, and abundant macro- and microconidia on synthetic nutrient-poor agar. Macroconidia were straight to slightly curved, typically with 2 to 3 septa, 16.6 to 23.3 × 1.7 to 3.7 µm (n = 30); microconidia were oval or kidney-shaped, and commonly hyaline, 6.7 to 16.4 × 2.5 to 4.7 µm (n = 40). Genomic DNA was extracted from isolate AFHP-100, then the ITS region and the TEF1 and RPB2 partial genes were amplified and sequenced (Accession numbers PP977433, OR437358, PP537149) following Gardes and Bruns (1993) and O'Donnell et al. (1998). We conducted a BLASTn search of ITS sequence against the NCBI "nr" database and local 'megablast' searches of TEF1 and RPB2 sequences against FUSARIUM-ID v.3.0 (Torres-Cruz et al. 2022). We found 100%, 98.19 to 99.84%, and 98.81 to 99.76% identities in ITS, TEF1 , and RPB2 sequences, respectively, to the ex-epitype and other reference strains of Fusarium oxysporum (CBS 144134, NRRL26406, among others). A maximum likelihood phylogenetic analysis with a TEF1 - RPB2 concatenated dataset with FUSARIUM-ID sequences also showed isolate AFHP-100 was F. oxysporum . A pathogenicity test was carried out by inoculating wounded healthy roots of three cuttings with submersion in a 5 × 10 6 conidia/ml suspension for 25 min. Then, the inoculated plants were planted in sterile soil. One cutting with wounded roots submerged in sterile water served as a control. In parallel, sterile soil was inoculated with 20 mL of the conidial suspension, and another three healthy cuttings were planted. A cutting planted in noninoculated soil also served as a control. Basal rot symptoms developed in all inoculated plants after 25 days. After re-isolation, the same fungus, corroborated based on micromorphology and TEF1 sequence (PP335689), was recovered, fulfilling Koch's postulates. The isolate was deposited in the KUELAP Herbarium (voucher KUELAP-3214), located and administered by the National University Toribio Rodriguez de Mendoza de Amazonas, in Chachapoyas, Peru. Fusarium oxysporum has been reported to cause basal stem rot in Bangladesh and Argentina (Mahmud et al. 2021; Wright et al. 2007), and stem blight in Malaysia (Mohd Hafifi et al. 2019) on dragon fruit. This is the first report of F. oxysporum causing basal rot in S. megalanthus in Peru. This fungus is among the most destructive plant pathogens, and the rapid expansion of the crop in Peru requires a comprehensive knowledge of the biotic factors influencing production. Therefore, this report is foundational to implementing proper control strategies.

ASCL1 is a transcription factor that directs neural progenitors towards lineage differentiation. Although many of the molecular mechanisms underlying its action have been described, several of its targets remain unidentified. We identified in the chick genome a putative enhancer (cE1) upstream of the transcription factor Scratch2 ( Scrt2 ) locus with a predicted heterodimerization motif for ASCL1 and POU3F2. In this study, we investigated the role of ASCL1 and this enhancer in regulating the expression of the Scrt2 in the embryonic spinal cord. We confirmed that cE1 region interacted with the Scrt2 promoter. cE1 was sufficient to mediate ASCL1-driven expression in the neural tube through the heterodimerization sites. Moreover, Scrt2 expression was inhibited when we removed cE1 from the genome. These findings strongly indicate that ASCL1 regulates Scrt2 transcription in the neural tube through cE1., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Goes, Botezelli, De La Cruz, Cruz, Azambuja, Simoes-Costa and Yan.)

Background and Objectives: Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer with poor response to systemic therapies, including immunotherapy. Given the immunotherapeutic potential of natural killer (NK) cells, we evaluated intratumoral NK cell infiltrates along with cytotoxic T cells in PDAC to determine their association with patient outcomes., Methods: We analyzed tumors from 93 PDAC patients treated from 2012 to 2020. Predictor variables included tumor-infiltrating lymphocytes (TILs), T-cell markers (CD3, CD8, CD45RO), NK marker (NKp46), and NK inhibitory marker (major histocompatibility complex class I [MHC-I]) by immunohistochemistry. Primary outcome variables were recurrence-free survival (RFS) and overall survival (OS)., Results: Mean TILs, CD3, and NKp46 scores were 1.3 ± 0.63, 20.6 ± 17.5, and 3.1 ± 3.9, respectively. Higher expression of CD3 and CD8 was associated with higher OS, whereas NK cell infiltration was not associated with either RFS or OS. There was a tight positive correlation between MHC-I expression and all T-cell markers, but not with NKp46., Conclusions: Overall NK cell infiltrates were low in PDAC and did not predict clinical outcomes, whereas T-cell infiltrates did. Further characterization of the immune infiltrate in PDAC, including inhibitory signals and suppressive cell types, may yield better biomarkers of prognosis and immune targeting in this refractory disease., (© 2024 Wiley Periodicals LLC.)

The Peruvian Amazonian native cacao faces ongoing challenges that significantly undermine its productivity. Among them, frosty pod rot disease and cadmium accumulation result in losses that need for effective and environmentally safe strategies, such as those based on bacteria. To explore the biological resources in the cacao soil, a descriptive study was conducted to assess the diversity of culturable bacteria across three production districts in the Amazonas region: La Peca, Imaza, and Cajaruro. The study also focused on the functional properties of these bacteria, particularly those related to the major issues limiting cacao cultivation. For this purpose, 90 native bacterial isolates were obtained from the cacao rhizosphere. According to diversity analysis, the community was composed of 19 bacterial genera, with a dominance of the Bacillaceae family and variable distribution among the districts. This variability was statistically supported by the PCoA plots and is related to the pH of the soil environment. The functional assessment revealed that 56.8% of the isolates showed an antagonism index greater than 75% after 7 days of confrontation. After 15 days of confrontation with Moniliophthora roreri, 68.2% of the bacterial population demonstrated this attribute. This capability was primarily exhibited by Bacillus strains. On the other hand, only 4.5% were capable of removing cadmium, highlighting the biocontrol potential of the bacterial community. In addition, some isolates produced siderophores (13.63%), solubilized phosphate (20.45%), and solubilized zinc (4.5%). Interestingly, these traits showed an uneven distribution, which correlated with the divergence found by the beta diversity. Our results revealed a diverse bacterial community inhabiting the Amazonian cacao rhizosphere, showcasing crucial functional properties related to the biocontrol of M. roreri . The information generated serves as a significant resource for the development of further biotechnological tools that can be applied to native Amazonian cacao., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Oliva-Cruz, Segundo Manuel reports financial support was provided by National Council of Science Technology and Technology Innovation., (© 2024 The Authors.)

The cancer stem cell (CSC) hypothesis postulates that heterogeneous human cancers harbor a population of stem-like cells which are resistant to cytotoxic therapies, thus providing a reservoir of relapse following conventional therapies like chemotherapy and radiation (RT). CSCs have been observed in multiple human cancers, and their presence has been correlated with worse clinical outcomes. Here, we sought to evaluate the impact of drug dosing of the multi-tyrosine kinase inhibitor, sorafenib, on CSC and non-CSCs in soft tissue sarcoma (STS) models, hypothesizing differential effects of sorafenib based on dose and target cell population. In vitro, human cancer cell lines and primary STS from surgical specimens were exposed to escalating doses of sorafenib to determine cell viability and expression of CSC marker aldehyde dehydrogenase (ALDH). In vivo, ALDH bright CSCs were isolated, exposed to sorafenib, and xenograft growth and survival analyses were performed. We observed that sarcoma CSCs appear to paradoxically respond to the tyrosine kinase inhibitor sorafenib at low doses with increased proliferation and stem-like function of CSCs, whereas anti-viability effects dominated at higher doses. Importantly, STS patients receiving neoadjuvant sorafenib and RT on a clinical trial (NCT00864032) showed increased CSCs post therapy, and higher ALDH scores post therapy were associated with worse metastasis-free survival. These data suggest that low-dose sorafenib may promote the CSC phenotype in STS with clinically significant effects, including increased tumor growth and higher rates of metastasis formation in sarcoma patients.

Introduction: Chia (Salvia hispanica L.) is a plant from the Lamiaceae family that has been used as ancestral food, medicine, and oil, with culinary, artistic, and religious purposes by most of the Mesoamerican civilizations. Native from Mesoamerica, introduced into South America, Australia, and Europe, it is presently consumed as a nutritional and functional food., Objective: This research aims to characterize ancient native cultivars from four provenances in Guatemala to recommend their direct consumption by the population as well as to establish its trade., Methods: Seed samples were collected in four places where they have been cultivated for several generations. The oil was obtained by expression and analyzed chemically by gas chromatography following standard qualitative and quantitative methods., Results: Variations in oil yield and some of the characteristic parameters of the phytochemical analysis were obtained. In general, the profile was similar to most of the reported data in the literature, with the saturated fatty acids (8.54-9.25%) relatively lower than the references (7.95-11.45%) but a higher concentration of unsaturated fatty oils, particularly of omega-3 (64.68-68.62%)., Conclusion: The oil from native cultivars contains high quantities of omega-3, which might help pregnant women during gestation and to control other conditions such as metabolic syndrome, particularly in low- and middle-income populations where these seeds are consumed regularly. The suggestion is made to encourage the cultivation and use of these ancestral seeds with the possibility of commercialization abroad with an appellation of origin label., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: The Litsea genus has four native species from Mesoamerica. Litsea guatemalensis Mez. is a native tree, traditionally used as a condiment and herbal medicine in the region. It has demonstrated antimicrobial, aromatic, anti-inflammatory and antioxidant activity. Bioactive fractionation attributed the anti-inflammatory and anti-hyperalgesic activities to pinocembrin, scopoletin, and 5,7,3´4´-tetrahydroxy-isoflavone. In silico analysis, these molecules were analyzed on receptors involved in the anti-inflammatory process to determine which pathways they interact., Objective: To analyze and evaluate 5,7,3',4'-tetrahydroxyisoflavone, pinocembrin, and scopoletin using the in silico analysis against selected receptors involved in the inflammatory pathway., Methods: Known receptors involved in the anti-inflammatory process found as protein-ligand complex in the Protein Data Bank (PDB) were used as references for each receptor and compared with the molecules of interest. The GOLD-ChemScore function, provided by the software, was used to rank the complexes and visually inspect the overlap between the reference ligand and the poses of the studied metabolites., Results: 53 proteins were evaluated, each one in five conformations minimized by molecular dynamics. The scores obtained for dihydroorotate dehydrogenase were greater than 80 for the three molecules of interest, scores for cyclooxygenase 1 and glucocorticoid receptor were greater than 50, and identified residues with interaction in binding sites overlap with the reference ligands in these receptors., Conclusion: The three molecules involved in the anti-inflammatory process of L. guatemalensis show in silico high affinity to the enzyme dihydroorotate dehydrogenase, glucocorticoid receptors and cyclooxygenase-1., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Introduction: Soft tissue sarcomas (STS) are rare, heterogenous malignancies with an unmet need for novel immunotherapies. Tumor infiltrating lymphocytes (TILs) have been linked with favorable outcomes in STS patients, though the contribution of natural killer (NK) cells and spatial relationships of TILs with MHC-I expressing cells lacks detailed characterization., Experimental Design: Using archived and prospectively collected specimens, we evaluated intratumoral NK cells by immunohistochemistry (IHC), flow cytometry, and immunofluorescence (IF). We assessed spatial localization of NK and T cells by multiplex IF, analyzing the effects of MHC-I expression status on NK and T cell clustering., Results: Both intratumoral NKp46 and CD56 dim expression were associated with significantly improved overall survival (P=0.05), while higher infiltrates of CD56bright NK cells predicted a worse prognosis (P=0.05). The presence of intratumoral NK cells was inversely proportional to CD3 + T cells. Spatial analyses showed NK cells preferentially clustering close to other NK cells with sparse CD3 + T and CD8 + T cells in range (P<0.0001). Additionally, CD3 + T and CD8 + T cells showed significantly greater co-localization with MHC-I + cells, compared to NK cells (P<0.0001). After neoadjuvant radiotherapy, there was greater CD8 clustering, while after neoadjuvant chemotherapy, there was overall lower TIL clustering., Conclusion: Intratumoral NK cells are prognostic in STS and localize closer to MHC-I- cells than T cells. Although both NK and T cells are associated with improved survival in STS, their differential distribution in the TME based on MHC-I expression status may serve as a biomarker for improved immunotherapy treatment selection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Cruz, Sholevar, Judge, Darrow, Iranpur, Farley, Lammers, Razmara, Dunai, Gingrich, Persky, Mori, Thorpe, Monjazeb, Murphy and Canter.)

Efforts by universities to reduce the spread of COVID-19 include health campaigns intended to encourage students to wear masks. While well-intended, these efforts may produce counter-persuasion (e.g., decrease masking) if they are seen as threatening individuals' freedom to choose. In a rolling cross-sectional study of one university campaign ( n = 681), we found that the presence of the campaign did instigate a form of resistance known as reactance and that reactance was negatively associated with masking behavior. Masking was also diminished by the frequency with which respondents observed others not wearing a mask (anti-masking descriptive norm) and the frequency with which respondents observed others expressing disdain for masking (anti-masking injunctive norm). Most of these findings were magnified among students who identified as politically conservative. There was no evidence that the frequency of seeing others speak in favor of masks (pro-masking injunctive norm) produced an increase in masking. The results provide valuable theoretical insights into the causes of reactance and empirical evidence of the risks associated with student-oriented COVID safety campaigns.

Background: Intake of sugar-sweetened beverages (SSBs) is associated with many negative health outcomes. Efforts to curb consumption generally take one of two approaches: (a) Individual change based on the communication of personal risk information or (b) policies that limit or disincentivize the behavior, such as restricting access or implementing taxes., Methods and Results: Using samples of 2347 persons and 139 pro- and anti-SSB messages, this study tested whether individual-level persuasion attempts would spill over to voting intentions and whether that spillover would amplify or attenuate policy preferences. The influence of anti-SSB messages was seen in (a) direct, positive effects on intention to vote for restrictive policies and in (b) indirect, positive effects where change in intended consumption mediated message impact on change in voting intentions. Anti-SSB messages were generally more powerful than pro-SSB advertising at producing direct and indirect change. Yet, there was evidence of boomerang effects among small numbers of both SSB drinkers and nondrinkers., Conclusions: Anti-SSB messages that target individual consumption bring about intentions to reduce consumption and increased receptivity to restrictive SSB policies., (Copyright © 2023. Published by Elsevier Ltd.)

Background: Coronavirus disease 2019 (COVID-19) disrupted access to food and adequate nutrition and the types of foods consumed. However, little empiric data exists on the changes in American's food and nutrition habits 2 y into the pandemic., Objectives: To assess current and altered food choices ∼2 y into the COVID-19 pandemic in the months after historic public pandemic relief., Methods: A national sample of 1878 United States adults balanced by age, sex, race/ethnicity, and income completed a one-time, online, semi-quantitative, 44-item questionnaire in Fall 2021 asking about the demographics, COVID-19 food choice changes (including free-text), and consumer priorities. This analysis investigates COVID-19 impacts on food security, healthfulness, and access., Results: More than 35% of respondents reported improved food security and >45% reported improved food healthfulness compared with prepandemic status. Improvement was reported in more than 30% of Black/African-American and Hispanic/Latinx adults, adults with lower annual income, and female sex, despite over 75% reporting reduced choice of where to eat or buy food. The pandemic offered occasion for many to improve diet, but a similar number expressed that the pandemic destabilized healthy habits., Conclusions: Our novel findings suggest that by late 2021, most Americans had improved food security and food choice healthfulness, despite reduced access to food service and retail, although with worsening among a meaningful proportion of Americans as well as heterogeneity in these changes. Vigorous federal, state, city, and community responses to the pandemic may have played a role in improving the food security and food choice healthfulness during the COVID-19 pandemic. Health crises differently impact health behaviors, but when accompanied by vigorous civic and community response, food security, and food healthfulness can be fortified., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Dariush Mozaffarian reports financial support was provided by Vail Innovative Global Research., (© 2023 The Authors.)

Ethnopharmacological Relevance: The prevalence of kidney disease has increased rapidly in recent years and has emerged as one of the leading causes of mortality worldwide. Natural products have been suggested as valuable nephroprotective agents due to their multi-target and synergistic effects on modulating important proteins involved in kidney injury. There is a large number of plant species that have been used traditionally for kidney-related conditions in Mesoamerican medicine by different cultural groups that could provide a valuable source of nephroprotective therapeutic candidates and could lead to potential drug discovery., Aim of Review: This review aims to provide an overview of the currently known efficacy of plant species used traditionally in Mesoamerica by Mayan groups to treat kidney-related conditions and to analyze the phytochemical, pharmacological, molecular, toxicological, and clinical evidence to contribute to public health efforts and for directing future research., Methods: Primary sources of plant use reports for traditional kidney-related disorders in Mesoamerica were searched systematically from library catalogs, theses, and scientific databases (PubMed, Google Scholar; and Science Direct), and were filtered according to usage frequency in Mayan groups and plant endemism. The database of traditional plants was further analyzed based on associations with published reports of the phytochemical, pharmacological, molecular, toxicological, and clinical evidence., Results: The most reported kidney-related conditions used traditionally in Mayan medicine involve reducing renal damage (a cultural interpretation that considers an inflammatory or infectious condition), cleaning or purifying the blood and kidney, reducing kidney pain, and eliminating kidney stones. A total of 208 plants used for kidney-related problems by 10 Mayan groups were found, representing 143 native species, where only 42 have reported pharmacological activity against kidney damage, mainly approached by in vitro and in vivo models of chemical- or drug-induced nephrotoxicity, diabetes nephropathy, and renal injury produced by hypertension. Nephroprotective effects are mainly mediated by reducing oxidative stress, inflammatory response, fibrosis mechanisms, and apoptosis in the kidney. The most common nephroprotective compounds associated with traditional Mayan medicine were flavonoids, terpenoids, and phenolic acids. The most widely studied traditional plants in terms of pharmacological evidence, bioactive compounds, and mechanisms of action, are Annona muricata L., Carica papaya L., Ipomoea batatas (L.) Lam., Lantana camara L., Sechium edule (Jacq.) Sw., Tagetes erecta L., and Zea mays L. Most of the plant species with reported pharmacological activity against kidney damage were considered safe in toxicological studies., Conclusion: Available pharmacological reports suggest that several herbs used in traditional Mayan medicine for renal-associated diseases may have nephroprotective effects and consistent pharmacological evidence, nephroprotective compounds, and mechanisms of action in different models of kidney injury. However, more research is required to fully understand the potential of traditional Mayan medicine in drug discovery given the limited ethnobotanical studies and data available for most species with regards to identification on bioactive components, pharmacological mechanisms, and the scarce number of clinical studies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Groundbreaking studies have linked the gut microbiome with immune homeostasis and antitumor immune responses. Mounting evidence has also demonstrated an intratumoral microbiome, including in soft tissue sarcomas (STS), although detailed characterization of the STS intratumoral microbiome is limited. We sought to characterize the intratumoral microbiome in patients with STS undergoing preoperative radiotherapy and surgery, hypothesizing the presence of a distinct intratumoral microbiome with potentially clinically significant microbial signatures., Methods: We prospectively obtained tumor and stool samples from adult patients with non-metastatic STS using a strict sterile collection protocol to minimize contamination. Metagenomic classification was used to estimate abundance using genus and species taxonomic levels across all classified organisms, and data were analyzed with respect to clinicopathologic factors., Results: Fifteen patients were enrolled. Most tumors were located at an extremity (67%) and were histologic grade 3 (87%). 40% were well-differentiated/dedifferentiated liposarcoma histology. With a median follow-up of 24 months, 4 (27%) patients developed metastases, and 3 (20%) died. Despite overwhelming human DNA (>99%) intratumorally, we detected a small but consistent proportion of bacterial DNA (0.02-0.03%) in all tumors, including Proteobacteria, Bacteroidetes, and Firmicutes , as well as viral species. In the tumor microenvironment, we observed a strong positive correlation between viral relative abundance and natural killer (NK) infiltration, and higher NK infiltration was associated with superior metastasis-free and overall survival by immunohistochemical, flow cytometry, and multiplex immunofluorescence analyses., Conclusions: We prospectively demonstrate the presence of a distinct and measurable intratumoral microbiome in patients with STS at multiple time points. Our data suggest that the STS tumor microbiome has prognostic significance with viral relative abundance associated with NK infiltration and oncologic outcome. Additional studies are warranted to further assess the clinical impact of these findings., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Cacao plant cadmium accumulation has become a major concern, especially for small Amazonian producers. A sustainable alternative to address its toxicity is the use of cadmium removal bacteria. In this regard, 138 rhizosphere isolates from cacao were examined. Supported by their phenotypic characterization and in vitro cadmium tolerance, three hypertolerant bacteria were selected and identified as members of the Bacillus (S1C2, R1C2) and Pseudomonas (V3C3) genera. They were able to grow normally and reduce the cadmium content under in vitro conditions. However, only S1C2 and R1C2 evidenced to employ intracellular Cd 2+ accumulation, suggesting the variability of bacterial detoxification mechanisms. Their bioremediation capacity for Theobroma cacao CCN51 was also analyzed. Surprisingly, we found high detectable levels of Cd 2+ in the non-cadmium supplemented control, suggesting an extra source of cadmium in the pot. Moreover, despite their cadmium reduction performance under in vitro conditions, they exerted highly variable outcomes on stem cadmium accumulation. While S1C2 and R1C2 showed a considerable reduction of Cd content in cacao stems, the strain V3C3 did not show any effect on Cd content. This highlights the complexity of the plant-bacteria interactions and the importance of the in vivo test for the selection of promising PGPR bacteria. Overall, our results suggest the cadmium alleviation potential and promising prospects of native Bacillus strains associated with Amazonian cacao., Competing Interests: The authors declare that they have no conflict of interest.

The microbiome has clearly been established as a cutting-edge field in tumor immunology and immunotherapy. Growing evidence supports the role of the microbiome in immune surveillance, self-tolerance, and response to immune checkpoint inhibitors such as anti PD-L1 and CTLA-4 blockade (1-6). Moreover, recent studies including those using fecal microbial transplantation (FMT) have demonstrated that response to checkpoint immunotherapies may be conferred or eliminated through gut microbiome modulation (7, 8). Consequently, studies evaluating microbiota-host immune and metabolic interactions remain an area of high impact research. While observations in murine models have highlighted the importance of the microbiome in response to therapy, we lack sufficient understanding of the exact mechanisms underlying these interactions. Furthermore, mouse and human gut microbiome composition may be too dissimilar for discovery of all relevant gut microbial biomarkers. Multiple cancers in dogs, including lymphoma, high grade gliomas, melanomas and osteosarcoma (OSA) closely resemble their human analogues, particularly in regard to metastasis, disease recurrence and response to treatment. Importantly, dogs with these spontaneous cancers also have intact immune systems, suggesting that microbiome analyses in these subjects may provide high yield information, especially in the setting of novel immunotherapy regimens which are currently expanding rapidly in canine comparative oncology (9, 10). Additionally, as onco-microbiotic therapies are developed to modify gut microbiomes for maximal responsiveness, large animal models with intact immune systems will be useful for trialing interventions and monitoring adverse events. Together, pre-clinical mechanistic studies and large animal trials can help fully unlock the potential of the microbiome as a diagnostic and therapeutic target in cancer., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Kleber, Iranpur, Perry, Cruz, Razmara, Culp, Kent, Eisen, Rebhun and Canter.)

Purpose: Clinical successes using current T-cell based immunotherapies have been limited in soft tissue sarcomas (STS), while pre-clinical studies have shown evidence of natural killer (NK) cell activity. Since tumor immune infiltration, especially tumor-infiltrating lymphocytes, is associated with improved survival in most solid tumors, we sought to evaluate the gene expression profile of tumor and blood NK and T cells, as well as tumor cells, with the goal of identifying potential novel immune targets in STS., Experimental Design: Using fluorescence-activated cell sorting, we isolated blood and tumor-infiltrating CD3 - CD56 + NK and CD3 + T cells and CD45 - viable tumor cells from STS patients undergoing surgery. We then evaluated differential gene expression (DGE) of these purified populations with RNA sequencing analysis. To evaluate survival differences and validate primary DGE results, we also queried The Cancer Genome Atlas (TCGA) database to compare outcomes stratified by bulk gene expression., Results: Sorted intra-tumoral CD3 + T cells showed significant upregulation of established activating (CD137) and inhibitory genes (TIM-3) compared to circulating T cells. In contrast, intra-tumoral NK cells did not exhibit upregulation of canonical cytotoxic genes (IFNG, GZMB), but rather significant DGE in mitogen signaling (DUSP4) and metabolic function (SMPD3, SLC7A5). Tumors with higher NK and T cell infiltration exhibited significantly increased expression of the pro-inflammatory receptor TLR4 in sorted CD45 - tumor cells. TCGA analysis revealed that tumors with high TLR4 expression ( P = 0.03) and low expression of STMN1 involved in microtubule polymerization ( P < 0.001) were associated with significantly improved survival., Conclusions: Unlike T cells, which demonstrate significant DGE consistent with upregulation of both activating and inhibiting receptors in tumor-infiltrating subsets, NK cells appear to have more stable gene expression between blood and tumor subsets, with alterations restricted primarily to metabolic pathways. Increased immune cell infiltration and improved survival were positively correlated with TLR4 expression and inversely correlated with STMN1 expression within tumors, suggesting possible novel therapeutic targets for immunotherapy in STS., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Judge, Bloomstein, Sholevar, Darrow, Stoffel, Vick, Dunai, Cruz, Razmara, Monjazeb, Rebhun, Murphy and Canter.)

Purpose: Although recombinant human interleukin-15 (rhIL-15) has generated much excitement as an immunotherapeutic agent for cancer, activity in human clinical trials has been modest to date, in part due to the risks of toxicity with significant dose escalation. Since pulmonary metastases are a major site of distant failure in human and dog cancers, we sought to investigate inhaled rhIL-15 in dogs with naturally occurring lung metastases from osteosarcoma (OSA) or melanoma. We hypothesized a favorable benefit/risk profile given the concentrated delivery to the lungs with decreased systemic exposure., Experimental Design: We performed a phase I trial of inhaled rhIL-15 in dogs with gross pulmonary metastases using a traditional 3+3 cohort design. A starting dose of 10 µg twice daily × 14 days was used based on human, non-human primate, and murine studies. Safety, dose-limiting toxicities (DLT), and maximum tolerated dose (MTD) were the primary objectives, while response rates, progression-free and overall survival (OS), and pharmacokinetic and immune correlative analyses were secondary., Results: From October 2018 to December 2020, we enrolled 21 dogs with 18 dogs reaching the 28-day response assessment to be evaluable. At dose level 5 (70 μg), we observed two DLTs, thereby establishing 50 µg twice daily × 14 days as the MTD and recommended phase 2 dose. Among 18 evaluable dogs, we observed one complete response >1 year, one partial response with resolution of multiple target lesions, and five stable disease for an overall clinical benefit rate of 39%. Plasma rhIL-15 quantitation revealed detectable and sustained rhIL-15 concentrations between 1-hour and 6 hour postnebulization. Decreased pretreatment lymphocyte counts were significantly associated with clinical benefit. Cytotoxicity assays of banked peripheral blood mononuclear cells revealed significant increases in peak cytotoxicity against canine melanoma and OSA targets that correlated with OS., Conclusions: In this first-in-dog clinical trial of inhaled rhIL-15 in dogs with advanced metastatic disease, we observed promising clinical activity when administered as a monotherapy for only 14 days. These data have significant clinical and biological implications for both dogs and humans with refractory lung metastases and support exploration of combinatorial therapies using inhaled rhIL-15., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Peru is the second largest producer of organic cocoa and one of the most important suppliers of fine aroma cocoa beans in the world (Sánchez et al. 2019). The fine aroma cocoa produced by smallholder farmers in the Bagua and Utcubamba Provinces, Amazonas Department, under the name of "Cacao Amazonas Peru", is protected by the Peruvian appellation rules (Díaz-Valderrama et al. 2020). Despite this importance, native diseases of the crop ( Theobroma cacao ) are poorly documented due to difficulty of access in this region. In November 2020 we conducted expeditions into Imaza District (4°47'09.4"S 78°16'51.6"W), a significant producer of fine aroma cocoa in terms of number of cultivated plots (4,651 out of 6,505 total in the Bagua Province) (INEI 2012). We visited 20 farms of < 2-ha in size; in 19 of these small farms, T. cacao trees were found infected with a white fungal thread blight and rhizomorphs covering branches and leaves. Disease incidence ranged from 90 to nearly 100%, and severity exceeded 80% on the eight farms with the most deficient phytosanitary management. Heavily infected leaves were hanging on branches by mycelial threads, harboring tiny (0.5 to 5.3 mm broad) white mushrooms. These symptoms and signs correspond to the thread blight disease constellation (TBD) of cacao caused by various species of Marasmius and Marasmiellus (Amoako-Attah et al. 2020). Mushrooms lacked a collarium, and their stipes were absent or rudimentary (< 2-mm long) and eccentric, consistent with Marasmius tenuissimus (Tan et al. 2009). Axenic cultures were obtained by surface sterilization of mycelium threads with 2% NaClO, rinsed three times in sterile water, plated on potato dextrose agar medium (PDA), and incubated for 7 days at 25°C. Hyphae was non-pigmented with clamp connections, consistent with the genus Marasmius . We extracted the DNA of isolate INDES-AFHP31 using the Wizard® Purification Kit (Promega Corp., Madison, Wisconsin) and sequenced the rDNA internal transcribed spacer 1 and 2 intervening the 5.8S subunit (ITS), and the 28S subunit (LSU) (Accession numbers: OM720123 and OM720135) according to Aime and Phillips-Mora (2005). The ITS and LSU sequences were 97.92 to 98.79% and 99.07 to 99.30% identical, respectively, with published sequences from M. tenuissimus from Ghana (Amoako-Attah et al. 2020). The pathogenicity test was conducted by inoculation of ten healthy cacao leaves with 7-day-old mycelium PDA discs of isolate INDES-AFHP31. An equal number of healthy cacao leaves were inoculated with PDA discs without mycelium as control. The observation of TBD symptoms and signs in the non-control set of cacao leaves starting at 3 days post inoculation, and the re-isolation of the same fungus from infected tissue confirmed its pathogenicity on cacao. Isolate INDES-AFHP31 was deposited as a dried culture into the herbarium Kuélap of the Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas (voucher KUELAP-2251). Marasmius tenuissimus was originally reported from dead and living twigs and leaves of unidentified dicotyledonous trees from Indonesia, Brazil, and Bolivia (Singer 1976). However, it was first associated with TBD of cacao in Ghana in 2020, being the most frequently TBD-causing fungus isolated in the country (Amoako-Attah et al. 2020). Its discovery in 19 of the 20 surveyed cacao farms in Imaza District, Amazonas, Peru, reveals its importance as a cacao pathogen in the Western hemisphere.

Peru is the ninth exporter of coffee ( Coffea arabica ) in the world, and Amazonas is among its most important producing departments (INIA 2019). In July 2021, in the nursery of the "Instituto de Investigación para el Desarrollo Sustentable de Ceja de Selva", in Huambo district (6° 26' 11.19'' S; 77° 31' 24.18'' W), four-month-old coffee seedlings cv. Catimor with 0.5-2.0 cm brown concentric leaf spots and rotten stems, bearing white mycelial tufts and black sporodochia, were observed at 30% incidence. Infected seedlings were collected. Foliar sections of 2-3 mm with infected tissue were surfaced disinfected in 2% NaClO and transferred onto Petri plates containing potato dextrose agar medium (PDA). The plates were incubated at 25° C for 7 days. We obtained three isolates (INDES-AFHP61, INDES-AFHP62 and INDES-AFHP66) with similar morphology from different seedlings. Colonies (16-17 mm diam.) formed concentric rings with white aerial mycelium, giving rise to viscous and olivaceous dark green sporodochial conidiomata. Conidia (4.82-5.77 × 1.34-1.65 µm; n = 30) were cylindric, hyaline, smooth, and aseptate. These morphological features correspond to Paramyrothecium spp. (Lombard et al. 2016). The DNA of isolates was extracted using the Wizard® Purification Kit (Promega Corp., Madison, Wisconsin), and the internal transcribed spacer 1 and 2 intervening the 5.8S subunit rDNA region (Accession numbers: OM892830 to OM892832), and part of the second-largest subunit of the RNA polymerase II, the calmodulin and the β-tubulin genes (OM919453 to OM919461) were sequenced following Lombard et al. (2016). All sequences had a percent identity greater than or equal to 99% to corresponding sequences of the P. roridum type specimen (CBS 357.89). Additionally, a multilocus Maximum Likelihood phylogenetic analysis incorporating sequence data from previous relevant studies (Lombard et al. 2016; Pinruan et al. 2022) grouped our three isolates together with the type and other specimens of P. roridum in a strongly supported clade, confirming the species identification. To evaluate pathogenicity, four-month-old coffee seedlings cv. Catimor were sprayed with 10 mL of conidial suspensions at 1 x 10 6 /mL. A set of control seedlings were inoculated with sterile water. Seedlings were maintained in a humidity chamber at 25 °C. After 15 days brown concentric foliar spots, stem rotting, mycelial tufts and sporodochia (same symptoms and signs observed originally at the nursery) arose in the non-control seedlings. The pathogen was re-isolated on PDA, confirming P. roridum was the causal agent of leaf spot and stem rot diseases of coffee. Paramyrothecium roridum has wide geographic distribution and host range (Lombard et al. 2016). This pathogen was reported to infect C. arabica in Mexico and Coffea sp. in Colombia (Pelayo-Sánchez et al. 2017; Lombard et al. 2016; Huaman et al. 2021). It was also reported in Africa infecting soybeans (Haudenshield et al. 2018), in Brazil infecting Tectona grandis (Borges et al. 2018), in Egypt infecting strawberries (Soliman 2020), and in Malaysia infecting Eichhornia crassipes (Hassan et al. 2021). To the best of our knowledge, this is the first time P. roridum is reported on coffee in Peru.

In this work, high purity graphite, a high chemically stable material, was effectively digested using a single method allowing compatible solutions for the further multitechnique determination of halogens by: ion chromatography (F and Cl), inductively coupled plasma mass spectrometry (Cl, Br and I) and by ion selective electrode (only for F). The recent system using microwave-induced self-ignition (MISI) is based on the strong interaction between microwave radiation and graphite in a closed system pressurized with oxygen (Maxwell-Wagner effect). Carbon-based materials present intense and specific interfacial polarization when exposed to microwave electromagnetic field resulting in a fast heating rate. This effect associated to a pressurized oxygen system, provides a quick self-ignition of carbon-based materials and consequent combustion/digestion of organic matrices. Under optimized conditions, sample masses up to 600 mg were fully digested in a quartz vessel under 20 bar of oxygen pressure and using just a diluted solution (100 mmol L -1  NH 4 OH) for the quantitative absorption of all the analytes. MISI method was validated, and the accuracy (better than 94%) was evaluated by comparison of results obtained by pyrohydrolysis for two coal certified reference materials as well as with subsequent analytes determination by the three techniques: IC, ICP-MS and ISE. It is important to point out that no filter paper disks, electrical connections or other ignition aids are required as in the case of previous or classical combustion methods. Moreover, just a diluted absorbing solution was used resulting in negligible blanks and relatively low limits of detection. The digestion efficiency was higher than 99%, making the proposed method a suitable and powerful alternative for the quasi complete digestion of graphite and determination of halogens virtually free of interferences., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Snakebite envenoming is a public health problem of high impact in Central America. Bothrops asper, known as barba amarilla, terciopelo, and equis, is the snake species responsible for most snakebites in Central America. In this region, there is a long-standing tradition on the use of plants in the management of snakebites, especially in indigenous communities. Ethnomedical use of Eryngium foetidum L., Neurolaena lobata (L.) Cass. and Pimenta dioica (L.) Merr. to treat snakebite envenoming has been reported in Belice, Guatemala, Nicaragua, and Costa Rica. Extracts of the leaves of these plants have shown anti-venom activities in in vitro assays in previous studies., Aim of the Study: To assess the ability of organic fractions from these three plants to inhibit enzymatic activities associated with toxicity of the venom of B. asper, and to study, by docking analysis, the interaction of metalloproteinase and phospholipases A 2 (PLA 2 ) from B. asper venom with secondary metabolites previously described in these plants., Materials and Methods: Organic fractions were obtained from these three plant species and their ability to neutralize proteolytic, PLA 2 and in vitro coagulant activities of B. asper venom was assessed. A phytochemical analysis was carried out in these fractions. The interaction of secondary metabolites previously described in these plants with three toxins from B. asper venom (a metalloproteinase, a PLA 2 and a PLA 2 homologue) was investigated by docking analysis., Results: The inhibitory activity of plants was mainly concentrated in their polar fractions. Acetonic fraction from P. dioica was the most active against PLA 2 activity, while the acetonic fraction of E. foetidum completely inhibited the proteolytic activity of the venom. Coagulant activity was partially inhibited only by the acetone and ethyl acetate fractions of P. dioica. Phytochemical analysis of the most bioactive fractions identified flavonoids, saponins, essential oils, coumarins, alkaloids, tannins and sesquiterpene lactones. Docking analysis revealed high affinity interactions of several secondary metabolites of these plants with residues in the vicinity of the catalytic site of these enzymes and, in the case of PLA 2 homologue myotoxin II, in the hydrophobic channel., Conclusions: Various fractions from these plants have inhibitory activity against enzymatic actions of B. asper venom which are directly associated with toxicological effects. Docking analysis showed structural evidence of the interaction of secondary metabolites with three toxins. These observations provide support to the potential of these plants to inhibit relevant toxic components of this snake venom., (Copyright © 2021 Elsevier B.V. All rights reserved.)

This document provides an update to the recommendations for physiotherapy management for adults with coronavirus disease 2019 (COVID-19) in the acute hospital setting. It includes: physiotherapy workforce planning and preparation; a screening tool for determining requirement for physiotherapy; and recommendations for the use of physiotherapy treatments and personal protective equipment. New advice and recommendations are provided on: workload management; staff health, including vaccination; providing clinical education; personal protective equipment; interventions, including awake proning, mobilisation and rehabilitation in patients with hypoxaemia. Additionally, recommendations for recovery after COVID-19 have been added, including roles that physiotherapy can offer in the management of post-COVID syndrome. The updated guidelines are intended for use by physiotherapists and other relevant stakeholders caring for adult patients with confirmed or suspected COVID-19 in the acute care setting and beyond., (Copyright © 2021 Australian Physiotherapy Association. Published by Elsevier B.V. All rights reserved.)

Background and Objectives: Although arterial involvement for advanced tumors is rare, vascular resection may be indicated to achieve complete tumor resection. Given the potential morbidity of this approach, we sought to evaluate perioperative outcomes, vascular graft patency, and survival among patients undergoing tumor excision with en bloc arterial resection and reconstruction., Methods: From 2010 to 2020, we identified nine patients with tumors encasing or extensively abutting major arterial structures for whom en bloc arterial resection and reconstruction was performed., Results: Mean age was 53 ± 20 years, and 89% were females. Diagnoses were primary sarcomas (5), recurrent gynecologic carcinomas (3), and benign retroperitoneal fibrosis (1). Tumors involved the infrarenal aorta (2), iliac arteries (6), and superficial femoral artery (1). Three patients (33%) had severe perioperative morbidity (Grade III + ) with no mortality. At a median follow-up of 23 months, eight patients (89%) had primary graft patency, and five patients (56%) had no evidence of disease., Conclusions: Arterial resection and reconstruction as part of the multimodality treatment of regionally advanced tumors is associated with acceptable short- and long-term outcomes, including excellent graft patency. In appropriately selected patients, involvement of major arterial structures should not be viewed as a contraindication to attempted curative surgery., (© 2021 Wiley Periodicals LLC.)

Metastatic osteosarcoma has a bleak prognosis in both humans and dogs, and there have been minimal therapeutic advances in recent decades to improve outcomes. Naturally occurring osteosarcoma in dogs is shown to be a highly suitable model for human osteosarcoma, and limited data suggest the similarities between species extend into immune responses to cancer. Studies show that immune infiltrates in canine osteosarcoma resemble those of human osteosarcoma, and the analysis of tumor immune constituents as predictors of therapeutic response is a promising direction for future research. Additionally, clinical studies in dogs have piloted the use of NK transfer to treat osteosarcoma and can serve as valuable precursors to clinical trials in humans. Cytotoxic lymphocytes in dogs and humans with osteosarcoma have increased activation and exhaustion markers within tumors compared with blood. Accordingly, NK and T cells have complex interactions among cancer cells and other immune cells, which can lead to changes in pathways that work both for and against the tumor. Studies focused on NK and T cell interactions within the tumor microenvironment can open the door to targeted therapies, such as checkpoint inhibitors. Specifically, PD-1/PD-L1 checkpoint expression is conserved across tumors in both species, but further characterization of PD-L1 in canine osteosarcoma is needed to assess its prognostic significance compared with humans. Ultimately, a comparative understanding of T and NK cells in the osteosarcoma tumor microenvironment in both dogs and humans can be a platform for translational studies that improve outcomes in both dogs and humans with this frequently aggressive disease., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Razmara, Judge, Gingrich, Cruz, Culp, Kent, Rebhun and Canter.)

Bisphenol A (BPA) is an endocrine-disruptor compound that exhibits estrogenic activity. BPA is used in the production of materials such as polycarbonate plastics, epoxy resins and dental sealants. Whereas, the endocrine modulating activity of BPA and its effects on reproductive health have been widely studied, its effects on the function of the immune system are poorly characterized. This might be attributable to the different BPA doses used in a diversity of animal models. Moreover, most studies of the effect of BPA on the immune response are limited to in vitro and in vivo studies that have focused primarily on the impact of BPA on the number and proportion of immune cell populations, without evaluating its effects on immune function in response to an antigenic challenge or infectious pathogens. In this review, we discuss the current literature on the effects of BPA on the function of immune system that potentially increases the susceptibility to infections by the virtue of acting as a pro-inflammatory molecule. Thus, it appears that BPA, while by such an impact might be useful in the control of certain disease states that are helped by an inflmmatory response, it can worsen the prognosis of diseases that are adversely affected by inflammation.

Scratch2 is a transcription factor expressed in a very restricted population of vertebrate embryonic neural cell precursors involved in their survival, differentiation, and migration. The mechanisms that control its expression remain unknown and could contribute towards our understanding of gene regulation during neural differentiation and evolution. Here we investigate the role of microRNAs (miRNAs) in the Scrt2 post-transcriptional regulatory mechanism. We identified binding sites for miR-125b and -200b in the Scrt2 3'UTR in silico. We confirmed the repressive-mediated activity of the Scrt2 3'UTR through electroporation of luciferase constructs into chick embryos. Further, both CRISPR/Cas9-mediated deletion of miR-125b/-200b responsive elements from chicken Scrt2 3'UTR and expression of miRNAs sponges increased Scrt2 expression field, suggesting a role for these miRNAs as post-transcriptional regulators of Scrt2. The biological effect of miR-125b titration was much more pronounced than that of miR-200b. Therefore, we propose that, after transcription, miR-125b fine-tunes the Scrt2 expression domain., (Copyright © 2020 Goes, Vieceli, De La Cruz, Simões-Costa and Yan.)

Growing concern about the seriousness of issues such as climate change has made the value of research on social and behavioral aspects of environmental problems clearer than ever. For authors studying environmental concern or attitudes, however, survey development can be a daunting task. A large number of scales measuring environmental concern have been developed, and it can be challenging to make informed decisions about which to use. To assist authors in navigating the literature, we present a review of existing scales, followed by two studies in which we examine the structural validity of five scales that are commonly implemented in this corpus and that adhere to classical test theory. These results have important implications for general issues with measurement in this area, and inform our recommendations for authors about key considerations when selecting and using environmental concern scales., (Copyright © 2020 Cruz and Manata.)

The processes that control water quality improvement during artificial recharge (filtering, degradation, and adsorption) can be enhanced by adding a reactive barrier containing different types of sorption sites and promoting diverse redox states along the flow path, which increases the range of pollutants degraded. While this option looks attractive for renaturazing reclaimed water, three issues have to be analyzed prior to broad scale application: (1) a fair comparison between the system with and without reactive barrier; (2) the role of plants in prevention of clogging and addition of organic carbon; and (3) the removal of pathogens. Here, we describe a pilot installation built to address these issues within a waste water treatment plant that feeds on water reclaimed from the secondary outflow. The installation consists of six systems of recharge basin and aquifer with some variations in the design of the reactive barrier and the heterogeneity of the aquifer. We report preliminary results after one year of operation. We find that (1) the systems are efficient in obtaining a broad range of redox conditions (at least iron and manganese reducing), (2) contaminants of emerging concern are significantly removed (around 80% removal, but very sensitive to the compound), (3) pathogen indicators (E. coli and Enterococci) drop by some 3-5 log units, and (4) the recharge systems maintained infiltration capacity after one year of operation (only the system without plants and the one without reactive barrier displayed some clogging). Overall, the reactive barrier enhances somewhat the performance of the system, but the gain is not dramatic, which suggests that barrier composition needs to be improved., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Introduction: Fetal magnetic resonance imaging (MRI) has been used to stratify severity of congenital diaphragmatic hernia (CDH) after ultrasound diagnosis. The purpose of this study was to determine if timing of MRI influenced prediction of severity of outcome in CDH., Methods: A single institution retrospective review of all CDH referred to our institution from February 2004 to May 2017 was performed. Patients were included if they underwent at least 2 fetal MRIs prior to delivery. Prenatal MRI indices including observed-to-expected total fetal lung volume (o/e TFLV) were evaluated. Indices were categorized by trimester, either 2nd (20-27 weeks gestation) or 3rd trimester (>28 weeks gestation) and further analyzed for outcome predictability. Primary outcomes were survival, extracorporeal membrane oxygenation (ECMO), and pulmonary hypertension (PAH). Student t test and logistic and linear regression were used for data analyses., Results: Of 256 fetuses evaluated for CDH, 197 were further characterized by MRI with 57 having both an MRI in the 2nd and 3rd trimesters. There was an average of 9.95 weeks (±4.3) between the 1st and 2nd MRI. Second trimester o/e TFLV was the only independent predictor of survival by logistic regression (OR 0.890, p < 0.01). Third trimester MRI derived lung volumes were associated with, and independent predictors of, severity of PAH and need for ECMO. Interval TFLV growth was a strong predictor of PAH postnatally (OR 0.361, p < 0.01). Overall cohort survival was 79%., Conclusion: Accuracy of MRI lung volumes to predict outcomes is dependent on the -gestational age at the time of exam. While MRI lung volumes at either the 2nd or 3rd trimester are predictive of morbidity, 2nd trimester lung volumes strongly correlated with mortality., (© 2019 S. Karger AG, Basel.)

Objective: Fetal endoscopic tracheal occlusion (FETO) is associated with increased perinatal survival and reduced need for extracorporeal membrane oxygenation (ECMO) in fetuses with severe congenital diaphragmatic hernia (CDH). This study evaluates the impact of FETO on the resolution of pulmonary hypertension (PH) in fetuses with isolated CDH., Methods: We reviewed retrospectively the medical records of all fetuses evaluated for CDH between January 2004 and July 2017 at a single institution. Fetuses with additional major structural or chromosomal abnormalities were excluded. CDH cases were classified retrospectively into mild, moderate and severe groups based on prenatal magnetic resonance imaging indices (observed-to-expected total fetal lung volume and percentage of intrathoracic liver herniation). Presence of PH was determined based on postnatal echocardiograms. Logistic regression analyses were performed to evaluate the relationship between FETO and resolution of PH by 1 year of age while controlling for side of the CDH, use of ECMO, gestational age at diagnosis, gestational age at delivery, fetal gender, sildenafil use at discharge and CDH severity. Resolution of PH by 1 year of age was compared between a cohort of fetuses with severe CDH that underwent FETO and a cohort that did not have the procedure (non-FETO). A subanalysis was performed restricting the analysis to isolated left CDH. Parametric and non-parametric tests were used for comparisons., Results: Of 257 CDH cases evaluated, 72% (n = 184) had no major structural or chromosomal anomalies of which 58% (n = 107) met the study inclusion criteria. The FETO cohort consisted of 19 CDH cases and the non-FETO cohort (n = 88) consisted of 31 (35%) mild, 32 (36%) moderate and 25 (28%) severe CDH cases. All infants with severe CDH, regardless of whether they underwent FETO, had evidence of neonatal PH. FETO (OR, 3.57; 95% CI, 1.05-12.10; P = 0.041) and ECMO (OR, 5.01; 95% CI, 2.10-11.96; P < 0.001) were independent predictors of resolution of PH by 1 year of age. A higher proportion of infants with severe CDH that underwent FETO had resolution of PH by 1 year after birth compared with infants with severe CDH in the non-FETO cohort (69% (11/16) vs 28% (7/25); P = 0.017). Similar results were observed when the analysis was restricted to cases with left-sided CDH (PH resolution in 69% (11/16) vs 28% (5/18); P = 0.032)., Conclusion: In infants with severe CDH, FETO and ECMO are independently associated with increased resolution of PH by 1 year of age. Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd., (Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd.)

Postnatal growth of lean mass is commonly blunted in preterm infants and may contribute to short- and long-term morbidities. To determine whether preterm birth alters the protein anabolic response to feeding, piglets were delivered at term or preterm, and fractional protein synthesis rates (K s ) were measured at 3 days of age while fasted or after an enteral meal. Activation of signaling pathways that regulate protein synthesis and degradation were determined. Relative body weight gain was lower in preterm than in term. Gestational age at birth (GAB) did not alter fasting plasma glucose or insulin, but when fed, plasma insulin and glucose rose more slowly, and reached peak value later, in preterm than in term. Feeding increased K s in longissimus dorsi (LD) and gastrocnemius muscles, heart, pancreas, and kidney in both GAB groups, but the response was blunted in preterm. In diaphragm, lung, jejunum, and brain, feeding increased K s regardless of GAB. Liver K s was greater in preterm than term and increased with feeding regardless of GAB. In all tissues, changes in 4EBP1, S6K1, and PKB phosphorylation paralleled changes in K s . In LD, eIF4E·eIF4G complex formation, phosphorylation of TSC2, mTOR, and rpS6, and association of mammalian target of rapamycin (mTOR1) complex with RagA, RagC, and Rheb were increased by feeding and blunted by prematurity. There were no differences among groups in LD protein degradation markers. Our results demonstrate that preterm birth reduces weight gain and the protein synthetic response to feeding in muscle, pancreas, and kidney, and this is associated with blunted insulin- and/or amino acid-induced translation initiation signaling.

Community engagement is a vital aspect of addressing environmental contamination and remediation. In the United States, the Superfund Research Program (SRP) forms groups of academic researchers from the social and physical sciences into Community Engagement Cores (CECs) and Research Translation Cores (RTCs), which focus on various aspects of informing and working with communities during and through the resolution of environmental crises. While this work typically involves engaging directly with members of affected communities, no two situations are the same. In some cases, alternative approaches to community engagement can be more appropriate for community improvement than traditional approaches. In particular, when research teams become involved in contamination crises at a late point in the process, their contributions can be better directed at supporting and reinforcing the work of institutional stakeholders charged with remediating pollution. Relevant factors include issue fatigue among a local population, and contamination that is due to a major employer. Supported by literature and experience, we offer several propositions that we believe lay out conditions that warrant such an approach by academic teams, rather than their direct engagement with unaffiliated individuals in communities.

The articles in this special issue on New Directions in Environmental Communication in the International Journal of Environmental Research and Public Health present new research and perspectives on engaging communities impacted by Superfund sites-the hazardous waste sites that have been identified by the U.S. Environmental Protection Agency (EPA) as needing cleanup. In particular, these articles focus on the community engagement cores (CECs) that work with affected communities as part the Superfund Research Program at the National Institute of Environmental Health Sciences (NIEHS). The purpose of this closing article is to highlight important themes evident across the eight articles in the special issue. When considered together, the findings reveal important lessons learned about community engagement and environmental communication, but also reveal that much more remains to be known. Recommendations are made for how these teams can continue to practice, reflect on, and research community engagement in ways that build toward a better understanding and implementation of best practices.

Alcohol consumption is often characterized by heavy episodic, or binge drinking, which has been on the rise. The aim of this study was to examine the neural dynamics of inhibitory control in demographically matched groups of young, healthy adults (N = 61) who reported engaging in binge (BD) or light drinking patterns (LD). Electroencephalography signal was recorded during a fast-paced visual Go/NoGo paradigm probing the ability to inhibit prepotent responses. No group differences were found in task performance. BDs showed attenuated event-related theta (4-7 Hz) on inhibition trials compared to LDs, which correlated with binge episodes and alcohol consumption but not with measures of mood or disposition including impulsivity. A greater overall decrease of early beta power (15-25 Hz) in BDs may indicate deficient preparatory "inhibitory brake" before deliberate responding. The results are consistent with deficits in the inhibitory control circuitry and are suggestive of allostatic neuroadaptive changes associated with binge drinking., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Purpose: The purpose of this study was to determine the accuracy of fetal echocardiogram (ECHO) for detecting cardiac structural anomalies that may impact Extracorporeal Membrane Oxygenation (ECMO) candidacy in infants with Congenital Diaphragmatic Hernia (CDH)., Methods: A retrospective review was performed on fetuses with CDH (January 2007-June 2017). Inclusion criteria were inborn and at least one prenatal and postnatal ECHO. ECHOs were evaluated for structural heart defects. Primary outcomes were accuracy of prenatal fetal ECHO and identify differences between prenatal and postnatal ECHO. Descriptive statistics and Chi-square analysis were performed., Results: 131 inborn patients were identified. Mean gestational age of fetal ECHO was 26.6 ± 5.5 weeks. The median time to postnatal ECHO was DOL 1 [0-30]. Fetal ECHO had 92% accuracy, 83% sensitivity, 93% specificity, PPV of 95%, NPV of 92%, and a 90% accuracy for visualization of at least one pulmonary vein into the left atrium on the contralateral (non-CDH) side. Thirty-five percent of patients received ECMO, and 26% had an associated cardiac anomaly. All ECMO patients had an accurate structural fetal ECHO., Conclusion: Fetal ECHO is sufficient for identifying major structural heart defects at large volume centers with trained pediatric cardiologists and may be used to guide clinical management, particularly in regards to ECMO candidacy., Level of Evidence: Level III., (Copyright © 2019. Published by Elsevier Inc.)

Objective: To evaluate the effect of investigational vaginal rings containing nomegestrol acetate (NOMAC) plus 17β-estradiol (E2) or etonogestrel (ENG) plus E2 in women with moderate to severe primary dysmenorrhea., Study Design: This was a Phase 2b randomized, placebo-controlled, multicenter, double-blind study. We randomized participants to one of five treatment groups: four hormonal rings and one placebo ring. The investigational vaginal rings released 300 μg of E2 daily along with 700 μg or 900 μg of NOMAC or 100 μg or 125 μg of ENG. Each participant received 2 identical rings and was to insert each for 21 days followed by a 7-day ring-free period. The primary endpoint, as assessed by a daily electronic diary (e-Diary), was the change in menstrual pain score from baseline to the second in-treatment withdrawal bleeding episode (Cycle 2). The pain score was the mean of the three highest scores for menstrual cramping pain (0-4 point scale) recorded from the day before menses to the third day of bleeding. The primary hypothesis was that at least one investigational vaginal ring would demonstrate a statistically significant larger reduction from baseline in pain score compared to placebo. Secondary endpoints included total mean impact score (which assessed the negative impact on work/school, physical activities, leisure/social activities) and the amount and days of rescue medication (ibuprofen) used., Clinical Trial Registration Number: NCT01670656., Results: We randomized 439 participants. The mean pain score decreased from baseline to Cycle 2 in all groups; the decrease in all four treatment groups compared to placebo was statistically significant (p-values ≤0.002). All treatment groups had greater reductions than placebo in ibuprofen intake and greater improvement in impact scores; these differences were statistically significant for both endpoints for the ENG-E2 100/300 μg/day group, while the other groups were not statistically significant for one or both endpoints., Conclusion: All four investigational rings produced a statistically significantly larger reduction from baseline in mean menstrual pain score compared to placebo while pain medication use decreased., Implications: This placebo-controlled study provides evidence that vaginal contraceptive rings containing NOMAC-E2 or ENG-E2 improve moderate to severe dysmenorrhea, across all of doses studied. This adds to the evidence that hormonal contraceptives are effective treatments for dysmenorrhea., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Studies demonstrating an association between anesthesia and brain cell death (neuroapoptosis) in young animals were performed without accompanying surgery. This study tests the hypothesis that fetal surgery decreases anesthesia-induced neuroapoptosis., Materials and Methods: Seventy-day-pregnant ewes received 2% isoflurane for 1 h (low dose [LD]) or 4% for 3 h (high dose [HD]) with or without fetal surgery (S). Unexposed fetuses served as controls (C). Fetal brains were processed for neuroapoptosis using anti-caspase-3 antibodies. Data were analyzed using ANOVA., Results: Twenty-eight fetal sheep were evaluated. Dentate gyrus neuroapoptosis was lower in the HD+S group (13.1 ± 3.76 × 105/mm3) than in the HD (19.1 ± 1.40 × 105/mm3, p = 0.012) and C groups (18.3 ± 3.55 × 105/mm3, p = 0.035). In the pyramidal layer of the hippocampus, neuroapoptosis was lower in the HD+S group (8.11 ± 4.88 × 105/mm3) than in the HD (14.8 ± 2.82 × 105/mm3, p = 0.006) and C groups (14.1 ± 4.54 × 105/mm3, p = 0.019). The LD+S group showed a trend towards a significant decrease in neuroapoptosis in the pyramidal layer (LD+S 7.51 ± 1.48 vs. LD 13.5 ± 1.87 vs. C 14.1 ± 4.54 × 105/mm3, p = 0.07) but not in the dentate gyrus. Fetal surgery did not affect neuroapoptosis in the frontal cortex or endplate., Conclusions: Fetal surgery decreases isoflurane-induced neuroapoptosis in the dentate gyrus and the pyramidal layer of mid-gestational fetal sheep. Long-term effects of these observations on memory and learning deserve further exploration., (© 2018 S. Karger AG, Basel.)

Purpose: To examine postsurgical outcomes of a consecutive series of children treated with elective operations for congenital lung malformations (CLM)., Methods: A retrospective review was performed on a prospectively collected dataset of all fetuses evaluated for a CLM between July 2001 and June 2016. Prenatal findings, operative treatment and postnatal outcomes were collected. Children having elective operations were divided in two groups based on age at time of surgery., Results: Of 220 fetuses, 143 had operations and follow-up at our center. Six had open fetal lobectomy, 17 had EXIT-to-resection, 16 infants had urgent resection for symptoms and 110 with asymptomatic lesions had elective resection. Of these 110, the median fetal maximum CVR was 0.8 [range 0.1-2.2], and median age at operation was 4 (1.5-60) months (58% had resection at ≤4 months). Overall complication rate, including air-leak and pleural effusion, was 15%. When comparing those having resection at ≤4 months to those >4 months, there were no significant differences in complication rates or length of stay. Operative time was shorter for patients with early resection (154 ± 59 vs 181 ± 89, p = 0.05). No infant having resection at ≤4 months was readmitted. Overall, children survived with good recovery., Conclusion: Early elective resection of congenital lung malformations prior to 4 months of age is feasible and not associated with increased operative risk., Type of Study: Restropective study., Level of Evidence: Level III., (Copyright © 2018. Published by Elsevier Inc.)

Necrotizing enterocolitis (NEC) is associated with low plasma arginine and vascular dysfunction. It is not clear whether low intestinal citrulline production, the precursor for arginine synthesis, occurs before and thus predisposes to NEC or if it results from tissue damage. This study was designed to test the hypothesis that whole body rates of citrulline, arginine, and nitric oxide synthesis are low in premature pigs and that they precede NEC. Piglets delivered by cesarean section at 103 days [preterm (PT)], 110 days [near-term (NT)], or 114 days [full-term (FT)] of gestation were given total parenteral nutrition and after 2 days orogastrically fed infant formula for 42 h to induce NEC. Citrulline and arginine fluxes were determined before and during the feeding protocol. Gross macroscopic and histological NEC scores and plasma fatty acid binding protein (iFABP) concentration were determined as indicators of NEC. Intestinal gene expression for enzymes of the arginine pathway were quantitated. A lower ( P < 0.05) survival rate was observed for PT (8/27) than for NT (9/9) and FT pigs (11/11). PT pigs had higher macroscopic gross ( P < 0.05) and histological NEC ( P < 0.05) scores and iFABP concentration ( P < 0.05) than pigs of more advanced gestational age. PT pigs had lower citrulline production and arginine fluxes ( P < 0.05) throughout and a reduced gene expression in genes of the citrulline-arginine pathway. In summary, intestinal enzyme expression and whole body citrulline and arginine fluxes were reduced in PT pigs compared with animals of more advance gestational age and preceded the development of NEC. NEW & NOTEWORTHY Arginine supplementation prevents necrotizing enterocolitis (NEC), the most common gastrointestinal emergency of prematurity. Citrulline (precursor for arginine) production is reduced during NEC, and this is believed to be a consequence of intestinal damage. In a swine model of NEC, we show that intestinal gene expression of the enzymes for citrulline production and whole body citrulline and arginine fluxes are reduced and precede the onset of NEC in premature pigs. Reduced citrulline production during prematurity may be a predisposition to NEC.

Prematurity and low birth weight have been exclusion criteria for extracorporeal membrane oxygenation (ECMO); however, these criteria are not evidence based. With advances in anticoagulation, improved technology, and surgical expertise, it is difficult to deny a potential therapy based on these criteria alone. We report the outcome of three neonates who were ineligible based on traditional criteria but were offered ECMO as a life-saving measure. We highlight the interdisciplinary nature of modern decision-making. All three neonates had severe congenital diaphragmatic hernia diagnosed prenatally, had normal fetal karyotypes, were born prematurely, and weighed less than 2 kg. All three neonates underwent cervical venoarterial cannulation, stabilization on ECMO, and repair of their congenital diaphragmatic hernia early in their ECMO courses. All three infants had long courses of respiratory support attributable to lung hypoplasia, but there were no short- or long-term complications attributable to ECMO support directly. All three are alive at 2 years of age and were making progress developmentally. In conclusion, with interdisciplinary collaboration and clinical guidelines uniformly implemented, low birth weight infants may benefit from ECMO and should not be denied the therapy arbitrarily based on gestational age or size alone. Further research is essential to determine appropriate patient selection in premature infants.

Heavy episodic alcohol consumption (also termed binge drinking) contributes to a wide range of health and cognitive deficits, but the associated brain-based indices are poorly understood. The current study used electroencephalography (EEG) to examine spontaneous neural oscillations in young adults as a function of quantity, frequency, and the pattern of their alcohol consumption. Sixty-one young adults (23.4 ± 3.4 years of age) were assigned to binge drinking (BD) and light drinking (LD) groups that were equated on gender, race/ethnic identity, age, educational background, and family history of alcoholism. EEG activity was recorded during eyes-open and eyes-closed resting conditions. Each participant's alpha peak frequency (APF) was used to calculate absolute power in individualized theta and alpha frequency bands, with a canonical frequency range used for beta. APF was slower by 0.7 Hz in BD, especially in individuals engaging in high-intensity drinking, but there were no changes in alpha power. BD also exhibited higher frontal theta and beta power than LD. Alpha slowing and increased theta power in BD remained after accounting for depression, anxiety, and personality characteristics, while elevated beta power covaried with sensation seeking. Furthermore, APF slowing and theta power correlated with various measures of alcohol consumption, including binge episodes and blackouts, but not with measures of working and episodic memory, cognitive flexibility, processing speed, or personality variables, suggesting that these physiological changes may be modulated by high-intensity alcohol intake. These results are consistent with studies of alcohol-use disorder (AUD) and support the hypothesis that binge drinking is a transitional stage toward alcohol dependence. The observed thalamocortical dysrhythmia may be indicative of an excitatory-inhibitory imbalance in BD and may potentially serve as an index of the progressive development of AUD, with a goal of informing possible interventions to minimize alcohol's deleterious effects on the brain., (Copyright © 2018 Elsevier Inc. All rights reserved.)