Linking intraspecies variability of Salmonella enterica isolates under acidic conditions to genotype.

There is a lack of information about Salmonella enterica strains under acidic conditions and their association with their genome. This study characterized intraspecies variability in the growth of 167 S. enterica isolates under two acid conditions (pH 4 and 5) and linked to the whole genome sequencing (WGS) data. A total of 1002 curves for each condition were obtained using turbidimetry measurements, and Baranyi and Roberts model was used to estimate the maximum rate of change (rcmax; OD600 nm h-1). Strains were categorized into slow, intermediate, and fast; and associations with their WGS data were performed. Huge variability in r c max ¯ $\overline {{\mathrm{r}}{{{\mathrm{c}}}_{{\mathrm{max}}}}} $ was observed at both conditions (pH 5 = 0.016-0.066 OD600nm h-1 and pH 4 = 0.003-0.028 OD600nm h-1). The majority of isolates was classified as intermediate r c max ¯ $\overline {{\mathrm{r}}{{{\mathrm{c}}}_{{\mathrm{max}}}}} $ (59.5% at pH 5 and 46.1% at pH 4). Strains classified as fast had a low frequency of allABCD genes at both pHs, and any of them having the presence of pefABCD, spvBCR, aadA2, dfrA12, and gyrA_D87G genes were linked to virulence or antimicrobial resistance. This study suggests that strains with fast capacity for growth under acidic conditions could have a fitness cost in their virulence or resistance potential. PRACTICAL APPLICATION: Data presented in this study could be used to select representative strains to evaluate the exposure assessment in different food items, mainly the growth and survival in acidic foods.

Effect of the Polyanion Structure on the Mechanism of Alcohol Oxidation with H2O2 Catalyzed by Zr-Substituted Polyoxotungstates.

Zr-monosubstituted polyoxometalates (Zr-POMs) of the Lindqvist (Bu4N)6[{W5O18Zr(µ-OH)}2] (1), Keggin (Bu4N)8[{PW11O39Zr(µ-OH)}2] (2), and Wells-Dawson (Bu4N)11.3K2.5H0.2[{P2W17O61Zr}2(µ-OH)2] (3) structures catalyze oxidation of alcohols using aqueous hydrogen peroxide as an oxidant. With 1 equiv of H2O2 and 1 mol % of Zr-POM, selectivity toward aldehydes and ketones varied from good to excellent, depending on the alcohol nature. Catalytic activity and attainable substrate conversions strongly depended on the Zr-POM structure and most often decreased in the order 1 > 2 ≫ 3. The reaction mechanism was probed using a test substrate, cyclobutanol, radical and 1O2 scavengers, and kinetic and spectroscopic (attenuated total reflectance-Fourier transform infrared (ATR-FT-IR), 31P NMR and electrospray ionization-mass spectrometry (ESI-MS)) tools. The results point to heterolytic alcohol oxidation in the presence of 1 and 2 and homolytic alcohol oxidation in the presence of 3. Kinetic and spectroscopic studies implicated an oxidation mechanism that involves both alcohol and peroxide binding to 2 followed by an inner-sphere heterolytic H-abstraction from the α-C-H bond by the Zr-hydroperoxo group, leading to a carbonyl compound. The unique capability of 1 to generate 1O2 upon interaction with H2O2 complicates the reaction kinetics and improves the product yield. Spectroscopic studies coupled with stoichiometric experiments unveiled that dimeric monoperoxo {Zr2(µ-η2:η2-O2)} and monomeric hydroperoxo {Zr(η2-OOH)} species accomplish the transformation of alcohols to carbonyl compounds.

Thermosensitive injectable fibrillar gels based on cellulose nanocrystals grafted with poly(N-isopropylacrylamide) as biocompatible brain implants.

Drug treatment of glioblastoma, the most aggressive and widespread form of brain cancer, is complicated due to the difficulty of penetration of chemotherapeutic drugs through the blood-brain barrier (BBB). Moreover, with surgical removal of tumors, in 90 % of cases they reappear near the original focus. To solve this problem, we propose to use hydrogel based on cellulose nanocrystals grafted with poly(N-isopropylacrylamide) (CNC-g-PNIPAM) as a promising material for filling postoperative cavities in the brain with the release of antitumor drugs. The CNC-g-PNIPAM is formed by "grafting to" method for precise control of molecular weight and grafting density. This colloidal system is liquid under injection conditions (at r. t.) and turns into a gel at human body temperature (when filling the postoperative area). It was shown for the first time that due to the rod-shaped of CNC, the gel has a fibrillar structure and, thus, mechanical properties similar to those of brain tissue, including nonlinear mechanics (strain-stiffening and compression softening). The biocompatibility of the hydrogel with primary brain cells is demonstrated. In addition, the release of the antitumor drug paclitaxel from the hydrogel and its antitumor activity is shown. The resulting nanocolloid system provides an innovative alternative approach to filling postoperative cavities and can be used for postoperative treatment due to the programmable release of drugs, as well as for in vitro modeling of tumor interaction with the BBB affecting drug transport in the brain.

Structural and Phylogenetic In Silico Characterization of Vitis vinifera PRR Protein as Potential Target for Plasmopara viticola Infection.

Fungi infection, especially derived from Plasmopara viticola, causes severe grapevine economic losses worldwide. Despite the availability of chemical treatments, looking for eco-friendly ways to control Vitis vinifera infection is gaining much more attention. When a plant is infected, multiple disease-control molecular mechanisms are activated. PRRs (Pattern Recognition Receptors) and particularly RLKs (receptor-like kinases) take part in the first barrier of the immune system, and, as a consequence, the kinase signaling cascade is activated, resulting in an immune response. In this context, discovering new lectin-RLK (LecRLK) membrane-bounded proteins has emerged as a promising strategy. The genome-wide localization of potential LecRLKs involved in disease defense was reported in two grapevine varieties of great economic impact: Chardonnay and Pinot Noir. A total of 23 potential amino acid sequences were identified, exhibiting high-sequence homology and evolution related to tandem events. Based on the domain architecture, a carbohydrate specificity ligand assay was conducted with docking, revealing two sequences as candidates for specific Vitis vinifera-Plasmopara viticola host-pathogen interaction. This study confers a starting point for designing new effective antifungal treatments directed at LecRLK targets in Vitis vinifera.

Darifenacin: a promising chitinase 3-like 1 inhibitor to tackle drug resistance in pancreatic ductal adenocarcinoma.

PURPOSE: Pancreatic ductal adenocarcinoma (PDAC) is among the most aggressive malignancies. Our previous work revealed Chitinase 3-like 1 (CHI3L1) involvement in PDAC resistance to gemcitabine, identifying it as a promising therapeutic target. Here, we aimed to identify putative CHI3L1 inhibitors and to investigate their chemosensitizing potential in PDAC. METHODS: Docking analysis for CHI3L1 identified promising CHI3L1 inhibitors, including darifenacin (muscarinic receptor antagonist). PDAC cell lines (BxPC-3, PANC-1) and primary PDAC cells were used to evaluate darifenacin's effects on cell growth (Sulforhodamine B, SRB), alone or in combination with gemcitabine or gemcitabine plus paclitaxel. Cytotoxicity against normal immortalized pancreatic ductal cells (HPNE) was assessed. Recombinant protein was used to confirm the impact of darifenacin on CHI3L1-induced PDAC cellular resistance to therapy (SRB assay). Darifenacin's effect on Akt activation was analysed by ELISA. The association between cholinergic receptor muscarinic 3 (CHRM3) expression and therapeutic response was evaluated by immunohistochemistry of paraffin-embedded tissues from surgical resections of a 68 patients' cohort. RESULTS: In silico screening revealed the ability of darifenacin to target CHI3L1 with high efficiency. Darifenacin inhibited PDAC cell growth, with a GI50 of 26 and 13.6 µM in BxPC-3 and PANC-1 cells, respectively. These results were confirmed in primary PDAC-3 cells, while darifenacin showed no cytotoxicity against HPNE cells. Importantly, darifenacin sensitized PDAC cells to standard chemotherapies, reverted CHI3L1-induced PDAC cellular resistance to therapy, and decreased Akt phosphorylation. Additionally, high CHMR3 expression was associated with low therapeutic response to gemcitabine. CONCLUSION: This work highlights the potential of darifenacin as a chemosensitizer for PDAC treatment.

Infants' and young children's dietary exposures to lead and cadmium: FDA total diet study 2018-2020.

Food can be a source of lead and cadmium exposure for infants and children. Employing a semi-probabilistic approach, dietary exposures to lead and cadmium were assessed for infants 0-11 months (excluding human milk-fed infants) and children 1-6 years using U.S. total diet study data from 2018 to 2020 and food consumption data from 2015 to 2018. Estimated mean lead and cadmium exposures range from 0.7-3.6 µg/day to 0.18-0.47 µg/kg bw/day, respectively, depending on the age group and method for handling non-detected values. Dietary exposures to lead and cadmium are slightly lower and slightly higher than our estimates published in 2019. In addition to the use of more recent datasets for consumption and contamination, differences may be due to the use of refined exposure assessment methodology, particularly a new system of mapping contamination data to intake data. The processed baby food and infant formula food group is the major contributor to lead and cadmium exposure, driven by intake, among infants who do not consume human milk. The food groups contributing most to children's lead and cadmium exposure are grains/baking, dairy and fruit and grains/baking and vegetables, respectively. This work will inform FDA initiatives such as closer to zero, including research needs and regulatory priorities.

The U.S. Food and Drug Administration (FDA) Food Disaggregation Database (FDA-FDD): a new tool for U.S. dietary exposure assessment.

Dietary exposure to a food chemical (e.g. contaminant, nutrient, or other natural constituent) is a function of the concentration of the chemical in foods and the quantity of each food consumed. Exposures to food chemicals can be estimated using intake data from What We Eat in America (WWEIA), the food consumption survey portion of the National Health and Nutrition Examination Survey (NHANES). To estimate exposures to chemicals in foods consumed by NHANES/WWEIA respondents, the consumption data must be mapped to chemical concentration data on the same or similar foods. However, food chemical data are generally not available on all the foods and food mixtures that are reported in NHANES/WWEIA. To address this, we developed the FDA Food Disaggregation Database (FDA-FDD), a 'recipe' database with estimates of ingredient percentages. FDA-FDD allows mapping to food chemical data based on ingredients in NHANES/WWEIA foods rather than on food mixtures, resulting in more accurate exposure estimates. Using FDA-FDD, FDA mapped over 11,000 NHANES/WWEIA foods to FDA's Total Diet Study (TDS) foods. FDA-FDD is available as part of a publicly available interactive application that also allows access to the TDS mapping.

Characterization of the Epileptogenic Phenotype and Response to Antiseizure Medications in Lissencephaly Patients.

BACKGROUND: Patients with lissencephaly typically present with severe psychomotor retardation and drug-resistant seizures. The aim of this study was to characterize the epileptic phenotype in a genotypically and radiologically well-defined patient cohort and to evaluate the response to antiseizure medication (ASM). Therefore, we retrospectively evaluated 47 patients of five genetic forms (LIS1/PAFAH1B1, DCX, DYNC1H1, TUBA1A, TUBG1) using family questionnaires, standardized neuropediatric assessments, and patients' medical reports. RESULTS: All but two patients were diagnosed with epilepsy. Median age at seizure onset was 6 months (range: 2.1-42.0), starting with epileptic spasms in 70%. Standard treatment protocols with hormonal therapy (ACTH or corticosteroids) and/or vigabatrin were the most effective approach for epileptic spasms, leading to seizure control in 47%. Seizures later in the disease course were most effectively treated with valproic acid and lamotrigine, followed by vigabatrin and phenobarbital, resulting in seizure freedom in 20%. Regarding psychomotor development, lissencephaly patients presenting without epileptic spasms were significantly more likely to reach various developmental milestones compared to patients with spasms. CONCLUSION: Classic lissencephaly is highly associated with drug-resistant epilepsy starting with epileptic spasms in most patients. The standard treatment protocols for infantile epileptic spasms syndrome lead to freedom from seizures in around half of the patients. Due to the association of epileptic spasms with an unfavorable course of psychomotor development, early and reliable diagnosis and treatment of spasms should be pursued. For epilepsies occurring later in childhood, ASM with valproic acid and lamotrigine, followed by vigabatrin and phenobarbital, appears to be most effective.

Sugar assimilation underlying dietary evolution of Neotropical bats.

Dietary specializations in animals lead to adaptations in morphology, anatomy and physiology. Neotropical bats, with their high taxonomic and trophic diversity, offer a unique perspective on diet-driven evolutionary adaptations. Here we assess the metabolic response to different dietary sugars among wild-caught bats. We found that insectivorous bats had a pronounced metabolic response to trehalose, whereas bats with nectar and fruit-based diets showed significantly higher blood glucose levels in response to glucose and sucrose, reaching levels over 750 mg dl-1. The genomic analysis of 22 focal species and two outgroup species identified positive selection for the digestive enzyme trehalase in insect eaters, while sucrase-isomaltase showed selection in lineages with omnivorous and nectar diets. By examining anatomical and cellular features of the small intestine, we discovered that dietary sugar proportion strongly impacted numerous digestive traits, providing valuable insight into the physiological implications of molecular adaptations. Using hybridization chain reaction (HCR) RNA fluorescence in situ hybridization, we observed unusually high expression in the glucose transporter gene Slc2a2 in nectar bats, while fruit bats increased levels of Slc5a1 and Slc2a5. Overall, this study highlights the intricate interplay between molecular, morphological and physiological aspects of diet evolution, offering new insights into the mechanisms of dietary diversification and sugar assimilation in mammals.

CRKL Enhances YAP Signaling through Binding and JNK/JUN Pathway Activation in Liver Cancer.

The Hippo pathway transducers yes-associated protein (YAP) and WW-domain containing transcription regulator 1 (WWTR1/TAZ) are key regulators of liver tumorigenesis, promoting tumor formation and progression. Although the first inhibitors are in clinical trials, targeting the relevant upstream regulators of YAP/TAZ activity could prove equally beneficial. To identify regulators of YAP/TAZ activity in hepatocarcinoma (HCC) cells, we carried out a proximity labelling approach (BioID) coupled with mass spectrometry. We verified CRK-like proto-oncogene adaptor protein (CRKL) as a new YAP-exclusive interaction partner. CRKL is highly expressed in HCC patients, and its expression is associated with YAP activity as well as poor survival prognosis. In vitro experiments demonstrated CRKL-dependent cell survival and the loss of YAP binding induced through actin disruption. Moreover, we delineated the activation of the JNK/JUN pathway by CRKL, which promoted YAP transcription. Our data illustrate that CRKL not only promoted YAP activity through its binding but also through the induction of YAP transcription by JNK/JUN activation. This emphasizes the potential use of targeting the JNK/JUN pathway to suppress YAP expression in HCC patients.

Diversification of the Rho transcription termination factor in bacteria.

Correct termination of transcription is essential for gene expression. In bacteria, factor-dependent termination relies on the Rho factor, that classically has three conserved domains. Some bacteria also have a functional insertion region. However, the variation in Rho structure among bacteria has not been analyzed in detail. This study determines the distribution, sequence conservation, and predicted features of Rho factors with diverse domain architectures by analyzing 2730 bacterial genomes. About half (49.8%) of the species analyzed have the typical Escherichia coli like Rho while most of the other species (39.8%) have diverse, atypical forms of Rho. Besides conservation of the main domains, we describe a duplicated RNA-binding domain present in specific species and novel variations in the bicyclomycin binding pocket. The additional regions observed in Rho proteins exhibit remarkable diversity. Commonly, however, they have exceptional amino acid compositions and are predicted to be intrinsically disordered, to undergo phase separation, or have prion-like behavior. Phase separation has recently been shown to play roles in Rho function and bacterial fitness during harsh conditions in one species and this study suggests a more widespread role. In conclusion, diverse atypical Rho factors are broadly distributed among bacteria, suggesting additional cellular roles.

Attentional Biases and Nonsuicidal Self-Injury Urges in Adolescents.

Importance: Nonsuicidal self-injury (NSSI) is a significant clinical concern among adolescents. Exposure to NSSI-related content on social media platforms has been suspected to potentially act as a trigger for NSSI. Objective: To use free-viewing eye-tracking and dot-probe paradigms to examine attentional bias and psychophysiological responses to NSSI-related pictorial and textual stimuli in adolescents with and without a history of NSSI. Design, Setting, and Participants: From June 2022 to April 2023, adolescent participants in Vienna, Austria with and without a history of NSSI were exposed to NSSI-related stimuli in this nonrandomized controlled trial. Data were analyzed from December 2023 to January 2024. Exposure: Exposure to NSSI-related stimuli. Main Outcomes and Measures: During both tasks, subjective arousal, NSSI urges, and autonomic nervous system activity were assessed. Results: A total of 50 adolescents in 2 groups, 25 who engaged in NSSI (mean [SD] age 15.86 [1.14] years; 19 female participants [76%]) and 25 who did not (mean [SD] age 16.40 [1.71] years; 19 female participants [76%]) were included. Adolescents with a history of NSSI-but not those without a history of NSSI-showed a clear attentional bias toward NSSI-related images during eye-tracking, as indicated by increased initial fixations (500 ms stimulus presentation mean difference, 28.64%; 95% CI, 18.31%-38.98%; P < .001; 1000 ms stimulus presentation mean difference, 18.50%; 95% CI, 9.05%-27.95%; P < .001) and longer fixation durations (500 ms mean difference, 29.51 ms; 95% CI, 4.3-54.72 ms; P < .001; 1000 ms mean difference, 39.83 ms; 95% CI, 6.90-72.76 ms; P < .001), regardless of stimulus duration. This bias was associated with a heightened urge to engage in NSSI (d = 1.22; 95% CI, 0.69-1.73; P < .001), a trend not seen in adolescents without a history of NSSI. Similarly, in the dot-probe task, only the NSSI group showed an attentional bias toward NSSI images but not toward trauma images, emphasizing the specificity of their attentional bias. Physiological measures revealed no significant differences, suggesting that viewing NSSI images is not associated with increased autonomic arousal. Textual NSSI content did not provoke an attentional bias or heighten NSSI urges in either group. Conclusions and Relevance: In this nonrandomized controlled trial of 50 adolescents, results highlighted a specific attentional bias toward NSSI-related pictorial stimuli in adolescents with a history of NSSI, particularly a difficulty in disengaging from NSSI images. These findings contribute to understanding maladaptive information processing in NSSI and suggest implications for clinical management and cognitive models addressing NSSI triggers. Trial Registration: German Clinical Trials Register identifier: DRKS00025905.

A second space age spanning omics, platforms and medicine across orbits.

The recent acceleration of commercial, private and multi-national spaceflight has created an unprecedented level of activity in low Earth orbit, concomitant with the largest-ever number of crewed missions entering space and preparations for exploration-class (lasting longer than one year) missions. Such rapid advancement into space from many new companies, countries and space-related entities has enabled a 'second space age'. This era is also poised to leverage, for the first time, modern tools and methods of molecular biology and precision medicine, thus enabling precision aerospace medicine for the crews. The applications of these biomedical technologies and algorithms are diverse, and encompass multi-omic, single-cell and spatial biology tools to investigate human and microbial responses to spaceflight. Additionally, they extend to the development of new imaging techniques, real-time cognitive assessments, physiological monitoring and personalized risk profiles tailored for astronauts. Furthermore, these technologies enable advancements in pharmacogenomics, as well as the identification of novel spaceflight biomarkers and the development of corresponding countermeasures. In this Perspective, we highlight some of the recent biomedical research from the National Aeronautics and Space Administration, Japan Aerospace Exploration Agency, European Space Agency and other space agencies, and detail the entrance of the commercial spaceflight sector (including SpaceX, Blue Origin, Axiom and Sierra Space) into aerospace medicine and space biology, the first aerospace medicine biobank, and various upcoming missions that will utilize these tools to ensure a permanent human presence beyond low Earth orbit, venturing out to other planets and moons.

Impact of device resistances in the performance of graphene-based terahertz photodetectors.

In recent years, graphene field-effect-transistors (GFETs) have demonstrated an outstanding potential for terahertz (THz) photodetection due to their fast response and high-sensitivity. Such features are essential to enable emerging THz applications, including 6G wireless communications, quantum information, bioimaging and security. However, the overall performance of these photodetectors may be utterly compromised by the impact of internal resistances presented in the device, so-called access or parasitic resistances. In this work, we provide a detailed study of the influence of internal device resistances in the photoresponse of high-mobility dual-gate GFET detectors. Such dual-gate architectures allow us to fine tune (decrease) the internal resistance of the device by an order of magnitude and consequently demonstrate an improved responsivity and noise-equivalent-power values of the photodetector, respectively. Our results can be well understood by a series resistance model, as shown by the excellent agreement found between the experimental data and theoretical calculations. These findings are therefore relevant to understand and improve the overall performance of existing high-mobility graphene photodetectors.

A one-step low-cost molecular test for SARS-CoV-2 detection suitable for community testing using minimally processed saliva.

The gold standard for coronavirus disease 2019 diagnostic testing relies on RNA extraction from naso/oropharyngeal swab followed by amplification through reverse transcription-polymerase chain reaction (RT-PCR) with fluorogenic probes. While the test is extremely sensitive and specific, its high cost and the potential discomfort associated with specimen collection made it suboptimal for public health screening purposes. In this study, we developed an equally reliable, but cheaper and less invasive alternative test based on a one-step RT-PCR with the DNA-intercalating dye SYBR Green, which enables the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) directly from saliva samples or RNA isolated from nasopharyngeal (NP) swabs. Importantly, we found that this type of testing can be fine-tuned to discriminate SARS-CoV-2 variants of concern. The saliva RT-PCR SYBR Green test was successfully used in a mass-screening initiative targeting nearly 4500 asymptomatic children under the age of 12. Testing was performed at a reasonable cost, and in some cases, the saliva test outperformed NP rapid antigen tests in identifying infected children. Whole genome sequencing revealed that the antigen testing failure could not be attributed to a specific lineage of SARS-CoV-2. Overall, this work strongly supports the view that RT-PCR saliva tests based on DNA-intercalating dyes represent a powerful strategy for community screening of SARS-CoV-2. The tests can be easily applied to other infectious agents and, therefore, constitute a powerful resource for an effective response to future pandemics.

Alcaligenes faecalis corrects aberrant matrix metalloproteinase expression to promote reepithelialization of diabetic wounds.

Chronic wounds are a common and costly complication of diabetes, where multifactorial defects contribute to dysregulated skin repair, inflammation, tissue damage, and infection. We previously showed that aspects of the diabetic foot ulcer microbiota were correlated with poor healing outcomes, but many microbial species recovered remain uninvestigated with respect to wound healing. Here, we focused on Alcaligenes faecalis, a Gram-negative bacterium that is frequently recovered from chronic wounds but rarely causes infection. Treatment of diabetic wounds with A. faecalis accelerated healing during early stages. We investigated the underlying mechanisms and found that A. faecalis treatment promotes reepithelialization of diabetic keratinocytes, a process that is necessary for healing but deficient in chronic wounds. Overexpression of matrix metalloproteinases in diabetes contributes to failed epithelialization, and we found that A. faecalis treatment balances this overexpression to allow proper healing. This work uncovers a mechanism of bacterial-driven wound repair and provides a foundation for the development of microbiota-based wound interventions.

Recent progress in the roles of microRNAs in pulmonary arterial hypertension associated with congenital heart disease.

Research on noncoding RNA, particularly microRNAs (miRNAs), is growing rapidly. Advances in genomic technologies have revealed the complex roles of miRNAs in pulmonary arterial hypertension (PAH) associated with congenital heart disease (CHD). It has been demonstrated that the progression of PAH associated with CHD is characterized by particular dysregulation of miRNAs and is related to cardiovascular remodeling, cell death, and right ventricle dysfunction. This review provides a comprehensive overview of the current state of knowledge regarding the involvement of miRNAs in the pathogenesis and progression of PAH associated with CHD. We commence by explaining the process of miRNA synthesis and its mode of action, as well as the role of miRNA in PAH associated with CHD. Moreover, the article delves into current breakthroughs in research, potential clinical implications, and prospects for future investigations. The review provides the insight into novel approaches for diagnosis, prognosis, and therapy of PAH associated with CHD.