Persistence phenotype and small colony variants (SCVs) can be part of a bacterial bet-hedging strategy for survival under environmental stresses, such as antimicrobial exposure. These phenotypes are ...of particular concern in persistent and relapsing infections, since cells resume to normal growth after cessation of the stressful condition. In this context, we found persisters and unstable SCVs as phenotypic variants of Salmonella enterica that were able to survive ciprofloxacin exposure. A high heterogeneity in persister levels was observed among S. enterica isolates grown under planktonic and biofilm conditions and exposed to ciprofloxacin or ceftazidime, which may indicate persistence as a non-multidrug-tolerant phenotype. Nevertheless, a comparable variability was not found in the formation of SCVs among the isolates. Indeed, similar proportions of SCV in relation to normal colony phenotype (NCP) were maintained even after three successive cycles of ciprofloxacin exposure testing colonies from both origins (SCV or NCP). Additionally, we found filamentous and dividing cells in the same scanning electron microscopy images from both SCV and NCP. These findings lead us to hypothesize that besides variability among isolates, a single isolate may generate distinct populations of persisters, where cells growing under distinct conditions may adopt different and perhaps complementary survival strategies.
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•DFT method was used to study six donor PCPDTBT derivatives based on ketone radicals.•All electronic properties of the derivatives are improved than pure PCPDTBT.•The results indicate ...new potential candidates to be used in the polymer solar cells.
Density functional theory (DFT) was used to study the donor PCPDTBT and its derivatives based on six small ketone radicals to be employed in polymer solar cells (PSCs). The optimized structures show that the ketone groups preserve the PCPDTBT skeleton planarity. For all PCPDTBT derivatives, the HOMO and LUMO energies, energy gap, and open-circuit voltage are improved in comparison to PCPDTBT. Time-dependence DFT calculations reveal a blue shift of ~37 nm in the absorption spectra for all derivatives. In general, herein six new compounds were studied and characterized from theoretical level, and they are potential candidates for use on PSCs.
Advanced-stage solid primary tumors and metastases often express mucin 16 (MUC16), carrying immature glycans such as the Tn antigen, resulting in specific glycoproteoforms not found in healthy human ...tissues. This presents a valuable approach for designing targeted therapeutics, including cancer glycovaccines, which could potentially promote antigen recognition and foster the immune response to control disease spread and prevent relapse. In this study, we describe an adjuvant-free poly(lactic-
-glycolic acid) (PLGA)-based nanoglycoantigen delivery approach that outperforms conventional methods by eliminating the need for protein carriers while exhibiting targeted and adjuvant properties. To achieve this, we synthesized a library of MUC16-Tn glycoepitopes through single-pot enzymatic glycosylation, which were then stably engrafted onto the surface of PLGA nanoparticles, generating multivalent constructs that better represent cancer molecular heterogeneity. These glycoconstructs demonstrated affinity for Macrophage Galactose-type Lectin (MGL) receptor, known to be highly expressed by immature antigen-presenting cells, enabling precise targeting of immune cells. Moreover, the glycopeptide-grafted nanovaccine candidate displayed minimal cytotoxicity and induced the activation of dendritic cells
, even in the absence of an adjuvant.
, the formulated nanovaccine candidate was also nontoxic and elicited the production of IgG specifically targeting MUC16 and MUC16-Tn glycoproteoforms in cancer cells and tumors, offering potential for precise cancer targeting, including targeted immunotherapies.
A methodology enabling the customization of shoes for comfort improvement is proposed and assessed. For this aim, 3D printed graded density inserts were placed in one of the critical plantar pressure ...zones of conventional insoles, the heel. A semi-automated routine was developed to design the 3D inserts ready for printing, which comprises three main stages: (i) the definition of the number of areas with different mesh density, (ii) the generation of 2D components with continuous graded mesh density, and (iii) the generation of a 3D component having the same 2D base mesh. The adequacy of the mesh densities used in the inserts was previously assessed through compression tests, using uniform mesh density samples. Slippers with different pairs of inserts embedded in their insoles were mechanically characterized, and their comfort was qualitatively assessed by a panel of users. All users found a particular pair, or a set, of prototype slippers more comfortable than the original ones, taken as reference, but their preferences were not consensual. This emphasizes the need for shoe customization, and the usefulness of the proposed methodology to achieve such a goal.
Acinetobacter baumannii is one of the main causes of healthcare-associated infections that threaten public health, and carbapenems, such as meropenem, have been a therapeutic option for these ...infections. Therapeutic failure is mainly due to the antimicrobial resistance of A. baumannii, as well as the presence of persister cells. Persisters constitute a fraction of the bacterial population that present a transient phenotype capable of tolerating supra-lethal concentrations of antibiotics. Some proteins have been suggested to be involved in the onset and/or maintenance of this phenotype. Thus, we investigated the mRNA levels of the adeB (AdeABC efflux pump component), ompA, and ompW (outer membrane proteins) in A. baumannii cells before and after exposure to meropenem.
We found a significant increase (p-value < 0.05) in the expression of ompA (> 5.5-fold) and ompW (> 10.5-fold) in persisters. However, adeB did not show significantly different expression levels when comparing treated and untreated cells. Therefore, we suggest that these outer membrane proteins, especially OmpW, could be part of the mechanism of A. baumannii persisters to deal with the presence of high doses of meropenem. We also observed in the Galleria mellonella larvae model that persister cells are more virulent than regular ones, as evidenced by their LD
values.
Taken together, these data contribute to the understanding of the phenotypic features of A. baumannii persisters and their relation to virulence, as well as highlight OmpW and OmpA as potential targets for drug development against A. baumannii persisters.
Muscle invasive bladder cancer (MIBC, stage ≥T2) is generally associated with poor prognosis, constituting the second most common cause of death among genitourinary tumours. Due to high molecular ...heterogeneity significant variations in the natural history and disease outcome have been observed. This has also delayed the introduction of personalized therapeutics, making advanced stage bladder cancer almost an orphan disease in terms of treatment. Altered protein glycosylation translated by the expression of the sialyl-Tn antigen (STn) and its precursor Tn as well as the activation of the PI3K/Akt/mTOR pathway are cancer-associated events that may hold potential for patient stratification and guided therapy. Therefore, a retrospective design, 96 bladder tumours of different stages (Ta, T1-T4) was screened for STn and phosphorylated forms of Akt (pAkt), mTOR (pmTOR), S6 (pS6) and PTEN, related with the activation of the PI3K/Akt/mTOR pathway. In our series the expression of Tn was residual and was not linked to stage or outcome, while STn was statically higher in MIBC when compared to non-muscle invasive tumours (p = 0.001) and associated decreased cancer-specific survival (log rank p = 0.024). Conversely, PI3K/Akt/mTOR pathway intermediates showed an equal distribution between non-muscle invasive bladder cancer (NMIBC) and MIBC and did not associate with cancer-specif survival (CSS) in any of these groups. However, the overexpression of pAKT, pmTOR and/or pS6 allowed discriminating STn-positive advanced stage bladder tumours facing worst CSS (p = 0.027). Furthermore, multivariate Cox regression analysis revealed that overexpression of PI3K/Akt/mTOR pathway proteins in STn+ MIBC was independently associated with approximately 6-fold risk of death by cancer (p = 0.039). Mice bearing advanced stage chemically-induced bladder tumours mimicking the histological and molecular nature of human tumours were then administrated with mTOR-pathway inhibitor sirolimus (rapamycin). This decreased the number of invasive lesions and, concomitantly, the expression of STn and also pS6, the downstream effector of the PI3K/Akt/mTOR pathway. In conclusion, STn was found to be marker of poor prognosis in bladder cancer and, in combination with PI3K/Akt/mTOR pathway evaluation, holds potential to improve the stratification of stage disease. Animal experiments suggest that mTOR pathway inhibition could be a potential therapeutic approach for this specific subtype of MIBC.
Xylene isomerization is an important step in the production of p‐xylene. The reaction is normally conducted in the gas phase; however, the liquid‐phase isomerization has attracted the attention of ...both industry and academia. Three MFI‐type, two MOR‐type, and one BEA‐type zeolites with different SiO2/Al2O3 ratios were used to study the conversion of aromatics in the liquid‐phase at 513 K. The catalysts were characterized through several techniques to assess their textural and acidic properties. The experiments consisted of 3 ml/min (5 × 10−8 m3/s) of m‐xylene fed to a fixed‐bed column at 2.1 MPa to maintain liquid‐phase conditions for 25–30 h. MFI‐type zeolites showed high selectivity towards p‐xylene and long stability, but m‐xylene conversions were below 5%. Large‐pore zeolites exhibited significantly higher conversion but faster deactivation due to secondary reactions. MOR‐type zeolite with a SiO2/Al2O3 ratio of 40 showed the best performance with a m‐xylene conversion of 45.3% with only 1.9% of secondary products.
Comparison of the catalyst on the conversion of m‐xylene to its isomers and on the deactivation caused by heavy aromatics formed by disproportionation.
Microplastics (MP), i.e., particles measuring less than 5 mm in size, are considered emerging pollutants. The ubiquity of MP is causing great concern among environmental and public health agencies. ...Anthropogenic activities are responsible for the extensive dispersal of MP in nature. Adverse effects on living organisms, interactions with other contaminants occurring in the environment, and the lack of effective degradation/removal techniques are significant issues related to MP. Most MP found in nature are fibrous (FMP). FMP originate from textile products, mainly synthetic fibers (e.g., polyester). Synthetic fibers are intensively used to produce countless goods due to beneficial characteristics such as high mechanical resistance and economic feasibility. FMP are ubiquitous on the planet and impart lasting adverse effects on biodiversity. Data on the consequences of long-term exposure to these pollutants are scarce in the literature. In addition, few studies address the main types of synthetic microfibers released from textiles, their occurrence, adverse effects on organisms, and remediation strategies. This review discusses the relevant topics about FMP and alerts the dangers to the planet. Furthermore, future perspectives and technological highlights for the FMP mitigation/degradation are presented.
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•Fibrous microplastics are released from textile products during manufacture and use.•Occurrence of fibrous microplastics in the textile industry were addressed.•The primary environmental impacts caused by microplastics were discussed.•Fibrous microplastics removal and degradation techniques were presented.
Muscle invasive bladder cancer (MIBC) remains amongst the deadliest genitourinary malignancies due to treatment failure and extensive molecular heterogeneity, delaying effective targeted ...therapeutics. Hypoxia and nutrient deprivation, oversialylation and O-glycans shortening are salient features of aggressive tumours, creating cell surface glycoproteome fingerprints with theranostics potential.
A glycomics guided glycoproteomics workflow was employed to identify potentially targetable biomarkers using invasive bladder cancer cell models. The 5637 and T24 cells O-glycome was characterized by mass spectrometry (MS), and the obtained information was used to guide glycoproteomics experiments, combining sialidase, lectin affinity and bottom-up protein identification by nanoLC-ESI-MS/MS. Data was curated by a bioinformatics approach developed in-house, sorting clinically relevant molecular signatures based on Human Protein Atlas insights. Top-ranked targets and glycoforms were validated in cell models, bladder tumours and metastases by MS and immunoassays. Cells grown under hypoxia and glucose deprivation disclosed the contribution of tumour microenvironment to the expression of relevant biomarkers. Cancer-specificity was validated in healthy tissues by immunohistochemistry and MS in 20 types of tissues/cells of different individuals.
Sialylated T (ST) antigens were found to be the most abundant glycans in cell lines and over 900 glycoproteins were identified potentially carrying these glycans. HOMER3, typically a cytosolic protein, emerged as a top-ranked targetable glycoprotein at the cell surface carrying short-chain O-glycans. Plasma membrane HOMER3 was observed in more aggressive primary tumours and distant metastases, being an independent predictor of worst prognosis. This phenotype was triggered by nutrient deprivation and concomitant to increased cellular invasion. T24 HOMER3 knockdown significantly decreased proliferation and, to some extent, invasion in normoxia and hypoxia; whereas HOMER3 knock-in increased its membrane expression, which was more pronounced under glucose deprivation. HOMER3 overexpression was associated with increased cell proliferation in normoxia and potentiated invasion under hypoxia. Finally, the mapping of HOMER3-glycosites by EThcD-MS/MS in bladder tumours revealed potentially targetable domains not detected in healthy tissues.
HOMER3-glycoforms allow the identification of patients' subsets facing worst prognosis, holding potential to address more aggressive hypoxic cells with limited off-target effects. The molecular rationale for identifying novel bladder cancer molecular targets has been established.
Inferring causal relationships from observational data is a key challenge in understanding the interpretability of Machine Learning models. Given the ever-increasing amount of observational data ...available in many areas, Machine Learning algorithms used for forecasting have become more complex, leading to a less understandable path of how a decision is made by the model. To address this issue, we propose leveraging ensemble models, e.g., Random Forest, to assess which input features the trained model prioritizes when making a forecast and, in this way, establish causal relationships between the variables. The advantage of these algorithms lies in their ability to provide feature importance, which allows us to build the causal network. We present our methodology to estimate causality in time series from oil field production. As it is difficult to extract causal relations from a real field, we also included a synthetic oil production dataset and a weather dataset, which is also synthetic, to provide the ground truth. We aim to perform causal discovery, i.e., establish the existing connections between the variables in each dataset. Through an iterative process of improving the forecasting of a target's value, we evaluate whether the forecasting improves by adding information from a new potential driver; if so, we state that the driver causally affects the target. On the oil field-related datasets, our causal analysis results agree with the interwell connections already confirmed by tracer information; whenever the tracer data are available, we used it as our ground truth. This consistency between both estimated and confirmed connections provides us the confidence about the effectiveness of our proposed methodology. To our knowledge, this is the first time causal analysis using solely production data is employed to discover interwell connections in an oil field dataset.
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