Wide Panel Research Articles

Systematic Profile of Oxylipins in Myocardial Infarction by Liquid Chromatography-Tandem Mass Spectrometry.

Oxylipins play an important role in inflammatory processes, accompanying the occurrence of myocardial infarction. Analyzing a wide panel of oxylipins derived from more polyunsaturated fatty acids may provide valuable information to elucidate the relationships between the signaling mediator profile and myocardial infarction comprehensively. An ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for simultaneously measuring 74 oxylipins in 50-μL human samples with a 10-min chromatographic run and easy operation has been developed and validated. Accuracy and precision were below 15% of the relative errors in 99% of quality control. Recoveries and matrix effect were considered acceptable. Potential effects of different collecting tubes were also assessed. We successfully utilized our approach to measure plasma samples obtained from 99 healthy individuals and 302 myocardial infarction patients. Profiles of oxylipins discovered potential biomarkers and clarified the pathological characteristics of oxylipin metabolism in myocardial infarction. Our approach was rapid, accurate, and precise, with high throughput, low sample volume, and easy operation, suitable for large-scale studies.

Longitudinal Cracking in Alternate Jointed Plain Concrete Panel Widths in Wisconsin

This paper presents the results of comparative analyses of longitudinal cracking occurring in wider concrete panels (>12ft) in Wisconsin. The analysis revealed that approximately 60% of 1,008 segments involved in the study experienced longitudinal cracking. For the same pavement thickness and joint spacing, panel widths of 12ft and 14ft that exhibited higher amounts of longitudinal cracking than the larger 15-ft panel, received corresponding higher levels of truck traffic than the 15-ft panel. Pavement segments with thickness of 9in at a joint spacing of 20ft experienced higher amounts of longitudinal cracking in 14ft and 15ft wide panels than in 12ft wide panels. For a 9-in pavement thickness at a joint spacing of 16ft, the best performance may be attained by using 12ft wide panels. For a 14-ft wide panel with 9-in or 10-in thickness, the best performance may be attained using a 15-ft joint spacing. In addition, models developed showed that longitudinal cracking variations in 10-in 14-ft wide panels at 16-ft or 20-ft joint spacing is dictated by the Annual Average Daily Traffic, while percent trucks is the foremost factor that explains variations in longitudinal cracking for 12-ft and 15-ft wide panels at specific joint spacing and thickness values.

Development of 1,2,3-triazole hybrids as multi-faced anticancer agents co-targeting EGFR/mTOR pathway and tubulin depolymerization.

Novel 1,2,3-triazole hybrids bearing various substituents have been synthesized as potential anticancer agents. Ligand-based approach has been adopted to design these compounds relying on the hybridization of 1,2,3-triazole with α,β-unsaturated carbonyl, 5- and 6-membered heterocyclic scaffolds. All synthesized members were investigated for their cytotoxic potency against nine types comprising 60 panels of human cancerous cells by the US National Cancer Institute: Development Therapeutic Program (US_NCI_DTP). Among the tested members, 4b, 4e, and 4h showed prominent cytotoxic effects (> 80% growth inhibition: GI) on a wide panel of tested cancer cell lines, mainly melanoma and colorectal cancer redeeming their selection for five dose testing. Presenting low nanomolar GI50 concentrations, two representative potent anticancer compounds 4b and 4e were subjected to cytotoxicity testing on colon normal cell (FHC) to investigate their safety window and they showed less toxicity to normal cells at the concentration required to produce anticancer effect. Furthermore, 4b and 4e were exposed to additional mechanistic studies in colorectal cancer cell HCT-116 suggesting multifaceted mechanisms of action. A study into the effects of cytotoxic chemicals 4b and 4e on cell cycle progression regulation showed triggered the arrest of cell cycles during the G1 and S phases. Moreover, 4b and 4e caused cell death mainly through apoptosis the thing that has been reinforced by the elevated Bax: Bcl2 ratio, as well as concentrations of caspases 3 and 9 within HCT-116. Further, both compounds showed prominent inhibition profiles against tubulin polymerization as well as EGFR catalytic activity reaching down to low-digit micromolar and sub-micromolar concentrations, respectively, as compared to positive reference controls. Compounds' impacts on gene expression of cancer-associated and EGFR-downstream signaling markers including TNFα, IL-6, and mTOR, were explored in HCT-116 highlighted significant downregulations versus the untreated cells. Docking studies demonstrated the specific fit of 4b and 4e into EGFR and the colchicine binding site of tubulin.

Binding mechanism of oligopeptides on solid surface: assessing the significance of single-molecule approach.

This paper addresses the complementarity and potential disparities between single-molecule and ensemble-average approaches to probe the binding mechanism of oligopeptides on inorganic solids. Specifically, we explore the peptide/gold interface owing to its significance in various topics and its suitability to perform experiments both in model and real conditions. Experimental results show that the studied peptide adopts a lying configuration upon adsorption on the gold surface and interacts through its peptidic links and deprotonated thiolate extremities, in agreement with theoretical predictions. Single-molecule force spectroscopy (SMFS) measurements revealed the existence of a wide panel of adhesion forces, resulting from the interaction between individual peptide moieties and the abundant surface sites. We therefore propose methodological developments for sorting the events of interest to understand the peptide adsorption mechanism. Thermodynamic and kinetic aspects of the peptide adsorption are probed using both static and dynamic force spectroscopy measurements. Specifically, we show the possibility of providing a reasonable estimate of the peptide free energy of adsorption ΔadsG° by exploring the fluctuations of the adhesion work, based on the Jarzynski equality, and by using a parametric Gamma estimator. The proposed approach offers a relevant method for studying the different factors influencing the peptide adsorption and evaluating their impact on ΔadsG° as an alternative to exploring adhesion forces that may lead to misinterpretations. This is illustrated by the comparison of the adsorption of two peptides with specific amino acids substitution. Our method provides insights into the overall mechanism by which peptides interact with the surface and allows an integration of the single-molecule versus ensemble-average points of view.

Evaluation of Reference Gene Stability for Investigations of Intracellular Signalling in Human Cancer and Non-Malignant Mesenchymal Stromal Cells.

Real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a powerful tool for analysing target gene expression in biological samples. To achieve reliable results by RT-qPCR, the most stable reference genes must be selected for proper data normalisation, particularly when comparing cells of different types. We aimed to choose the least variable candidate reference genes among eight housekeeping genes tested within a set of human cancer cell lines (HeLa, MCF-7, SK-UT-1B, A549, A431, SK-BR-3), as well as four lines of normal, non-malignant mesenchymal stromal cells (MSCs) of different origins. The reference gene stability was evaluated using four algorithms (BestKeeper, NormFinder, geNorm and the comparative ΔCt method) and ranked with the RefFinder web-based tool. We found increased variability in the housekeeping genes' expression in the cancer cell lines compared to that in normal MSCs. POP4 and GAPDH were identified as the most suitable reference genes in cancer cells, while 18S and B2M were the most suitable in MSCs. POP4 and EIF2B1 were shown to be the least variable genes when analysing normal and cancer cell lines together. Epidermal growth factor receptor (EGFR) mRNA relative expression was normalised by the three most stable or three least stable reference genes to demonstrate the reliability of reference genes validation. We analysed and selected stable reference genes for RT-qPCR analysis in the wide panel of cancer cell lines and MSCs. The study provides a reliable tool for future research concerning the expression of genes involved in various intracellular signalling pathways and emphasises the need for careful selection of suitable references before analysing target gene expression.

Construct validity and responsiveness of ASAS Health Index assessed in two longitudinal studies of tumour necrosis factor alpha inhibitor initiation and dose reduction in patients with axial spondyloarthritis

BackgroundThe Assessment of SpondyloArthritis international Society Health Index (ASAS HI) is a novel questionnaire of global functioning for patients with axial spondyloarthritis (SpA).ObjectiveThe objective was to assess the construct validity,...

Uncovering the arsenal of class II bacteriocins in salivarius streptococci

Facing the antibiotic resistance crisis, bacteriocins are considered as a promising alternative to treat bacterial infections. In the human commensal Streptococcus salivarius, the production of unmodified bacteriocins (or salivaricins) is directly controlled at the transcriptional level by quorum-sensing. To discover hidden bacteriocins, we harnessed here the unique molecular signatures of salivaricins not yet used in available computational pipelines and performed genome mining followed by orthogonal reconstitution and expression. From 100 genomes of S. salivarius, we identified more than 50 bacteriocin candidates clustered into 21 groups. Strain-based analysis of bacteriocin combinations revealed significant diversity, reflecting the plasticity of seven independent loci. Activity tests showed both narrow and broad-spectrum bacteriocins with overlapping activities against a wide panel of Gram-positive bacteria, including notorious multidrug-resistant pathogens. Overall, this work provides a search-to-test generic pipeline for bacteriocin discovery with high impact for bacterial ecology and broad applications in the food and biomedical fields.

Analytical and clinical evaluation of a duplex RT-qPCR assay for the detection and identification of o’nyong-nyong and chikungunya virus

ABSTRACT The mosquito-borne alphavirus o’nyong-nyong virus (ONNV) has proven its potential to cause major human outbreaks. On the African continent, ONNV causes unspecific febrile illness and co-circulates with the close relative chikungunya virus (CHIKV). The true scale of ONNV burden is poorly understood in Africa, because of the scarce availability of molecular in-house and commercial assays, strong cross-reactivity between ONNV and CHIKV in serological assays and a lack of surveillance. We designed a new RT-qPCR assay targeting the E1 gene for the detection of ONNV that can be used in monoplex or in duplex format, combined with a previously published CHIKV monoplex assay targeting the nsp1. The lower limit of detection with 95% positivity rate was determined to be <10 RNA copies/µL in monoplex and duplex format for both ONNV and CHIKV. Both monoplex assays and the duplex assay proved to be linear within the tested range of approximately 108 to 102 RNA copies/µl, and showed 100% specificity against a wide panel of arbovirus supernatants as well as several other pathogens in clinical samples. Testing of CHIKV positive serum and ONNV-spiked plasma samples confirmed the suitability of the assays in a clinical setting. The new assays provide a robust tool for molecular ONNV as well as ONNV/CHIKV simultaneous detection and may contribute to clarify the true burden of the two viruses, to improve arbovirus surveillance and to strengthen epidemics preparedness in Africa.

A diagnostic journey: Unraveling von Willebrand disease in a child with inconclusive initial testing

Background: Epistaxis attacks in childhood are an important reason for pediatric emergency visits, most of which are minor and infrequent. However, recurrent epistaxis can be a sign of an underlying bleeding disorder such as von Willebrand disease. This case report highlights controversies and obstacles that may arise during the diagnostic process, hindering a clear confirmation. Case presentation: An eight-year-old boy, who had a history of recurrent epistaxis and easy bruising since infancy, was presented to the Pediatric Emergency Department of Al Khor Hospital for a repeat episode of nosebleed. Local examination showed dilated capillaries in addition to nasal bleeding signs. Family history revealed similar past complaints in both his brother and his father. Previous epistaxis attacks were mostly left-sided. Blood workups were not diagnostic. Later, repeated blood tests led by high clinical suspicion resulted in the diagnosis of von Willebrand disease. The new diagnosis had a direct effect on the patient’s bleeding management. Nose bleeds peaked in frequency and severity over the next two years and decreased remarkably thereafter, the same pattern as before in the father and older brother. Discussion: The heterogeneous nature of von Willebrand disease, overlapping with the healthy population, creates a diagnostic dilemma. Therefore, balanced diagnostic approach is crucial. Bleeding history and family history provide invaluable clues but are not definitive. The blood workups may struggle with the variations in function and fluctuating levels of the von Willebrand factor molecule. Although genetic testing is helpful in some cases, it has its limitations. Conclusion: The diagnosis of von Willebrand’s disease generally follows the traditional approach. It begins with a detailed bleeding history combined with a focused family history that pinpoints potential cases, followed by blood testing. However, interpretation of blood workups can be challenging due to the patient-to-patient and within-patient variability of the disease, the wide panel of investigations, and the different cutoff levels in different guidelines. Despite all this, testing is irreplaceable. Management of bleeding should begin immediately and should not be affected by inconclusive blood workups.

BlaGES-producing ST654 comprises a quarter of all carbapenem-resistant Pseudomonas aeruginosa in blood isolates from 15 hospitals.

Carbapenem-resistant Pseudomonas aeruginosa (CRPA) are of major clinical concern. We analyzed 85 P. aeruginosa blood isolates non-susceptible to carbapenems collected during 2021-2023 from 15 medical centers in Israel. We aimed to determine the prevalence of high-risk clones, examine clonality, test antibiotic susceptibility, and assess the presence of acquired resistance genes, including carbapenemases. Whole-genome sequencing was performed using Illumina sequencing technology. Susceptibly was determined using the broth microdilution method. In the entire sample, 43.5% were high-risk clones. A main clade (27.1% of isolates) found in multiple hospitals comprised 19 isolates belonging to the high-risk ST654 clone and four closely related isolates. The isolates in this main clade harbored a broad set of resistance genes, including GES-type genes, and 91% had a mutated outer membrane protein (OprD). Isolates in the main clade were uniformly tobramycin (TOB) resistant and 83% were ceftolozane/tazobactam resistant. In the entire sample, we found high resistance to most antipseudomonal agents, including new beta-lactam/beta-lactamase inhibitor combinations. No uniform susceptibility to an antipseudomonal agent was found. Carbapenemases were carried by 9.4% of isolates (5.9% blaGES-5 and 3.5% blaNDM-1) and oprD was mutated in 67% of isolates. Thus, the epidemiology of CRPA is explained by a combination of clonal expansion of a dominant high-risk clade and sporadic occurrence of mutated strains. Our findings highlight the importance of susceptibility testing using a wide panel of antibiotics when CRPA is detected. Prevention measures tracking and controlling emerging high-risk clades and clones are crucial to limit the spread of CRPA.

A-247 Performance and Compatibility Assessment of Serially Diluted Microbix QAPTM Controls with Seegene NovaplexTM Assays

Abstract Background Choosing high-quality external (third-party) controls like Microbix QAPTM PROCEEDxTM Controls and REDxTM Controls is crucial during test development, validation of Laboratory Developed Tests (LDTs), performance verification, proficiency testing, instrument qualifications, operator training and R&amp;D testing. Laboratories may prefer to include multiple positive controls at varying concentrations to achieve reliable performance across a broad range of detection concentrations. In this study, we assess the performance and compatibility of Microbix QAPTM PROCEEDxTM liquid Controls and REDxTM swabs with Seegene’s real time qPCR assays that utilize TOCE and MuDTTM technologies. TOCE and MuDTTM technologies achieve multiple reads in a single channel and are the backbone of Seegene’s wide coverage panel assays. Methods The controls below were reconstituted and/or diluted in Copan UTM and extracted on the Seegene STARlet liquid handler using Seegene’s STARmag Universal extraction kit. After extraction, PCR was run on the Bio-Rad CFX 96 Opus Dx and analyzed using the Seegene Viewer software. Results Microbix QAPsTM liquid formulations were tested at various dilutions from stock up to 1/2048 with N=5per dilution. The VP-13-01 Flu A control was detected up to a concentration of1/2048 and at this dilution had Ct values of 30 + 0.3 for the Novaplex Respiratory Panel 1A (RP1A) Assay, 28.6 + 0.3 cycles for the Seegene SARS-CoV-2/FluA/FluB/RSV (Quad) Assay, and 27.9 + 0.3 for the Respiratory Virus Master (RVM) Assay. The Microbix VP-14-01 Flu B control was detected up to 1/2048 with Ct values of 34.3 + 0.3 for the RP1A Assay, 30 + 0.5 for the Quad Assay, and 30.3 + 0.3 for the RVM Assay. The VP-07-01 RSV A control was also detected at a 1/2048 dilution with Ct values of 32.7 + 0.3 for RP1A, 30.6 + 0.6 for Quad Assay, and 29.2 + 0.4 for RVM. The VP-61-01 Trichomonas vaginalis (TV) control was detected up to a dilution of 1/128 with a Ct value of 37.3 + 0.4 when tested with the Seegene Sexually Transmitted Infection Essential (STI) Assay. The VP-02-M1 HSV1 control was detected up to a dilution of 1/128 with a Ct value of 31.3 + 0.7 for the Seegene Genital Ulcer (GU) Assay. The VP-23-M1 HSV2 control was detected p to a dilution of 1/128 with a Ct value of 31.9 + 0.5 for the GU Assay. Microbix also makes a dry swab formulation product line that we tested in 3mL of UTM. Of these we tested the RED-S-10-M1 swab covering the SARS-CoV-2, Flu A, Flu B, and RSV controls. The VP-S-12-M5 swab covering the Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), and TV controls. We also tested the VP-S-03-M1 swab covering HSV1, HSV2, VZV and syphilis controls. When tested with various assays all swabs were detected at the stock formulation. Conclusions Microbix QAPsTM are inactivated whole-organism controls, mimicking patient samples, and meet the stringent demand for quality or performance assessments. Their compatibility with Seegene’s Assays over a range of dilutions and formulations makes them ideal for ensuring reliable detection across multiple diagnostic needs.

Bioactive Compound Diversity in a Wide Panel of Sweet Potato (Ipomoea batatas L.) Cultivars: A Resource for Nutritional Food Development.

This study provides an overview of the composition of the raw root flesh of a panel of 22 sweet potato (Ipomoea batatas L.) cultivars, with a focus on bioactive compounds. The large diversity of the proximate and phytochemical compositions observed between cultivars and within and between different flesh colors pointed out the importance of composition analysis and not only color choice for the design of foods with nutritional benefits. The nutritional composition (starch, protein, total dietary fibers) and bioactive compound composition of 22 cultivars from Reunion Island, maintained in the Vatel Biological Resource Center, were investigated. Orange and purple cultivars stood out from white and yellow cultivars for their higher nutritional composition. Purple sweet potatoes were notable for their high contents of anthocyanins (55.7 to 143.4 mg/g dry weight (DW)) and phenolic compounds, in particular chlorogenic acid and ferulic acid, contributing to antioxidant activities, as well as their fiber content (14.1 ± 2.1% DW). Orange cultivars were rich in β-carotene (47.2 ± 0.7 mg/100 g DW) and to a lesser extent α-carotene (4.8 ± 1.2 mg/100 g DW). In contrast, certain white cultivars demonstrated suboptimal nutritional properties, rendering them less relevant even for applications where the lack of coloration in food is desired. Those characteristics enable the selection of sweet potato varieties to design food products ensuring optimal nutritional benefits and culinary versatility.

Genome-wide association study of lignin trait in elite spring wheat against spot blotch disease

Spot blotch disease in wheat is one of major yield-limiting factor in warm humid conditions of South Asia. Developing the disease-resistant variety through molecular methods is an economically and environmentally eco-friendly approach. Diseased wheat leaves associated with lignin content increased by two or three times at 72 h compared with control. The genomic region associated with lignin content was investigated in a set of 289 diverse spring wheat genotypes. The genome - wide association studies panel employed a 90K Illumina SNP array and phenotyped in four environments. The GWAS analysis showed a total of 86 marker-trait associations on 1A, 1B, 2A, 2B, 2D, 3A, 3D, 4A, 5A, 5B, 6A, 6D, 7A, 7B, 7D were linked lignin trait. Insilco analysis revealed that significant SNPs were located on import putative candidate genes thirteen in the Transmembrane domain of the protein-kinase family and were involved in the host-pathogen interaction. The identified significant MTAs for further validation and useful for the wheat breeding programs to develop the spot blotch disease resistance.

Illuminating insights: Exploring the effect of 16/8 intermittent fasting on serum cytokine levels in overweight adults.

The immune system's pivotal role extends to numerous diseases, and maintaining a balance between dietary and consumed energy is vital for preventing chronic illnesses and increasing life expectancy. Intermittent fasting (IF), a dietary approach typically implemented through time restrictions, exerts positive effects on the immune system and shows promising outcomes in managing various diseases. To evaluate the effectiveness of IF on the immune system with a wide cytokine panel. A total of 21 volunteers with body mass index (BMI) between 25 and 30 were included in the study. Fasting was applied for 16 h in a day to the volunteers, and they were free to consume food for the rest of the day. The weight, BMI, interleukin (IL)-1β, interferon (IFN)-α2, IFN-γ, tumor necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1, IL-6, IL-8, IL-10, IL-12p70, IL-17A, IL-18, IL-23, and IL-33 values were measured using flow cytometry and compared before and after 21 days follow-up. The mean age of study participants was 37.76 ±8.06 years and weight loss of the volunteers was 3.35 percentile compared to the values obtained before fasting. The pro-inflammatory cytokines decreased, while anti-inflammatory cytokines increased after fasting; there was a significant difference in TNF-α, MCP-1, IL-6, IL-8, and IL-33 values. Also, IL-1β, IL-8 and IL-12p70 had moderately positive, IL-33 had strongly negative, and IL-10 had moderately negative correlation with the BMI change over time. Intermittent fasting has positive effects on obesity-induced inflammation and promotes decrease in proinflammatory cytokines and increase in IL-33 values, which is known to have a protective effect on fat-associated inflammation.

A generalized geometric mechanics theory for multi-curve-fold origami: Vertex constrained universal configurations

Folding paper along curves leads to spatial structures that have curved surfaces meeting at spatial creases, defined as curve-fold origami. In this work, we provide an Eulerian framework focusing on the mechanics of arbitrary curve-fold origami, especially for multi-curve-fold origami with vertices. We start with single-curve-fold origami that has wide panels. Wide panel leads to different domains of mechanical responses induced by various generator distributions of the curved surface. The theories are then extended to multi-curve-fold origami, involving additional geometric correlations between creases. As an illustrative example, the deformation and equilibrium configuration of origami with annular creases are studied both theoretically and numerically. Afterward, single-vertex curved origami theory is studied as a special type of multi-curve-fold origami. We find that the extra periodicity at the vertex strongly constrains the configuration space, leading to a region near the vertex that has a striking universal equilibrium configuration regardless of the mechanical properties. Both theories and numerics confirm the existence of the universality in the near-field region. In addition, the far-field deformation is obtained via energy minimization and validated by finite element analysis. Our generalized multi-curve-fold origami theory, including the vertex-contained universality, is anticipated to provide a new understanding and framework for the shape programming of the curve-fold origami system.

Exploring the therapeutic potential of a novel series of imidazothiadiazoles targeting focal adhesion kinase (FAK) for pancreatic cancer treatment: synthesis, mechanistic insights and promising antitumor and safety profile

Focal Adhesion Kinase (FAK) is a non-receptor protein tyrosine kinase that plays a crucial role in various oncogenic processes related to cell adhesion, migration, proliferation, and survival. The strategic targeting of FAK represents a burgeoning approach to address resistant tumours, such as pancreatic ductal adenocarcinoma (PDAC). Herein, we report a new series of twenty imidazo[2,1-b][1, 3, 4]thiadiazole derivatives assayed for their antiproliferative activity against the National Cancer Institute (NCI-60) panel and a wide panel of PDAC models. Lead compound 10l exhibited effective antiproliferative activity against immortalised (SUIT-2, CAPAN-1, PANC-1, PATU-T, BxPC-3), primary (PDAC-3) and gemcitabine-resistant clone (PANC-1-GR) PDAC cells, eliciting IC50 values in the low micromolar range (1.04–3.44 µM), associated with a significant reduction in cell-migration and spheroid shrinkage in vitro. High-throughput kinase arrays revealed a significant inhibition of the FAK signalling network, associated to induction of cell cycle arrest in G2/M phase, suppression of tumour cell invasion and apoptosis induction. The high selectivity index/toxicity prompted studies using PDAC mouse xenografts, demonstrating significant inhibition of tumour growth and safety. In conclusion, compound 10l displayed antitumor activity and safety in both in vitro and in vivo models, emerging as a highly promising lead for the development of FAK inhibitors in PDAC.

A comprehensive in vitro characterization of non-crosslinked, diverse tissue-derived collagen-based membranes intended for assisting bone regeneration.

Collagen-based membranes are class III-medical devices widely used in dental surgical procedures to favour bone regeneration. Here, we aimed to provide biophysical and biochemical data on this type of devices to support their optimal use and design/manufacturing. To the purpose, four commercial, non-crosslinked collagen-based-membranes, obtained from various sources (equine tendon, pericardium or cortical bone tissues, and porcine skin), were characterized in vitro. The main chemical, biophysical and biochemical properties, that have significant clinical implications, were evaluated. Membranes showed similar chemical features. They greatly differed in morphology as well as in porosity and density and showed a diverse ranking in relation to these latter two parameters. Samples highly hydrated in physiological medium (swelling-ratio values in the 2.5-6.0 range) and, for some membranes, an anisotropic expansion during hydration was, for the first time, highlighted. Rheological analyses revealed great differences in deformability (150-1500kPa G') also alerting about the marked variation in membrane mechanical behaviour upon hydration. Samples proved diverse sensitivity to collagenase, with the cortical-derived membrane showing the highest stability. Biological studies, using human-bone-derived cells, supported sample ability to allow cell proliferation and to prompt bone regeneration, while no relevant differences among membranes were recorded. Prediction of relative performance based on the findings was discussed. Overall, results represent a first wide panel of chemical/biophysical/biochemical data on collagen-based-membranes that 1) enhances our knowledge of these products, 2) aids their optimal use by providing clinicians with scientific basis for selecting products based on the specific clinical situation and 3) represents a valuable reference for optimizing their manufacturing.

The Combined Immunohistochemical Expression of GLI1 and BCOR in Synovial Sarcomas for the Identification of Three Risk Groups and Their Prognostic Outcomes: A Study of 52 Patients.

Synovial sarcoma (SS) is a rare soft-tissue tumor characterized by a monomorphic blue spindle cell histology and variable epithelial differentiation. Morphologically, SSs may be confused with other sarcomas. Systemic treatment is more effective for patients with high-risk SSs, patients with advanced disease, and younger patients. However, further studies are required to find new prognostic biomarkers. Herein, we describe the morphological, molecular, and clinical findings, using a wide immunohistochemical panel, of a series of SS cases. We studied 52 cases confirmed as SSs by morphological diagnosis and/or molecular studies. Clinical data (gender, age, tumor size, tumor location, resection margins, adjuvant treatment, recurrences, metastasis, and survival) were also retrieved for each patient. All the available H&E slides were examined by four pathologists. Three tissue microarrays (TMAs) were constructed for each of the tumors, and a wide immunohistochemical panel was performed. For time-to-event variables, survival analysis was performed using Kaplan-Meier curves and log-rank testing, or Cox regression. Statistical significance was considered at p < 0.05. The mean age of our patients was 40.33, and the median was 40.5 years. We found a predominance of males versus females (1.7:1). The most frequent morphological subtype was monophasic. TRPS1, SS18-SSX, and SSX-C-terminus were positive in 96% of cases. GLI1 expression was strong in six and focal (cytoplasmic) in twenty patients. Moreover, BCOR was expressed in more than half of SSs. Positive expression of both proteins, BCOR and GLI1, was correlated with a worse prognosis. Multivariate analysis was also performed, but only BCOR expression appeared to be significant. The combination of GLI1 and BCOR antibodies can be used to group SSs into three risk groups (low, intermediate, and high risk). We hypothesize that these findings could identify which patients would benefit from receiving adjuvant treatment and which would not. Moreover, these markers could represent therapeutic targets in advanced stages. However, further, larger series of SSs and molecular studies are necessary to corroborate our present findings.

New Insights in the Computational pKb Determination of Primary Amines and Anilines.

Extensive research has already provided reliable methods for the in silico prediction of pKa, while a trustworthy strategy for pKb determination is still being sought. Indeed, the approaches previously exploited for computing pKa have shown their weakness in predicting pKb. In the light of the exceptional reliability demonstrated in the pKa calculation of a wide panel of organic acids, in this work, we exploited our "easy to use methodology", based on the direct approach, to predict the pKb of primary amines. Herein, CAM-B3LYP was compared to WB97XD and B3PW91, exploring the solvation model based on density (SMD) and the polarizable continuum model (PCM), in the presence of two explicit water molecules. Noteworthy, CAM-B3LYP and WB97XD returned completely different solvent accessible surfaces (SAS) and electron potential maps (EPM) for the bases and the conjugated acids, independently from the nature of the substituents. Once again, CAM-B3LYP/SMD/2H2O method confirmed its remarkable reliability, leading to a minimum average error (MAE) lower than 0.3. This outstanding result strengthens the trustworthiness of our method, already successfully applied to predict the pKa of different substituted phenols and carboxylic acids. Thus, our "easy-to-use" process can predict also the pKb of primary ammines and anilines, always ensuring consistent outputs.

Antibacterial Interactions of Ethanol-Dispersed Multiwalled Carbon Nanotubes with Staphylococcus aureus and Pseudomonas aeruginosa.

Infectious diseases are acknowledged as one of the leading causes of death worldwide. Statistics show that the annual death toll caused by bacterial infections has reached 14 million, most of which are caused by drug-resistant strains. Bacterial antibiotic resistance is currently regarded as a compelling problem with dire consequences, which motivates the urgent identification of alternative ways of fighting bacteria. Various types of nanomaterials have been reported to date as efficient antibacterial solutions. Among these, carbon-based nanomaterials, such as carbon nanodots, carbon graphene oxide, and carbon nanotubes (CNTs), have been shown to be effective in killing a wide panel of pathogenic bacteria. With this study, we aim to provide additional insights into this topic of research by investigating the antibacterial activity of a specific type of multiwalled CNTs, with diameters from 50 to 150 nm, against two representative opportunistic pathogens, i.e., the Gram-positive bacterium Staphylococcus aureus and the Gram-negative bacterium Pseudomonas aeruginosa, both included among the top antibiotic-resistant pathogens. We also test the synergistic effect of CNTs with different antibiotics commonly used in the treatment of infections caused by S. aureus and/or P. aeruginosa. Additionally, a novel approach for quantitatively analyzing bacterial aggregation in brightfield microscopy images was implemented. This method was utilized to assess the effectiveness of CNTs, either alone or in combination with antibiotics, in dispersing bacterial aggregates. Finally, atomic force microscopy coupled with a newly devised image analysis pipeline was used to examine any potential morphological changes in bacterial cells following exposure to CNTs and antibiotics.