Novel microenvironment-based classification of intrahepatic cholangiocarcinoma with therapeutic implications.

OBJECTIVE: The diversity of the tumour microenvironment (TME) of intrahepatic cholangiocarcinoma (iCCA) has not been comprehensively assessed. We aimed to generate a novel molecular iCCA classifier that incorporates elements of the stroma, tumour and immune microenvironment ('STIM' classification). DESIGN: We applied virtual deconvolution to transcriptomic data from ~900 iCCAs, enabling us to devise a novel classification by selecting for the most relevant TME components. Murine models were generated through hydrodynamic tail vein injection and compared with the human disease. RESULTS: iCCA is composed of five robust STIM classes encompassing both inflamed (35%) and non-inflamed profiles (65%). The inflamed classes, named immune classical (~10%) and inflammatory stroma (~25%), differ in oncogenic pathways and extent of desmoplasia, with the inflammatory stroma showing T cell exhaustion, abundant stroma and KRAS mutations (p<0.001). Analysis of cell-cell interactions highlights cancer-associated fibroblast subtypes as potential mediators of immune evasion. Among the non-inflamed classes, the desert-like class (~20%) harbours the lowest immune infiltration with abundant regulatory T cells (p<0.001), whereas the hepatic stem-like class (~35%) is enriched in 'M2-like' macrophages, mutations in IDH1/2 and BAP1, and FGFR2 fusions. The remaining class (tumour classical: ~10%) is defined by cell cycle pathways and poor prognosis. Comparative analysis unveils high similarity between a KRAS/p19 murine model and the inflammatory stroma class (p=0.02). The KRAS-SOS inhibitor, BI3406, sensitises a KRAS-mutant iCCA murine model to anti-PD1 therapy. CONCLUSIONS: We describe a comprehensive TME-based stratification of iCCA. Cross-species analysis establishes murine models that align closely to human iCCA for the preclinical testing of combination strategies.

Effects of out-of-school physical activity interventions based on self-determination theory in children and adolescents: A systematic review and meta-analysis.

OBJECTIVE: This systematic review and meta-analysis aimed to examine the effects of out-of-school physical activity (PA) interventions, based on Self-Determination Theory (SDT), on basic psychological needs (BPN), motivation toward PA, and PA levels in youths. DESIGN: Systematic review and meta-analyses. METHOD: We searched for intervention studies examining the effects of PA interventions based on SDT implemented outside the school published in English and Spanish in six electronic databases up to January 2022. RESULTS: Outcomes of interest were BPN, motivation, and PA levels. In total, nine studies were included in this review. Seven individual meta-analyses were conducted for each variable, revealing nonsignificant clustered effects for the outcomes autonomy satisfaction (g = 0.12, 95% CI [-0.31, 0.55]), competence satisfaction (g = 0.02, 95% CI [-0.28, 0.32]), relatedness satisfaction (g = 0.13, 95% CI [-0.43, 0.68]), autonomous motivation (g = 0.15, 95% CI [-0.38, 0.67]), controlled motivation (g = 0.12, 95% CI [-0.32, 0.55]), amotivation (g = -0.36, 95% CI [-0.88, 0.16]), and PA behavior (g = 0.02, 95% CI [-0.08, 0.12]). CONCLUSION: Meta-analyses suggest that out-of-school PA interventions based on SDT are not effective in increasing levels of needs satisfaction, types of motivation, and PA levels.

ACUTE RETINAL DAMAGE AFTER USING A TOXIC PERFLUORO-OCTANE FOR VITREO-RETINAL SURGERY.

PURPOSE: To describe a series of retinal acute toxicity cases with severe visual loss after intraocular use of a toxic perfluoro-octane (PFO). The clinical presentation is described, and the likely causes are analyzed. New biological methods for testing safety of intraocular medical devices are proposed. METHODS: Information regarding a series of eyes suffering acute severe events after intraocular use of a toxic PFO was analyzed. Four types of spectroscopy, nuclear magnetic resonance, and chromatography were used to identify the potential PFO contaminants. Cultures of human retinal pigment epithelial cells (ARPE-19) and porcine neuroretina were used to quantify the toxicity of the suspect PFO lots. RESULTS: Of 117 cases of intraocular toxicity, 96 were considered clearly related to the use of PFO. Fifty-three cases had no light perception, and 97 had no measurable visual acuity. Retinal necrosis (n = 38) and vascular occlusion (n = 33) were the most characteristic findings. Two hydroxyl compounds, perfluorooctanoic acid and dodecafluoro-1-heptanol, and benzene derivatives were identified as the suspected toxic agents. While existing toxicity testing failed, we proposed new tests that demonstrated clear toxicity. CONCLUSION: Protocols to determine cytotoxicity of intraocular medical devices should be revised to assure safety. Acute toxic events should be reported to health authorities and scientific media.

KIDMED 2.0, An update of the KIDMED questionnaire: Evaluation of the psychometric properties in youth.

Background and aims: As children and adolescents' eating patterns have changed over the last few years, researchers have found inconsistencies in the current questionnaires. Therefore, this research aims to (i) update the 2019 KIDMED questionnaire; and (ii) test the psychometric properties of this new questionnaire. Method: A study with 419 children and adolescents in southwestern Spain was conducted in 2021. The new version of the KIDMED 2.0 was tested, which measures adherence to the Mediterranean diet through 16 items, of which 12 are positive, and 4 are negative. Content validation involved consultation with nutritionists, experts, and adolescents to assess whether the questionnaire was reliable and valid regarding dietary patterns associated with the Mediterranean diet. The expert assessment provided content validity indices for the clarity and representativeness of the questionnaire. Construct validity and test-retest reliability involved 419 students (M age = 14.40 ± 2.00) from southwestern Spain. Students responded twice (one week apart) to the KIDMED developed in the previous stage and completed a 7-day dietary record. Results: Regarding validity, results show a moderate agreement for 10 items (ranging between 0.21 and 0.47) of the KIDMED and the 7-day dietary record. Concerning Items 3, 4, 5, and 6, the agreement was slight (ranging between 0.08 and 0.17), whereas the agreement for Item 8 was low. Cohen's kappa showed that most items had moderate to substantial test-retest reliability. Also, kappa showed significant test-retest values for all items (p < 0.001). Conclusion: The new version of the KIDMED 2.0 was shown to be a reliable and valid instrument to measure adherence to the Mediterranean diet in children and adolescents.

Approaches to the Mechanical Properties of Threaded Studs Welded to RHS Columns.

The use of Rectangular Hollow Sections (RHS) as columns in steel construction includes important advantages like higher mechanical strength and fire resistance. However, the practical demountable bolted joints between beams and columns are not easy to execute, due to impossibility of access to the inner part of the tube. The use of threaded studs welded to the face of the tube and bolted to the beam by means of angle cleats is one of the cheaper and most efficient solutions to obtain beam-column joints with a semi-rigid behavior, as is usually sought in building structures. Nevertheless, it is important to point out that the stud-diameter and the stud-class selection may affect the mechanical properties of the welded parts of the joint. In this paper, 8MnSi7 (with a commercial designation K800) and 4.8 threaded studs were welded to RHS steel tubes and mechanical properties on the weld, the Heat Affected Zones (HAZ), and the base metal were obtained in two different ways: through a correlation with the Vickers hardness and by means of the Small Punch Test (SPT). A study of the microstructure and tensile tests on the threaded studs and in the columns was also carried out. The research involved different types of stud qualities, tube wall thicknesses, and stud diameters. The work presented in this paper proved that in most cases, the welded joint between these studs and the RHS steel tubes present a reasonable static behavior that fulfils the requirements for the beam-column joints under static loading.

Is Perceived Athlete Leadership Quality Related to Inside Sacrifice and Perceived Performance in Team Sports? The Mediating Role of Team Identification.

The study aimed to analyze the relationship between athletes' perceptions of athlete leadership quality, team identification, inside sacrifice, and performance. A total of 299 players of collective sports (soccer, beach soccer, basketball, volleyball; M age 19.05, SD = 5.10) participated through a cross-sectional design survey. Data were analyzed using structural equation modeling. Results highlight the positive relationships between perceived quality of athlete leaders, inside sacrifice, and perceived performance, and between inside sacrifice and perceived performance. Furthermore, inside sacrifice perceived by the athletes was a positive mediator between perceived athlete leadership quality and perceived performance. Also, team identification was a positive mediator in the association between inside sacrifice and perceived performance. These findings extend knowledge about the athlete leadership quality context. These results can also be useful for further research and implications in team sports' performance, as coaches and sports psychologists would have more information about their teams' perceptions of leadership quality to achieve positive outcomes in players' inside sacrifice and performance. The findings also highlight the importance of developing team identification to improve the relationships between perceived athlete leadership quality, inside sacrifice, and perceived performance.

The Influence of the Heat-Affected Zone Mechanical Properties on the Behaviour of the Welding in Transverse Plate-to-Tube Joints.

Eurocode 3 establishes the component method to analytically characterize the structural joints between beam and columns. When one of the members involved in the joint is a hollow section (i.e., a tube) there is a lack of information for the specific components present in the joint. There are two different ways to bridge the gap: experimental testing on the actual beam column joints involving tubular sections; or numerical modelization, typically by means of finite element analysis. For this second option, it is necessary to know the actual mechanical properties of the material. As long as the joint implies a welding process, there is a concern related to how the mechanical properties in the heat-affected zone (HAZ) influence the behavior of the joint. In this work, some coupons were extracted from the HAZ of the beam-column joint. The coupons were tested and the results were implemented in the numerical model of the joint, in an attempt to bring it closer to the experimental results of the tested joints.

Effectiveness evaluation of whole-body electromyostimulation as a postexercise recovery method.

BACKGROUND: Whole-body electromyostimulation (WB-EMS) devices are now being used in health and sports training, although there are few studies investigating their benefits. The objective of this research was to evaluate the effectiveness of WB-EMS as a postexercise recovery method and compare it with other methods like active and passive recovery. METHODS: The study included nine trained men (age =21±1 years, height =1.77±0.4 m, mass =62±7 kg). Three trials were performed in three different sessions, 1 week apart. Each trial, the participants completed the same exercise protocol and a different recovery method each time. A repeated measures design was used to check the basal reestablishing on several physiological variables (lactate, heart rate, percentage of tissue hemoglobin saturation, temperature, and neuromuscular fatigue) and to evaluate the quality of recovery. The non-parametric Wilcoxon and Friedman ANOVA tests were used to examine the differences between recovery methods. RESULTS: The results showed no differences between methods in the physiological and psychological variables analyzed. Although, the blood lactate concentration showed borderline statistical significance between methods (P=0.050). Likewise, WB-EMS failed to recover baseline blood lactate concentration (P=0.021) and percentage of tissue hemoglobin saturation (P=0.023), in contrast to the other two methods. CONCLUSIONS: These findings suggest that WB-EMS is not a good recovery method because the power of reestablishing of several physiological and psychological parameters is not superior to other recovery methods like active and passive recovery.

p38γ and p38δ Are Involved in T Lymphocyte Development.

p38 mitogen-activated protein kinase (MAPK) signal transduction pathways are essential regulators of the immune response. Particularly, p38γ and p38δ regulate many immune cell functions such as cytokine production, migration, or T cell activation; however, their involvement in immune cell development is largely unknown. Here, we analysed the role of p38 MAPK isoforms p38γ and p38δ in T cell differentiation in the thymus and in lymph nodes, using mice deficient in p38γ, p38δ, or in both. We found that the T cell differentiation program in the thymus was affected at different stages in p38γ-, p38δ-, and p38γ/δ-deficient mice, and also peripheral T cell homaeostasis was compromised. Particularly, p38δ deletion affects different stages of early CD4-CD8- double-negative thymocyte development, whereas lack of p38γ favours thymocyte positive selection from CD4+CD8+ double-positive to CD4+ or CD8+ single-positive cells. Our results identify unreported functions for p38γ and p38δ in T cells.

Myeloid cell deficiency of p38γ/p38δ protects against candidiasis and regulates antifungal immunity.

Candida albicans is a frequent aetiologic agent of sepsis associated with high mortality in immunocompromised patients. Developing new antifungal therapies is a medical need due to the low efficiency and resistance to current antifungal drugs. Here, we show that p38γ and p38δ regulate the innate immune response to C. albicans We describe a new TAK1-TPL2-MKK1-ERK1/2 pathway in macrophages, which is activated by Dectin-1 engagement and positively regulated by p38γ/p38δ. In mice, p38γ/p38δ deficiency protects against C. albicans infection by increasing ROS and iNOS production and thus the antifungal capacity of neutrophils and macrophages, and by decreasing the hyper-inflammation that leads to severe host damage. Leucocyte recruitment to infected kidneys and production of inflammatory mediators are decreased in p38γ/δ-null mice, reducing septic shock. p38γ/p38δ in myeloid cells are critical for this effect. Moreover, pharmacological inhibition of p38γ/p38δ in mice reduces fungal burden, revealing that these p38MAPKs may be therapeutic targets for treating C. albicans infection in humans.

Simplified Models for the Material Characterization of Cold-Formed RHS.

It is well known that the cold-forming process used to manufacture tubes causes an increase in both the yielding stress and the ultimate strength of the corner material in rectangular steel hollow sections. This may have a significant effect on the resistance of any structure built with those profiles. However, the mentioned material hardening can be difficult to take into account in the calculations for member design or to evaluate the connection resistance through the design formulation or when using numerical simulation models. As an attempt to face the above-mentioned problem, the present paper presents a comparison among simplified approaches that consider homogeneous material properties for the whole section. It has been carried out by comparing the results obtained from the finite element modelling of stub column tests in which the material properties based on the flat faces were considered for the whole profile.

Importance of self-efficacy in psychoendocrine responses to competition and performance in women.

Cognitive appraisal before competition includes self-efficacy, traditionally defined as motivation and the perceived ability to perform well; presumably, both dimensions would affect the biological response to a contest. We aimed to analyze the role of self-efficacy in the psychobiological response to a competition in women. Forty university students were confronted in pairs on a laboratory competition while hormonal and emotional changes were measured. Our results indicated that self-efficacy was positively related to testosterone levels and positive mood, and also to better performance. These results empirically support the importance of main dimensions of the cognitive appraisal in androgenic and emotional responses to competition. In addition, they emphasize the importance of cognitive processes in this response. In conclusion, the relationship between androgens and self-efficacy may play an important role as a facilitator of performance in competitive settings.

Exposure to phenanthrene and depuration: Changes on gene transcription, enzymatic activity and lipid peroxidation in gill of scallops Nodipecten nodosus.

Understanding the mechanism of phenanthrene (PHE) biotransformation and related cellular responses in bivalves can be an important tool to elucidate the risks of polycyclic aromatic hydrocarbons (PAHs) to aquatic organisms. In the present study it was analyzed the transcriptional levels of 13 biotransformation genes related to cytochrome P450 (CYP), glutathione S-transferase (GST), sulfotransferase (SULT), flavin-containing monooxygenase and fatty acid-binding proteins by qPCR in gill of scallops Nodipecten nodosus exposed for 24 or 96h to 50 or 200µgL(-1) PHE (equivalent to 0.28 and 1.12µM, respectively), followed by depuration in clean water for 96h (DEP). Likewise, it was quantified the activity of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PDH), GST and levels of lipid peroxidation. Increased transcriptional levels of CYP2UI-like, CYP2D20-like, CYP3A11-like, GSTomega-like, SULT1B1-like genes were detected in organisms exposed to PHE for 24 or 96h. In parallel, GR and GPX activities increased after 96h exposure to 200µgL(-1) PHE and G6PDH activity increased after 24h exposure to 50µgL(-1) PHE. This enhancement of antioxidant and phase I and II biotransformation systems may be related to the 2.7 and 12.5 fold increases in PHE bioaccumulation after 96h exposure to 50 and 200µgL(-1) PHE, respectively. Interestingly, DEP caused reestablishment of GPX and GR activity, as well as to the transcript levels of all upregulated biotransformation genes (except for SULT1B1-like). Bioaccumulated PHE levels decreased 2.5-2.9 fold after depuration, although some biochemical and molecular modifications were still present. Lipid peroxidation levels remained lower in animals exposed to 200µgL(-1) PHE for 24h and DEP. These data indicate that N. nodosus is able to induce an antioxidant and biotransformation-related response to PHE exposure, counteracting its toxicity, and DEP can be an effective protocol for bivalve depuration after PHE exposure.

Effect of linear alkylbenzene mixtures and sanitary sewage in biochemical and molecular responses in pacific oyster Crassostrea gigas.

Urban effluents are rich in nutrients, organic matter, pharmaceuticals and personal care products (PPCPs), pesticides, hydrocarbons, surfactants, and others. Previous studies have shown that oysters Crassostrea gigas accumulate significant levels of linear alkylbenzenes (LABs) in sanitary sewage contaminated sites, but there is little information about its toxicological effects in marine bivalves. The aim of this study was to analyze the transcription of genes in two tissues of C. gigas exposed for 12, 24, and 36 h to LABs or sanitary sewage. Likewise, the activity of antioxidant and biotransformation enzymes was measured in oysters exposed for 36 h in all groups. Oysters exposed to LABs and oysters exposed to sanitary sewage showed different patterns of transcriptional responses. LAB-exposed oysters showed lower level of biological responses than the oysters exposed to sanitary sewage. Despite the ability of the oyster C. gigas to accumulate LABs (28-fold), the data indicate that these contaminants are not the cause for the transcriptional responses observed in oysters exposed to sanitary sewage. Possibly, the biological changes observed in the sanitary sewage-exposed oysters are associated with the presence of other contaminants, which might have caused synergistic, additive, or antagonistic effects. The results show that FABP-like and GST-ω-like messenger RNAs (mRNAs) have a rapid response in tissues of oyster C. gigas exposed to sanitary sewage, suggesting a possible protective response and a role in maintaining homeostasis of these organisms.

Implication of Akt, ERK1/2 and alternative p38MAPK signalling pathways in human colon cancer cell apoptosis induced by green tea EGCG.

We investigated apoptosis induced by the green tea component the epigallocatechin-3-gallate (EGCG) and the pathways underlying its activity in a colon cancer cell line. A complete understanding of the mechanism(s) and molecules targeted by green tea polyphenols could be useful in developing novel therapeutic approaches for cancer treatment. EGCG, which is the major polyphenol in green tea, has cytotoxic effects and induced cell death in HT-29 cell death. In this study, we evaluated the effect EGCG on mitogen-activated protein kinase (MAPK) and Akt pathways. EGCG treatment increased phospho-ERK1/2, -JNK1/2 and -p38α, -p38γ and -p38δ, as well as phospho-Akt levels. Using a combination of kinase inhibitors, we found that EGCG-induced cell death is partially blocked by inhibiting Akt, ERK1/2 or alternative p38MAPK activity. Our data suggest that these kinase pathways are involved in the anti-cancer effects of EGCG and indicate potential use of this compound as chemotherapeutic agent for colon cancer treatment.

Differential gene transcription, biochemical responses, and cytotoxicity assessment in Pacific oyster Crassostrea gigas exposed to ibuprofen.

Pharmaceuticals, such as anti-inflammatory nonsteroidal drugs, are frequently detected in aquatic ecosystems. Studies about the effects of these substances in nontarget organisms, such as bivalves, are relevant. The aim of this study was to evaluate the effects on antioxidant status caused by ibuprofen (IBU) in oysters Crassostrea gigas exposed for 1, 4, and 7 days at concentrations 1 and 100 µg L(-1). Levels of IBU in tissues of oysters, as well as cell viability of hemocytes, were measured. The transcription of cytochrome P450 genes (CYP2AU2, CYP356A1, CYP3071A1, CYP30C1), glutathione S-transferase isoforms (GST-ω-like and GST-π-like), cyclooxygenase-like (COX-like), fatty acid binding protein-like (FABP-like), caspase-like, heat shock protein-like (HSP70-like), catalase-like (CAT-like), and the activity of catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S-transferase (GST) were also evaluated in the gills of oysters. The highest levels of IBU were observed in animals exposed to 100 µg L(-1). A significant upregulation of CYP2AU1, CYP356A1, CYP3071A1, GST-ω-like, GST-π-like, COX-like, and FABP-like was observed in oysters exposed to IBU under different experimental conditions. Oysters exposed to 1 µg L(-1) for 7 days showed a significantly higher transcription of CYP2AU2, CYP356A1, CYP3071A1, GST-ω-like, and GST-π-like but lower GR activity. In conclusion, C. gigas exposed to environmentally relevant concentrations of IBU (1 µg L(-1)) exhibited increased transcription of certain genes and alterations on antioxidant and auxiliary enzymes, which could, in the the long term, cause damages to exposed organisms.

Use of ceramic monoliths as stationary phase in affinity chromatography.

The use of coated ceramic monoliths as support for affinity chromatography is described. Ceramic monoliths are robust active matrix supports and present a very small pressure drop. Monoliths are coated with a very thin agarose gel layer and activated using a standard activation process for agarose beads. Experiments demonstrate that enzyme adsorption occurs exclusively on the outside surface of the agarose coating since enzyme molecules are too large to fit into the porous matrix. Adsorption and desorption rates are large and production of enzyme per unit monolith volume justifies further exploring this separation process for large throughput operation.

Modeling of monolith-supported affinity chromatography.

Ceramic monoliths have been used successfully as active support for affinity chromatography (1). A mathematical model was developed to simulate the adsorption-elution experimental behavior of asparaginase in an agarose-coated monolith support. The computer-based model allows precise determination of experimental parameters. Because of the simple geometry of ceramic monoliths used as support, the mathematical model can be used to design adsorption/elution cycles for the large throughput separation of biomolecules.