Analysis of Amyloid Fibrillation of Two Family 1 Glycoside Hydrolases.

The formation and analysis of amyloid fibers by two ß-glucosidases, BglA and BglB, belonging to the GH1 enzyme family, are reported. Both proteins have the (ß/α)8 TIM-barrel fold, which is characteristic of this family and is also the most common protein structure. BglA is an octamer, whereas BglB is a monomer. Amyloid fibrillation using pH and temperature as perturbing agents was investigated using fluorescence spectroscopy as a preliminary approach and corroborated using wide-field optical microscopy, confocal microscopy, and field-emission scanning electron microscopy. These analyses showed that both enzymes fibrillate at a wide range of acidic and alkaline conditions and at several temperature conditions, particularly at acidic pH (3-4) and at temperatures between 45 and 65 °C. Circular dichroism spectroscopy corroborated the transition from an α-helix to a ß-sheet secondary structure of both proteins in conditions where fibrillation was observed. Overall, our results suggest that fibrillation is a rather common phenomenon caused by protein misfolding, driven by a transition from an α-helix to a ß-sheet secondary structure, that many proteins can undergo if subjected to conditions that disturb their native conformation.

Genotypic and phenotypic characterization of resistance to fenhexamid, carboxin, and, prochloraz, in Botrytis cinerea isolates collected from cut roses in Colombia.

Gray mold, caused by Botrytis sp., is a significant disease in Colombian rose crops and its control depends primarily on the intensive use of chemically synthesized fungicides. Despite the importance of this pathogen, there is limited information in Colombian floriculture about molecular taxonomy of species, fungicide resistance of populations and their genetic mechanism of resistance. In this study, we analyze 12 isolates of this fungus collected from rose-producing crops in the Department of Cundinamarca and conducted phylogenetic analysis using HSP60, G3PDH, and RPB2 gene sequences. Additionally, we realize phenotypic and genotypic characterization of resistance to the fungicides fenhexamid, carboxin, and prochloraz, evaluating the in vitro EC50 and presence of mutations of target genes of each isolate. All isolates were characterized as Botrytis cinerea in the phylogenetic analysis and presents different levels of resistance to each fungicide. These levels are related to mutations in target genes, with predominancy of L195F and L400F in the ERG27 gene to fenhexamid resistance, H272R/Y in the SDHB gene for carboxin resistance, and Y136F in the CYP51 gene for prochloraz resistance. Finally, these mutations were not related to morphological changes. Collectively, this knowledge, presented for the first time to the Colombian floriculture, contribute to a better understanding of the genetic diversity and population of B. cinerea from rose-producing crops in the department of Cundinamarca, and serve as a valuable tool for making informed decisions regarding disease management, future research, and improving crop management and sustainability in the Colombian floriculture industry.

[Childhood overweight and obesity: A biomedical challenge in Mexico]. / Sobrepeso y obesidad infantil: un reto biomédico en México.

In Mexico, 1 out of 3 schoolchildren aged 5 to 11 years is overweight or obese, which represents one of the main public health concerns, due to the fact that this condition in the child population is highly associated with the development of metabolic complications in adults. To date, dietary and physical activity interventions to prevent this problem have shown modest results worldwide. Biomedical studies in Mexico have shown that the pathophysiology of childhood overweight and obesity presents different molecular patterns, inflammation and oxidative stress, possibly associated with specific variants in the genome. However, the challenge is to achieve a secure characterization of this evidence so that it can be used in intervention studies aimed to improve the ability to predict and treat childhood overweight and obesity in Mexico. The biomedical challenge is to make knowledge a prevention strategy in families, in society and in the country, in order to fight the serious problem of obesity and its consequences.

En México 1 de cada 3 escolares de 5 a 11 años presenta sobrepeso u obesidad, lo cual representa una de las principales preocupaciones de salud pública, debido a que en la población infantil este padecimiento se asocia altamente con el desarrollo de complicaciones metabólicas en el adulto. Hasta el momento las intervenciones dietéticas y de actividad física para prevenir este problema han mostrado resultados modestos a nivel mundial. Los estudios biomédicos en México han demostrado que la fisiopatología del sobrepeso y la obesidad infantil presenta diferentes patrones moleculares, de inflamación y de estrés oxidativo, posiblemente asociados a variantes específicas en el genoma. Sin embargo, el reto es lograr la caracterización segura de estas evidencias para que sea posible emplearlas en los estudios de intervención encaminados a mejorar la capacidad de predicción y tratamiento del sobrepeso y la obesidad infantil en México. El reto biomédico es hacer del conocimiento una estrategia de prevención en las familias, en la sociedad y en el país, a fin de combatir el grave problema de la obesidad y sus consecuencias.

Ceramides in the central control of metabolism.

Central ceramides regulate energy metabolism by impacting hypothalamic neurons. This allows ceramides to integrate endocrine signals - such as leptin, ghrelin, thyroid hormones, or estradiol - and to modulate the central control of puberty. In this forum article we discuss recent evidence suggesting that specific ceramide species and neuronal populations are involved in these effects.

Indicators to assess temporal variability in marine connectivity processes: A semi-theoretical approach.

Oceanographic connectivity in an effective network of protected areas is crucial for restoring and stabilising marine populations. However, temporal variability in connectivity is rarely considered as a criterion in designing and evaluating marine conservation planning. In this study, indicators were defined to characterise the temporal variability in occurrence, flux, and frequency of connectivity in a northwestern Mediterranean Sea area. Indicators were tested on semi-theoretically-estimated connections provided by the runs of a passive particle transport model in a climatological year and in three years between 2006-2020, showing large deviation from the climatological year. The indicators allowed comparing the temporal variability in connectivity of four zones, highlighted differences in connectivity due to their locations and the mesoscale hydrodynamics, and identified areas that require further investigation. The three indicators also showed that the temporal variability in connectivity was influenced by the duration and depth of particle transport, although no consistent pattern was observed in the indicator variations of the compared zones. Provided that specific objectives will be given when parameterising transport models (i.e., selection of focus species and time period), indicators of temporal variability in connectivity have potential to support spatial conservation planning, prioritise the protection of marine resources, and measure the effectiveness of Marine Protected Areas, in line with a long-term vision of ocean management.

Specific Detection of Uropathogenic Escherichia coli via Fourier Transform Infrared Spectroscopy Using an Optical Biosensor.

Urinary tract infections (UTIs) are caused mainly by uropathogenic Escherichia coli (UPEC), accounting for both uncomplicated (75%) and complicated (65%) UTIs. Detecting UPEC in a specific, rapid, and timely manner is essential for eradication, and optical biosensors may be useful tools for detecting UPEC. Recently, biosensors have been developed for the selective detection of antigen-antibody-specific interactions. In this study, a methodology based on the principle of an optical biosensor was developed to identify specific biomolecules, such as the PapG protein, which is located at the tip of P fimbriae and promotes the interaction of UPEC with the uroepithelium of the human kidney during a UTI. For biosensor construction, recombinant PapG protein was generated and polyclonal anti-PapG antibodies were obtained. The biosensor was fabricated in silicon supports because its surface and anchor biomolecules can be modified through its various properties. The fabrication process was carried out using self-assembled monolayers (SAMs) and an immobilized bioreceptor (anti-PapG) to detect the PapG protein. Each stage of biosensor development was evaluated by Fourier transform infrared (FTIR) spectroscopy. The infrared spectra showed bands corresponding to the C-H, C=O, and amide II bonds, revealing the presence of the PapG protein. Then, the spectra of the second derivative were obtained from 1600 to 1700 cm-1 to specifically determine the interactions that occur in the secondary structures between the biological recognition element (anti-PapG antibodies) and the analyte (PapG protein) complex. The analyzed secondary structure showed ß-sheets and ß-turns during the detection of the PapG protein. Our data suggest that the PapG protein can be detected through an optical biosensor and that the biosensor exhibited high specificity for the detection of UPEC strains. Furthermore, these studies provide initial support for the development of more specific biosensors that can be applied in the future for the detection of clinical UPEC samples associated with ITUs.

Hemospray® (hemostatic powder TC-325) as monotherapy for acute gastrointestinal bleeding: a multicenter prospective study.

Background: Hemostatic powders are used as second-line treatment in acute gastrointestinal (GI) bleeding (AGIB). Increasing evidence supports the use of TC-325 as monotherapy in specific scenarios. This prospective, multicenter study evaluated the performance of TC-325 as monotherapy for AGIB. Methods: Eighteen centers across Europe and USA contributed to a registry between 2016 and 2022. Adults with AGIB were eligible, unless TC-325 was part of combined hemostasis. The primary endpoint was immediate hemostasis. Secondary outcomes were rebleeding and mortality. Associations with risk factors were investigated (statistical significance at P≤0.05). Results: One hundred ninety patients were included (age 51-81 years, male: female 2:1), with peptic ulcer (n=48), upper GI malignancy (n=79), post-endoscopic treatment hemorrhage (n=37), and lower GI lesions (n=26). The primary outcome was recorded in 96.3% (95% confidence interval [CI]: 92.6-98.5) with rebleeding in 17.4% (95%CI 11.9-24.1); 9.9% (95%CI 5.8-15.6) died within 7 days, and 21.7% (95%CI 15.6-28.9) within 30 days. Regarding peptic ulcer, immediate hemostasis was achieved in 88% (95%CI 75-95), while 26% (95%CI 13-43) rebled. Higher ASA score was associated with mortality (OR 23.5, 95%CI 1.60-345; P=0.02). Immediate hemostasis was achieved in 100% of cases with malignancy and post-intervention bleeding, with rebleeding in 17% and 3.1%, respectively. Twenty-six patients received TC-325 for lower GI bleeding, and in all but one the primary outcome was achieved. Conclusions: TC-325 monotherapy is safe and effective, especially in malignancy or post-endoscopic intervention bleeding. In patients with peptic ulcer, it could be helpful when the primary treatment is unfeasible, as bridge to definite therapy.

Evaluation of video-based rPPG in challenging environments: Artifact mitigation and network resilience.

Video-based remote photoplethysmography (rPPG) has emerged as a promising technology for non-contact vital sign monitoring, especially under controlled conditions. However, the accurate measurement of vital signs in real-world scenarios faces several challenges, including artifacts induced by videocodecs, low-light noise, degradation, low dynamic range, occlusions, and hardware and network constraints. In this article, a systematic and comprehensive investigation of these issues is conducted, measuring their detrimental effects on the quality of rPPG measurements. Additionally, practical strategies are proposed for mitigating these challenges to improve the dependability and resilience of video-based rPPG systems. Methods for effective biosignal recovery in the presence of network limitations are detailed, along with denoising and inpainting techniques aimed at preserving video frame integrity. Compared to previous studies, this paper addresses a broader range of variables and demonstrates improved accuracy across various rPPG methods, emphasizing generalizability for practical applications in diverse scenarios with varying data quality. Extensive evaluations and direct comparisons demonstrate the effectiveness of these approaches in enhancing rPPG measurements under challenging environments, contributing to the development of more reliable and effective remote vital sign monitoring technologies.

Environment driven changes in type 2 diabetes, overweight and obesity in an isolated Mixe community in the Valley of Oaxaca, southern Mexico.

BACKGROUND: This study focused on type 2 diabetes mellitus (T2DM) in a group of adult Mixe, an Indigenous population from Oaxaca, Mexico. Mixe comprised an estimated 9.4% (n ≅ 90 000) of the Indigenous population in Oaxaca. Mexico. OBJECTIVE: This study focused on a group of adult Mixe, an Indigenous population from Oaxaca, Mexico. To compare the prevalence of T2DM, overweight (OW), obesity (OB), and hypertension (HTN) between 2007 and 2017 for a small, isolated Mixe community in the Valley of Oaxaca, Mexico. We test whether or not environmental changes have affected T2DM prevalence. METHODS AND MATERIALS: Demographic and medical record data were collected in the community in 2007 and 2017 from the medical clinic and the mayor's office. T2DM was medically diagnosed among adults (>34 years old), in 2007 (n = 730) and in 2017 (n = 829). RESULTS: T2DM crude prevalence increased from 6.7% to 12.1% (p < .001) from 2007 to 2017. The mean age of the sample analyzed was 60.6 (SD = 9.7). Age-adjusted T2DM prevalence increased from 6.7% to 10.8% (p < .002). T2DM was 5.7%-5.5% among males (p < .53) and 7.1%-13.6% among females (p < .001). Sex-specific OW and OB simulation studies indicate females had 7% less OW in 2007, and males were unchanged compared with 2017. OB among males and females was significantly higher in 2017 compared with 2007 (increased by 15.2% and 8.3%, males and females, respectively). Sexes combined OW + OB increased 12.7% among males but was unchanged in females (-0.5%). In the sexes combined analysis, OW prevalence increased 12.7% to 27.1% (p < .001) and OB prevalence increased 10.7%-27.9% (p < .001) from 2007 to 2017. HTN did not change significantly from 2007 to 2017 (15.4% and 14.6%, respectively) (p = .63) in adults. Among T2DM individuals, the frequency of HTN was not significantly different in 2007 and 2017 (57.1% and 37%, respectively) (p = .65). Transition to a Western diet consisting of high-carbohydrate foods occurred at the same time as increased T2DM from 2007 to 2017, with a higher prevalence of T2DM noted among females in 2017. CONCLUSIONS: An increased prevalence of T2DM, OW, and OB but not HTN was observed in the Mixe community from 2007 to 2017 and was associated with the adoption of a high-carbohydrate Western diet.

Reliability of Tear Meniscus Height Measurements in Contact Lens Wearers and Its Relationship With Discomfort Symptoms.

OBJECTIVES: To evaluate the reliability and agreement of tear meniscus height (TMH) measurements performed with a corneal analyzer and optical coherence tomography (OCT) technology in contact lens (CL) wearers and its correlation with contact lens discomfort symptoms. METHODS: Asymptomatic and symptomatic CL wearers classified through the Contact Lens Dry Eye Questionnaire-8 were evaluated with the Corneal Analyzer (Topcon CA-800) and OCT technology (Topcon 3D OCT-2000). The repeatability and intraclass correlation coefficient (ICC) were calculated. The agreement between devices was calculated using the Bland-Altman method. The relationship between TMH measurements and the Contact Lens Dry Eye Questionnaire-8 and Contact Lens Discomfort Index scores was assessed through the Spearman correlation coefficient. RESULTS: Seventy-nine asymptomatic and 42 symptomatic CL wearers aged 34.24±12.50 years were enrolled. The repeatability values obtained for the CA-800 were 0.07 mm in all cases, and the ICC was 0.93 for the whole sample. The CA-800 provided significantly (P<0.01) higher TMH values than the OCT for the whole sample (0.22±0.08 vs. 0.17±0.06 mm). A weak indirect correlation (ρ=-0.22) between the OCT TMH measurement and Contact Lens Discomfort Index scores was found (P≤0.04). CONCLUSION: The CA-800 provides reliable TMH measurements during CL wear; however, they might not be interchangeable with OCT ones. Tear meniscus height measurements might be useful as a complementary sign to detect CL discomfort, but it cannot be used alone as a diagnostic tool.

Regulation of dye-decolorizing peroxidase gene expression in Pleurotus ostreatus grown on glycerol as the carbon source.

Dye-decolorizing peroxidases (DyPs) (E.C. 1.11.1.19) are heme peroxidases that catalyze oxygen transfer reactions similarly to oxygenases. DyPs utilize hydrogen peroxide (H2O2) both as an electron acceptor co-substrate and as an electron donor when oxidized to their respective radicals. The production of both DyPs and lignin-modifying enzymes (LMEs) is regulated by the carbon source, although less readily metabolizable carbon sources do improve LME production. The present study analyzed the effect of glycerol on Pleurotus ostreatus growth, total DyP activity, and the expression of three Pleos-dyp genes (Pleos-dyp1, Pleos-dyp2 and Pleos-dyp4), via real-time RT-qPCR, monitoring the time course of P. ostreatus cultures supplemented with either glycerol or glucose and Acetyl Yellow G (AYG) dye. The results obtained indicate that glycerol negatively affects P. ostreatus growth, giving a biomass production of 5.31 and 5.62 g/L with respective growth rates (micra; m) of 0.027 and 0.023 h-1 for fermentations in the absence and presence of AYG dye. In contrast, respective biomass production levels of 7.09 and 7.20 g/L and growth rates (µ) of 0.033 and 0.047 h-1 were observed in equivalent control fermentations conducted with glucose in the absence and presence of AYG dye. Higher DyP activity levels, 4,043 and 4,902 IU/L, were obtained for fermentations conducted on glycerol, equivalent to 2.6-fold and 3.16-fold higher than the activity observed when glucose is used as the carbon source. The differential regulation of the DyP-encoding genes in P. ostreatus were explored, evaluating the carbon source, the growth phase, and the influence of the dye. The global analysis of the expression patterns throughout the fermentation showed the up- and down- regulation of the three Pleos-dyp genes evaluated. The highest induction observed for the control media was that found for the Pleos-dyp1 gene, which is equivalent to an 11.1-fold increase in relative expression (log2) during the stationary phase of the culture (360 h), and for the glucose/AYG media was Pleos-dyp-4 with 8.28-fold increase after 168 h. In addition, glycerol preferentially induced the Pleos-dyp1 and Pleos-dyp2 genes, leading to respective 11.61 and 4.28-fold increases after 144 h. After 360 and 504 h of culture, 12.86 and 4.02-fold increases were observed in the induction levels presented by Pleos-dyp1 and Pleos-dyp2, respectively, in the presence of AYG. When transcription levels were referred to those found in the control media, adding AYG led to up-regulation of the three dyp genes throughout the fermentation. Contrary to the fermentation with glycerol, where up- and down-regulation was observed. The present study is the first report describing the effect of a less-metabolizable carbon source, such as glycerol, on the differential expression of DyP-encoding genes and their corresponding activity.

Risk of recurrence after discontinuing anticoagulation in patients with COVID-19- associated venous thromboembolism: a prospective multicentre cohort study.

Background: The clinical relevance of recurrent venous thromboembolism (VTE) after discontinuing anticoagulation in patients with COVID-19-associated VTE remains uncertain. We estimated the incidence rates and mortality of VTE recurrences developing after discontinuing anticoagulation in patients with COVID-19-associated VTE. Methods: A prospective, multicenter, non-interventional study was conducted between March 25, 2020, and July 26, 2023, including patients who had discontinued anticoagulation after at least 3 months of therapy. All patients from the registry were analyzed during the study period to verify inclusion criteria. Patients with superficial vein thrombosis, those who did not receive at least 3 months of anticoagulant therapy, and those who were followed for less than 15 days after discontinuing anticoagulation were excluded. Outcomes were: 1) Incidence rates of symptomatic VTE recurrences, and 2) fatal PE. The rate of VTE recurrences was defined as the number of patients with recurrent VTE divided by the patient-years at risk of recurrent VTE during the period when anticoagulation was discontinued. Findings: Among 1106 patients with COVID-19-associated VTE (age 62.3 ± 14.4 years; 62.9% male) followed-up for 12.5 months (p25-75, 6.3-20.1) after discontinuing anticoagulation, there were 38 VTE recurrences (3.5%, 95% confidence interval [CI]: 2.5-4.7%), with a rate of 3.1 per 100 patient-years (95% CI: 2.2-4.2). No patient died of recurrent PE (0%, 95% CI: 0-7.6%). Subgroup analyses showed that patients with diagnosis in 2021-2022 (vs. 2020) (Hazard ratio [HR] 2.86; 95% CI 1.45-5.68) or those with isolated deep vein thrombosis (vs. pulmonary embolism) (HR 2.31; 95% CI 1.19-4.49) had significantly higher rates of VTE recurrences. Interpretation: In patients with COVID-19-associated VTE who discontinued anticoagulation after at least 3 months of treatment, the incidence rate of recurrent VTE and the case-fatality rate was low. Therefore, it conceivable that long-term anticoagulation may not be required for many patients with COVID-19-associated VTE, although further research is needed to confirm these findings. Funding: Sanofi and Rovi, Sanofi Spain.

Presence and Persistence of ESKAPEE Bacteria before and after Hospital Wastewater Treatment.

The metagenomic surveillance of antimicrobial resistance in wastewater has been suggested as a methodological tool to characterize the distribution, status, and trends of antibiotic-resistant bacteria. In this study, a cross-sectional collection of samples of hospital-associated raw and treated wastewater were obtained from February to March 2020. Shotgun metagenomic sequencing and bioinformatic analysis were performed to characterize bacterial abundance and antimicrobial resistance gene analysis. The main bacterial phyla found in all the samples were as follows: Proteobacteria, Bacteroides, Firmicutes, and Actinobacteria. At the species level, ESKAPEE bacteria such as E. coli relative abundance decreased between raw and treated wastewater, but S. aureus, A. baumannii, and P. aeruginosa increased, as did the persistence of K. pneumoniae in both raw and treated wastewater. A total of 172 different ARGs were detected; blaOXA, blaVEB, blaKPC, blaGES, mphE, mef, erm, msrE, AAC(6'), ant(3″), aadS, lnu, PBP-2, dfrA, vanA-G, tet, and sul were found at the highest abundance and persistence. This study demonstrates the ability of ESKAPEE bacteria to survive tertiary treatment processes of hospital wastewater, as well as the persistence of clinically important antimicrobial resistance genes that are spreading in the environment.

Tear Cytokines as Predictive Biomarkers of Success in Contact Lens Discomfort Management.

PURPOSE: To analyze changes in tear levels of inflammatory mediators in symptomatic contact lens (CL) wearers after refitting with daily disposable CLs and to identify potential biomarkers of success in CL discomfort (CLD) management. METHODS: Symptomatic CL wearers (CLDEQ-8 ≥ 12) were refitted (V1) with daily disposable CLs (Delefilcon A). After one month (V2), participants were classified into the post-fitting non-symptomatic (CLDEQ <12) and symptomatic (CLDEQ ≥12) groups. At each visit, the participants were clinically evaluated, tears were collected, and 20 inflammatory mediators and substance P (SP) were measured using multiplex immunobead analysis and ELISA, respectively. The detection rates and concentrations were compared between visits and groups, and logistic regression models were performed. RESULTS: Forty-three subjects (32 women/11 men; mean age: 23.2 ± 4.9 years) were enrolled. The IL-1ß and IL-9 detection rates were higher at V2 (p ≤ 0.044). The detection rates of IL-1ß, IL-9, MIP-1α/CCL3, and MMP-9 at V1 (p ≤ 0.045) and IL-17A at V2 (p ≤ 0.014) were higher in the post-fitting symptomatic group. The tear IL-9 concentration was increased at V2 (p = 0.018). The tear concentrations of fractalkine/CX3CL1, IL-2, IL-6, IL-10, MCP-3/CCL7, MIP-1ß, NGF, RANTES/CCL5, and TNF-α were higher in the post-fitting symptomatic group (p ≤ 0.044). Additionally, levels of fractalkine/CX3CL1, IL-2, IL-6, IL-10, RANTES/CCL5, and TNF-α at V1 were significantly associated with the post-fitting grouping (p ≤ 0.044). CONCLUSIONS: Low tear concentrations of specific inflammatory mediators may be used as a predictive biomarker of success for refitting symptomatic CL wearers with daily disposable CLs. However, complementary treatments might be required for symptomatic CL wearers with higher levels of these inflammatory molecules.

Combined Supplementation of Two Selenium Forms (Organic and Inorganic) and Iodine in Dairy Cows' Diet to Obtain Enriched Milk, Cheese, and Yogurt.

This study evaluated the effects of dietary supplementation in dairy cows with two Se forms (organic and inorganic) and I at the maximum levels permitted in the European Union, with the aim to obtain naturally enriched milk and derived products. A total of 20 Holstein Friesian cows in lactation were fed 2 diets for 64 days: a control diet with a supply of 0.57 mg of inorganic Se and 0.57 mg of I per kg of ration in dry matter (DM), and an experimental diet (SeI) with a supply of 0.34 mg of inorganic Se, 0.23 mg of organic Se, and 5.68 mg of I per kg of ration in DM. The SeI diet did not modify the performance or, in general, the metabolic profile of cows. Se and I levels in milk were affected by diet type and time of measurement (p < 0.01). Thus, a marked increase of both microminerals was evident between the beginning and the end of the test, when the SeI diet was administered. For Se, this increase ranged from 1.95 to 3.29 µg/100 g of milk; and for I, from 19.69 to 110.06 µg/100 g of milk. The SeI diet increased (p < 0.01) the Se and I content in the cheese, reaching levels of 16.4 µg/100 g for Se and 269.7 µg/100 g for I. An increase in I was observed in yogurt from the SeI diet (p < 0.001). The supplementation of two forms of Se and I in the cows' ration, at the levels evaluated, produced milk and dairy products enriched in these microelements without altering their quality parameters. However, a responsible intake of these products is necessary to avoid risks of deficiencies or excesses that could negatively affect the health of consumers.

Mitochondrial antiviral signaling protein enhances MASLD progression through the ERK/TNFα/NFκß pathway.

BACKGROUND AND AIMS: Mitochondrial antiviral signaling protein (MAVS) is a critical regulator that activates the host's innate immunity against RNA viruses, and its signaling pathway has been linked to the secretion of proinflammatory cytokines. However, the actions of MAVS on inflammatory pathways during the development of metabolic dysfunction-associated steatotic liver disease (MASLD) have been little studied. APPROACH AND RESULTS: Liver proteomic analysis of mice with genetically manipulated hepatic p63, a transcription factor that induces liver steatosis, revealed MAVS as a target downstream of p63. MAVS was thus further evaluated in liver samples from patients and in animal models with MASLD. Genetic inhibition of MAVS was performed in hepatocyte cell lines, primary hepatocytes, spheroids, and mice. MAVS expression is induced in the liver of both animal models and people with MASLD as compared with those without liver disease. Using genetic knockdown of MAVS in adult mice ameliorates diet-induced MASLD. In vitro, silencing MAVS blunts oleic and palmitic acid-induced lipid content, while its overexpression increases the lipid load in hepatocytes. Inhibiting hepatic MAVS reduces circulating levels of the proinflammatory cytokine TNFα and the hepatic expression of both TNFα and NFκß. Moreover, the inhibition of ERK abolished the activation of TNFα induced by MAVS. The posttranslational modification O -GlcNAcylation of MAVS is required to activate inflammation and to promote the high lipid content in hepatocytes. CONCLUSIONS: MAVS is involved in the development of steatosis, and its inhibition in previously damaged hepatocytes can ameliorate MASLD.

Role of Volatile Organic Compounds Produced by Kosakonia cowanii Cp1 during Competitive Colonization Interaction against Pectobacterium aroidearum SM2.

The competitive colonization of bacteria on similar ecological niches has a significant impact during their establishment. The synthesis speeds of different chemical classes of molecules during early competitive colonization can reduce the number of competitors through metabolic effects. In this work, we demonstrate for the first time that Kosakonia cowanii Cp1 previously isolated from the seeds of Capsicum pubescens R. P. produced volatile organic compounds (VOCs) during competitive colonization against Pectobacterium aroidearum SM2, affecting soft rot symptoms in serrano chili (Capsicum annuum L.). The pathogen P. aroidearum SM2 was isolated from the fruits of C. annuum var. Serrano with soft rot symptoms. The genome of the SM2 strain carries a 5,037,920 bp chromosome with 51.46% G + C content and 4925 predicted protein-coding genes. It presents 12 genes encoding plant-cell-wall-degrading enzymes (PCDEWs), 139 genes involved in five types of secretion systems, and 16 genes related to invasion motility. Pathogenic essays showed soft rot symptoms in the fruits of C. annuum L., Solanum lycopersicum, and Physalis philadelphica and the tubers of Solanum tuberosum. During the growth phases of K. cowanii Cp1, a mix of VOCs was identified by means of HS-SPME-GC-MS. Of these compounds, 2,5-dimethyl-pyrazine showed bactericidal effects and synergy with acetoin during the competitive colonization of K. cowanii Cp1 to completely reduce soft rot symptoms. This work provides novel evidence grounding a better understanding of bacterial interactions during competitive colonization on plant tissue, where VOC synthesis is essential and has a high potential capacity to control pathogenic microorganisms in agricultural systems.

dsRNAi-mediated silencing of PIAS2beta specifically kills anaplastic carcinomas by mitotic catastrophe.

The E3 SUMO ligase PIAS2 is expressed at high levels in differentiated papillary thyroid carcinomas but at low levels in anaplastic thyroid carcinomas (ATC), an undifferentiated cancer with high mortality. We show here that depletion of the PIAS2 beta isoform with a transcribed double-stranded RNA-directed RNA interference (PIAS2b-dsRNAi) specifically inhibits growth of ATC cell lines and patient primary cultures in vitro and of orthotopic patient-derived xenografts (oPDX) in vivo. Critically, PIAS2b-dsRNAi does not affect growth of normal or non-anaplastic thyroid tumor cultures (differentiated carcinoma, benign lesions) or cell lines. PIAS2b-dsRNAi also has an anti-cancer effect on other anaplastic human cancers (pancreas, lung, and gastric). Mechanistically, PIAS2b is required for proper mitotic spindle and centrosome assembly, and it is a dosage-sensitive protein in ATC. PIAS2b depletion promotes mitotic catastrophe at prophase. High-throughput proteomics reveals the proteasome (PSMC5) and spindle cytoskeleton (TUBB3) to be direct targets of PIAS2b SUMOylation at mitotic initiation. These results identify PIAS2b-dsRNAi as a promising therapy for ATC and other aggressive anaplastic carcinomas.