Genomic variants association with selective hypersensitivityreactions to amoxicillin and clavulanic acid

In summary, there are rare variants found in our patients that could be related to the development of AX or CLV- induced IDHRs. Interestingly, some of these rare variants could be associated with selective response to AX, such as the proapoptotic endonuclease ENDOG and neutrophil degranulation pathway, previously associ- ated with drug- induced anaphylaxis,6 or to CLV, like immunoprote- asome component PSMD8. Differences could be related to nature and metabolic characteristics of each drug; however, more research is needed. These findings represent a first step to identify potential genetic biomarkers to predict the susceptibility of suffering AX and CLV IDHRs.This work has been supported by Institute of Health ‘Carlos III’ (ISCIII) of the Ministry of Economy and Competitiveness (MINECO), (grants co- funded by European Regional Development Fund: PI15/01206, PI17/01237, PI18/00095, PI20/01734, PI21/0329, RETICS ARADYAL RD16/0006/0001, RICORS REI, RD21/0002/0008); Andalusian Regional Ministry of Health (grants PI- 0241- 2016, PE- 0172- 2018, PI- 0127- 2020). GB is a Clinical Investigator (B- 0007- 2022) re- search contract by Andalusian Regional Ministry Health. PDE holds a Sara Borrell research contract by Institute of Health ‘Carlos III’ (CD22/00112). CM holds a ‘Nicolas Monardes’ research contract by Andalusian Regional Ministry Health (RC- 0004- 2021). Funding for open access charge: Univesidad de Malaga/CBUA

Solanum aethiopicum L. from the Basilicata region as a source of specialized metabolites with promising anti-obesity effects: phytochemical characterization and in vivo investigation in high fat diet-fed mice

: Introduction: Solanum aethiopicum L., commonly known as scarlet eggplant (Solanaceae family) is one of the most traditionally cultivated vegetables in Basilicata, a southern region of Italy. Although multiple uses have been given to this vegetable, data about its anti-obesogenic activity are still limited. Methods: This study focuses on testing two different extracts obtained either from the peel or from the whole fruit of the Lucanian Solanum aethiopicum. Their ability to inhibit certain enzymatic activities was tested in vitro and then, the one that showed the better outcomes was tested on an experimental model of High-Fat Diet (HFD) induced obesity. Results: Spectrophotometric assays demonstrated that the peel extract possessed the highest ability to inhibit the selected enzymatic activities and so, its phytochemical profile was obtained through LC-MS chromatography. The oral administration of this extract (25 mg/kg) to HFD-fed mice reduced body weight gain and improved glucose and lipid metabolism. Similarly, the extract ameliorated the obesity-induced inflammatory status by reducing the expression of pro-inflammatory cytokines in both adipose and hepatic tissues. Interestingly, these effects were associated with the improvement of vascular dysfunction. Discussion: Lucanian Solanum aethiopicum extract may represent a new strategic approach for managing obesity and its associated diseases

The Antioxidant Activity of Thymus serpyllum Extract Protects against the Inflammatory State and Modulates Gut Dysbiosis in Diet-Induced Obesity in Mice

Nowadays, there is an increasing interest in alternative therapies in the treatment of metabolic syndrome that combine efficacy and safety profiles. Therefore, this study aimed to evaluate the effect of an extract of Thymus serpyllum, containing rosmarinic acid, on high-fat diet (HFD)- induced obesity mice, highlighting the impact of its antioxidant activity on the inflammatory status and gut dysbiosis. The extract was administered daily (50, 100 and 150 mg/kg) in HFD-fed mice. The treatment reduced body weight gain, glucose and lipid metabolic profiles. Moreover, the extract ameliorated the inflammatory status, with the c-Jun N-terminal kinases (JUNK) pathway being involved, and showed a significant antioxidant effect by the reduction of radical scavenging activity and the mitigation of lipid peroxidation. Moreover, the extract was able to modulate the altered gut microbiota, restoring microbial richness and diversity, and augmenting the counts of short-chain fatty acid producing bacteria, which have been associated with the maintenance of gut permeability and weight regulation. In conclusion, the antioxidant activity of Thymus serpyllum extract displayed a positive impact on obesity and its metabolic alterations, also reducing systemic inflammation. These effects may be mediated by modulation of the gut microbiota.Junta de Andalucia CTS 164Instituto de Salud Carlos III European Commission PI19.01058Spanish Government AGL201567995-C3-3-REuropean CommissionInstituto de Salud Carlos II

Comparative Study of the Antioxidant and Anti-Inflammatory E ects of Leaf Extracts from Four Di erent Morus alba Genotypes in High Fat Diet-Induced Obesity in Mice

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.Increased levels of reactive oxygen species (ROS) and a low-grade chronic inflammation in multiple organs have been demonstrated in obesity. Morus alba leaves extracts (MAEs) have been used in traditional medicine as anti-inflammatory agents. In this work, the bioactive compounds of di erent genotypes of M. alba L. (Filipina, Valenciana Temprana, Kokuso, and Italia) were analyzed not only by reverse phase high performance liquid chromatography–electrospray ionization-time of flight-mass spectrometry (RP-HPLC-ESI-TOF-MS) and hydrophilic interaction chromatography–electrospray ionization-time of flight-mass spectrometry (HILIC-ESI-TOF-MS), but also screened for in vitro and in vivo antioxidant activity by means of DPPH radical scavenging assay and Caenorhabditis elegans model. These MAEs were administered daily in a model of diet-induced obesity in mice. Filipina and Italia genotypes significantly reduced weight gain, the glycemic levels in high fat diet, as well as, levels of LDL-cholesterol and triglycerides. Filipina and Italia MAEs also reduced the expression of proinflammatory mediators such as Tnf- , Il-1 , Il-6 and increased the levels of adiponectin and AMPK, which exert anti-inflammatory e ects. Moreover, Italia genotype ameliorated the intestinal barrier function. In conclusion, Filipina and Italia methanolic extracts show the highest antioxidant and anti-inflammatory e ect, due to the presence of compounds such as protocatechuic acid or quercetin-3-glucoside, and they could be developed as a complementary treatment for obesity and metabolic disorders.Junta de Andalucia CTS 164Spanish Ministry of Economy and Competitiveness AGL2015-67995-C3-3-REuropean Commission (FEDER/ERDF)ERDF/FEDER Operational Programme of the Region of Murcia 2007ES161PO001 14-20/20Instituto de Salud Carlos II

Probiotic and Functional Properties of Limosilactobacillus reuteri INIA P572

Limosilactobacillus reuteri INIA P572 is a strain able to produce the antimicrobial compound reuterin in dairy products, exhibiting a protective effect against some food-borne pathogens. In this study, we investigated some probiotic properties of this strain such as resistance to gastrointestinal passage or to colonic conditions, reuterin production in a colonic environment, and immunomodulatory activity, using different in vitro and in vivo models. The results showed a high resistance of this strain to gastrointestinal conditions, as well as capacity to grow and produce reuterin in a human colonic model. Although the in vitro assays using the RAW 264.7 macrophage cell line did not demonstrate direct immunomodulatory properties, the in vivo assays using a Dextran Sulphate Sodium (DSS)-induced colitic mice model showed clear immunomodulatory and protective effects of this strain.This work was supported by project no. RTA2017-00002-00-00 from the Spanish Ministry of Science and Innovation, by the Junta de Andalucía (CTS 164) and Instituto de Salud Carlos III (PI19/01058) with funds from the European Union.Ye

The relationship between gut and nasopharyngeal microbiome composition can predict the severity of COVID-19

Background: Coronavirus disease 2019 (COVID-19) is a respiratory illness caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that displays great variability in clinical phenotype. Many factors have been described to be correlated with its severity but no specific determinants of infection outcome have been identified yet, maybe due the complex pathogenic mechanisms. The microbiota could play a key role in the infection and in the progression and outcome of the disease. Hence, SARS-CoV-2 infection has been associated with nasopharyngeal and gut dysbiosis and higher abundance of opportunistic pathogens. Methods: To identify new prognostic markers for the disease, a multicenter prospective observational cohort study was carried out in COVID-19 patients that were divided in three cohorts according to their symptomatology: mild (n=24), moderate (n=51) and severe/critical (n=31). Faecal and nasopharyngeal samples were taken and the microbiota was analysed. Results: Microbiota composition could be associated with the severity of the symptoms and the linear discriminant analysis identified the genera Mycoplasma and Prevotella as severity biomarkers in nasopharyngeal samples, and Allistipes, Enterococcus and Escherichia in faecal samples. Moreover, M. salivarium was defined as a unique microorganism in COVID-19 patients' nasopharyngeal microbiota while P. bivia and P. timonensis were defined in faecal microbiota. A connection between faecal and nasopharyngeal microbiota in COVID-19 patients was also identified as a strong positive correlation between P. timonensis (faeces) towards P. dentalis and M. salivarium(nasopharyngeal) was found in critically ill patients. Conclusions: This ratio could be used as a novel prognostic biomarker for severe COVID-19 patients.The research project was sup-ported by Government of Andalucia (Spain) (CV20-99908).N

Anti-inflammatory bowel effect of industrial orange by-products in DSS-treated mice

This work addresses the role of different by-products derived from the industrial extraction of orange juice in a possible anti-inflammatory effect in mice with colitis induced by dextran sulfate sodium (DSS). Fresh orange residue (FOR), dry orange residue (DOR), orange liqueur (OL) and animal feed (AF), as well as commercial citrus pectin (CP), were administered to C57BL/6J mice for 15 days before starting the DSS treatment. Analysis of macroscopic parameters such as the Disease Activity Index (DAI) and the colonic weight/length ratio revealed an anti-inflammatory effect following intake of FOR, AF or CP. Moreover, q-PCR of RNA from colonic tissue indicated measurable changes in the expression of TNF-α, IL-1β, iNOS, and intercellular adhesion molecules ICAM I, as well as in intestinal barrier proteins such as MUC-3, occludin, and ZO-1. Pectin, phenolic compounds and/or Maillard reaction products formed at initial steps were identified as relevant components exerting the ascribed beneficial effects. Our findings could open up the further application of a variety of orange by-products as food supplements in the potential amelioration of inflammatory bowel diseases.The authors are grateful to the García Carrión SA for providing samples and funding projects AGL2014-53445-R (MINECO) and ALIBIRD-CM S-2013/ABI-272 (CAM). M. T. P. thanks the National Secretary of Higher Education, Science, Technology and Innovation of Ecuador (SENESCYT).Peer reviewe

Effect of orange by-products on inflammatory indicators in a DSS mouse model

Trabajo presentado al XII Congreso Internacional Nutrición, Alimentación y Dietética, celebrado en Madrid el 11 y 12 de abril de 2018.[Introducción] Inflammation is a response triggered by damage to living tissues. The mechanisms involved in the initiation and perpetuation of inflammatory bowel disease (IBD) are not known exactly; however, there are mediators released in the early stages and may act as indicators of the disease. Colonic cytokines production and macroscopic indicators such as weight loss, reduction of food consumption, soft stools, among others, allow to evaluate the degree of IBD. Orange by-products (OBP) namely: fresh residue (FOR), dry residue (DOR), liquor (OL) and animal feed (AF), analysed in a previous study contain: phenols, pectin and/or furosine (Maillard reaction product). These compounds can show anti-oxidant, anti-bacterial and anti-cancer effect, but its anti-inflammatory activity is still little studied.[Objetivos] In consequence the aim of this work was to study the effect of OBP ingestion on IBD, using a DSS mice model; which exhibits similar characteristics to those present in human Crohn´s disease.[Métodos] 2-Furoylmethyl-Amino Acids (2-FM-AA) were analyzed by ionpair RP-HPLC-UV, and monosaccharides through hydrolysis with 2 M trifluoroacetic acid and subsequent formation of trimethylsilyloximes derivatives, by GC-FID. Male C57BL/6 mice (Janvier, St Berthevin Cedex, France) were distributed in 7 groups: healthy, DSS control, FOR, DOR, OL, AF and Citrus pectin (CP) (CEAMSA, O Porriño, Spain) was used as standard. Animals pre-treatment start 15 days before induction of colitis with DSS (2,7%) on drinking water. On day 22, animals were euthanized and colon samples were obtained. Macroscopic indicators were observed every day (body weigth, food consumption, disease activity index (DAI)), and tissue pro-inflammatory cytoquines and intestinal protective proteins were determined by Real-Time quantitative PCR (EcoTM Real-Time PCR System, Illumina, CA, USA).[Resultados] Weights lost was high for DSS control group and CP, AF and FOR significantly reduced the drop. In addition, FOR and CP groups, presented low disease activity index (DAI) values vs. DSS control group (p < 0.05). On the contrary, DOR and OL groups showed a DAI value similar to non-treated groups. Colonic weight/length ratio was significantly less in all the OBP or CP treated mice vs the DSS group. Colonic inflammatory cytokines expression as TNF-α, IL-1β, IL-6 and NOS were significantly reduced in AF and CP group and expression of protein with intestinal barrier functions as ICAM I, MUC 3, occludin, and ZO-1 were also positively modified in mice treated with FOR, AF and CP (p < 0.05).[Discusión] Analyzing the consumption effect of AF vs. CP; CP intake allowed to obtain, a better anti-inflammatory response, which could be due to its greatest content of soluble dietary fiber SDF and N-fructosyl-lysine.[Conclusiones] In general, CP and AF intake allowed to obtain a better anti-inflammatory response in DSS model of IBD compared with other OBP products assayed, which could be due to its greatest content of furosine. The product namely FOR also show a beneficial effect in some parameters measured probably due at its biggest phenols content. The evaluated indicators underline the potential role of pectin and Maillard reaction compounds in ameliorating some IBD symptoms. However, in order to reach more accurate conclusions, the analysis of additional biochemical indicators should be conducted.Peer reviewe

Exopolysaccharide Producing Bifidobacterium animalis subsp. lactis Strains Modify the Intestinal Microbiota and the Plasmatic Cytokine Levels of BALB/c Mice According to the Type of Polymer Synthesized

Bacteria-host interactions are mediated by different microbial associated molecular patterns which are most often surface structures such as, among others, exopolysaccharides (EPSs). In this work, the capability of two isogenic EPS-producing Bifidobacterium animalis subsp. lactis strains to modulate the gut microbiota of healthy mice, was assessed. Each strain produces a different type of polymer; the ropy strain S89L synthesized a rhamnose-rich, high-molecular weight EPS in highest abundance than the non-ropy DMS10140 one. BALB/c mice were orally fed for 10 days with milk-bifidobacterial suspensions and followed afterward for 7 post-intervention days (wash-out period). The colonic content of mice was collected in several sampling points to perform a metataxonomic analysis. In addition, the influence of specific microbial clades, apparently stimulated by the ropy and non-ropy strains, on mouse plasmatic cytokine levels was investigated through hierarchical association testing. Analysis of 16S rRNA gene sequences showed that the abundance of Firmicutes phylum significantly increased 7 days after cessing the treatment with both strains. The relative abundance of Alloprevotella genus also rose, but after shorter post-treatment times (3 days for both DMS10140 and S89L strains). Some bacterial clades were specifically modulated by one or another strain. As such, the non-ropy DMS10140 strain exerted a significant influence on Intestinomonas genus, which increased after 4 post-administration days. On the other hand, feeding with the ropy strain S89L led to an increase in sequences of Faecalibaculum genus at 4 post-treatment days, while the abundance of Erysipelotrichaceae and Lactobacillaceae families increased for prolonged times. Association testing revealed that several lactobacilli and bifidobacterial significantly stimulated by ropy S89L strain were positively associated with the levels of certain cytokines, including IL-5 and IL-27. These results highlight relevant changes in mice gut microbiota produced after administration of the ropy S89L strain that were associated to a potential immune modulation effect.The MicroHealth group acknowledge the projects AGL2015- 64901-R (AEI/FEDER, UE) and RTI2018-096339-B-I00 (MCIU/AEI/FEDER, UE), as well as the grant IDI/2018/000236 from the “Plan for Research, Development and Innovation of the Principado de Asturias 2018–2020” co-financed by the European Regional Development Funds (FEDER). NC-B thanks her FPI fellowship to the MINECO (BES2013-063984). CS acknowledges his Postdoctoral research contract funded by the “Instituto de Investigación Sanitaria del Principado de Asturias” (ISPA). The CIBER-EHD was funded by the Instituto de Salud Carlos III. PDE and LH-G are predoctoral fellows from University of Granada of “Programa de Doctorado: Medicina Clínica y Salud Pública.” We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI)

Administration of intestinal mesenchymal stromal cells reduces colitis-associated cancer in C57BL/6J mice modulating the immune response and gut dysbiosis

Background: Patients with inflammatory bowel disease (IBD) have a higher risk of developing colitis-associated colorectal cancer (CAC) with poor prognosis. IBD etiology remains undefined but involves environmental factors, genetic predisposition, microbiota imbalance (dysbiosis) and mucosal immune defects. Mesenchymal stromal cell (MSC) injections have shown good efficacy in reducing intestinal inflammation in animal and human studies. However, their effect on tumor growth in CAC and their capacity to restore gut dysbiosis are not clear. Methods: The outcome of systemic administrations of in vitro expanded human intestinal MSCs (iMSCs) on tumor growth in vivo was evaluated using the AOM/DSS model of CAC in C57BL/6J mice. Innate and adaptive immune responses in blood, mesenteric lymph nodes (MLNs) and colonic tissue were analyzed by flow cytometry. Intestinal microbiota composition was evaluated by 16S rRNA amplicon sequencing. Results: iMSCs significantly inhibited colitis and intestinal tumor development, reducing IL-6 and COX-2 expression, and IL-6/STAT3 and PI3K/Akt signaling. iMSCs decreased colonic immune cell infiltration, and partly restored intestinal monocyte homing and differentiation. iMSC administration increased the numbers of Tregs and IFN-γ+CD8+ T cells in the MLNs while decreasing the IL-4+Th2 response. It also ameliorated intestinal dysbiosis in CAC mice, increasing diversity and Bacillota/Bacteroidota ratio, as well as Akkermansia abundance, while reducing Alistipes and Turicibacter, genera associated with inflammation. Conclusion: Administration of iMSCs protects against CAC, ameliorating colitis and partially reverting intestinal dysbiosis, supporting the use of MSCs for the treatment of IBD