Gut Microbiota Composition Can Predict Colonization by Multidrug-Resistant Bacteria in SARS-CoV-2 Patients in Intensive Care Unit: A Pilot Study

The SARS-CoV-2 infection has increased the number of patients entering Intensive Care Unit (ICU) facilities and antibiotic treatments. Concurrently, the multi-drug resistant bacteria (MDRB) colonization index has risen. Considering that most of these bacteria are derived from gut microbiota, the study of its composition is essential. Additionally, SARS-CoV-2 infection may promote gut dysbiosis, suggesting an effect on microbiota composition. This pilot study aims to determine bacteria biomarkers to predict MDRB colonization risk in SARS-CoV-2 patients in ICUs. Seventeen adult patients with an ICU stay >48 h and who tested positive for SARS-CoV-2 infection were enrolled in this study. Patients were assigned to two groups according to routine MDRB colonization surveillance: non-colonized and colonized. Stool samples were collected when entering ICUs, and microbiota composition was determined through Next Generation Sequencing techniques. Gut microbiota from colonized patients presented significantly lower bacterial diversity compared with non-colonized patients (p < 0.05). Microbiota in colonized subjects showed higher abundance of Anaerococcus, Dialister and Peptoniphilus, while higher levels of Enterococcus, Ochrobactrum and Staphylococcus were found in non-colonized ones. Moreover, LEfSe analysis suggests an initial detection of Dialister propionicifaciens as a biomarker of MDRB colonization risk. This pilot study shows that gut microbiota profile can become a predictor biomarker for MDRB colonization in SARS-CoV-2 patients.Junta de Andalucía (CTS 164; CV20-77708)Instituto de Salud Carlos III (PI19/01058; PI20/01447Fundación Andaluza de Farmacia Hospitalaria” (3095/2020)

Anti-Inflammatory and Chemopreventive Effects of Bryophyllum pinnatum (Lamarck) Leaf Extract in Experimental Colitis Models in Rodents

Inflammatory bowel diseases, mainly ulcerative colitis and Crohn’s disease are characterized by chronic inflammation in the intestine. Currently several therapeutic strategies available to treat inflammatory bowel diseases. Though, most treatments can be associated with serious adverse effects what justifies the search for new treatments. In this sense, we highlight the interest in herbal products rich in bioactive compounds which immunomodulatory and antioxidant properties as is the case of Bryophyllum pinnatum (Crassulaceae). This plant is used in traditional medicine in Brazil for treating inflammatory diseases. We hypothesized that hydroethanolic B. pinnatum leaf extract has intestinal anti-inflammatory effects on two experimental colitis models: 2.4-dinitrobenzene sulfonic acid (DNBS) in rats, and dextran sulfate sodium (DSS) in mice. Ultra-fast liquid chromatography method used for the quantification of the main compounds indicated good linearity, specificity, selectivity, precision, robustness and accuracy. The major flavonoids (mg/g of the extract) quantified were: quercetin 3-O-a-L-arabinopyranosyl-(1!2)-a-L-rhamnopyranoside (35.56 ± 0.086 mg/g), kaempferol 3-O-a-L-arabinopyranosyl-(1!2)-a-L-rhamnopyranoside (4.66 ± 0.076 mg/g) and quercetin-3-O-rhamnopyranoside (4.56 ± 0.026 mg/g). The results obtained in the DNBS and DSS models indicate that extract has both chemopreventive and antiinflammatory effects, observing a significant reduction in the disease activity index score, and less macroscopic and microscopic damage. The extract promoted downregulation of Tolllike receptor and kappa B p65 nuclear factor gene expression, leading to a reduction in pro-inflammatory and oxidative mediators, chemokines, and cell adhesion molecules. This immunomodulatory property was proposed that one of the possible action mechanisms of extract. An improvement in intestinal damage was also associated with a reduction in oxidative stress and infiltration of leukocytes, as evidenced by the reduction in malonaldialdehyde and myeloperoxidase activity and increase in total glutathione in the colonic tissue. Moreover, the extract improved the cytoarchitecture of the colonic tissue and the integrity of the intestinal epithelial barrier by restoring the expression of the proteins associated with mucosa protection. In view of the beneficial effects showed by the B. pinnatum leaf extract in preclinical rodent models of colitis there is the potential to conduct some future clinical studies to ensure safe and effective development of a phytotherapeutic treatment for human inflammatory bowel diseases.CAPESJunta de Andalucia CTS 164Spanish Ministry of Economy and Competitiveness AGL2015-67995-C3-3-REuropean Union (EU)Instituto de Salud Carlos II

Tigecycline reduces tumorigenesis in colorectal cancer via inhibition of cell proliferation and modulation of immune response

Junta de Andalucía (CTS 164)Instituto de Salud Carlos III (Spain)Fondo Europeo de Desarrollo Regional (FEDER)European Union, through the research grants PI18/00826, P18-RT-4930, PI0206–2016, PIE16/00045 and PI19/01058Spanish Ministry of Science and Innovation (MCIN/AEI/ 10.13039/501100011033/FEDER)RTI2018–101309-BC22Chair “Doctors Galera-Requena in cancer stem cell research” (CMC-CTS963Spanish Ministry of Science and Innovation (“Programa de Doctorado: Medicina Clínica y Salud Pública” B12.56.1).Instituto de Salud Carlos III (FI17/00176)Junta de Andalucía (P18-RT-4930)University of GranadaCIBER-EHD is funded by the Instituto de Salud Carlos II

Intestinal mesenchymal cells regulate immune responses and promote epithelial regeneration in vitro and in dextran sulfate sodium-induced experimental colitis in mice

This work was funded by the Junta de Andalucia (CTS 164) and by the Instituto de Salud Carlos III (Spain) and Fondo Europeo de Desarrollo Regional (FEDER), from the European Union, through the research grants PI18/00826, PI0206-2016 and PI19/01058. L.H-G and A.J.R-M are predoctoral fellows funded by the Spanish Ministry of Science and Innovation ("Programa de Doctorado: Medicina Clinica y Salud Publica" B12.56.1). J.A.M-T is a predoctoral fellow from the Instituto de Salud Carlos III (FI17/00176). P.A is supported by the Consejeria de Salud, Junta de Andalucia through the contract "Nicolas Monardes" (C-0013-2018). A. R-N is a postdoctoral fellow from the Instituto de Salud Carlos III (Miguel Servet program [CP19/00191]). CIBER-EHD is funded by the Instituto de Salud Carlos III.We would like to express our gratitude to Dr E. Aksoy and Dr L. Medrano Gonzalez (William Harvey Research Institute, Queen Mary University of London, London, UK) for providing us the cell line NCM356. Additionally, we thank Juan N. Moliz, Ana Santos and Mohamed Tassi of the Centre for Scientific Instrumentation (CIC, University of Granada) for their technical guidance and assistance.Aim Disruption of the intestinal mucosal tolerance, that is, the immunological unresponsiveness to innocuous food antigens and the commensal microbiota, in the colon is associated with several chronic diseases including inflammatory bowel disease (IBD). Understanding the mechanisms responsible for intestinal mucosal tolerance has potential translational value for its therapy and management. Human intestinal mesenchymal cells (iMCs) play important roles in colonic mucosal tolerance, but further studies on their tissue regenerative and immunomodulatory capacities are necessary in order to fully understand their function in health and disease. Methods In this study, we have isolated and analysed the capacity of human iMCs to promote wound healing and modulate immune responses in vitro and in vivo, using the dextran sulfate sodium (DSS)-induced colitis model. Results Cultured iMCs were CD45(-)CD73(+)CD90(+)CD105(+) and accelerated the wound closure in a normal colon mucosa (NCM) 356 human epithelial cell wound healing assay. Furthermore, iMCs blocked the LPS-mediated induction of TNF-alpha in THP-1 macrophages and inhibited the proliferation of peripheral blood mononuclear cells, partly through the induction of indoleamine-2,3-dioxygenase. In DSS colitic mice, iMCs administration reduced the disease activity index and ameliorated intestinal tissue damage and permeability. Furthermore, iMCs reduced intestinal inflammation, evidenced by a decreased mRNA expression of pro-inflammatory cytokines, reduced IL-1 beta secretion by intestinal explants and inhibited colonic iNOS protein expression. Conclusions Our data show that human iMCs isolated from the noninflamed intestine possess tissue-regenerative and immunomodulatory capacities that could potentially be harnessed/restored in order to reduce IBD severity.Junta de Andalucia CTS 164Instituto de Salud Carlos III European CommissionFondo Europeo de Desarrollo Regional (FEDER), from the European Union PI18/00826 PI0206-2016 PI19/01058Spanish Ministry of Science and Innovation ("Programa de Doctorado: Medicina Clinica y Salud Publica") B12.56.1Junta de Andalucia C-0013-2018Miguel Servet program CP19/00191Instituto de Salud Carlos III European Commissio

Silk fibroin nanoparticles enhance quercetin immunomodulatory properties in DSS-induced mouse colitis

This work has been supported from the European Commission ERDF/FEDER Operational Programme `Murcia' CCI N. 2007ES161PO001 (Project No. 14-20/20), the Junta de Andalucia (CTS164), Instituto de Salud Carlos III (PI19/01058) and the Spanish MINECO (Ref. CTQ201787708-R). P.D.-E. is a postdoctoral from Junta de Andalucia (European Commission FEDER); A.J.R.-M and L.H.-G. are predoctoral fellows from University of Granada ("Programa de Doctorado: Medicina Clinica y Salud Publica"); A.A.L.-P's research contract was supported by the ERDF/FEDER Operational Programme `Murcia' CCI N 2007ES161PO001 (Project No. 14-20/20); A.R.-N. is a postdoctoral fellow of Instituto de Salud Carlos III (Miguel Servet Program); T.V. is a postdoctoral fellow from Instituto de Investigación Biosanitaria de Granada.Inflammatory bowel disease (IBD) is a chronic and idiopathic inflammatory disorder affecting the gastrointes-tinal tract. The pharmacological treatments used currently for its treatment lack efficacy, so new therapeutic strategies should be developed. In this context, flavonoids loaded in biopolymeric nanoparticles can be considered as novel promising candidates. The aim of the present study was to evaluate the intestinal anti-inflammatory effects of quercetin when is administered loaded in silk fibroin nanoparticles (QSFN) in the dextran sulphate sodium experimental model of mouse colitis, which displays some similarities to human IBD. Previously characterized quercetin-loaded silk fibroin nanoparticles (QSFN). QSFN showed a reversible aggre-gation profile induced by the acidification of the solution but did not affect the loaded quercetin. Daily administration of QSFN significantly reduced disease activity index values compared to the control colitic group. This beneficial effect was not only corroborated by the histological examination of the colonic specimens but also the improvement of the colonic expression of the different proinflammatory cytokines (Tnf-alpha, Il-1 beta, Il-6, Mcp-1, Icam-1, Nlrp3 and iNOS). Therefore, these data suggest that QSFN could be a promising alternative to current treatments as a drug delivery system for IBD treatment.European Commission ERDF/FEDER Operational Programme `Murcia' CCI 2007ES161PO001- 14-20/20Junta de Andalucia CTS164Instituto de Salud Carlos III European Commission PI19/01058Spanish MINECO CTQ201787708-RERDF/FEDER Operational Programme 'Murcia' CCI 2007ES161PO001- 14-20/2

The Antioxidant Properties of Lavandula multifida Extract Contribute to Its Beneficial Effects in High-Fat Diet-Induced Obesity in Mice

Obesity is a worldwide public health problem whose prevalence rate has increased steadily over the last few years. Therefore, it is urgent to improve the management of obesity and its comorbidities, and plant-based treatments are receiving increasing attention worldwide. In this regard, the present study aimed to investigate a well-characterized extract of Lavandula multifida (LME) in an experimental model of obesity in mice and explore the underlying mechanisms. Interestingly, the daily administration of LME reduced weight gain as well as improved insulin sensitivity and glucose tolerance. Additionally, LME ameliorated the inflammatory state in both liver and adipose tissue by decreasing the expression of various proinflammatory mediators (Il-6, Tnf-a, Il-1b, Jnk-1, Ppara, Pparg, and Ampk) and prevented increased gut permeability by regulating the expression of mucins (Muc-1, Muc-2, and Muc-3) and proteins implicated in epithelial barrier integrity maintenance (Ocln, Tjp1, and Tff-3). In addition, LME showed the ability to reduce oxidative stress by inhibiting nitrite production on macrophages and lipid peroxidation. These results suggest that LME may represent a promising complementary approach for the management of obesity and its comorbidities.Junta de Andalucía (CTS 164), by the Instituto de Salud Carlos III (ISCIII) (PI19.01058)Spanish Ministry of Economy and Competitiveness (AGL2015-67995-C3-3-R)European Commission (FEDER/ERDF). The CIBEREHDInstituto de Salud Carlos II

Beneficial Effects of Limosilactobacillus fermentum in the DCA Experimental Model of Irritable Bowel Syndrome in Rats

Limosilactobacillus fermentum CECT5716, a probiotic strain isolated from human milk, has reported beneficial effects on different gastrointestinal disorders. Moreover, it has shown its ability to restore altered immune responses, in association with microbiome modulation in different pathological conditions. Therefore, our aim was to assess the effects of a Limosilacbacillus fermentum CECT5716 in a rat experimental model of irritable bowel syndrome (IBS) that resembles human IBS. The experimental IBS was induced by deoxycholic acid (DCA) in rats and then, Limosilactobacillus fermentum CECT5716 (109 CFU/day/rat) was administered. Behavioral studies, hyperalgesia and intestinal hypersensitivity determinations were performed and the impact of the probiotic on the inflammatory and intestinal barrier integrity was evaluated. Additionally, the gut microbiota composition was analyzed. Limosilactobacillus fermentum CECT5716 attenuated the anxiety-like behavior as well as the visceral hypersensitivity and referred pain. Moreover, this probiotic ameliorated the gut inflammatory status, re-establishing the altered intestinal permeability, reducing the mast cell degranulation and re-establishing the gut dysbiosis in experimental IBS. Therefore, our results suggest a potential use of Limosilactobacillus fermentum CECT5716 in clinical practice for the management of IBS patients.Junta de Andalucia A-CTS-447-UGR18 CTS 164Instituto de Salud Carlos IIIEuropean Commission PI19/01058 PI20/0144

The melatonergic agonist agomelatine ameliorates high fat diet-induced obesity in mice through the modulation of the gut microbiome

Background and purpose: Melatonin has shown beneficial effects on obesity, both in humans and experimental models, via regulating the altered circadian rhythm and thus ameliorating the gut dysbiosis associated with this metabolic condition. However, its clinical use is limited, mostly due to its short half-life. Agomelatine is an agonist of the melatonin receptors that could be used to manage obesity and offer a better profile than melatonin. Experimental approach: Male C57BL/6 mice were fed a high fat diet and orally treated for five weeks with agomelatine, or melatonin or metformin, used as control drugs. Metabolic profile, inflammatory status, vascular dysfunction and intestinal microbiota composition were assessed. Key results: Agomelatine lessened body weight gain, enhanced glucose and lipid metabolisms, and improved insulin resistance. It also reduced the obesity-associated inflammatory status and endothelial dysfunction, probably linked to its effect on gut dysbiosis, consisting of the restoration of bacterial populations with key functions, such as short chain fatty acid production. Conclusions and implications: Agomelatine can be considered as a novel therapeutic tool for the management of human obesity and its associated comorbidities.Junta de Andalucia CTS 164Instituto de Salud Carlos IIIEuropean Commission PI19/01058 European Commissio

The Prebiotic Effects of an Extract with Antioxidant Properties from Morus alba L. Contribute to Ameliorate High-Fat Diet-Induced Obesity in Mice

Obesity is a global health issue, in which modifications in gut microbiota composition have a key role. Different therapeutic strategies are being developed in combination with diet and exercise, including the use of plant extracts, such as those obtained from Morus alba L. leaves. Recent studies have revealed their anti-inflammatory and antioxidant properties. The aim of the present work was to evaluate whether the beneficial effects of M. alba L. leaf extract in high-fat diet-induced obesity in mice is correlated with its impact on gut microbiota. The extract reduced body weight gain and attenuated lipid accumulation, as well as increased glucose sensitivity. These effects were associated with an amelioration of the obesity-associated inflammatory status, most probably due to the described antioxidant properties of the extract. Moreover, M. alba L. leaf extract mitigated gut dysbiosis, which was evidenced by the restoration of the Firmicutes/Bacteroidota ratio and the decrease in plasma lipopolysaccharide (LPS) levels. Specifically, the extract administration reduced Alistipes and increased Faecalibaculum abundance, these effects being correlated with the beneficial effects exerted by the extract on the obesity-associated inflammation. In conclusion, anti-obesogenic effects of M. alba L. leaf extract may be mediated through the amelioration of gut dysbiosis.Junta de Andalucia CTS 164Instituto de Salud Carlos III PI19.01058Spanish Government AGL2015-67995-C3-3-Rperational Programme of the Region of Murcia (CCI) 2007ES161PO001 14-20/20European CommissionInstituto de Salud Carlos IIIi-pFIS, Instituto de Salud Carlos III; Programa de Doctorado BiomedicinapFIS, Instituto de Salud Carlos III; Programa de Doctorado Nutricio

Beneficial effects of Hibiscus sabdariffa and Lippia citriodora extracts on diet-induced obesity in mice: potential treatments for metabolic syndrome

El síndrome metabólico, enfermedad que afecta a un 25% de la población adulta mundial (1), es un conjunto de alteraciones metabólicas como son la hiperglucemia asociada a la resistencia a la insulina, obesidad (adiposidad visceral), dislipidemia con niveles reducidos de colesterol unido a lipoproteínas de alta densidad (colesterol HDL) y niveles elevados de triglicéridos en plasma, e hipertensión (2). Entre todos, la obesidad es considerada el eje central, y se trata de una enfermedad compleja y multifactorial, que se desarrolla como consecuencia de un desequilibro en el balance energético. Como consecuencia, hay un exceso de acumulación de energía en forma de grasa, sobre todo en el tejido adiposo, lo cual hace que los adipocitos aumenten en tamaño y/o número. El presente trabajo de tesis doctoral pretende evaluar el efecto de un extracto del cáliz de H. sabdariffa, y otro extracto de las hojas de L. citriodora, bien caracterizados desde un punto de vista químico, en un modelo experimental de síndrome metabólico en ratones, y evaluar los posibles mecanismos responsables de los efectos beneficiosos La administración de ambos extractos mostró una mejora en la disminución de obesidad inducida por una dieta rica en grasa, mejorando la resistencia a la insulina, y por lo tanto una mejora en el metabolismo glucídico y lipídico. Estos efectos beneficiosos están relacionados con una reducción en la respuesta inflamatoria sistémica y la restauración de la función de la permeabilidad intestinal alterada que se asocian a la obesidad. La capacidad de modular la microbiota intestinal por parte de los extractos ensayados puede tener un papel determinante en las acciones descritas. Por lo tanto, ambos extractos pueden ser candidatos para su uso futuro como complementos nutricionales en el manejo del síndrome metabólico.Tesis Univ. Granada