Human Microbiota of the Argentine Population- A pilot study

The human microbiota is the collection of microorganisms living in or on the human body. An imbalance or dysbiosis in these microbial communities can be associated with a wide variety of human diseases (Petersen and Round, 2014; Pham and Lawley, 2014; Zaura et al., 2014). Moreover, when the microbiota of the same body sites is compared between different healthy individuals, specific microbial community features are apparent (Li et al., 2012; Yatsunenko et al., 2012; Oh et al., 2014; Relman, 2015). In addition, specific selective pressures are found at distinct body sites leading to different patterns in microbial community structure and composition (Costello et al., 2009; Consortium, 2012b; Zhou et al., 2013). Because of these natural variations, a comprehensive characterization of the healthy microbiota is critical for predicting alterations related to diseases. This characterization should be based on a broad healthy population over time, geography, and culture (Yatsunenko et al., 2012; Shetty et al., 2013; Leung et al., 2015; Ross et al., 2015). The study of healthy individuals representing different ages, cultural traditions, and ethnic origins will enable to understand how the associated microbiota varies between populations and respond to different lifestyles. It is important to address these natural variations in order to later detect variations related to disease.Fil: Carbonetto, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Agrobiotecnología de Rosario; ArgentinaFil: Fabbro Frías, Mónica Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Agrobiotecnología de Rosario; ArgentinaFil: Sciara, Mariela Ines. Centro de Diagnostico Medico de Alta Complejidad; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Serevalle, Analia. Centro de Diagnostico Medico de Alta Complejidad; ArgentinaFil: Mejico, Guadalupe. Centro de Diagnostico Medico de Alta Complejidad; ArgentinaFil: Revale, Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Agrobiotecnología de Rosario; ArgentinaFil: Romero, Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Agrobiotecnología de Rosario; ArgentinaFil: Brun, Bianca. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Agrobiotecnología de Rosario; ArgentinaFil: Fay, Marcelo. Centro de Diagnostico Medico de Alta Complejidad; ArgentinaFil: Fay, Fabian. Centro de Diagnostico Medico de Alta Complejidad; ArgentinaFil: Vazquez, Martin Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Agrobiotecnología de Rosario; Argentin

Unsaturated long-chain free fatty acids are input signals of the Salmonella enterica PhoP/PhoQ regulatory system

The Salmonella enterica serovar Typhimurium PhoP/PhoQ system has largely been studied as a paradigmatic two-component regulatory system not only to dissect structural and functional aspects of signal transduction in bacteria but also to gain knowledge about the versatile devices that have evolved allowing a pathogenic bacterium to adjust to or counteract environmental stressful conditions along its life cycle. Mg2+ limitation, acidic pH, and the presence of cationic antimicrobial peptides have been identified as cues that the sensor protein PhoQ can monitor to reprogram Salmonella gene expression to cope with extra- or intracellular challenging conditions. In this work, we show for the first time that long chain unsaturated free fatty acids (LCUFAs) present in Salmonella growth medium are signals specifically detected by PhoQ. We demonstrate that LCUFAs inhibit PhoQ autokinase activity, turning off the expression of the PhoP-dependent regulon. We also show that LCUFAs exert their action independently of their cellular uptake and metabolic utilization by means of the β-oxidative pathway. Our findings put forth the complexity of input signals that can converge to finely tune the activity of the PhoP/PhoQ system. In addition, they provide a new potential biochemical platform for the development of antibacterial strategies to fight against Salmonella infections.Fil: Viarengo, Gastón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Sciara, Mariela Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Salazar, Mario Oscar. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Kieffer, Pablo M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Furlan, Ricardo Luis Eugenio. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Garcia Vescovi, Eleonora. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentin

Biological activities of Castela coccinea Griseb. extracts

Infusions of leaf and bark of Castela coccinea Griseb. (Simaroubaceae) were traditionally used against dysentery. In this work, anti-bacterial and antioxidant activity were studied in C. coccinea extracts. The higest DPPH scavenging activity was detected in ethanolic leaf extracts with an IC50= 0.015 mg/ml. Bark ethanolic extracts showed inhibition of lipoperoxidation and free radical scavenging. However, a previous work evidenced that this extract is toxic at a concentration needed to show antioxidant activity. It might be consider that wood could be used instead of bark extract as the first one showing higher scavenging activity (Wood 55 %, Bark 33 % of DPPH scavenging) and good inhibition of lipoperoxidation. It is a very interesting fact the high tannin and mucilage content, which are localized particularly in leaves. The high antioxidant activity and abundant mucilage and tannin contents could justify, at least in part, its popular use.Colegio de Farmacéuticos de la Provincia de Buenos Aire

A Thin-layer Chromatography Autographic Method for the Detection of Inhibitors of the Salmonella PhoP-PhoQ Regulatory System

Introduction The PhoP–PhoQ system from Salmonella enterica serovar Typhimurium controls the expression of factors that are critical for the bacterial entry into host cells and the bacterial intramacrophage survival. Therefore it constitutes an interesting target to search for compounds that would control Salmonella virulence. Localisation of such compounds in complex matrixes could be facilitated by thin-layer chromatography (TLC) bioautography. Objective To develop a TLC bioautography to detect inhibitors of the PhoP–PhoQ regulatory system in complex matrixes. Methods The TLC plates were covered by a staining solution containing agar, Luria-Bertani medium, 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal), kanamycin and a S. typhimurium strain that harbours a reporter transcriptional lacZ-fusion to an archetypal PhoP-activated gene virK. After solidification, the plate was incubated at 37°C for 16 h. Results A bioautographic assay suitable for the localisation of inhibitors of the PhoP–PhoQ system activity in S. enterica serovar Typhimurium present in a complex matrix is described. The assay was used to analyse a series of hydrolysed extracts prepared by alkaline treatment of crude plant extracts. Bioassay-guided analysis of the fractions by NMR spectroscopy and MS led to the identification of linolenic and linoleic acids as inhibitory input signals of the PhoP–PhoQ system. Conclusion A practical tool is introduced that facilitates detection of inhibitors of the Salmonella PhoP–PhoQ regulatory system. The assay convenience is illustrated with the identification of the first naturally occurring organic compounds that down-regulate a PhoP–PhoQ regulatory system from a hydrolysed extract.Fil: Salazar, Mario Oscar. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Viarengo, Gastón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Sciara, Mariela Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Kieffer, Pablo M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Garcia Vescovi, Eleonora. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Furlan, Ricardo Luis Eugenio. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentin

A different gut microbiome linked to inflammation found in cirrhotic patients with and without hepatocellular carcinoma

Introduction and aim: A pro-oncogenic intestinal microbiome was observed in murine models; however, no specific microbiome in patients with hepatocellular carcinoma (HCC) has been reported. We aimed to compare the gut microbiome found in cirrhotic patients with or without HCC. Materials and methods: From 407 patients with Child Pugh A/B cirrhosis prospectively followed, 25 with HCC (cases) were matched with 25 without HCC (wo-HCC) in a 1:1 ratio according to age, gender, etiology, Child Pugh and severity of portal hypertension. In addition, results were also compared with 25 healthy subjects. Fecal stool samples were sequenced for the V3?V4 region of the microbial 16S rRNA (Illumina MiSeq Platform). Plasma cytokines were quantified including interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Results: We found a differential abundance in family members of Firmicutes with a 3-fold increase of Erysipelotrichaceae and a 5-fold decrease in family Leuconostocaceae in HCC when compared to wo-HCC controls. Genus Fusobacterium was found to be 5-fold decreased in HCC vs wo-HCC. The ratio bacteriodes/prevotella was increased in HCC. Three operational taxonomic units (OTUs), genus Odoribacter and Butyricimonas were more abundant in HCC, whereas a decreased abundance in Lachnospiraceae family genus Dorea was observed in HCC patients. A Random Forest model trained with differential abundant taxa correctly classified HCC individuals. This pattern was associated with an inflammatory milieu with a putative increased activation of NOD-like receptor pathways. Conclusion: We found a pattern of microbiome linked to inflammation that could be potentially useful as HCC biomarker after follow-up validation studies.Fil: Piñero, Federico. Red Latinoamericana de Investigación, Educación y Conciencia del Hígado; Argentina. Universidad Austral; ArgentinaFil: Vazquez, Martín. Heritas; ArgentinaFil: Baré, Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Rohr, Cristian Oscar. Heritas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mendizabal, Manuel. Universidad Austral; Argentina. Red Latinoamericana de Investigación, Educación y Conciencia del Hígado; ArgentinaFil: Sciara, Mariela Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas; ArgentinaFil: Alonso, Cristina. Universidad Austral; Argentina. Red Latinoamericana de Investigación, Educación y Conciencia del Hígado; ArgentinaFil: Fay, Fabián. Heritas; ArgentinaFil: Silva, Marcelo. Universidad Austral; Argentina. Red Latinoamericana de Investigación, Educación y Conciencia del Hígado; Argentin

Multivariate curve resolution-based data fusion approaches applied in 1H NMR metabolomic analysis of healthy cohorts

Metabolomics plays a critical role in deciphering metabolic alterations within individuals, demanding the use of sophisticated analytical methodologies to navigate its intricate complexity. While many studies focus on single biofluid types, simultaneous analysis of multiple matrices enhances understanding of complex biological mechanisms. Consequently, the development of data fusion methods enabling multiblock analysis becomes essential for comprehensive insights into metabolic dynamics.The authors are grateful to CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), UNR (Universidad Nacional de Rosario) and ASaCTeI (Agencia Santafesina de Ciencia, Tecnología e Innovación). The authors thanks Andrea Valeria Coscia for NMR data acquisition. The authors would like to extend their heartfelt gratitude to Dr. Alejandro J. Vila for his exceptional management efforts at the outset of this project. A.R.M.B. thanks CONICET for his PhD. fellowship and UNR for the AVE travel grant received.Peer reviewe