Nod2 Deficiency in mice is Associated with Microbiota Variation Favouring the Expansion of mucosal CD4+ LAP+ Regulatory Cells

Nucleotide-binding Oligomerization Domain-2 (NOD2) mutations are associated with an increased risk to develop Crohn's Disease. In previous studies, we have shown that Nod2-/- mice manifest increased proportion of Lamina Propria (LP) CD4+ LAP+ Foxp3- regulatory cells, when compared with Nod2+/+ mice, while CD4+ Foxp3 + regulatory cells were not affected. Here, we investigated the Nod2 gut microbiota, by 16S rRNA pyrosequencing, at steady state and after TNBS-colitis induction in mice reared separately or in cohousing, correlating the microbial profiles with LP regulatory T cells proportion and tissue cytokines content. We found that enrichment of Rikenella and Alistipes (Rikenellaceae) in Nod2-/- mice at 8 weeks of age reared separately was associated with increased proportion of CD4+ LAP+ Foxp3- cells and less severe TNBS-colitis. In co-housed mice the acquisition of Rickenellaceae by Nod2+/+ mice was associated with increased CD4+ LAP+ Foxp3- proportion and less severe colitis. Severe colitis was associated with enrichment of gram-negative pathobionts (Escherichia and Enterococcus), while less severe colitis with protective bacteria (Barnesiella, Odoribacter and Clostridium IV). Environmental factors acting on genetic background with different outcomes according to their impact on microbiota, predispose in different ways to inflammation. These results open a new scenario for therapeutic attempt to re-establish eubiosis in Inflammatory Bowel Disease patients with NOD2 polymorphisms

Estimasi Kapasitas dan Titik Berat pada Pembuatan Kapal Jaring Tradisional di Galangan Kapal Bagan SiapiApi Kabupaten Rokan Hilir Menggunakan Software Autodesk Inventor 2010

Jaring ship building in Bagan Siapiapi Kabupaten Rokan Hilir Riau in general is traditionally. One example that still use the traditional way is how to determine the capacity and center of gravity. In estimating the capacity and gravity in Bagan Siapiapi Kabupaten Rokan Hilir Riau is not accurate so as to anticipate it, Bagan Siapiapi community change the shape and size of the vessel at the time of testing The purpose of the study to estimate the capacity and center of gravity in Bagan Siapiapi. This research is a study case method that uses a single fishing vessel industry is in Bagan Siapiapi. Jaring's ships measured ship's main dimensions, construction Linggi, keel construction, joist construction, foundation construction machinery, building construction on the upper deck, and wood density testing, subsequent construction of ships drawn using Autodesk Inventor 2010 software. Capacity and center of gravity jaring ship calculations using Autodesk Inventor 2010 software. From the data processing capacity of jaring Bagan Siapiapi Chart for 7.093 kg and an emphasis on jaring ship Bagan Siapiapi is at position: (1) unloading condition (0,9 mm; 888,5 mm ;-162,5 mm) the front view with the tilt angle of the ship experienced a shaky 89,24o and -84o side view (2) maximum load condition ( 0,7 mm; 976,7 mm; -600,4 mm) the front view with the tilt angle of the ship experienced a shaky -89,8o and side view 80,03o

Application of Sustainable Remediation Techniques for Heavy Metal Reduction in Polluted Rivers in Mining Zones: Study Area Ponce Enriquez

Mining activity in Ecuador is primarily artisanal and small-scale. Rivers near the mining county of Camilo Ponce Enriquez have been severely affected by this type of mining. A field study was conducted encompassing 29 sites on the rivers Siete, Fermin, and Pagua during sampling campaigns that took place in both dry (December) and wet seasons (February). Physiochemical parameters and heavy metal concentrations were measured to enable environmental characterization of the rivers. Results revealed that most of the sampled sites had bad water quality and exceeded the environmental limits of heavy metal concentrations set by the Ecuadorian government. A parallel pilot investigation was performed for reducing the heavy metal concentrations in water ecosystems polluted by mining activities using bioremediation techniques. The bioremediation technique applied in the study used bacteria from a wastewater digestor and sugarcane bagasse as a carbon source to support the bacterial community. Sulfide was measured to monitor bacterial activity in experimental bioreactors containing local acid mine drainage (AMD). The results obtained were encouraging, as a significant decrease in heavy metal concentration was observed to nearly the level of Ecuadorian environmental regulations for Al, Cd, Cu, Fe, Pb, and Zn concentrations in freshwater in a set of bioreactors. Keywords: acid drainage, artisanal mining, bioremediation, heavy metals, pollution. Resumen La actividad minera en el Ecuador es principalmente artesanal y de pequeña escala. Los ríos cercanos al área minera de Camilo Ponce Enríquez se han visto severamente afectados por este tipo de minería. Se realizó un estudio de campo en 29 sitios sobre los ríos Siete, Fermín y Pagua durante campañas de muestreo que se realizaron tanto en la época seca (diciembre) como en época húmeda (febrero). Se midieron parámetros fisicoquímicos y concentraciones de metales pesados para permitir la caracterización ambiental de los ríos. Los resultados revelaron que la mayoría de los sitios muestreados tienen agua de mala calidad y excedieron los límites ambientales de concentración de metales pesados establecidos por el gobierno ecuatoriano. También se realizó una investigación piloto paralela destinada a reducir las concentraciones de metales pesados en ecosistemas acuáticos contaminados por actividades mineras utilizando técnicas de biorremediación. La técnica de biorremediación aplicada en el estudio utilizó bacterias de un digestor de aguas residuales y bagazo de caña de azúcar como fuente de carbono para apoyar a la comunidad bacteriana. Se midió el sulfuro para controlar la actividad bacteriana en biorreactores experimentales que contenían drenaje ácido minero (AMD) local. Los resultados obtenidos son alentadores ya que en un conjunto de biorreactores se observó una disminución significativa de las concentraciones de algunos metales pesados alcanzando casi en su mayoría el nivel permisible según las regulaciones ambientales ecuatorianas para las concentraciones de Al, Cd, Cu, Fe, Pb y Zn en agua dulce. Palabras Clave: drenajes ácidos, minería artesanal, biorremediación, metales pesados, contaminación

Metabarcoding protocol: Analysis of Bacteria (including Cyanobacteria) using the 16S rRNA gene and a DADA2 pipeline (Version 1)

This protocol has been prepared as part of the Interreg Alpine Space project Eco-AlpsWater (ASP569) - Innovative Ecological Assessment and Water Management Strategy for the Protection of Ecosystem Services in Alpine Lakes and Rivers, Activity A.T1.3, Deliverable D.T1.3.2 – 1, https://www.alpine-space.eu/projects/eco-alpswater/en/hom

Preliminary results on the evolution of proglacial ponds in the deglaciating Alps

Deglaciation is one of the most evident effects of the ongoing climatic changes on the Alpine environment. One of its common consequences is the formation of new water bodies in the proglacial area, where proglacial lakes and ponds are increasingly relevant ecosystems for the mountain landscape. The EVERLAKE project focuses on a recent system of proglacial ponds that originated from the retreat of the Zufall/Cevedale Glacier (Plima catchment, Central/Eastern Italian Alps). The aims of the project are to: (i) provide a first hydroecological characterisation of these pond ecosystems from a physical, chemical and biological point of view; (ii) understand their evolutionary trend during the process of deglaciation, with a space-for-time substitution approach. Here, we present data collected during the ice-free season 2022, showing the seasonal development occurring in three ponds located along a gradient of distance from the Cevedale Glacier terminus (i.e., at 2700-2900 m a.s.l.). We monitored water level, temperature and electrical conductivity and assessed the origin of water through analyses of stable isotopes (δ2H, δ18O). Bathymetric measurements were performed to estimate the residence time of each waterbody. We also analysed basic water chemistry, concentrations of trace elements, benthic and planktonic chlorophyll-a and organic content. The biological communities of these poorly known aquatic ecosystems were characterised by adopting an integrated approach combining morphological observations of microalgae and 16S and 18S rRNA metabarcoding of eDNA from both benthic and planktonic samples. The ponds showed different ecological conditions related to their distance from the glacier margins

Metabarcoding protocol: Analysis of protists using the 18S rRNA gene and a DADA2 pipeline (Version 1)

This protocol has been prepared as part of the Interreg Alpine Space project Eco-AlpsWater (ASP569) - Innovative Ecological Assessment and Water Management Strategy for the Protection of Ecosystem Services in Alpine Lakes and Rivers, Activity A.T1.3, Deliverable D.T1.3.2 – 2, https://www.alpine-space.eu/projects/eco-alpswater/en/hom

A cost-effective strategy for marker assisted selection (MAS) in apple (M. pumila Mill.): the experience from the Fondazione Edmund Mach programme for resistance and quality traits

7III International Symposium on Molecular Markers in Horticulture, Riva del Garda (TN), Italy, September 25-27, 2013partially_openInternationalMarker assisted selection (MAS) permits the selection of plants carrying genes that control interesting agronomic traits through the screening of associated markers; this allows the breeders to retain only germplasm conferring desirable traits from an early stage. Usually MAS involves the screening of hundreds or thousands of seedlings and thus the main challenge faced is the development of a pipeline that starts with a “lab friendly” and unambiguous seedling disposition in greenhouse, allows the rapid collection of samples avoiding cross contamination, implements a fast, cost-effective, high-throughput method of DNA extraction to be used for a small number of PCRs, and utilizes robust, reliable, and easy to interpret molecular markers. In this study ~6,500 seedlings from 24 apple crosses were genotyped in order to select seedlings conferring resistance traits, fruit quality traits or both. Lab work was performed using the KAPA 3G Plant PCR kit. The main problems encountered were associated with dirty extractions; however, following optimization less than 10% of the seedlings had to be resampled and in total about 70% of the samples tested carried desirable traitspartially_openBanchi, E.; Poles, L.; Magnago, P.; Pindo, M.; Costa, F.; Velasco, R.; Sargent, D.J.Banchi, E.; Poles, L.; Magnago, P.; Pindo, M.; Costa, F.; Velasco, R.; Sargent, D.J

Early melanoma invasivity correlates with gut fungal and bacterial profiles

7openInternationalItalian coauthor/editorBackground The microbiome is emerging as a crucial player of the immune checkpoint in cancer. Melanoma is a highly immunogenic tumour, and the composition of the gut microbiome has been correlated to prognosis and evolution of advanced melanoma and proposed as a biomarker for immune checkpoint therapy. Objectives We investigated the gut fungal and bacterial compositions in early-stage melanoma and correlated microbial profiles with histopathological features. Methods Sequencing of bacterial 16S rRNA and the fungal internal transcribed spacer region was performed on faecal samples of patients with stage I and II melanoma, and healthy controls. A meta-analysis with gut microbiota data from patients with metastatic melanoma was also carried out. Results We found a combination of gut fungal and bacterial profiles significantly discriminating patients with melanoma from controls. In patients with melanoma, we observed an abundance of Prevotella copri and yeasts belonging to the order Saccharomycetales. We found that the bacterial and fungal community correlated to melanoma invasiveness, whereas the specific fungal profile correlated to melanoma regression. Bacteroides was identified as general marker of immunogenicity, being shared by regressive and invasive melanoma. In addition, the bacterial communities in patients with stage I and II melanoma were different in structure and richer than those from patients with metastatic melanoma. Conclusions The composition of the gut microbiota in early-stage melanoma changes along the gradient from in situ to invasive (and metastatic) melanoma. Changes in the microbiota and mycobiota are correlated to the histological features of early-stage melanoma, and to the clinical course and response to immune therapies of advanced-stage melanoma, through direct or indirect immunomodulation.openVitali, F.; Colucci, R.; Di Paola, M.; Pindo, M.; De Filippo, C.; Moretti, S.; Cavalieri, D.Vitali, F.; Colucci, R.; Di Paola, M.; Pindo, M.; De Filippo, C.; Moretti, S.; Cavalieri, D