Colorectal cancer cells increase the production of short chain fatty acids by Propionibacterium freudenreichii impacting on cancer cells survival

Propionibacterium freudenreichii is a commercially relevant bacterium with probiotic potential. This bacterium can exert protective effects particularly against colorectal cancer (CRC), via the production of short chain fatty acids (SCFA), namely acetate and propionate. In this work, we aimed to evaluate the performance and adaptation capacity of P. freudenreichii to a simulated digestive stress using different culture media, namely YEL, Basal medium, Mimicking the Content of the Human Colon medium (MCHC) and DMEM. The effect of the fermented culture broth on CRC cells survival and of CRC cells conditioned media on the bacteria performance was also evaluated. Basal medium was found to be the best for P. freudenreichii to produce SCFA. MCHC medium, despite being the medium in which lower amounts of acetate and propionate were produced, showed higher acetate and propionate yields as compared to other media. We also observed that the presence of lactate in CRC cells conditioned growth medium resulting from cell metabolism, leads to an increased production of SCFA by the bacteria. The bacterial fermented broth successfully inhibited CRC cells proliferation and increased cell death. Our results showed for the first time that P. freudenreichii performance might be stimulated by extracellular lactate produced by CRC metabolic switch also known as Warburg effect, where cancer cells ferment glucose into lactate. Additionally, our results suggest that P. freudenreichii could be potentially used as a probiotic in CRC prevention at early stages of the carcinogenesis process and might help in CRC therapeutic approaches.The authors acknowledge the financial support from the FEDER through POFC—COMPETE and by FCT through project PEst-OE/BIA/UI4050/2014. This work was supported by FCT I.P. through the strategic funding of UID/BIO/04469/2013 unit and project ref. RECI/BBB-EBI/0179/2012 (project number FCOMP-01-0124- FEDER-027462). This work was also supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI). This article is a result of the project EcoAgriFood NORTE-01-0145-FEDER-00009, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF).info:eu-repo/semantics/publishedVersio

Distance-Based Methods: Ripley’s K function vs. K density function

In this paper, we propose an analytical and methodological comparison between two of the most known distance-based methods in the evaluation of the geographic concentration of economic activity. These two methods are Ripley’s K function, a cumulative function popularised by Marcon and Puech (2003) that counts the average number of neighbours of each point within a circle of a given radius, and K density function, a probability density function of point-pair distances introduced by Duranton and Overman (2005), which considers the distribution of distances between pairs of points. To carry out this comparison, we first apply both methodologies to an exhaustive database containing Spanish manufacturing establishments and we evaluate the spatial location patterns obtained from both analysis. After an initial analysis, we realise that although these functions have always been treated as substitutes they should be considered as complementary, as both cumulative function and probability density function provide relevant and necessary information about the distribution of activity in space. Therefore, our next step will be to assess what are the advantages and disadvantages of each methodology from a descriptive and analytical way.

Improved production of acetate and propionate by Propionibacterium freudenreichii

Propionibacterium freudenreichii is a commercially important bacterium that is well-known for its role as ripening starter in the cheese industry and its probiotic potential. These bacteria may beneficially modulate the intestinal ecosystem and can exert anti-neoplastic effects via the production of short chain fatty acids (SCFAs), acetate and propionate. Several studies have demonstrated that the SCFA production by P. freudenreichii is responsible for its probiotic abilities. The aim of this work was to optimize the acetate and propionate production by P. freudenreichii towards its future use as a nutraceutical agent. In order to optimize the production of the abovementioned SCFAs in a minimal synthetic media the different composition of the several components were evaluated. Characterization of the acetate and propionate production in a medium mimicking the content of the human colon (MCHC) and a medium used by colorectal carcinoma cell lines (DMEM) was performed. The basal medium (BM) was found to be the most promising regarding the production of the SCFAs, showing 0.530 ± 0.011 g L-1 of biomass; high acetate and propionate yields (0.216 ± 0.001 g g-1 and 0.572 ± 0.002 g g-1, respectively), as well as high productivities (0.031 ± 0.000 g L-1 h-1 and 0.010 ± 0.000 g L-1 h-1, respectively). In the MCHC and DMEM media, it was possible to observe microbial growth (0.234 ± 0.006 g L-1 and 1.54 ± 0.00 g L-1, respectively); however the amounts of acetate and propionate were lower than the ones produced in BM medium. The results suggest that acetate and propionate production depends not only on the substrate type, but also on the medium constituents, being the simplest medium the one that show higher productivities as P. freudenreichii show low SCFA production when grown in MCHC and DMEM media. Future work will be conducted in order the increase bacteria growth and SCFA production in those media as this represents an essential feature for its use as a nutraceutical

Spatial location patterns of Spanish manufacturing firms

In this paper, we evaluate the spatial location patterns of Spanish manufacturing firms in each industry. We use a distance-based method, Ripley’s K function, which allows us to treat space as continuous. With ‘complete spatial randomness’ as benchmark, every sector presents significant concentration whatever the distance considered. If we use the locations of all manufacturing firms as benchmark, we find dispersion in some sectors and concentration in others, finding also differences in the spatial scale at which clustering occurs. Finally, the use of cities as benchmark reveals that not only is ‘first nature’ among the location determinants of manufacturing firms but they also include the self-reinforcing advantages of ‘second nature’

Chondrogenic differentiation induced by extracellular vesicles bound to a nanofibrous substrate

Extracellular vesicles (EVs) are being increasingly studied owing to its regenerative potential, namely EVs derived from human bone marrow mesenchymal stem cells (hBM-MSCs). Those can be used for controlling inflammation, repairing injury, and enhancing tissue regeneration. Differently, the potential of EVs derived from human articular chondrocytes (hACs) to promote cartilage regeneration has not been thoroughly investigated. This work aims to develop an EVs immobilization system capable of selectively bind EVs present in conditioned medium obtained from cultures of hACs or hBM-MSC. For that, an anti-CD63 antibody was immobilized at the surface of an activated and functionalized electrospun nanofibrous mesh. The chondrogenic potential of bound EVs was further assessed by culturing hBM-MSCs during 28 days under basal conditions. EVs derived from hACs cultured under differentiation medium or from chondrogenically committed hBM-MSCs induced a chondrogenic phenotype characterized by marked induction of SOX9, COMP, Aggrecan and Collagen type II, and matrix glycosaminoglycans synthesis. Indeed, both EVs immobilization systems outperformed the currently used chondroinductive strategies. These data show that naturally secreted EVs can guide the chondrogenic commitment of hBM-MSCs in the absence of any other chemical or genetic chondrogenic inductors based in medium supplementation.The authors would like to acknowledge the Portuguese Foundation for Science and Technology (FCT) for the PhD grant to M.R.C. (PD/BD/113797/2015) financed by the Doctoral Program on Advanced Therapies for Health (PATH) (FSE/POCH/PD/169/2013) and the project Cells4_IDs (PTDC/BTM-SAL/28882/2017). This work was also financed by the Portuguese Mass Spectrometry Network, integrated in the National Roadmap of Research Infrastructures of Strategic Relevance (ROTEIRO/0028/2013; LISBOA-01-0145-FEDER-022125)

Biomimetic surface topography as a potential modulator of macrophages inflammatory response to biomaterials

The implantation of biomaterial devices can negatively impact the local microenvironment through several processes including the injury incurred during the implantation process and the associated host inflammatory response. Immune cell responses to implantable biomaterial devices mediate host-material interactions. Indeed, the immune system plays a central role in several biological processes required for the integration of biomaterials such as wound healing, tissue integration, inflammation, and foreign body reactions. The implant physicochemical properties such as size, shape, surface area, topography, and chemistry have been shown to provide cues to the immune system. Its induced immune-modulatory responses towards inflammatory or wound healing phenotypes can determine the success of the implant. In this work, we aim to evaluate the impact of some biomimetic surface topographies on macrophages' acute inflammatory response. For that, we selected 4 different biological surfaces to replicate through soft lithography on spin casting PCL membranes. Those topographies were: the surface of E. coli, S.eppidermidis and L929 cells cultured in polystyrene tissue culture disks, and an Eggshell membrane. We selected a model based on THP-1-derived macrophages to study the analysis of the expression of both pro-inflammatory and anti-inflammatory markers. Our results revealed that depending on the surface where these cells are seeded, they present different phenotypes. Macrophages present a M1-like phenotype when they are cultured on top of PCL membranes with the surface topography of E. coli and S. epidermidis. When cultured on membranes with L929 monolayers or Eggshell membrane surface topography, the macrophages present a M2-like phenotype. These results can be a significant advance in the development of new implantable biomaterial devices since they can help to modulate the inflammatory responses to implanted biomaterials by controlling their surface topography.FCT -Fundação para a Ciência e a Tecnologia(PD/59/2013

The biomimetic surface topography of Rubus fruticosus leaves stimulate the induction of osteogenic differentiation of rBMSCs

The interaction between cells and biomaterials is essential for the success of biomedical applications in which the implantation of biomaterials in the human body is necessary. It has been demonstrated that material's chemical, mechanical, and structural properties can influence cell behaviour. The surface topography of biomaterials is a physical property that can have a major role in mediating cellâ material interactions. This interaction can lead to different cell responses regarding cell motility, proliferation, migration, and even differentiation. The combination of biomaterials with mesenchymal stem cells (MSCs) for bone regeneration is a promising strategy to avoid the need for autologous transplant of bone. Surface topography was also associated with the capacity to control MSCs differentiation. Most of the topographies studied so far involve machine-generated surface topographies. Herein, our strategy differentiates from the above mentioned since we selected natural surface topographies that can modulate cell functions for regenerative medicine strategies. Rubus fruticosus leaf was the selected topography to be replicated in polycaprolactone (PCL) membranes through polydimethylsiloxane moulding and using soft lithography. Afterwards, rat bone marrow stem cells (rBMSCs) were seeded at the surface of the imprinted PCL membranes to characterize the bioactive potential of our biomimetic surface topography to drive rBMSCs differentiation into the osteogenic lineage. The selected surface topography in combination with the osteogenic inductive medium reveals having a synergistic effect promoting osteogenic differentiation.This work is a result of the project FROnTHERA (NORTE-01-0145-FEDER-000023), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and Portuguese Foundation for Science and Technology under the doctoral programme in Tissue Engineering, Regenerative Medicine and Stem Cells (PD/59/2013), (PD/BD/128087/2016) (COVID/BD/151599/2020) and by the project Cells4_IDs (PTDC/BTM-SAL/28882/2017)

Insuficiencia renal en un varón de 23 años con colitis ulcerosa tratada con 5-aminosalicilato

Caso clínico Se presenta el caso de un varón de 23 años, diagnosticado de colitis ulcerosa (CU) en agosto 2016 durante un brote moderado y recibió 5-aminosalicilato (5-ASA; mesalazina) 4800 mg/día, rápidamente logró la remisión de la enfermedad. Cuando se realizó el diagnóstico su función renal era normal (creatinina 0, 8 mg/dl). Después de la remisión sintomática, la dosis de 5-ASA se redujo a 1500 mg/día. Un año después, en una consulta programada, tuvo otro brote leve-moderado de CU, lo que llevó a un aumento en la dosis de 5-ASA (4500 mg/día). En este momento, los resultados analíticos demostraron un aumento en la creatinina sérica a 1, 8 mg/dl, esto se interpretó como consecuencia de la deshidratación por diarrea durante la actividad de la CU. Tres semanas después, la enfermedad alcanzó rápidamente la remisión clínica y se reanudó la dosis anterior de 5-ASA. Cuatro meses después, en remisión clínica completa presentó una creatinina sérica de 1, 5 mg/dl, por tanto, la dosis de mesalazina se redujo (1000 mg/día) para mantener la creatinina estable. Ocho meses después, su creatinina aumentó dos veces a 3 mg/dl y el paciente estaba asintomático. En la exploración física, se encontraba eupneico, normocoloreado, consciente y orientado. La presión arterial era 100/60 mmHg, la frecuencia cardiaca de 80 lpm, la SaO2 del 99% y la temperatura de 36 °C. La exploración abdominal era normal y la puño-percusión renal era negativa..

Effects of suspension versus traditional resistance training on explosive strength in elementary school-aged boys

The aim of this study was to determine the effects of an 8-week program of resistance training (RT) or suspension training (ST) on explosive strength in prepubescent boys. Methods: Fifty-seven boys aged 10–11 years were assigned to 2 training groups, RT or ST or a control group (no training program). Boys trained twice weekly for 8 weeks. Results: A significant interaction was reported with a large (P < .001, η2p = .463), medium (P < .001, η2p = .395), and small effect sized (P ≤ .001, η2p = .218) in the 1-kg ball throw, 3-kg ball throw, and time-at-20-m test, respectively. There was no significant interaction in the countermovement vertical jump or the standing long jump. Changes from preintervention to postintervention for the 1-kg ball throw were 5.94% and 5.82% for the ST and RT, respectively, and 8.82% and 8.14% in the 3-kg ball throw for the ST and RT, respectively. The improvement in the 20-m sprint was 1.19% for the ST and 2.33% for the RT. Conclusion: Traditional RT and ST seem to be effective methods for improving explosive strength in prepubescent boys. ST could be considered as an alternative modality to optimize explosive strength training in school-based programs