Cell-specific pattern of berberine pleiotropic effects on different human cell lines

The natural alkaloid berberine has several pharmacological properties and recently received attention as a potential anticancer agent. In this work, we investigated the molecular mechanisms underlying the anti-Tumor effect of berberine on glioblastoma U343 and pancreatic carcinoma MIA PaCa-2 cells. Human dermal fibroblasts (HDF) were used as non-cancer cells. We show that berberine differentially affects cell viability, displaying a higher cytotoxicity on the two cancer cell lines than on HDF. Berberine also affects cell cycle progression, senescence, caspase-3 activity, autophagy and migration in a cell-specific manner. In particular, in HDF it induces cell cycle arrest in G2 and senescence, but not autophagy; in the U343 cells, berberine leads to cell cycle arrest in G2 and induces both senescence and autophagy; in MIA PaCa-2 cells, the alkaloid induces arrest in G1, senescence, autophagy, it increases caspase-3 activity and impairs migration/invasion. As demonstrated by decreased citrate synthase activity, the three cell lines show mitochondrial dysfunction following berberine exposure. Finally, we observed that berberine modulates the expression profile of genes involved in different pathways of tumorigenesis in a cell line-specific manner. These findings have valuable implications for understanding the complex functional interactions between berberine and specific cell types

A GIS Referenced Methodological Approach for the Brownfield Redevelopment

In the perspective of a sustainable development of the urban territory, the aim of this research is to define a methodology for the characterization of a brownfield inventory, accessible by the government and the private investors to facilitate decision making, by reaching the needed information upon land capability, development incentives, public goals, interests and preferences, environmental concerns. This issue is very current, taking into account the numerous researches on the urban development of the cities and how the urban sprawl caused the industrial areas that initially existed on the city outskirts to become a part of the city center. This rapid growth motivates the governments and private investors to look at these abandoned industrial buildings as important assets. The research discusses a decision support framework and the benefits of access to geospatial databases in the decision-making process

Myelin-Associated MAL and PLP Are Unusual among Multipass Transmembrane Proteins in Preferring Ordered Membrane Domains

Eukaryotic membranes can be partitioned into lipid-driven membrane microdomains called lipid rafts, which function to sort lipids and proteins in the plane of the membrane. As protein selectivity underlies all functions of lipid rafts, there has been significant interest in understanding the structural and molecular determinants of raft affinity. Such determinants have been described for lipids and single-spanning transmembrane proteins; however, how multipass transmembrane proteins (TMPs) partition between ordered and disordered phases has not been widely explored. Here we used cell-derived giant plasma membrane vesicles (GPMVs) to systematically measure multipass TMP partitioning to ordered membrane domains. Across a set of 24 structurally and functionally diverse multipass TMPs, the large majority (92%) had minimal raft affinity. The only exceptions were two myelin-associated four-pass TMPs, myelin and lymphocyte protein (MAL), and proteo lipid protein (PLP). We characterized the potential mechanisms for their exceptional raft affinity and observed that PLP requires cholesterol and sphingolipids for optimal association with ordered membrane domains and that PLP and MAL appear to compete for cholesterol-mediated raft affinity. These observations suggest broad conclusions about the composition of ordered membrane domains in cells and point to previously unrecognized drivers of raft affinity for multipass transmembrane proteins

Myelin-Associated MAL and PLP Are Unusual among Multipass Transmembrane Proteins in Preferring Ordered Membrane Domains

Eukaryotic membranes can be partitioned into lipid-driven membrane microdomains called lipid rafts, which function to sort lipids and proteins in the plane of the membrane. As protein selectivity underlies all functions of lipid rafts, there has been significant interest in understanding the structural and molecular determinants of raft affinity. Such determinants have been described for lipids and single-spanning transmembrane proteins; however, how multipass transmembrane proteins (TMPs) partition between ordered and disordered phases has not been widely explored. Here we used cell-derived giant plasma membrane vesicles (GPMVs) to systematically measure multipass TMP partitioning to ordered membrane domains. Across a set of 24 structurally and functionally diverse multipass TMPs, the large majority (92%) had minimal raft affinity. The only exceptions were two myelin-associated four-pass TMPs, myelin and lymphocyte protein (MAL), and proteo lipid protein (PLP). We characterized the potential mechanisms for their exceptional raft affinity and observed that PLP requires cholesterol and sphingolipids for optimal association with ordered membrane domains and that PLP and MAL appear to compete for cholesterol-mediated raft affinity. These observations suggest broad conclusions about the composition of ordered membrane domains in cells and point to previously unrecognized drivers of raft affinity for multipass transmembrane proteins

Analisi dell'effetto protettivo di Bacteroides fragilis nell'infezione indotta da Bartonella henselae.

Bartonella henselae è un batterio gram-negativo intracellulare facoltativo responsabile della malattia da graffito del gatto (cat-scratch disease), un infezione caratterizzata da linfoadenopatia o da un decorso asintomatico in pazienti immunocompetenti. Tuttavia, l'angiomatosi bacillare e la peliosi epatica, nonché l'endocardite in pazienti con malattie valvolari, la batteriemia con febbre, e malattie epatiche o spleniche, sono tra le manifestazioni più comuni dell'infezione in pazienti immuno-compromessi. Bacteroides fragilis è un batterio anaerobio gram-negativo appartenente alla microflora intestinale, di recente è stato dimostrato che protegge da colite sperimentale indotta da Helicobacter hepaticus attraverso il polisaccaride A (PSA). Lo scopo del presente studio è stato stabilire se la colonizzazione di B. fragilis può proteggere dall'infezione da B. henselae in un modello animale di coinfezione. Analisi morfologiche e ultrastrutturali mediante colorazione ematossilina-eosina e microscopia elettronica sui tessuti murini hanno rivelato che i danni da B. henselae possono essere ridotti nella coinfezione con B. fragilis, ma che tale effetto non si verifica con il ceppo mutante difettivo nella capacità di produrre PSA (B. fragili delta PSA). Analisi immunoistochimiche con anticorpi anti-Bartonella hanno, inoltre, mostrato che il numero di cellule positive per campo è ridotto di almeno il 50% nel fegato (20±4 vs 50±8), aorta (5±1 vs 10±2) e milza (25±3 vs 40±6) dei topi coinfettati rispetto agli animali infettati solo con B. henselae. Questa diminuzione è meno evidente nella coinfezione con il ceppo DeltaPSA (35±6 nel fegato, 5±1 nell'aorta e 30±5 nella milza). Tali dati suggeriscono che la colonizzazione di B. fragilis possa essere in grado di prevenire i danni indotti da B. henselae tramite il PSA

Cyclosporin promotes neurorestoration and cell replacement therapy in pre-clinical models of Parkinson's disease.

The early clinical trials using fetal ventral mesencephalic (VM) allografts in Parkinson's disease (PD) patients have shown efficacy (albeit not in all cases) and have paved the way for further development of cell replacement therapy strategies in PD. The preclinical work that led to these clinical trials used allografts of fetal VM tissue placed into 6-OHDA lesioned rats, while the patients received similar allografts under cover of immunosuppression in an α-synuclein disease state. Thus developing models that more faithfully replicate the clinical scenario would be a useful tool for the translation of such cell-based therapies to the clinic

El defensor : Diario Liberal independiente, protector de los intereses materiales y morales de esta provincia: Año II Epoca 2ª Número 217 - 1904 noviembre 4

Copia digital. Madrid : Ministerio de Cultura. Subdirección General de Coordinación Bibliotecaria, 200

Evidence of <em>Bacteroides fragilis</em> Protection from <em>Bartonella henselae</em>-Induced Damage

<div><p><em>Bartonella henselae</em> is able to internalize endothelial progenitor cells (EPCs), which are resistant to the infection of other common pathogens. <em>Bacteroides fragilis</em> is a gram-negative anaerobe belonging to the gut microflora. It protects from experimental colitis induced by <em>Helicobacter hepaticus</em> through the polysaccharide A (PSA). The aim of our study was to establish: 1) whether <em>B. fragilis</em> colonization could protect from <em>B. henselae</em> infection; if this event may have beneficial effects on EPCs, vascular system and tissues. Our <em>in vitro</em> results establish for the first time that <em>B. fragilis</em> can internalize EPCs and competes with <em>B. henselae</em> during coinfection. We observed a marked activation of the inflammatory response by Real-time PCR and ELISA in coinfected cells compared to <em>B. henselae-</em>infected cells (63 <em>vs</em> 23 up-regulated genes), and after EPCs infection with mutant <em>B. fragilis</em> ΔPSA (≅90% up-regulated genes) compared to <em>B. fragilis</em>. Interestingly, in a mouse model of coinfection, morphological and ultrastructural analyses by hematoxylin-eosin staining and electron microscopy on murine tissues revealed that damages induced by <em>B. henselae</em> can be prevented in the coinfection with <em>B. fragilis</em> but not with its mutant <em>B. fragilis</em> ΔPSA. Moreover, immunohistochemistry analysis with anti-Bartonella showed that the number of positive cells per field decreased of at least 50% in the liver (20±4 <em>vs</em> 50±8), aorta (5±1 <em>vs</em> 10±2) and spleen (25±3 <em>vs</em> 40±6) sections of mice coinfected compared to mice infected only with <em>B. henselae</em>. This decrease was less evident in the coinfection with ΔPSA strain (35±6 in the liver, 5±1 in the aorta and 30±5 in the spleen). Finally, <em>B. fragilis</em> colonization was also able to restore the EPC decrease observed in mice infected with <em>B. henselae</em> (0.65 <em>vs</em> 0.06 media). Thus, our data establish that <em>B. fragilis</em> colonization is able to prevent <em>B. henselae</em> damages through PSA.</p> </div

<i>B. fragilis</i> and <i>B. henselae</i> internalize EPCs. A.

<p>Confocal images of human early EPCs infected with <i>B. henselae, B. fragilis</i> and <i>B. fragilis</i> ΔPSA at 100 MOI. Cells were stained with anti-<i>Bacteroides</i> (red) or with anti-<i>Bartonella</i> (green) specific antibodies after 24 h from infection. <b>B.</b> EPCs coinfected with <i>B. henselae,</i> and <i>B. fragilis</i> or <i>B. fragilis</i> ΔPSA as indicated. A MOI of 100 was used for all bacteria strains. Cells were stained with anti-<i>Bacteroides</i> (red) and with anti-<i>Bartonella</i> (green) specific antibodies after 24 h from infection.</p

EPC number of infected mice.

<p>EPC relative number of infected mice <i>versus</i> control mice was measured by FACS analysis. Sca-I/Flk-I double positive cells were considered as murine EPCs. Bars indicate standard errors.</p