Targeting Helicobacter pylori urease activity and maturation: In-cell high-throughput approach for drug discovery

Background: Helicobacter pylori is a bacterium strongly associated with gastric cancer. It thrives in the acidic environment of the gastric niche of large portions of the human population using a unique adaptive mechanism that involves the catalytic activity of the nickel-dependent enzyme urease. Targeting urease represents a key strategy for drug design and H. pylori eradication. Method: Here, we describe a novel method to screen, directly in the cellular environment, urease inhibitors. A ureolytic Escherichia coli strain was engineered by cloning the entire urease operon in an expression plasmid and used to test in-cell urease inhibition with a high-throughput colorimetric assay. A two-plasmid system was further developed to evaluate the ability of small peptides to block the protein interactions that lead to urease maturation. Results: The developed assay is a robust cellular model to test, directly in the cell environment, urease inhibitors. The efficacy of a co-expressed peptide to affect the interaction between UreF and UreD, two accessory proteins necessary for urease activation, was observed. This event involves a process that occurs through folding upon binding, pointing to the importance of intrinsically disordered hot spots in protein interfaces. Conclusions: The developed system allows the concomitant screening of a large number of drug candidates that interfere with the urease activity both at the level of the enzyme catalysis and maturation. General significance: As inhibition of urease has the potential of being a global antibacterial strategy for a large number of infections, this work paves the way for the development of new candidates for antibacterial drugs

Canine Placenta Histological Findings and Microvascular Density: The Histological Basis of a Negative Neonatal Outcome?

Placenta is essential for the development of the fetus, and its impaired function can lead to a negative outcome (i.e., neonatal mortality). In dogs, investigations on placenta histology and neonatal outcome in healthy bitches are lacking, and a contribution is provided in this study to emphasize the use of placenta histology in practice. Fifty-one placentas from 11 litters were collected during cesarean section, classified according to the litter size (large (L) or small (S)) and the outcome, this latter as healthy (Group 1) or dead within 7 days (Group 2). The placenta/puppy weight ratio (PPR) was calculated, and specimens were formalin-fixed and paraffin-wax embedded, and on the resulting histological slides, capillary density (CD) was quantified. Among necrosis, calcification, and intravascular leucocytes, only the presence of multifocal-confluent necrosis (significantly more frequent in Group 2) was associated with a higher risk of death within 7 days (odds ratio = 30.7). Mixed logistic regression ruled out the effect on death both of a bitch and cesarean type (programmed vs. emergency). PPR and CD values were associated with litter size; large litters had lower PPR (p < 0.01) and higher CD (p < 0.05) than small litters. The relationship between PPR and CD was negative and significant (p < 0.01). Necrosis was a frequent finding in canine placentas, but only when multifocal-confluent was it associated with a poor outcome. The litter size influenced PPR (lower in L) and CD (higher in L), and this is likely due to the plasticity of placenta adaptation

Binary Copolymerization of p-Methylstyrene with Butadiene and Isoprene Catalyzed by Titanium Compounds Showing Different Stereoselectivity

The synthesis of p-methylstyrene–butadiene and p-methylstyrene–isoprene binary copolymers promoted by the titanium complexes Ti(η5-C5H5)-(κ2-MBMP)Cl (1) (MBMP = 2,2′-methylenebis(6-tert-butyl-4-methylphenoxo)) and chloro{1,4-dithiabutanediyl-2,2′-bis(4,6-di-tert-butylphenoxy)}titanium (2) activated by methylaluminoxane (MAO) is reported. Syndiotactic poly(p-methylstyrene)-co-cis-1,4-poly(butadiene) and syndiotactic poly(p-methylstyrene)-co-cis-1,4-polyisoprene were obtained using catalyst 1, whereas isotactic poly(p-methylstyrene)-co-trans-1,4-poly(butadiene) and isotactic poly(p-methylstyrene)-co-trans-1,4-polyisoprene were obtained using the catalyst 2. 13C NMR analysis of the copolymer microstructure allowed to assess the monomer block lengths and distribution in the polymer chain, revealing a blocky distribution of the two monomers along the polymer chain in the presence of the catalyst 1 and a random distribution with the catalyst 2 for both binary copolymers

Il sistema agro-alimentare dell’Emilia-Romagna, Rapporto 2014

Il Rapporto 2014 sul Sistema Agroalimentare dell'Emilia-Romagna rappresenta un importante contributo alla conoscenza di un settore fondamentale dell'economia regionale, un utile strumento per gli operatori e una guida per le politiche degli enti locali. Il Rapporto si apre con due capitoli che descrivono da un lato, lo scenario internazionale e, dall'altro le politiche comunitarie e nazionali, per il settore agroalimentare, che modificano lo scenario nel quale gli operatori saranno chiamati a muoversi dal 2014 al 2020. I principali cambiamenti congiunturali del sistema agroalimentare regionale occupano la parte central del Rapporto, con Quattro capitol dedicati all'agricoltura. Successivamente vengono affrontati gli altri aspetti rilevanti del sistema agro-alimentare regionale partendo dall'industria alimentare, con le dinamiche congiunturali e alcuni approfondimenti strutturali dell'occupazione

White Paper of Italian Gastroenterology: Delivery of services for digestive diseases in Italy: Weaknesses and strengths

In 2011 the three major Italian gastroenterological scientific societies (AIGO, the Italian Society of Hospital Gastroenterologists and Endoscopists; SIED, the Italian Society of Endoscopy; SIGE, the Italian Society of Gastroenterology) prepared their official document aimed at analysing medical care for digestive diseases in Italy, on the basis of national and regional data (Health Ministry and Lombardia, Veneto, Emilia-Romagna databases) and to make proposals for planning of care. Digestive diseases were the first or second cause of hospitalizations in Italy in 1999-2009, with more than 1,500,000 admissions/year; however only 5-9% of these admissions was in specialized Gastroenterology units. Reported data show a better outcome in Gastroenterology Units than in non-specialized units: shorter average length of stay, in particular for admissions with ICD-9-CM codes proxying for emergency conditions (6.7 days versus 8.4 days); better case mix (higher average diagnosis-related groups weight in Gastroenterology Units: 1 vs 0.97 in Internal Medicine units and 0.76 in Surgery units); lower inappropriateness of admissions (16-25% versus 29-87%); lower in-hospital mortality in urgent admissions (2.2% versus 5.1%); for patients with urgent admissions due to gastrointestinnal haemorrhage, in-hospital mortality was 2.3% in Gastroenterology units versus 4.0% in others. The present document summarizes the scientific societies' official report, which constitutes the "White paper of Italian Gastroenterology". © 2014 Editrice Gastroenterologica Italiana S.r.l

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field