Use of larvae of the wax moth Galleria mellonella as an in vivo model to study the virulence of Helicobacter pylori

BACKGROUND: Helicobacter pylori is the first bacterium formally recognized as a carcinogen and is one of the most successful human pathogens, as over half of the world’s population is colonized by the bacterium. H. pylori-induced gastroduodenal disease depends on the inflammatory response of the host and on the production of specific bacterial virulence factors. The study of Helicobacter pylori pathogenic action would greatly benefit by easy-to-use models of infection. RESULTS: In the present study, we examined the effectiveness of the larvae of the wax moth Galleria mellonella as a new model for H. pylori infection. G. mellonella larvae were inoculated with bacterial suspensions or broth culture filtrates from either different wild-type H. pylori strains or their mutants defective in specific virulence determinants, such as VacA, CagA, CagE, the whole pathogenicity island (PAI) cag, urease, and gamma-glutamyl transpeptidase (GGT). We also tested purified VacA cytotoxin. Survival curves were plotted using the Kaplan-Meier method and LD(50) lethal doses were calculated. Viable bacteria in the hemocoel were counted at different time points post-infection, while apoptosis in larval hemocytes was evaluated by annexin V staining. We found that wild-type and mutant H. pylori strains were able to survive and replicate in G. mellonella larvae which underwent death rapidly after infection. H. pylori mutant strains defective in either VacA, or CagA, or CagE, or cag PAI, or urease, but not GGT-defective mutants, were less virulent than the respective parental strain. Broth culture filtrates from wild-type strains G27 and 60190 and their mutants replicated the effects observed using their respective bacterial suspension. Also, purified VacA cytotoxin was able to kill the larvae. The killing of larvae always correlated with the induction of apoptosis in hemocytes. CONCLUSIONS: G. mellonella larvae are susceptible to H. pylori infection and may represent an easy to use in vivo model to identify virulence factors and pathogenic mechanisms of H. pylori. The experimental model described can be useful to screen a large number of clinical H. pylori strain and to correlate virulence of H. pylori strains with patients’ disease status

MOVIO: A Toolkit for Creating Curated Digital Exhibitions

AbstractIn 2011, the Italian Ministry for Cultural Heritage and Tourism (MiBACT) published a guideline reference book analysing the state of the art and best practices of digital exhibitions made available on-line and offered a handbook successfully translated in English and even in Arabic. To satisfy the needs expressed by museum curators (but not limiting to them) GruppoMeta has implemented the MOVIO platform under the coordination of ICCU: MOVIO is a semantic CMS which provides tools to support the development of virtual/digital exhibitions, touristic and didactic applications. MOVIO supports the creation of a media archive and ‘non-scaring’ ontology builder for a storytelling approach and it allows cultural content publishing (it includes the creation of visit paths, up to mapping, time-line, galleries and social tools). The MOVIO open source SCMS platform is an easy and ready to use toolkit to build online and mobile virtual/digital exhibitions and narrations. It has begun to be experimented by several Italian institutions and several European partners from the AthenaPlus consortium

Effects of Dietary Supplementation of Lactobacillus acidophilus on Blood Parameters and Gut Health of Rabbits

SIMPLE SUMMARY: Gastrointestinal diseases are one of the most common causes of death in rabbits. Thus, maintaining a proper gut health is fundamental to guarantee adequate growth performance and welfare of the animals. Probiotics (e.g., Lactobacillus acidophilus) have been proposed as valuable alternatives to positively modulate gut health. The aim of this study was to evaluate the effects of Lactobacillus acidophilus D2/CSL on biochemical parameters, faecal score, cecal pH, gut histomorphometry, microbiota composition and faecal short-chain fatty acids in rabbits. Overall, the dietary inclusion of 1 × 10(9) cfu/kg feed once a day of Lactobacillus acidophilus D2/CSL did not impair rabbit productive performance, blood biochemical parameters, faecal score, gut morphometry, cecal pH, microbiota and short-chain fatty acids concentration. However, it reduced disease incidence and animal death, suggesting that it could improve disease resistance in rabbits. ABSTRACT: This study aimed to evaluate the effects of Lactobacillus acidophilus D2/CSL (L-1 × 10(9) cfu/kg feed/day) on biochemical parameters, faecal score (FS), cecal pH, gut morphometry, microbiota and cecal short-chain fatty acid (SCFAs) in rabbits. Three zootechnical trials were performed and in each trial 30 rabbits were allotted to two groups; a probiotic group (L) and a control group (C). At slaughter (day 45), samples of blood, duodenum, jejunum, ileum, liver and spleen were collected and submitted to histomorphometric analyses. Blood biochemical analyses, cecal microbiota and SCFAs determination were also performed. In trial 1 and 3, L. acidophilus D2/CSL did not affect productive parameters (p > 0.05). However, L group of trial 1 showed a lower morbidity and mortality compared to the control. In trial 2, C group showed a higher daily feed intake (p = 0.018) and a positive statistical tendency for live weight and average daily gain (p = 0.068). On the contrary, albumin was higher and ALFA-1 globulin was lower in the C group compared to L (p < 0.05). In all the trials, FS, cecal pH, histomorphometry, microbiota and SCFAs were unaffected. In conclusion, L. acidophilus D2/CSL did not impair growth performances, gut and rabbit’s health, reducing morbidity and mortality

Superinfections caused by carbapenem-resistant Enterobacterales in hospitalized patients with COVID-19: a multicentre observational study from Italy (CREVID Study)

Objectives To describe clinical characteristics and outcomes of COVID-19 patients who developed secondary infections due to carbapenem-resistant Enterobacterales (CRE). Methods Retrospective observational study including COVID-19 patients admitted to 12 Italian hospitals from March to December 2020 who developed a superinfection by CRE. Superinfection was defined as the occurrence of documented bacterial infection &gt;48 h from admission. Patients with polymicrobial infections were excluded. Demographic, clinical characteristics and outcome were collected. Isolates were classified as KPC, metallo-beta-lactamase (MBL) and OXA-48-producing CRE. A Cox regression analysis was performed to identify factors independently associated with 30 day mortality. Results Overall, 123 patients (median age 66 years, IQR 59-75) were included. The majority of infections occurred in the ICU (81, 65.9%), while 42 (34.1%) in medical wards. The most common types of infection were bloodstream infections (BSI) (n = 64, 52%), followed by urinary-tract infections (UTI) (n = 28, 22.8%), hospital-acquired/ventilator-associated pneumonia (HAP/VAP) (n = 28, 22.8%), intra-abdominal infections (n = 2, 1.6%) and skin infections (n = 1, 0.8%). Sixty-three (51.2%) infections were caused by KPC-, 54 (43.9%) by MBL-, and 6 (4.8%) by OXA-48-producing CRE. Thirty-day mortality was 33.3% (41/123). On Cox regression analysis, HAP/VAP compared with UTI (HR 7.23, 95% CI 2.09-24.97, P = 0.004), BSI compared with UTI (HR 3.96, 95% CI, 1.33-11.77, P = 0.004), lymphopenia on admission (HR 3, 95% CI 1.44-6.26, P = 0.003) and age (HR 1.05, 95% CI 1.02-1.08, P = 0.002) were predictors of 30 day mortality. Conclusions Superinfections by CRE were associated with high risk of 30 day mortality in patients with COVID-19. HAP/VAP was the strongest predictor of death in these patients

Il ruolo della Corte di giustizia nella definizione della politica economica e monetaria europea

Il ruolo giocato dalla Corte di giustizia nel campo della politica economica e della politica monetaria è stato importante quanto sottostimato. Le due competenze dell’Unione sono profondamente differenti - ciascuna dotata di significative peculiarità - e tuttavia strettamente interconnesse. Definire i confini incerti tra politica economica e politica monetaria è stato il ruolo più importante svolto dalla Corte in questo campo. Questo ha comportato una precisazione dei ruoli delle istituzioni chiamate ad agire - Commissione, Consiglio, Banca centrale - così come dei limiti all’indipendenza dell’istituto di emissione, ma anche della ripartizione di competenze tra Stati e Unione. Dal 2012, un’ulteriore funzione è stata esercitata dalla Corte: la verifica della legittimità dell’intervento straordinario della BCE nell’economia per gestire le crisi. L’assenza nei trattati europei di disposizioni specifiche che contemplassero un ruolo della Banca come prestatore di ultima istanza o nella gestione delle crisi economiche e finanziarie spiega bene le contestazioni dell’ultimo decennio, così come la necessità di pronunce autorevoli da parte della Suprema Corte europea. Queste si collocano a buon diritto nel solco della giurisprudenza in tema di rule of law e di garanzia del rispetto dei principi generali nell’ordinamento europeo

Common variants in Alzheimer’s disease and risk stratification by polygenic risk scores

Funder: Funder: Fundación bancaria ‘La Caixa’ Number: LCF/PR/PR16/51110003 Funder: Grifols SA Number: LCF/PR/PR16/51110003 Funder: European Union/EFPIA Innovative Medicines Initiative Joint Number: 115975 Funder: JPco-fuND FP-829-029 Number: 733051061Genetic discoveries of Alzheimer's disease are the drivers of our understanding, and together with polygenetic risk stratification can contribute towards planning of feasible and efficient preventive and curative clinical trials. We first perform a large genetic association study by merging all available case-control datasets and by-proxy study results (discovery n = 409,435 and validation size n = 58,190). Here, we add six variants associated with Alzheimer's disease risk (near APP, CHRNE, PRKD3/NDUFAF7, PLCG2 and two exonic variants in the SHARPIN gene). Assessment of the polygenic risk score and stratifying by APOE reveal a 4 to 5.5 years difference in median age at onset of Alzheimer's disease patients in APOE ɛ4 carriers. Because of this study, the underlying mechanisms of APP can be studied to refine the amyloid cascade and the polygenic risk score provides a tool to select individuals at high risk of Alzheimer's disease

Neutralizing antibodies to Omicron after the fourth SARS-CoV-2 mRNA vaccine dose in immunocompromised patients highlight the need of additional boosters

IntroductionImmunocompromised patients have been shown to have an impaired immune response to COVID-19 vaccines.MethodsHere we compared the B-cell, T-cell and neutralizing antibody response to WT and Omicron BA.2 SARS-CoV-2 virus after the fourth dose of mRNA COVID-19 vaccines in patients with hematological malignancies (HM, n=71), solid tumors (ST, n=39) and immune-rheumatological (IR, n=25) diseases. The humoral and T-cell responses to SARS-CoV-2 vaccination were analyzed by quantifying the anti-RBD antibodies, their neutralization activity and the IFN-γ released after spike specific stimulation.ResultsWe show that the T-cell response is similarly boosted by the fourth dose across the different subgroups, while the antibody response is improved only in patients not receiving B-cell targeted therapies, independent on the pathology. However, 9% of patients with anti-RBD antibodies did not have neutralizing antibodies to either virus variants, while an additional 5.7% did not have neutralizing antibodies to Omicron BA.2, making these patients particularly vulnerable to SARS-CoV-2 infection. The increment of neutralizing antibodies was very similar towards Omicron BA.2 and WT virus after the third or fourth dose of vaccine, suggesting that there is no preferential skewing towards either virus variant with the booster dose. The only limited step is the amount of antibodies that are elicited after vaccination, thus increasing the probability of developing neutralizing antibodies to both variants of virus.DiscussionThese data support the recommendation of additional booster doses in frail patients to enhance the development of a B-cell response directed against Omicron and/or to enhance the T-cell response in patients treated with anti-CD20