Structure-activity exploration of a small-molecule allosteric inhibitor of T790M/L858R double mutant EGFR

EGFR is a protein kinase whose aberrant activity is frequently involved in the development of non-small lung cancer (NSCLC) drug resistant forms. The allosteric inhibition of this enzyme is currently one among the most attractive approaches to design and develop anticancer drugs. In a previous study, we reported the identification of a hit compound acting as type III allosteric inhibitor of the L858R/T790M double mutant EGFR. Herein, we report the design, synthesis and in vitro testing of a series of analogues of the previously identified hit with the aim of exploring the structure-activity relationships (SAR) around this scaffold. The performed analyses allowed us to identify two compounds 15 and 18 showing improved inhibition of double mutant EGFR with respect to the original hit, as well as interesting antiproliferative activity against H1975 NSCLC cancer cells expressing double mutant EGFR. The newly discovered compounds represent promising starting points for further hit-to-lead optimisation

Whole Genome Sequencing of a Chlamydia trachomatis Strain Responsible for a Case of Rectal Lymphogranuloma Venereum in Italy

Lymphogranuloma venereum (LGV) is a systemic sexually transmitted infection caused by Chlamydia trachomatis serovars L1 to L3. The current LGV cases in Europe are mainly characterized by an anorectal syndrome, spreading within men who have sex with men (MSM). Whole-genome sequencing of LGV strains is crucial to the study of bacterial genomic variants and to improve strategies for contact tracing and prevention. In this study, we described the whole genome of a C. trachomatis strain (LGV/17) responsible for a case of rectal LGV. LGV/17 strain was isolated in 2017 in Bologna (North of Italy) from a HIV-positive MSM, presenting a symptomatic proctitis. After the propagation in LLC-MK2 cells, the strain underwent whole-genome sequencing by means of two platforms. Sequence type was determined using the tool MLST 2.0, whereas the genovariant was characterized by an ompA sequence evaluation. A phylogenetic tree was generated by comparing the LGV/17 sequence with a series of L2 genomes, downloaded from the NCBI website. LGV/17 belonged to sequence type ST44 and to the genovariant L2f. Nine ORFs encoding for polymorphic membrane proteins A-I and eight encoding for glycoproteins Pgp1-8 were detected in the chromosome and in the plasmid, respectively. LGV/17 was closely related to other L2f strains, even in the light of a not-negligible variability. The LGV/17 strain showed a genomic structure similar to reference sequences and was phylogenetically related to isolates from disparate parts of the world, indicative of the long-distance dynamics of transmission

Non-pathogenic Neisseria species of the oropharynx as a reservoir of antimicrobial resistance: a cross-sectional study

Commensal Neisseria species of the oropharynx represent a significant reservoir of antimicrobial resistance determinants that can be transferred to Neisseria gonorrhoeae. This aspect is particularly crucial in ‘men having sex with men’ (MSM), a key population in which pharyngeal co-colonization by N. gonorrhoeae and non-pathogenic Neisseria species is frequent and associated with the emergence of antimicrobial resistance. Here, we explored the antimicrobial susceptibility of a large panel of non-pathogenic Neisseria species isolated from the oropharynx of two populations: a group of MSM attending a ‘sexually transmitted infection’ clinic in Bologna (Italy) (n=108) and a group of males representing a ‘general population’ (n=119). We collected 246 strains, mainly belonging to N. subflava (60%) and N. flavescens (28%) species. Their antimicrobial susceptibility was evaluated assessing the minimum inhibitory concentrations (MICs) for azithromycin, ciprofloxacin, cefotaxime, and ceftriaxone using E-test strips. Overall, commensal Neisseria spp. showed high rates of resistance to azithromycin (90%; median MICs: 4.0 mg/L), and ciprofloxacin (58%; median MICs: 0.12 mg/L), whereas resistance to cephalosporins was far less common (<15%). Neisseria strains from MSM were found to have significantly higher MICs for azithromycin (p=0.0001) and ciprofloxacin (p<0.0001) compared to those from the general population. However, there was no significant difference in cephalosporin MICs between the two groups. The surveillance of the antimicrobial resistance of non-pathogenic Neisseria spp. could be instrumental in predicting the risk of the spread of multi-drug resistant gonorrhea. This information could be an early predictor of an excessive use of antimicrobials, paving the way to innovative screening and prevention policies

Years of life that could be saved from prevention of hepatocellular carcinoma

BACKGROUND: Hepatocellular carcinoma (HCC) causes premature death and loss of life expectancy worldwide. Its primary and secondary prevention can result in a significant number of years of life saved. AIM: To assess how many years of life are lost after HCC diagnosis. METHODS: Data from 5346 patients with first HCC diagnosis were used to estimate lifespan and number of years of life lost after tumour onset, using a semi-parametric extrapolation having as reference an age-, sex- and year-of-onset-matched population derived from national life tables. RESULTS: Between 1986 and 2014, HCC lead to an average of 11.5 years-of-life lost for each patient. The youngest age-quartile group (18-61 years) had the highest number of years-of-life lost, representing approximately 41% of the overall benefit obtainable from prevention. Advancements in HCC management have progressively reduced the number of years-of-life lost from 12.6 years in 1986-1999, to 10.7 in 2000-2006 and 7.4 years in 2007-2014. Currently, an HCC diagnosis when a single tumour <2 cm results in 3.7 years-of-life lost while the diagnosis when a single tumour 65 2 cm or 2/3 nodules still within the Milan criteria, results in 5.0 years-of-life lost, representing the loss of only approximately 5.5% and 7.2%, respectively, of the entire lifespan from birth. CONCLUSIONS: Hepatocellular carcinoma occurrence results in the loss of a considerable number of years-of-life, especially for younger patients. In recent years, the increased possibility of effectively treating this tumour has improved life expectancy, thus reducing years-of-life lost

Whole-Genome Analysis of Diversity and SNP-Major Gene Association in Peach Germplasm

Peach was domesticated in China more than four millennia ago and from there it spread world-wide. Since the middle of the last century, peach breeding programs have been very dynamic generating hundreds of new commercial varieties, however, in most cases such varieties derive from a limited collection of parental lines (founders). This is one reason for the observed low levels of variability of the commercial gene pool, implying that knowledge of the extent and distribution of genetic variability in peach is critical to allow the choice of adequate parents to confer enhanced productivity, adaptation and quality to improved varieties. With this aim we genotyped 1,580 peach accessions (including a few closely related Prunus species) maintained and phenotyped in five germplasm collections (four European and one Chinese) with the International Peach SNP Consortium 9K SNP peach array. The study of population structure revealed the subdivision of the panel in three main populations, one mainly made up of Occidental varieties from breeding programs (POP1OCB), one of Occidental landraces (POP2OCT) and the third of Oriental accessions (POP3OR). Analysis of linkage disequilibrium (LD) identified differential patterns of genome-wide LD blocks in each of the populations. Phenotypic data for seven monogenic traits were integrated in a genome-wide association study (GWAS). The significantly associated SNPs were always in the regions predicted by linkage analysis, forming haplotypes of markers. These diagnostic haplotypes could be used for marker-assisted selection (MAS) in modern breeding programs

Dallo stilo allo schermo

Il volume propone una panoramica che segue il pensiero traduttologico in cronologia «verticale» nella prima parte, cui corrispondono tre capitoli:1. a traduzione dalle origini a Lutero; 2.Seicento e Settecento; 3. Dall’età romantica al primo Novecento. I nove capitoli della seconda parte illustrano in «orizzontale» l’evoluzione degli studi di traduzione a partire dalla svolta linguistica degli anni Trenta del secolo scorso poi

Valutazione del sistema BD ProbeTec CT/GC Qx DNA Amplified assay su piattaforma BD Viper LT per la diagnosi molecolare di infezioni da Chlamydia trachomatis e Neisseria gonorrhoeae

INTRODUZIONE: Chlamydia trachomatis e Neisseria gonorroheae sono i principali patogeni ad eziologia batterica coinvolti nelle infezioni sessualmente trasmesse. Sebbene siano microorganismi dalle caratteristiche molto diverse, portano a manifestazioni cliniche simili e spesso confondenti. Negli ultimi anni, per la diagnosi di laboratorio di tali batteri, si \ue8 ampiamente diffuso l\u2019uso di metodiche di amplificazione degli acidi nucleici, caratterizzate da sensibilit\ue0 e specificit\ue0 maggiori rispetto alle metodiche tradizionali di coltura e/o identificazione di antigeni. In particolare, si \ue8 affermato l\u2019uso di test multiplex, che permettono di determinare parallelamente pi\uf9 patogeni a partire dallo stesso campione biologico. Lo scopo di questo studio \ue8 stato quello di valutare le performance del BD ProbeTec CT/GC Qx DNA Amplified assay su piattaforma BD Viper LT per la diagnosi delle infezioni da clamidia e gonococco, in comparazione ai dati ottenuti con il test VERSANT CT/GC 1.0 (Real-time PCR) (Siemens). METODI: Sono stati analizzati, in maniera retrospettiva, 352 campioni progressivi pervenuti al laboratorio di Microbiologia del Policlinico Sant\u2019Orsola-Malpighi di Bologna per la ricerca di acidi nucleici di C. trachomatis e N. gonorrhoeae. In totale, 232 campioni erano stati raccolti da soggetti recatisi in poliambulatori presenti sul territorio comunale e provinciale di Bologna (26 tamponi e 141 urine), mentre 120 urine e 65 tamponi provenivano da pazienti dell\u2019ambulatorio di Malattie a Trasmissione Sessuale (MTS) del Policlinico. I campioni sono stati testati con il kit VERSANT CT/GC 1.0, attualmente utilizzato nella routine diagnostica e successivamente con il BD ProbeTec CT/GC Qx DNA Amplified assay su piattaforma BD Viper LT. Le due metodiche si avvalgono di sistemi altamente automatizzati per le fasi di estrazione, allestimento piastre e rivelazione. RISULTATI: Le due metodiche hanno mostrato ottimi valori di concordanza tra loro; in particolare, la concordanza \ue8 stata del 98.4% per la determinazione delle infezioni da clamidia sulle urine e del 98.9% sui tamponi. Invece, la concordanza per le infezioni da gonococco \ue8 stata del 100% sulle urine e del 98.9% sui tamponi. La prevalenza di casi da N. gonorrhoeae nella popolazione afferente all\u2019ambulatorio MTS \ue8 risultata del 4%, mentre \ue8 stata pari allo 0.8% nella popolazione a minor rischio di infezioni sessualmente trasmesse. Infine, la prevalenza di C. trachomatis nella popolazione a pi\uf9 alto rischio \ue8 risultata pari al 9.1%, mentre nella la popolazione generale \ue8 stata del 2.4%. CONCLUSIONI: Il nostro studio conferma le ottime performance diagnostiche dei test BD ProbeTec CT/GC Qx DNA Amplified assay su piattaforma BD Viper LT e VERSANT CT/GC DNA 1.0 Assay, entrambi altamente automatizzati e di facile impego per gli utilizzatori

Long-term cognitive sequelae in a case of Wernicke’s encephalopathy after allogeneic stem cell transplantation

We describe the case of a non-alcoholic patient with chronic myeloid leukemia who developed iatrogenic Wernicke’s encephalopathy (WE) following stem cell transplantation. Four years after the WE acute event, the patient’s cognitive profile was mainly characterized by moderate memory impairment, and functional and daily-living difficulties. Our report sustains the hypothesis that a iatrogenic form of WE may produce long-term cognitive sequelae even when thiamine therapy is administered in the acute phase until the resolution of the neurological signs