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A CRISPR/Cas9 gene editing strategy has been remarkable in excising segments of integrated HIV-1 DNA sequences from the genome of latently infected human cell lines and by introducing InDel mutations, suppressing HIV-1 replication in patient-derived CD4+ T-cells, ex vivo . Here, we employed a short version of the Cas9 endonuclease, saCas9, together with a multiplex of guide RNAs (gRNAs) for targeting the viral DNA sequences within the 5′-LTR and the Gag gene for removing critically important segments of the viral DNA in transgenic mice and rats encompassing the HIV-1 genome. Tail-vein injection of transgenic mice with a recombinant Adeno-associated virus 9 (rAAV 9 ) vector expressing saCas9 and the gRNAs, rAAV:saCas9/gRNA, resulted in the cleavage of integrated HIV-1 DNA and excision of a 978 bp DNA fragment spanning between the LTR and Gag gene in the spleen, liver, heart, lung and kidney as well as in the circulating lymphocytes. Retro-orbital inoculation of rAAV 9 :saCas9/gRNA in transgenic rats eliminated a targeted segment of viral DNA and substantially decreased the level of viral gene expression in circulating blood lymphocytes. The results from the proof-of-concept studies, for the first time, demonstrate the in vivo eradication of HIV-1 DNA by CRISPR/Cas9 on delivery by an rAAV 9 vector in a range of cells and tissues that harbor integrated copies of viral DNA.

A set of 31 isolates were obtained from apricot (Prunus armeniaca L.) orchards showing symptoms of blast and gummosis in orchards located in north-eastern Italy, mostly during three subsequent growing seasons (2014–2016). These isolates were identified by means of their colony morphology, pathogenicity (hypersensitive reaction in tobacco leaves and the detection of both syringomycin and coronatine coding genes), genetic features (BLASTn analysis of the rpoD gene) and physiological tests (GATTa) as Pseudomonas syringae pv. syringae and P. syringae pv. morsprunorum race 1 and race 2 strains, respectively. Phylogenetic analysis based on the rpoD gene sequence showed that the two pathovars and two races are well separated and highlighted high intrapathovar/race variation for both P. syringae pv. syringae and P. syringae pv. morsprunorum race 2. Pseudomonas syringae pv. morsprunorum race 1 formed a unique clade that had high sequence homology. Repetitive element-based polymerase chain reaction fingerprinting analysis using the ERIC, REP and BOX primers again showed both high intra-pathovar variation within the P. syringae pv. syringae isolates, which grouped into six distinct clusters, and notable intra-race genetic diversity within both P. syringae pv. morsprunorum races. All 31 strains showed tolerance to copper at 200 ppm in vitro. Among these isolates, 13 were highly resistant (up to 500 ppm) and revealed the presence of the cusCBA gene cluster. All P. syringae pv. syringae strains showed ice nucleation activity, and nine were markedly active (up −3 °C) and characterised by the presence of the INAz gene. Finally, this study is the first report of the isolation of P. syringae pv. morsprunorum race 2 strains from apricot in Italy.

A total of 40 Pseudomonas syringae pv. actinidiae (Psa) strains isolated from past and current epidemics of kiwifruit bacterial canker worldwide were compared using repetitive-sequence PCR (rep-PCR) fingerprinting with BOX, ERIC and REP primer sets. The strains were also assessed for the presence of 23 type III secretion system effector genes, tolerance to sodium arsenite, the presence of coronatine and phaseolotoxin and for growth trends in Actinidia deliciosa leaves. Rep-PCR revealed the occurrence of 11 different Psa lineages among the strains and indicated a relevant genetic variability within the strains isolated in Japan during 1984–2011, whereas all strains obtained from the current pandemic showed the same banding pattern. All lineages showed the same growth trend when inoculated into A. deliciosa leaves. The difference between Psa strains from past and current epidemics was confirmed by the detection of different repertoires of type III effector protein genes. Not all Psa strains isolated in Japan during past epidemics of kiwifruit bacterial canker amplify genes of the argK-tox cluster of phaseolotoxin, suggesting their absence or gene mutation. The results of an arsenic tolerance assay indicated that almost all strains isolated in Italy during the current epidemic of kiwifruit bacterial canker were relatively tolerant to 0.15–0.35 mM sodium arsenite, whereas those isolated in Chile and New Zealand were very sensitive. Remarkably, the two strains from China were tolerant or very sensitive. Collectively, these data indicate a composite population structure of this pathogen, which was able to diversify in Japan during 27 years of recurrent infections to A. deliciosa. The current naming of Psa populations based on their numbering and presence/absence of phytotoxins should be reconsidered.

Recent studies based on multiple, comparative taxonomic analyses, including MLSA and average nucleotide identity (ANI) approaches, revealed the clear existence of strains associated with bacterial canker and decline of Corylus avellana trees belonging to two different genomospecies: Pseudomonas avellanae and Pseudomonas syringae. Strains of the latter were tentatively named P. s. pv. avellanae. The distinctiveness of the two phytopathogens, both associated with the same symptoms to C. avellana, requires the formal emended description of P. s. pv. avellanae. In fact, the strain BPIC 631 = NCPPB 3487, originally described as P. s. pv. avellanae and subsequently elevated to species/genomospecies level, was confirmed as the pathotype strain of P. avellanae. In this paper, the emended description of Pseudomonas syringae pv. avellanae, causal agent of European hazelnut bacterial canker and decline, is provided. P. s. pv. avellanae DSMZ 100307 = CRAPAV 013 is designed as the pathotype strain of this pathovar which infect solely C. avellana trees.

Mango (Mangifera indica) is grown throughout a wide range of frost-free climates and is one of the world's most important fruit crops. During 2010-2014, severe symptoms of bacterial apical necrosis (BAN) caused by Pseudomonas syringae pv. syringae (Pss) were observed on mango plants throughout all mango-growing areas in Sicily (southern Italy). The causal agent was identified based on phenotypic, genotypic and pathogenicity tests. The genetic variability of strains obtained from different areas and cultivars were assessed by rep-PCR and allowed clear differentiation of the Pss strains isolated from mango from other representative strains of the pathovar. In addition, the respective production of syringomycin, syringopeptin and mangotoxin was checked by in vitro tests and PCR detection. All tested strains showed the presence of mgoA and mgoB of the mangotoxin operon. Some copper-resistant Pss strains showing cusCBA genes were also found in some orchards. This feature could explain the failure of the disease control using copper compounds. Among 71 strains tested using in vitro assay, no copper-sensitive strains were detected, whereas 44 strains (62%) had high resistance, 16 strains (22.5%) were resistant and 11 strains (15.5%) had low resistance to copper sulfate. To our knowledge, this is the first report of copper resistance among Pss strains causing BAN on mango in Sicily.

Aim:A retrospective comparative case series was studied to determine whether the use of prostaglandin (PG) analogues to treat raised intraocular pressure (IOP) in patients with uveitis resulted in an increase in the frequency of anterior uveitis or cystoid macular oedema (CMO).Methods:163 eyes of 84 consecutive patients with uveitis and raised IOP treated with a PG analogue at two tertiary referral uveitis clinics were identified over a 3-month period. Control eyes were selected as those uveitic eyes of the same patients, which were treated with topical IOP-lowering agent(s) other than a PG analogue. Pretreatment IOP was compared with the mean IOP during PG analogue treatment. The frequency of anterior uveitis and CMO during PG analogue treatment was compared with the frequency of these complications in the control eyes during non-PG IOP-lowering treatment.Results:Significant IOP reductions were observed during PG analogue treatment. There was no significant difference in the frequency of anterior uveitis in those eyes treated with PG analogues and those treated with non-PG agents (p = 0.87, Fisher exact test). None of the 69 uveitic eyes without a previous history of CMO developed this complication. There was no increase in the frequency of visually significant CMO during PG treatment compared with that during non-PG treatment (p = 0.19, Fisher exact test).Conclusion:This study demonstrates that PG analogues are potent topical medications for lowering raised IOP in patients with uveitis and are not associated with increased risk of CMO or anterior uveitis.

Background: Wood dust has been classified as carcinogenic to humans and the association with nasal cancer risk has been observed in a large number of epidemiological studies. Objectives: The aim of this study is to summarise data about occupational exposure levels to wood dust in Italy and to examine some exposure determinants. Methods: Exposure measurements on wood dust were extracted from the SIREP (Italian Information System on Occupational Exposure to Carcinogens) database between 1996–2006. Descriptive statistics were calculated for exposure-related variables using univariate analyses. The prevalence of elevated exposure levels was estimated overall and for some industrial sectors. A multifactorial analysis of variance was performed to determine which factors influenced exposure levels to wood dust. Results: The total number of exposure measurements (n) reported is 10 837, which refer to 10 528 workers and 1181 companies. The overall arithmetic mean is 1.44 mg/m3 and the geometric mean is 0.97 mg/m3. Industrial sectors at high risk are “manufacture of wood and wood products” (n = 5539) as well as “manufacture of furniture” (n = 4347). About 74% of exposure measurements report a value <2 mg/m3. In the multifactorial analysis, it has been found that job category, industrial sector, company size and geographical location of the company influence the exposure levels. Conclusions: This study confirms the previous findings about occupational exposure to wood dust (mainly in wood industry and among woodworking machine operators) and suggests further investigations on other risk sectors (building and repairing of ships and boats). The potential of the occupational exposure database as a source of data for exposure assessment and surveillance is also confirmed.

Some Pseudomonas fluorescens strains, consistently isolated from the rhizosphere of wild plants grown in a soil that was highly polluted with illegal waste of smelter residues, were utilised for Mirabilis jalapa seed bioinoculation to verify their effects on seed germination and on promoting a higher heavy metal accumulation in the plant rhizosphere and/or uptake in the leaves. The high content of heavy metals in the soil induced a decrease in either the leaf dry weight or photosynthetic pigment concentration during all vegetative phase of M . jalapa . Bioinoculation with P . fluorescen s strains significantly increased the germination of seeds and the root length in the contaminated soil. In some bacterial strain/seed combination, bioinoculation significantly increased the accumulation of heavy metals in M . jalapa rhizosphere. For Cd, the concentration of this metal in the rhizospheres of bioinoculated plants ranged from 270 to 910 μg g −1 of dry weight compared with 200 μg g −1 of dry weight for the non-coated plants. Two P . fluorescens strains, AA27 and MO49, which were isolated from Artemisia annua and Melilotus officinalis , respectively, induced a significantly higher rhizosphere availability also for Cr, Cu, Ni and Zn. However, despite the relevant accumulation of the heavy metals in the plant rhizosphere, generally the metal uptake into the leaves was rather low. Both analysis of variance and principal component analysis confirmed this finding. However, one P . fluorescens strain, CD1, which was isolated from the multi-metal accumulator Cynodon dactylon , significantly promoted the M . jalapa leaf uptake for Cr, Cu and Zn. The plant metal uptake assessment, confirmed the per se capability of M . jalapa to effectively uptake Cd (30 %) and Cu (12.72 %) from the rhizosphere to the leaves, whereas the uptake for the other metals was low: Ni (2.66 %), Zn (2.46 %), Cr (1.75 %), Pb (0.73 %).

An ecological study, based on a data set containing all lung and pleural cancer deaths in each Italian municipality in the period 1980–2001, was performed. The pleural to lung cancer ratio was estimated to be 1 : 1 and 3% (around 700) of all male lung cancer deaths were found to be asbestos-related.