Occurrence and viability of Anisakis spp. larvae in ready to eat products made of herring (Clupea harengus) sold in Italy

The study aimed to assess the occurrence and viability of Anisakis spp. larvae in ready to eat products made of herring, a common host of Anisakis spp. and the third most commercialized fish species in the EU. A total of 135 products consisting of 50 smoked whole specimens and 85 filleted products (25 smoked, 30 marinated, 30 canned) were sampled from 2016 to 2018. Viscera and muscle of whole herrings were visually inspected and separately digested. Filleted products were also visually inspected and digested. Larvae viability was assessed, then they were counted and microscopically identified to genus level. A subsample was molecularly identified. At least one Anisakis spp. larva was found in 56 products (41.5%), for a total of 1715 larvae collected (0-172 larvae/product). Most of the larvae (91%) were found in the viscera of 49 of the 50 whole herrings (98%). A highly significant difference was observed between the positivity rate and larval density at muscle level, as 149 larvae were found in the muscle of 31 whole herrings (positivity rate 62%, 0.022 larval density/g), while only 7 larvae were found in the 85 filleted products (positivity rate 7%, 0.001 larval density/g). Larvae were molecularly identified as A. simplex. The study showed that obviously contaminated products are commercialized. All the larvae were dead, proving a negligible risk of developing anisakiasis. However, their allergenic potential is debated and the significant difference between muscle infection levels in whole and filleted herrings may result in a different risk of exposure to antigens

Variation of microsporogenesis in sexual, apomictic and recombinant plants of Poa pratensis L.

Apomixis is a rather widespread phenomenon in plants. It is defined as the asexual formation of a seed from the maternal tissues of the ovule, avoiding the processes of meiosis and fertilization. Some species are facultative apomicts and form seeds by means of sexual and apomictic pathways to different extents. This is the case of Poa pratensis, the Kentucky bluegrass, which reproduces by aposporous pseudogamous facultative apomixis. This grass is one of the most studied apomictic systems, however some aspects, such as the male meiotic behavior, have not been so far investigated. In this study the process of microsporogenesis in genotypes of P. pratensis with a different mode of reproduction was investigated. The analysis revealed an almost regular meiosis in the sexual plants whereas apomictic genotypes exhibited different levels of meiotic irregularities, mainly due to cell fusion and irregular segregation in I and II division. Our data did not reveal evident connections between the extent and types of abnormalities and the components of apomixis, apomeiosis and parthenogenesis. The meiotic behavior of the examined plants was discussed in the light of their origin

Response rate as a potential surrogate for survival and efficacy in patients treated with novel immune checkpoint inhibitors: A meta-regression of randomised prospective studies

INTRODUCTION: To assess the role of the tumour response rate (RR) after immune checkpoint inhibitors-based therapy as a potential surrogate end-point of progression-free survival (PFS) and overall survival (OS) in patients with solid tumours, we performed a trial-based meta-regression of randomised studies comparing different immune checkpoint inhibitors-based treatments. METHODS: The systematic literature search included the electronic databases and the proceedings of oncologic meetings. Treatment effects on PFS and OS were expressed as hazard ratios (HRs); treatment effects on RR were expressed as odds ratios (ORs). A weighted regression analysis was performed on log-transformed treatment effect estimates to test the association between treatment effects on the surrogate outcome and treatment effects on the clinical outcome. RESULTS: Twenty-four trials, for a total of 11,894 patients, were included in the analysis. Using the complete set of data, the regression of either the log(HR) for PFS or the log(HR) for OS on the log(OR) for RR demonstrated weak associations (R2 = 0.47; 95% confidence interval [CI], 0.03-0.77; P = 0.001; and R2 = 0.32; 95% CI, 0.02-0.76; P = 0.01, respectively). The pre-planned analyses stratifying trials according to different type of disease and different mechanism of action of immune checkpoint inhibitors showed a very weak association of the RR with the OS for non-small cell lung cancer indicated and a modest association of the RR with the PFS for cytotoxic T lymphocyte-associated antigen 4 checkpoint inhibitors. CONCLUSION: The results of the trial-based meta-regression analysis indicated a weak correlation between RR and OS, supporting future investigations to assess the surrogacy of RR in the patient treated with immune checkpoint inhibitors

An Insight into T-DNA Integration Events in Medicago sativa

The molecular mechanisms of transferred DNA (T-DNA) integration into the plant genome are still not completely understood. A large number of integration events have been analyzed in different species, shedding light on the molecular mechanisms involved, and on the frequent transfer of vector sequences outside the T-DNA borders, the so-called vector backbone (VB) sequences. In this work, we characterized 46 transgenic alfalfa (Medicago sativa L.) plants (events), generated in previous works, for the presence of VB tracts, and sequenced several T-DNA/genomic DNA (gDNA) junctions. We observed that about 29% of the transgenic events contained VB sequences, within the range reported in other species. Sequence analysis of the T-DNA/gDNA junctions evidenced larger deletions at LBs compared to RBs and insertions probably originated by different integration mechanisms. Overall, our findings in alfalfa are consistent with those in other plant species. This work extends the knowledge on the molecular events of T-DNA integration and can help to design better transformation protocols for alfalfa

Perspectives of alfalfa in Europe

Sustainable agriculture shall reconcile high productivity with preservation of the environment, and at the same time compete in a global market. Alfalfa can help to meet the challenges of climate change and pollution reduction, by favoring crop rotations capable of better preserving the soil and controlling weeds. The utilization of alfalfa through grazing may also reduce risks of pollution and production costs. Alfalfa will keep an important place in Europe agriculture, because it sustainably supports high quality dairy productions. These unique features can be further improved through technical and varietal innovation

Barley Genes as Tools to Confer Abiotic Stress Tolerance in Crops

WOS: 000380895100001PubMed ID: 27536305Barley is one of the oldest cultivated crops in the world with a high adaptive capacity. The natural tolerance of barley to stress has led to increasing interest in identification of stress responsive genes through small/large-scale omics studies, comparative genomics, and overexpression of some of these genes by genetic transformation. Two major categories of proteins involved in stress tolerance are transcription factors (TFs) responsible from the re-programming of the metabolism in stress environment, and genes encoding Late Embryogenesis Abundant (LEA) proteins, antioxidant enzymes, osmolytes, and transporters. Constitutive overexpression of several barley TFs, such as C-repeat binding factors (HvGBF4), dehydration-responsive element-binding factors (HyDREB1), and WRKYs (HvWRKY38), in transgenic plants resulted in higher tolerance to drought and salinity, possibly by effectively altering the expression levels of stress tolerance genes due to their higher DNA binding affinity. Na+/H+ antiporters, channel proteins, and lipid transporters can also be the strong candidates for engineering plants for tolerance to salinity and low temperatures.Scientific Research Projects Coordination Unit of Istanbul University (BAP) [4712]This work was supported by the Scientific Research Projects Coordination Unit of Istanbul University (BAP No. 4712)

Efficient, antibiotic marker-free transformation of a dicot and a monocot crop with glutamate 1-semialdehyde aminotransferase selectable marker genes

Antibiotic-free, efficient in vitro selection in plant genetic engineering can improve risk perception and speed up pre-market scrutiny of genetically modified crops. We provide a protocol for genetic transformation of two important crops, durum wheat and alfalfa, using a bacterial and a plant-derived selectable marker gene encoding mutated, gabaculine-insensitive glutamate 1-semialdehyde aminotransferase (GSA) enzymes. These methods can potentially be applied, with minor adaptations, to many other monocot and dicot crop plants

Copy Number Estimation of a Plant-Derived Selectable Marker Gene by High Resolution Melting Analysis: A Tool to Simplify Transgenic Plant Breeding

Transferring a transgenic trait from a lab genotype into a cultivated plant variety requires selection of single-locus T0 plants, a backcrossing breeding program with transgenic progeny selection at each cycle and, generally, attaining the homozygous state for the transgene locus. Polyploidy and outcrossing complicate this process, as in the case of alfalfa, an autotetraploid, cross-fertilizing forage legume. Recently, a new, efficient selectable marker gene, MsGSAgr, derived from the alfalfa glutamate 1-semialdehyde aminotransferase (GSA) gene was described. Since MsGSAgr differs from the wildtype GSA by one nucleotide, high resolution melting (HRM) analysis could be used to screen transgenic plants for MsGSAgr, and linked transgene(s), copy number. An HRM assay was developed by simulating different copy numbers with mixes of plasmids containing mutated and wild-type GSA sequences. The assay was validated by analyzing transgenic alfalfa plants containing one or multiple MsGSAgr loci. HRM enabled us to clearly discriminate transgenic from non-transgenic plants, and the single copy from the multicopy state, thus providing a tool to streamline molecular plant breeding