112 research outputs found
Diagnostica molecolare per la sicurezza alimentare
No full textLe zoonosi sono infezioni che possono essere trasmesse dagli animali all'uomo. Le zoonosi di tipo alimentare vengono trasmesse attraverso alimenti contaminati e rappresentano una seria e diffusa minaccia per la salute pubblica
Lo scopo di questo progetto è lo sviluppo di test diagnostici basati sul rilevamento di sequenze di DNA specifiche per ciascun agente zoonotico. Fasi del progetto
1) Ottimizzazione del protocollo di estrazione del DNA di agenti patogeni da campioni di carne di maiale, manzo e pollo
2) Ottimizzazione dei test di PCR per l’ identificazione di agenti zoonotici quali Campylobacter, Listeria monocytogenes, Escherichia coli e Salmonella
3) Assemblaggio e ottimizzazione di kit per l'identificazione di specifici agenti zoonotici da utilizzarsi con un dispositivo di PCR portatile.
Lo strumento è composto da un termoblocco per permettere di impostare temperature costanti (in fase di estrazione) o cicli termici (durante l’amplificazione). Lo strumento di PCR portatile contiene inoltre un fluorimetro in grado di leggere l’intensità di fluorescenza prima e dopo la reazione di PCR.
L’aumento di fluorescenza è proporzionale alla quantità di agente infettivo presente nel campione.
L’analisi è effettuabile agevolmente e rapidamente in qualsiasi luogo, il dispositivo infatti può essere alimentato anche da un semplice accendisigari per automobile.The zoonotic deseases are infections that can be passed from animals to humans. The Zoonotic deseases can be transferred by eating contaminated food.
Tha aim of this project is to develop diagnostic tests based on identification of specific sequences of DNA for each pathogen. The different phases of this project are :
1)Optimization of protocol for DNA extraction from samples of pig, bovine and chicken
2)Optimization of PCR tests to identify pathogenics like Campylobacter, Listeria monocytogenes, Escherichia coli and Salmonella
3) development of a kit to identify the pathogenics.
This kit includes a portable device that allow to perform all diagnostic phases (DNA extraction, PCR and collecting fluorescence). The device includes termal cycler that allow to reach the correct temperature to perform the DNA extraction and PCR. The device also include an instrument that allow to collect fluorescence and allow to compare the fluoroscence of the sample before and after the PCR raection. The increase of fluotrecence is proprortional to concentration of pathogenic into the sample.
The analysis it’s easy and it can be perfermed outside the laboratory by everyone
Nel cantiere degli ‘Scritti’ di Alberto Mario: l’asse Carducci-Jessie White.
No full textIl saggio mette in luce la collaborazione fra Carducci e la White nella pubblicazione degli Scritti diAlberto Mario
Toward a resolution of the cosmopolitan Botryllus schlosseri species complex (Ascidiacea, Styelidae): mitogenomics and morphology of clade E (Botryllus gaiae)
No full textBotryllus schlosseri is a model colonial ascidian and a marine invader. It is currently recognized as a species complex comprising five genetically divergent clades, with clade A globally distributed and clade E found only in Europe. This taxon has also been recently redescribed by designation of a clade A specimen as the neotype. To clarify the taxonomic status of clade E and its relationship to clade A, we examine the entire mitochondrial genome and study the morphology of clade E. The mitogenome of clade E has an identical gene order to clade A, but substantially differs in the size of several non-coding regions. Remarkably, the nucleotide divergence of clade A-clade E is incompatible with the intraspecies ascidian divergence, but similar to the congeneric one and almost identical to the divergence between species once considered morphologically indistinguishable (e.g. the pair Ciona intestinalis (Linnaeus, 1767)-Ciona robusta Hoshino & Tokioka, 1967, and the pair Botrylloides niger Herdman, 1886-Botrylloides leachii (Savigny, 1816)). Clade E differs morphologically from the Botryllusschlosseri neotype mainly in the number and appearance of the stomach folds, and the shape of the anal opening, the first intestinal loop and the typhlosole. Our integrative taxonomical approach clearly distinguishes clade E as a species separate from Botryllus schlosseri, with unique morphological and molecular characters. Therefore, we here describe clade E as the new species Botryllus gaiae sp. nov
Immunotherapeutic strategies in chronic lymphocytic leukemia: advances and challenges
No full textImmune-based therapeutic strategies have drastically changed the landscape of hematological disorders, as they have introduced the concept of boosting immune responses against tumor cells. Anti-CD20 monoclonal antibodies have been the first form of immunotherapy successfully applied in the treatment of CLL, in the context of chemoimmunotherapy regimens. Since then, several immunotherapeutic approaches have been studied in CLL settings, with the aim of exploiting or eliciting anti-tumor immune responses against leukemia cells. Unfortunately, despite initial promising data, results from pilot clinical studies have not shown optimal results in terms of disease control - especially when immunotherapy was used individually - largely due to CLL-related immune dysfunctions hampering the achievement of effective anti-tumor responses. The growing understanding of the complex interactions between immune cells and the tumor cells has paved the way for the development of new combined approaches that rely on the synergism between novel agents and immunotherapy. In this review, we provide an overview of the most successful and promising immunotherapeutic modalities in CLL, including both antibody-based therapy (i.e. monoclonal antibodies, bispecific antibodies, bi- or tri- specific killer engagers) and adoptive cellular therapy (i.e. CAR T cells and NK cells). We also provide examples of successful new combination strategies and some insights on future perspectives
CAR-modified Cellular Therapies in Chronic Lymphocytic Leukemia: Is the Uphill Road Getting Less Steep?
No full textThe clinical development of chimeric antigen receptor (CAR) T-cell therapy has been more challenging for chronic lymphocytic leukemia (CLL) compared to other settings. One of the main reasons is the CLL-associated state of immune dysfunction that specifically involves patient-derived T cells. Here, we provide an overview of the clinical results obtained with CAR T-cell therapy in CLL, describing the identified immunologic reasons for the inferior efficacy. Novel CAR T-cell formulations, such as lisocabtagene maraleucel, administered alone or in combination with the Bruton tyrosine kinase inhibitor ibrutinib, are currently under investigation. These approaches are based on the rationale that improving the quality of the T-cell source and of the CAR T-cell product may deliver a more functional therapeutic weapon. Further strategies to boost the efficacy of CAR T cells should rely not only on the production of CAR T cells with an improved cellular composition but also on additional changes. Such alterations could include (1) the coadministration of immunomodulatory agents capable of counteracting CLL-related immunological alterations, (2) the design of improved CAR constructs (such as third- and fourth-generation CARs), (3) the incorporation into the manufacturing process of immunomodulatory compounds overcoming the T-cell defects, and (4) the use of allogeneic CAR T cells or alternative CAR-modified cellular vectors. These strategies may allow to develop more effective CAR-modified cellular therapies capable of counteracting the more aggressive and still incurable forms of CLL
Trade-off between sexual activities and parental care: an experimental test using handicapped mates
No full textElecting to invest in parental care is an adaptive decision thought to involve a trade-off between remating and continuing parental effort. The rock sparrow, Petronia petronia, is an unusual species in which parental investment is highly variable and both sexes may desert the brood. Males contemporaneously engage in parental care, mate guarding, and courting their current or new females. In this study we experimentally handicapped male rock sparrows during the nestling period by increasing their body mass in order to study the effects on male behaviour and the female response. Handicapped males exhibited lower sexual activity than control males but handicapped males did not reduce their offspring feeding rates. Females with a handicapped partner significantly increased the number of sexual soliciting postures towards their mates compared to females paired with control males. The females' behaviour is probably a response to the sexual behaviour change of their partners. Our results suggest that with choices involving a trade-off between mating investment and parental investment, handicapped males chose the parental investment option
The mitochondrial genome of <it>Phallusia mammillata </it>and <it>Phallusia fumigata </it>(Tunicata, Ascidiacea): high genome plasticity at intra-genus level
No full textAbstract Background Within Chordata, the subphyla Vertebrata and Cephalochordata (lancelets) are characterized by a remarkable stability of the mitochondrial (mt) genome, with constancy of gene content and almost invariant gene order, whereas the limited mitochondrial data on the subphylum Tunicata suggest frequent and extensive gene rearrangements, observed also within ascidians of the same genus. Results To confirm this evolutionary trend and to better understand the evolutionary dynamics of the mitochondrial genome in Tunicata Ascidiacea, we have sequenced and characterized the complete mt genome of two congeneric ascidian species, Phallusia mammillata and Phallusia fumigata (Phlebobranchiata, Ascidiidae). The two mtDNAs are surprisingly rearranged, both with respect to one another and relative to those of other tunicates and chordates, with gene rearrangements affecting both protein-coding and tRNA genes. The new data highlight the extraordinary variability of ascidian mt genome in base composition, tRNA secondary structure, tRNA gene content, and non-coding regions (number, size, sequence and location). Indeed, both Phallusia genomes lack the trnD gene, show loss/acquisition of DHU-arm in two tRNAs, and have a G+C content two-fold higher than other ascidians. Moreover, the mt genome of P. fumigata presents two identical copies of trnI, an extra tRNA gene with uncertain amino acid specificity, and four almost identical sequence regions. In addition, a truncated cytochrome b, lacking a C-terminal tail that commonly protrudes into the mt matrix, has been identified as a new mt feature probably shared by all tunicates. Conclusion The frequent occurrence of major gene order rearrangements in ascidians both at high taxonomic level and within the same genus makes this taxon an excellent model to study the mechanisms of gene rearrangement, and renders the mt genome an invaluable phylogenetic marker to investigate molecular biodiversity and speciation events in this largely unexplored group of basal chordates.</p
Tick-Box for 3′-End Formation of Mitochondrial Transcripts in Ixodida, Basal Chelicerates and Drosophila
No full textAccording to the tRNA punctuation model, the mitochondrial genome (mtDNA) of mammals and arthropods is transcribed as large polycistronic precursors that are maturated by endonucleolytic cleavage at tRNA borders and RNA polyadenylation. Starting from the newly sequenced mtDNA of Ixodes ricinus and using a combination of mitogenomics and transcriptional analyses, we found that in all currently-sequenced tick lineages (Prostriata, Metastriata and Argasidae) the 3′-end of the polyadenylated nad1 and rrnL transcripts does not follow the tRNA punctuation model and is located upstream of a degenerate 17-bp DNA motif. A slightly different motif is also present downstream the 3′-end of nad1 transcripts in the primitive chelicerate Limulus polyphemus and in Drosophila species, indicating the ancient origin and the evolutionary conservation of this motif in arthropods. The transcriptional analyses suggest that this motif directs the 3′-end formation of the nad1/rrnL mature RNAs, likely working as a transcription termination signal or a processing signal of precursor transcripts. Moreover, as most regulatory elements, this motif is characterized by a taxon-specific evolution. Although this signal is not exclusive of ticks, making a play on words it has been named "Tick-Box", since it is a check mark that has to be verified for the 3′-end formation of some mt transcripts, and its consensus sequence has been here carefully characterized in ticks. Indeed, in the whole mtDNA of all ticks, the Tick-Box is always present downstream of nad1 and rrnL, mainly in non-coding regions (NCRs) and occasionally within trnL(CUN). However, some metastriates present a third Tick-Box at an intriguing site - inside the small NCR located at one end of a 3.4 kb translocated region, the other end of which exhibits the nad1 Tick-Box - hinting that this motif could have been involved in metastriate gene order rearrangement
The genome assembly of the fungal pathogen <i>Pyrenochaeta lycopersici</i> from Single-Molecule Real-Time sequencing sheds new light on its biological complexity
No full textThe first draft genome sequencing of the non-model fungal pathogen Pyrenochaeta lycopersici showed an expansion of gene families associated with heterokaryon incompatibility and lacking of mating-type genes, providing insights into the genetic basis of this “imperfect” fungus which lost the ability to produce the sexual stage. However, due to the Illumina short-read technology, the draft genome was too fragmented to allow a comprehensive characterization of the genome, especially of the repetitive sequence fraction. In this work, the sequencing of another P. lycopersici isolate using long-read Single Molecule Real-Time sequencing technology was performed with the aim of obtaining a gapless genome. Indeed, a gapless genome assembly of 62.7 Mb was obtained, with a fraction of repetitive sequences representing 30% of the total bases. The gene content of the two P. lycopersici isolates was very similar, and the large difference in genome size (about 8 Mb) might be attributable to the high fraction of repetitive sequences detected for the new sequenced isolate. The role of repetitive elements, including transposable elements, in modulating virulence effectors is well established in fungal plant pathogens. Moreover, transposable elements are of fundamental importance in creating and re-modelling genes, especially in imperfect fungi. Their abundance in P. lycopersici, together with the large expansion of heterokaryon incompatibility genes in both sequenced isolates, suggest the presence of possible mechanisms alternative to gene re-assorting mediated by sexual recombination. A quite large fraction (~9%) of repetitive elements in P. lycopersici, has no homology with known classes, strengthening this hypothesis. The availability of a gapless genome of P. lycopersici allowed the in-depth analysis of its genome content, by annotating functional genes and TEs. This goal will be an important resource for shedding light on the evolution of the reproductive and pathogenic behaviour of this soilborne pathogen and the onset of a possible speciation within this species.</div
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