22 research outputs found

    Transcriptome of pig muscle assessed by erial analysis of gene expression (SAGE)

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    Serial analysis of gene expression (SAGE) is a molecular biology technique applied to measure the global gene expression levels, characterise transcriptomes, compare the transcript levels between tissues and uncover new molecules within defined signal transduction pathways (Tutela and Tuteja, 2004)

    Substitution of a commercial diet with raw meat complemented with vegetable foods containing chickpeas or peas affects faecal microbiome in healthy dogs

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    The aim of the study was to investigate if the inclusion of chickpeas or peas in the diet can modify faecal microbiome in dogs. Eight healthy adult Border collie, fed a commercial extruded diet as reference diet (RD), were divided in two groups of four individuals. At the beginning of the trial, one group received a diet based mainly of raw meat, rice and chickpeas (CP) and in the other group this pulse was substituted with peas (PE). After 14 days, the dogs with CP diet shifted to the PE and those with PE shifted to the CP diet, for another 14 days. Faeces were col- lected at the beginning (T0), after 14 days (T14) and at the end of the study (T28). Faeces were analysed for 16S rRNA, short chain fatty acids (SCFA), lactate, pH and faecal score was also eval- uated. The SCFA and lactate in the faeces were not affected by the inclusion of pulses, with the only exception of isovalerate, which was higher in CP and PE diets in comparison with RD diet (p < .05). The abundances of Erysipelotrichaceae incertae sedis, Eubacterium, Anaerobacter and Sarcina significantly differed in CP and PE in comparison with RD. Moreover, the genera Prevotella, Lactobacillus, Alloprevotella, Suttarella varied significantly between CP and PE diets. The observed modifications of faecal microbioma were related not only to the change from RD to CP or PE, but also to the type of pulse, chickpeas or peas. However, long-term studies are required to investigate the implications that pulses can have for gut health

    Tissue reconstruction of abdominal wall with butyric acid-based nets: preliminary in vitro test using tissue engineering strategies

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    OBJECTIVE: A hernia of the abdominal wall is an opening of the muscles in the abdominal wall, which is frequently treated via the application of a surgical mesh. The purpose of this research is to study how human adipose-derived stem cells (hADSCs) interact with Phasix™ Mesh, a commercially available mesh for hernia repair. Studying how cells derived from the abdominal region behave with Phasix™ Mesh is crucial to improve the state of the art of current surgery and achieve effective tissue restoration. MATERIALS AND METHODS: hADSCs were seeded onto Phasix™ Mesh, a fully resorbable surgical mesh of poly (4-hydroxybutyric acid) (P4HB). Cell viability was assessed through MTT assay, and cell growth and adhesion were evaluated via multiple imaging techniques and gene imaging profiling. RESULTS: Results confirm that the nets support cells proliferation, extracellular matrix production and increasing of angiogenetic factor. CONCLUSIONS: Butyric acid-based nets are promising scaffolds for abdominal wall reconstruction

    The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance

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    INTRODUCTION Investment in Africa over the past year with regard to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing has led to a massive increase in the number of sequences, which, to date, exceeds 100,000 sequences generated to track the pandemic on the continent. These sequences have profoundly affected how public health officials in Africa have navigated the COVID-19 pandemic. RATIONALE We demonstrate how the first 100,000 SARS-CoV-2 sequences from Africa have helped monitor the epidemic on the continent, how genomic surveillance expanded over the course of the pandemic, and how we adapted our sequencing methods to deal with an evolving virus. Finally, we also examine how viral lineages have spread across the continent in a phylogeographic framework to gain insights into the underlying temporal and spatial transmission dynamics for several variants of concern (VOCs). RESULTS Our results indicate that the number of countries in Africa that can sequence the virus within their own borders is growing and that this is coupled with a shorter turnaround time from the time of sampling to sequence submission. Ongoing evolution necessitated the continual updating of primer sets, and, as a result, eight primer sets were designed in tandem with viral evolution and used to ensure effective sequencing of the virus. The pandemic unfolded through multiple waves of infection that were each driven by distinct genetic lineages, with B.1-like ancestral strains associated with the first pandemic wave of infections in 2020. Successive waves on the continent were fueled by different VOCs, with Alpha and Beta cocirculating in distinct spatial patterns during the second wave and Delta and Omicron affecting the whole continent during the third and fourth waves, respectively. Phylogeographic reconstruction points toward distinct differences in viral importation and exportation patterns associated with the Alpha, Beta, Delta, and Omicron variants and subvariants, when considering both Africa versus the rest of the world and viral dissemination within the continent. Our epidemiological and phylogenetic inferences therefore underscore the heterogeneous nature of the pandemic on the continent and highlight key insights and challenges, for instance, recognizing the limitations of low testing proportions. We also highlight the early warning capacity that genomic surveillance in Africa has had for the rest of the world with the detection of new lineages and variants, the most recent being the characterization of various Omicron subvariants. CONCLUSION Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve. This is important not only to help combat SARS-CoV-2 on the continent but also because it can be used as a platform to help address the many emerging and reemerging infectious disease threats in Africa. In particular, capacity building for local sequencing within countries or within the continent should be prioritized because this is generally associated with shorter turnaround times, providing the most benefit to local public health authorities tasked with pandemic response and mitigation and allowing for the fastest reaction to localized outbreaks. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

    Transcriptome of pig muscle assessed by erial analysis of gene expression (SAGE)

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    Serial analysis of gene expression (SAGE) is a molecular biology technique applied to measure the global gene expression levels, characterise transcriptomes, compare the transcript levels between tissues and uncover new molecules within defined signal transduction pathways (Tutela and Tuteja, 2004)

    Serial Analysis of Gene Expression (SAGE) in the skeletal muscle of pig.

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    Skeletal muscle growth represents one of the main economic traits in pig production. To gain a better understanding of expressions profile in pig muscle, serial analysis of gene expression (SAGE) was performed in Longissimus dorsi of two pigs at 3 and 9 months of age. A total of 53,120 long tags were obtained and sequenced from the four muscle SAGE libraries, representing 17,902 different tags, or putative transcripts, 0.64% (+0.09) of which had a relative expression level higher than 1\u2030. Overall, a total of 218 tags were highly expressed and 31 had a frequency higher than 3\u2030. Functional characterisation of the expression profiles was performed using Kyoto Encyclopedia of Genes and Genomes metabolic maps and 139 pathways were identified for swine skeletal muscle. Focal adhesion, Mitogen-Activated Protein Kinase signalling, oxidative phosphorylation, ribosomal proteins, regulation of actin cytoskeleton and insulin signalling pathways showed an abundance of genes greater than 1.5% of all the expressed transcripts. A comparison with human SAGE library indicated no statistical differences for the frequency of genes involved in these metabolic pathways

    Learning machine approach reveals microbial signatures of diet and sex in dog

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    The characterization of the microbial population of many niches of the organism, as the gastrointestinal tract, is now possible thanks to the use of high-throughput DNA sequencing technique. Several studies in the companion animals field already investigated faecal microbiome in healthy or affected subjects, although the methodologies used in the different laboratories and the limited number of animals recruited in each experiment does not allow a straight comparison among published results. In the present study, we report data collected from several in house researches carried out in healthy dogs, with the aim to seek for a variability of microbial taxa in the faeces, caused by factors such as diet and sex. The database contains 340 samples from 132 dogs, collected serially during dietary intervention studies. The procedure of samples collection, storage, DNA extraction and sequencing, bioinformatic and statistical analysis followed a standardized pipeline. Microbial profiles of faecal samples have been analyzed applying dimensional reduction discriminant analysis followed by random forest analysis to the relative abundances of genera in the feces as variables. The results supported the responsiveness of microbiota at a genera taxonomic level to dietary factor and allowed to cluster dogs according this factor with high accuracy. Also sex factor clustered dogs, with castrated males and spayed females forming a separated group in comparison to intact dogs, strengthening the hypothesis of a bidirectional interaction between microbiota and endocrine status of the host. The findings of the present analysis are promising for a better comprehension of the mechanisms that regulate the connection of the microorganisms living the gastrointestinal tract with the diet and the host. This preliminary study deserves further investigation for the identification of the factors affecting faecal microbiome in dogs

    Nanotechnology-Based Cisplatin Intracellular Delivery to Enhance Chemo-Sensitivity of Ovarian Cancer

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    Background: Platinum resistance is a major challenge in the management of ovarian cancer. Even low levels of acquired resistance at the cellular level lead to impaired response to cisplatin. In ovarian cancer intraperitoneal therapy, nanoparticle formulation can improve the cisplatin's pharmacokinetics and safety profile.Purpose: This work aimed to investigate the chemo-sensitivity of ovarian cancer SKOV3 cells upon short-term (72h) single treatment of cisplatin and cisplatin-loaded biodegradable nanoparticles (Cis-NP). The aim was then to determine the therapeutic properties of Cis-NP in vivo using a SKOV3-luc cells' xenograft model in mice.Methods: Cell cytotoxicity was assessed after the exposure of the cell culture to cisplatin or Cis-NP. The effect of treatments on EMT and CSC-like phenotype was studied by analyzing a panel of markers by flow cytometry. Intracellular platinum concentration was determined by inductively coupled plasma mass spectrometry (ICS-MS), and gene expression was evaluated by RNAseq analysis. The efficacy of intraperitoneal chemotherapy was evaluated in a SKOV3-luc cells' xenograft model in mice, through a combination of bioluminescence imaging, histological, and immunohistochemical analyses.Results: We observed in vitro that short-term treatment of cisplatin has a critical role in determining the potential induction of chemoresistance, and a nanotechnology-based drug delivery system can modulate it. The RNAseq actg 3analysis underlines a protective effect of nanoparticle system according to their ability to down-regulate several genes involved in chemoresistance, cell proliferation, and apoptosis. The highest intracellular platinum concentration obtained with Cis-NP treatment significantly improved the efficacy. Consistent with in vitro results, we found that Cis-NP treatment in vivo can significantly reduce tumor burden and aggressiveness compared to the free drug.Conclusion: Nanoparticle-mediated cisplatin delivery may serve as an intracellular depot impacting the cisplatin pharmacodynamic performance at cellular levels. These features may contribute to improving the drawbacks of conventional intraperitoneal therapy, and therefore will require further investigations in vivo
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