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1.
Food Microbiol ; 79: 123-131, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30621867

ABSTRACT

The microbiota of different types of Italian high-moisture Mozzarella cheese produced using cow or buffalo milk, acidified with natural or selected cultures, and sampled at the dairy or at the mass market, was evaluated using a Next Generation Sequencing approach, in order to identify possible drivers of the bacterial diversity. Cow Mozzarella and buffalo Mozzarella acidified with commercial cultures were dominated by Streptococcus thermophilus, while buffalo samples acidified with natural whey cultures showed similar prevalence of L. delbrueckii subsp. bulgaricus, L. helveticus and S. thermophilus. Moreover, several species of non-starter lactic acid bacteria were frequently detected. The diversity in cow Mozzarella microbiota was much higher than that of water buffalo samples. Cluster analysis clearly separated cow's cheeses from buffalo's ones, the former having a higher prevalence of psychrophilic taxa, and the latter of Lactobacillus and Streptococcus. A higher prevalence of psychrophilic species and potential spoilers was observed in samples collected at the mass retail, suggesting that longer exposures to cooling temperatures and longer production-to-consumption times could significantly affect microbiota diversity. Our results could help in detecting some kind of thermal abuse during the production or storage of mozzarella cheese.


Subject(s)
Bacteria/isolation & purification , Cheese/microbiology , Food Microbiology , Microbiota/genetics , Animals , Bacteria/classification , Bacteria/genetics , Biodiversity , Buffaloes , Cattle , Cheese/analysis , Cluster Analysis , DNA, Bacterial/genetics , Metagenomics , Milk/microbiology , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
2.
PLoS One ; 10(4): e0125259, 2015.
Article in English | MEDLINE | ID: mdl-25915065

ABSTRACT

Abnormalities of intracellular Ca2+ homeostasis and signalling as well as the down-regulation of neurotrophic factors in several areas of the central nervous system and in peripheral tissues are hallmarks of Huntington's disease (HD). As there is no therapy for this hereditary, neurodegenerative fatal disease, further effort should be made to slow the progression of neurodegeneration in patients through the definition of early therapeutic interventions. For this purpose, molecular biomarker(s) for monitoring disease onset and/or progression and response to treatment need to be identified. In the attempt to contribute to the research of peripheral candidate biomarkers in HD, we adopted a multiplex real-time PCR approach to analyse the mRNA level of targeted genes involved in the control of cellular calcium homeostasis and in neuroprotection. For this purpose we recruited a total of 110 subjects possessing the HD mutation at different clinical stages of the disease and 54 sex- and age-matched controls. This study provides evidence of reduced transcript levels of sarco-endoplasmic reticulum-associated ATP2A2 calcium pump (SERCA2) and vascular endothelial growth factor (VEGF) in peripheral blood mononuclear cells (PBMCs) of manifest and pre-manifest HD subjects. Our results provide a potentially new candidate molecular biomarker for monitoring the progression of this disease and contribute to understanding some early events that might have a role in triggering cellular dysfunctions in HD.


Subject(s)
Calcium/metabolism , Huntington Disease/diagnosis , Sarcoplasmic Reticulum Calcium-Transporting ATPases/metabolism , Vascular Endothelial Growth Factor A/metabolism , Adult , Aged , Aged, 80 and over , Biomarkers/metabolism , Disease Progression , Female , Homeostasis , Humans , Huntington Disease/genetics , Huntington Disease/metabolism , Leukocytes, Mononuclear/metabolism , Male , Middle Aged , RNA, Messenger/metabolism , Sarcoplasmic Reticulum Calcium-Transporting ATPases/genetics , Vascular Endothelial Growth Factor A/genetics
3.
Mol Cell Probes ; 24(5): 310-4, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20659551

ABSTRACT

Spinal muscular atrophy (SMA) is an autosomal recessive disease caused in about 95% of SMA patients by homozygous deletion of the survival motor neuron 1 (SMN1) gene or its conversion to the highly homologous SMN2 gene. In the majority of cases, disease severity correlates inversely with increased SMN2 copy number. Because of the comparatively high incidence of healthy carriers and severity of the disease, detection of sequence alterations and quantification of SMN1 and SMN2 copy numbers are essential for exact diagnosis and genetic counselling. Several assays have been developed for this purpose. Multiplex ligation-dependent probe amplification (MLPA) is a versatile technique for relative quantification of different nucleic acid sequences in a single reaction. Here, we establish a quick MLPA-based assay for the detection of SMN1 and SMN2 copy numbers with high specificity and low complexity.


Subject(s)
Gene Dosage , Nucleic Acid Amplification Techniques/methods , Survival of Motor Neuron 1 Protein/genetics , DNA/genetics , DNA Probes , Female , Genetic Carrier Screening/methods , Genetic Testing , Heterozygote , Humans , Male , Muscular Atrophy, Spinal/diagnosis , Muscular Atrophy, Spinal/genetics , Reproducibility of Results , Sensitivity and Specificity , Survival of Motor Neuron 2 Protein/genetics
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