Quantitative Evaluation of Very Low Levels of HIV-1 Reverse Transcriptase by a Novel Highly Sensitive RT-qPCR Assay

Based on previous experience in our laboratory, we developed a real-time reverse transcriptase (RT) quantitative PCR (RT-qPCR) assay for the assessment of very low levels of HIV-1 RT activity. The RNA, acting as a template for reverse transcription into cDNA by HIV-1 RT, consisted of a synthetic RNA ad hoc generated by in vitro transcription and included a coding sequence for HSV-1 gD (gD-RNA-synt). Different conditions of variables involved in the RT-qPCR reaction, notably different amounts of gD-RNA-synt, different mixes of the reaction buffer, and different dNTP concentrations, were tested to optimize the assay. The results indicated that the gD-RNA-synt-based RT assay, in its optimized formulation, could detect a specific cDNA reverse transcription even in the presence of 1 x 10(-9) U of HIV RT. This achievement greatly improved the sensitivity of the assay over previous versions. In summary, this constructed RT-qPCR assay may be considered a promising tool for providing accurate information on very low HIV-1 RT activity

Has COVID-19 Delayed the Diagnosis and Worsened the Presentation of Type 1 Diabetes in Children?

Objective: To evaluate whether the diagnosis of pediatric type 1 diabetes or its acute complications changed during the early phase of the coronavirus disease 2019 (COVID-19) pandemic in Italy. Research design and methods: This was a cross-sectional, Web-based survey of all Italian pediatric diabetes centers to collect diabetes, diabetic ketoacidosis (DKA), and COVID-19 data in patients presenting with new-onset or established type 1 diabetes between 20 February and 14 April in 2019 and 2020. Results: Fifty-three of 68 centers (77.9%) responded. There was a 23% reduction in new diabetes cases in 2020 compared with 2019. Among those newly diagnosed patient who presented in a state of DKA, the proportion with severe DKA was 44.3% in 2020 vs. 36.1% in 2019 (P = 0.03). There were no differences in acute complications. Eight patients with asymptomatic or mild COVID-19 had laboratory-confirmed severe acute respiratory syndrome coronavirus 2. Conclusions: The COVID-19 pandemic might have altered diabetes presentation and DKA severity. Preparing for any "second wave" requires strategies to educate and reassure parents about timely emergency department attendance for non-COVID-19 symptoms

Diabetic ketoacidosis at the onset of disease during a national awareness campaign: a 2-year observational study in children aged 0-18 years

After a previous survey on the incidence of diabetic ketoacidosis (DKA) at onset of type 1 diabetes in children in 2013-2014 in Italy, we aimed to verify a possible decline in the incidence of DKA at onset during a national prevention campaign

The Silent Epidemic of Diabetic Ketoacidosis at Diagnosis of Type 1 Diabetes in Children and Adolescents in Italy During the COVID-19 Pandemic in 2020

To compare the frequency of diabetic ketoacidosis (DKA) at diagnosis of type 1 diabetes in Italy during the COVID-19 pandemic in 2020 with the frequency of DKA during 2017-2019

Species substitution in goat yoghurt supply chain using melting-curve analysis

Mislabelling of dairy products has been widely reported worldwide but there are relatively few data available to help assess species substitution in goat yogurt. The purpose of this study was to investigate the presence of bovine and sheep milk in yogurt labeled as being made exclusively from goat milk, using qualitative real-time PCR. The results confirm that fermented products are targets for food species substitution, highlighting the undeclared presence of cow and/or sheep milk in 39.6% of samples analysed. Moreover, the study shows that the qualitative real-time PCR used here is a rapid, sensitive (LODabs of 0.015 ng/mu L), specific (no primer-dimers or non-specific products in the reactions), straightforward and low-cost molecular tool for discriminating adulterants in the dairy supply chain. Therefore, melt-curve analysis is a promising tool for facilitating routine safety, quality and authenticity checks in the dairy industry, and for implementing mitigation strategies against fraudulent activities. Application of DNA-based approaches is crucial for designing an innovative food safety management system and for helping to implement both a Food Fraud Vulnerability Assessment and a Food Fraud Prevention Strategy as common practices in the dairy sector

Tuning Nursing Educational in an Italian academic context

BackgroundThe European Union Bologna Process has laid the foundation for a common European competence-based educational framework. In many countries, nursing education is in transition from vocational to higher education, with many diverse systems. The competence-based approach provided by the project Tuning Educational Structures offers a common and coherent framework able to facilitate the implementation of the principles underpinning the Bologna Process reform.ObjectivesThis study aimed to ascertain the relevance that Italian nursing university lecturers attributed to the 40 competences of the Italian version of the nursing Bachelor's and Master's Degrees. These competences were developed through adoption of the Tuning Methodology in the nursing context.SettingThe study was conducted in the 4 universities of one region of Italy which offer nursing Bachelor's and Master's Degrees.ParticipantsA total of 164 Italian university nursing lecturers.MethodsUsing a four point scale, a cross sectional survey was conducted from March 2011 to April 2012. Participants evaluated each competence according to its relevance for Bachelor's or Master's Education. Frequency analysis was conducted.ResultsThe significance for each competence of Tuning was rated very high by Italian lecturers and appeared to overlap partially with the original European study. In Italy, the most relevant competences for Bachelor's Degree were the skills associated with the use of appropriate interventions, activities and skills in nursing and the skills associated with nursing practice and clinical decision-making. For Master's Degree, leadership, management and team competences were the most important.ConclusionsThe Tuning Nursing Project was accepted by the Italian lecturers. The competence-based approach was considered by Italian lectures as a support enabling to reflect on the current Italian nursing education cycles of study and to ensure shared visions and common approaches between Italian and European lecturers

Decoding Seafood: Multi-Marker Metabarcoding for Authenticating Processed Seafood

Given the recognized nutritional value of fish and shifting consumer lifestyles, processed seafood has become increasingly prevalent, comprising a significant portion of global food production. Although current European Union labeling regulations do not require species declaration for these products, food business operators often voluntarily provide this information on ingredient lists. Next Generation Sequencing (NGS) approaches are currently the most effective methods for verifying the accuracy of species declarations on processed seafood labels. This study examined the species composition of 20 processed seafood products, each labeled as containing a single species, using two DNA metabarcoding markers targeting the mitochondrial cytochrome c oxidase I (COI) and 16S rRNA genes. The combined use of these markers revealed that the majority of the products contained multiple species. Furthermore, two products were found to be mislabeled, as the declared species were not detected. These findings underscore that NGS is a robust technique that could be adopted to support routine food industry activities and official control programs, thereby enhancing the ‘From Boat to Plate’ strategy and combating fraudulent practices in the complex fisheries supply chain

Decoding Seafood: Multi-Marker Metabarcoding for Authenticating Processed Seafood

Given the recognized nutritional value of fish and shifting consumer lifestyles, processed seafood has become increasingly prevalent, comprising a significant portion of global food production. Although current European Union labeling regulations do not require species declaration for these products, food business operators often voluntarily provide this information on ingredient lists. Next Generation Sequencing (NGS) approaches are currently the most effective methods for verifying the accuracy of species declarations on processed seafood labels. This study examined the species composition of 20 processed seafood products, each labeled as containing a single species, using two DNA metabarcoding markers targeting the mitochondrial cytochrome c oxidase I (COI) and 16S rRNA genes. The combined use of these markers revealed that the majority of the products contained multiple species. Furthermore, two products were found to be mislabeled, as the declared species were not detected. These findings underscore that NGS is a robust technique that could be adopted to support routine food industry activities and official control programs, thereby enhancing the ‘From Boat to Plate’ strategy and combating fraudulent practices in the complex fisheries supply chain

Microbiome-based study in wild-caught Scomber scombrus fish products at the end of the supply chain

Fresh fish remain the dominant seafood forms and preservation technologies have enabled them to access ever more distant markets. In this study, we used metabarcoding of the 16S rRNA gene to generate gill microbiomes from Scomber scombrus bought at fishmonger’s stores as fresh products but whose labels showed they had been harvested in the Atlantic or Mediterranean FAO fishing areas. Microbial data were analysed with the aim of evaluating their ability to maintain signals from their different geographical origins and the presence of taxa which can potentially act as spoilers, foodborne pathogens, or histamine producers. Results revealed that microbiota, at the end of the wild fish supply chain, had differences related to the two FAO fishing areas (Atlantic vs Mediterranean). Despite the presence of microbial genera potentially associated with spoilage, histamine- production or foodborne pathogens, their patterns confirmed that low-temperature storage is a traditional but effective method of preservation. However, the ongoing spoilage processes were more evident in fresh non-local specimens, dominated by psychrophilic Gram-negative bacteria, whereas fresh local specimens contained Planctomycetes taxa. Therefore, despite the current limitations mainly related to time and cost of the method, our study highlighted that microbiome-based applications are an emergent tool for food system transformation