Challenge Test Improvement: Analytical Costs and Time Optimization

The study’s aim was to develop a quantitative risk assessment model of Listeria monocytogenes in liver chicken paté. The model was performed using the Integrated Challenge Test (Italian Journal of Food Safety, Vol. 1 N. 6 2012) with the objective to reduce the analytical cost and time. The challenge test was carried out on 3 different batches stored at 12°C and inoculated with a mix of Listeria monocytogenes strains. Lactobacillus spp. and Listeria monocytogenes plate counts were performed daily on each sample until the stationary phase was reached by both populations. The challenge test results at 12°C were input in the Combase DMfit software to determine the growth parameters of Listeria monocytogenes and lactic flora which showed mutual interaction. Then, using the Combase Predictor for Listeria monocytogenes and the FSSP (Food Spoilage and Safety Predictor) software for lactic flora, the growth parameters were extrapolated at 4°C and 8°C. The growth parameters of both populations at 4°C, 8°C and 12°C were then used to apply the model in order to predict the maximum daily concentration of Listeria monocytogenes. Model results were assessed against the results of an additional challenge test conducted with the same strain mix inoculum in 3 different batches stored for 4 days at 4°C, 4 days at 8°C and then 4 days at 12°C. The proposed model represents a reliable quantitative risk evaluation which provides realistic results with limited cost

The ability to learn new written words is modulated by language orthographic consistency

Introduction It is well known that a difficulty in forming lexical representations is a strong predictor of reading and spelling difficulties even after controlling for the effects of other cognitive skills. Our study had two main interrelated aims. First, we wanted to examine whether the ability to learn new written words (lexical learning) varies as a function of the orthographic consistency of the language of the learner. Second, we wanted to evaluate the cognitive abilities involved in orthographic lexical learning and whether they differed as a function of language consistency. Method 163 Italian children and 128 English children performed a lexical learning task as well as tasks assessing several cognitive skills potentially related to the ability to establish orthographic representations. Results We found that children learning an orthographic inconsistent orthography (English) were better able to learn novel written words presented in association with pictures than children learning a consistent orthography (Italian). This was true for both younger and older primary school children and also when children were matched for school grade. Lexical learning may be better in English children because the many irregularities of this language promote storing in memory whole-word representations and processing larger orthographic units. In Italian, instead, reading can be accomplished successfully on the basis of grapheme-phoneme conversion rules and on processing smaller orthographic units. This interpretation was supported by the pattern of cognitive skills associated with lexical learning skills in the two languages. Variations in lexical learning were explained by spatial visual memory and phonological awareness tasks in both languages, but phonological STM explained further variance in Italian, while a task tapping visuo-attentional capacity explained further variance in English. Conclusion Learning a language with inconsistent orthography is associated with better lexical learning skills in children at different stages of primary school; the pattern of cognitive skills associated with lexical learning skills is also partially modulated by orthographic consistency

Finding the junction between claudins and endometrial carcinoma

: Endometrial carcinoma (EC) defines a heterogeneous group of neoplastic diseases originating from the transformation of endometrial cells that constitute the internal lining of the uterus. To date several molecular targets have been analysed to describe the natural course of the disease, claudins being among these. Claudins are the main components of tight junctions (TJs), and their main functions are ascribed to the compartmentalization of tissues and cell-cell communication by means of intracellular ions diffusion: these features are typical of epithelial cells. Their overexpression, mis-localization or loss contribute to the malignancy of EC cells. This review collected all available data regarding the expression, regulation and claudin-related signaling pathways to provide a comprehensive view on the influence of claudin in EC progression. Further, the translational potential of claudin differential expression was explored, indicating that their role in personalized medicine could also contribute to EC therapy besides their employment for diagnosis and prognosis

Simple and efficient approach for shelf-life test on frozen spinach and parsley

open6noA simple test for shelf-life assessment of frozen spinach and parsley is presented. A specific shelf-life test that considers three storage temperatures is proposed to accelerate the rate of quality decay in frozen spinach and parsley. The scope was to provide a reliable and rapid way (one month vs years) to predict shelf-life by using a simple experimental approach and mathematical models based on some physical quality product attributes. Physical properties were evaluated at three storage temperatures: -5 degrees C, -10 degrees C and -26 degrees C, to simulate a possible thermal abuse. Mechanical and thermal indexes were defined measuring maximum compression force (N) and latent heat involved in ice melting (J/g). A zeroorder kinetic model was used to properly fit experimental data and thus to obtain related reaction rates. The determination coefficient indicates that there is a strong linear relation between kinetic parameters at -10 degrees C or -5 degrees C and -26 degrees C. This suggests a reliable procedure for shelf-life estimation, carrying out a test at -10 degrees C or -5 degrees C for one month and extending values to data acquired at 26 degrees C for the same period of time. The relations obtained from this research have led to a simple practical approach: one day at -10 degrees C could be considered roughly equivalent to 30 days at -26 degrees C. Accordingly, it could be possible to obtain a shelf-life estimation in short time, also considering other similar products.openIaccheri, Eleonora; Cevoli, Chiara; Romani, Santina; Dalla Rosa, Marco; Molari, Giovanni; Fabbri, AngeloIaccheri, Eleonora; Cevoli, Chiara; Romani, Santina; Dalla Rosa, Marco; Molari, Giovanni; Fabbri, Angel

Stability of circulating miRNA in saliva: The influence of sample associated pre-analytical variables

Background and aims: Increasing evidence supports the practicability of salivary cell-free (cf) miRNA as liquid biopsy markers in cancers. Its successful translation in the clinical setting requires reproducible approaches for saliva manipulation, in order to control for pre-analytical variables influencing miRNA stability. This study aims to define the optimal conditions to maintain the integrity of saliva during collection, transport and processing with respect to cf-miRNA quantification. Materials and methods: Saliva was collected from 20 healthy subjects and 8 oral cancer patients. Two sampling methods were tested and different storage temperatures and times were evaluated. Salivary expression level of target miRNAs was quantified by qPCR. Comparison between group mean values at specific conditions were performed using paired t-tests. Agreement between measurements was evaluated using a Bland-Altman plot. Results: Different collection methods revealed comparable levels of salivary miR-484 and miR-106b-5p in both subject cohorts. MiRNAs were stable for up to 48 h at 4 °C in saliva supernatant, showing significant alteration after 96 h. Mid-term storage of supernatant at -20 °C decreased miRNA stability significantly compared to standard -80 °C. Conclusions: Cf-miRNA in saliva were slightly altered by collection methods and storage conditions, both in healthy and in pathological contexts, and remained stable for a period of time compatible with main clinical routine needs

Health Effects of Phenolic Compounds Found in Extra-Virgin Olive Oil, By-Products, and Leaf of Olea europaea L.

Olea europaea L. fruit is a peculiar vegetal matrix containing high levels of fatty acids (98-99% of the total weight of extra-virgin olive oil, EVOO) and low quantities (1-2%) of phenolics, phytosterols, tocopherols, and squalene. Among these minor components, phenolics are relevant molecules for human health. This review is focused on their beneficial activity, in particular of hydroxytyrosol (HT), oleuropein (OLE), oleocanthal (OLC), and lignans found in EVOO, olive oil by-products and leaves. Specifically, the cardioprotective properties of the Mediterranean diet (MD) related to olive oil consumption, and the biological activities of polyphenols recovered from olive oil by-products and leaves were described. Recent European projects such as EPIC (European Prospective Investigation into Cancer and Nutrition) and EPICOR (long-term follow-up of antithrombotic management patterns in acute coronary syndrome patients) have demonstrated the functional and preventive activities of EVOO showing the relation both between cancer and nutrition and between consumption of EVOO, vegetables, and fruit and the incidence of coronary heart disease. The data reported in this review demonstrate that EVOO, one of the pillars of the MD, is the main product of Olea europaea L. fruits; leaves and by-products are secondary but precious products from which bioactive compounds can be recovered by green technologies and reused for food, agronomic, nutraceutical, and biomedical applications according to the circular economy strategy

Oleuropein, the Main Polyphenol of Olea europaea Leaf Extract, Has an Anti-Cancer Effect on Human BRAF Melanoma Cells and Potentiates the Cytotoxicity of Current Chemotherapies

Oleuropein (Ole), a secoiridoid glucoside present in Olea europaea leaves, gained scientific interest thanks to its several biological properties, including the anticancer one. We verified whether Ole might potentiate the cytotoxicity of conventional drugs used to treat melanoma, disclosing a potentially new therapeutic strategy. We tested the cytotoxic action of Ole alone or in combination with chemotherapeutics on A375 human melanoma cells. We found that Ole was able, at a dose of 500 µM, to stimulate apoptosis, while at a non-toxic dose of 250 µM, it affected cell proliferation and induced the downregulation of the pAKT/pS6 pathway. A dose of 250 µM Ole did not potentiate the effect of Vemurafenib (PLX4032), but it succeeded in increasing the cytotoxic effect of Dacarbazine (DTIC). The major effect was found in the association between Ole and Everolimus (RAD001), also on PLX4032-resistant BRAF melanoma cells, which possibly cooperate in the inhibition of the pAKT/pS6 pathway. Of interest, an olive leaf extract enriched in equimolar Ole was more effective and able to further improve DTIC and RAD001 efficacy on BRAF melanoma cells with respect to Ole alone. Therefore, Ole represents a natural product able to potentiate a wide array of chemotherapeutics against BRAF melanoma cells affecting the pAKT/pS6 pathway

Identification of stably expressed reference small non-coding RNAs for microRNA quantification in high-grade serous ovarian carcinoma tissues

MicroRNAs (miRNAs) belong to a family of small non‐coding RNAs (sncRNAs) playing important roles in human carcinogenesis. Multiple investigations reported miRNAs aberrantly expressed in several cancers, including high‐grade serous ovarian carcinoma (HGS‐OvCa). Quantitative PCR is widely used in studies investigating miRNA expression and the identification of reliable endogenous controls is crucial for proper data normalization. In this study, we aimed to experimentally identify the most stable reference sncRNAs for normalization of miRNA qPCR expression data in HGS‐OvCa. Eleven putative reference sncRNAs for normalization (U6, SNORD48, miR‐92a‐3p, let‐7a‐5p, SNORD61, SNORD72, SNORD68, miR‐103a‐3p, miR‐423‐3p, miR‐191‐5p, miR‐16‐5p) were analysed on a total of 75 HGS‐OvCa and 30 normal tissues, using a highly specific qPCR. Both the normal tissues considered to initiate HGS‐OvCa malignant transformation, namely ovary and fallopian tube epithelia, were included in our study. Stability of candidate endogenous controls was evaluated using an equivalence test and validated by geNorm and NormFinder algorithms. Combining results from the three different statistical approaches, SNORD48 emerged as stably and equivalently expressed between malignant and normal tissues. Among malignant samples, considering groups based on residual tumour, miR‐191‐5p was identified as the most equivalent sncRNA. On the basis of our results, we support the use of SNORD48 as best reference sncRNA for relative quantification in miRNA expression studies between HGS‐OvCa and normal controls, including the first time both the normal tissues supposed to be HGS‐OvCa progenitors. In addition, we recommend miR‐191‐5p as best reference sncRNA in miRNA expression studies with prognostic intent on HGS‐OvCa tissues

Characterization of reading errors in languages with different orthographic regularity: an Italian–English comparison

The study examined whether a classification of errors based on Hendriks and Kolk’s (1997) proposal would effectively characterize the reading profile of children learning two orthographies varying for regularity, such as Italian and English. The study considered both an age-match and a grade-match comparison. Offline analysis of error production was carried out for two lists of stimuli: List 1 including regular words varying for frequency and matched non-words and List 2 including low-frequency words varying for regularity. In List 1, Italian-reading children made more multiple attempts characterized by a slow and progressive approach to the target (sounding-out behavior) than English-reading children, while the latter made relatively more word substitutions and non-word lexicalizations. As for List 2, Italian-reading children made relatively more multiple attempts and progressive approaches to the target compared to the English-reading children (with more sounding-out behaviors and syllabications), while the opposite occurred for phonological-visual errors, word substitutions, morphological, and semantic errors. Both groups showed a high proportion of phonological-visual and regularization errors (stress assignment in the case of Italian-reading children). Overall, the use of an error coding system specifically tuned to the characteristics of the orthographies investigated allowed a more comprehensive identification of reading difficulties which allowed the different strategies used by children of different languages to emerge more clearly (more reliance on sub-lexical routines in Italian readers and on lexical routines in English readers). These results call for more attention to error patterns in the identification of reading difficulties in children of different languages including those learning a transparent orthography where error analyses have largely been ignored