Hygienic quality of dehydrated aromatic herbs marketed in Southern Portugal

Dehydrated aromatic herbs are highly valued ingredients, widely used at home level and by food processing industry, frequently added to a great number of recipes in the Mediterranean countries. Despite being considered low-moisture products and classified as GRAS, during pre and post-harvesting stages of production they are susceptible of microbial contamination. In Europe an increasing number of food recalls and disease outbreaks associated with dehydrated herbs have been reported in recent years. In this study the microbial quality of 99 samples of aromatic herbs (bay leaves, basil, coriander, oregano, parsley, Provence herbs, rosemary and thyme) collected from retails shops in the region of Algarve (Southern Portugal) was assessed. All the samples were tested by conventional methods and were assayed for the total count of aerobic mesophilic microorganisms, Salmonella spp., Escherichia coli, coagulase-positive staphylococci and filamentous fungi. Almost 50 % of the herbs did not exceed the aerobic mesophilic level of 104 CFU/g. The fungi count regarded as unacceptable (106 CFU/g) was not found in any of the tested herbs, while 84 % of the samples ranged from ≤102 to 104 CFU/g. No sample was positive for the presence of Salmonella spp., Escherichia coli and staphylococci. The results are in compliance with the European Commission criteria although they point out to the permanent need of surveillance on the good standards of handling/cooking practices as well as the importance of avoiding contamination at production, retailing and distribution. The microbiological hazards associated with the pathogenic and toxigenic microbiota of dried herbs remain as a relevant public health issue, due to the fact that they are added to foods not submitted to any following lethal procedure. Control measures should be adopted in order to ensure that all phases of their supply chain respect the food safety standards.FCT: UID/BIA/04325/2019.info:eu-repo/semantics/publishedVersio

A mouse mammary tumor virus env-like exogenous sequence is strictly related to progression of human sporadic breast carcinoma

A viral etiology of human breast cancer (HBC) has been postulated for decades since the identification of mouse mammary tumor virus (MMTV). The detection of MMTV env-like exogenous sequences (MMTVels) in 30% to 40% of invasive HBCs increased attention to this hypothesis. Looking for MMTVels during cancer progression may contribute to a better understanding of their role in HBC. Herein, we analyzed HBC preinvasive lesions for the presence of MMTVels. Samples were obtained by laser microdissection of FFPE tissues: 20 usual-type ductal hyperplasias, 22 atypical ductal hyperplasias (ADHs), 49 ductal carcinomas in situ (DCISs), 20 infiltrating ductal carcinomas (IDCs), and 26 normal epithelial cells collateral to a DCIS or an IDC. Controls included reductive mammoplastic tissue, thyroid and colon carcinoma, and blood samples from healthy donors. MMTVels were detected by fluorescence-nested PCR. DNA samples from the tissues of nine patients were analyzed by real-time quantitative PCR, revealing a different viral load correlated with stage of progression. Furthermore, as never previously described, the presence of MMTVels was investigated by chromogenic in situ hybridization. MMTVels were found in 19% of normal epithelial cells collateral to a DCIS or an IDC, 27% of ADHs, 82% of DCISs, and 35% of IDCs. No MMTVels were found in the control samples. Quantitative PCR and chromogenic in situ hybridization confirmed these results. These data could contribute to our understanding of the role of MMTVels in HBC. (Am J Pathol 2011, 179:2083-2090; DOI: 10.1016/j.ajpath.2011.06.046

Copy number variations residing outside the SHOX enhancer region are involved in Short Stature and Léri-Weill dyschondrosteosis

Background: SHOX enhancer CNVs, affecting one or more of the seven recognized evolutionary conserved non-coding elements (CNEs) represent one of the most frequent cause of SHOX-haploinsufficiency. During the diagnostic workflow deletions/duplications have been identified downstream SHOX not including any of the these CNEs. Methods: Fine tiling aCGH and breakpoint PCR were used to characterize the critical interval and to search for novel alterations in a cohort of selected patients. Results: Screening of 252 controls provided evidence that duplications in this area represent likely benign variants whereas none of the deletions were detected. These findings suggested that other alterations relevant for SHOX-haploinsufficiency might be missed by the standard diagnostic methods. To identify such undisclosed elements, the aCGH was used to reanalyze 52 unresolved cases with clinical features strongly suggestive of SHOX-haploinsufficiency. This analysis followed by the screening of 210 patients detected two partially overlapping small deletions of ~12 and ~8 kb in four unrelated individuals, approximately 15 kb downstream SHOX, that were absent in 720 normal stature individuals. Conclusion: Our results strengthen the hypothesis that alterations of yet unidentified cis-regulatory elements residing outside those investigated through conventional methods, might explain the phenotype in ISS/LWD patients thus enlarging the spectrum of variants contributing to SHOX-haploinsufficiency

Regulatory T Cell Extracellular Vesicles Modify T-Effector Cell Cytokine Production and Protect Against Human Skin Allograft Damage

Regulatory T cells (Tregs) are a subpopulation of CD4⁺ T cells with a fundamental role in maintaining immune homeostasis and inhibiting unwanted immune responses using several different mechanisms. Recently, the intercellular transfer of molecules between Tregs and their target cells has been shown via trogocytosis and the release of small extracellular vesicles (sEVs). In this study, CD4⁺CD25⁺CD127ˡᵒ human Tregs were found to produce sEVs capable of inhibiting the proliferation of effector T cells (Teffs) in a dose dependent manner. These vesicles also modified the cytokine profile of Teffs leading to an increase in the production of IL-4 and IL-10 whilst simultaneously decreasing the levels of IL-6, IL-2, and IFNγ. MicroRNAs found enriched in the Treg EVs were indirectly linked to the changes in the cytokine profile observed. In a humanized mouse skin transplant model, human Treg derived EVs inhibited alloimmune-mediated skin tissue damage by limiting immune cell infiltration. Taken together, Treg sEVs may represent an exciting cell-free therapy to promote transplant survival

Human saliva as route of inter-human infection for mouse mammary tumor virus

Etiology of human breast cancer is unknown, whereas the Mouse Mammary Tumor Virus (MMTV) is recognized as the etiologic agent of mouse mammary carcinoma. Moreover, this experimental model contributed substantially to our understanding of many biological aspects of the human disease. Several data strongly suggest a causative role of MMTV in humans, such as the presence of viral sequences in a high percentage of infiltrating breast carcinoma and in its preinvasive lesions, the production of viral particles in primary cultures of breast cancer, the ability of the virus to infect cells in culture. This paper demonstrates that MMTV is present in human saliva and salivary glands. MMTV presence was investigated by fluorescent PCR, RT-PCR, FISH, immunohistochemistry, and whole transcriptome analysis. Saliva was obtained from newborns, children, adults, and breast cancer patients. The saliva of newborns is MMTV-free, whereas MMTV is present in saliva of children (26.66%), healthy adults (10.60%), and breast cancer patients (57.14% as DNA and 33.9% as RNA). MMTV is also present in 8.10% of salivary glands. RNA-seq analysis performed on saliva of a breast cancer patient demonstrates a high expression of MMTV RNA in comparison to negative controls. The possibility of a contamination by murine DNA was excluded by murine mtDNA and IAP LTR PCR. These findings confirm the presence of MMTV in humans, strongly suggest saliva as route in inter-human infection, and support the hypothesis of a viral origin for human breast carcinoma

Identification of RNF168 as a PML nuclear body regulator

Promyelocytic leukemia (PML) protein forms the basis of PML nuclear bodies (PML NBs), which control many important processes. We have screened an shRNA library targeting ubiquitin pathway proteins for effects on PML NBs, and identified RNF8 and RNF168 DNA-damage response proteins as negative regulators of PML NBs. Additional studies confirmed that depletion of either RNF8 or RNF168 increased the levels of PML NBs and proteins, whereas overexpression induced loss of PML NBs. RNF168 partially localized to PML NBs through its UMI/MIU1 ubiquitin-interacting region and associated with NBs formed by any PML isoform. The association of RNF168 with PML NBs resulted in increased ubiquitylation and SUMO2 modification of PML. In addition, RNF168 was found to associate with proteins modified by SUMO2 and/or SUMO3 in a manner dependent on its ubiquitin-binding sequences, suggesting that hybrid SUMO–ubiquitin chains can be bound. In vitro assays confirmed that RNF168, preferentially, binds hybrid SUMO2–K63 ubiquitin chains compared with K63–ubiquitin chains or individual SUMO2. Our study identified previously unrecognized roles for RNF8 and RNF168 in the regulation of PML, and a so far unknown preference of RNF168 for hybrid SUMO–ubiquitin chains

Mycoflora and Ochratoxin A Control in Wheat Grain Using Natural Extracts Obtained from Wine Industry By-Products

The aim of this study was to evaluate the effect of some natural extracts obtained from grape pomace (GPE) and grape seeds (GSE) as compared to a synthetic food, antioxidant-butylated hydroxytoluene (BHT), in order to control fungal population and ochratoxin A (OTA) production in naturally contaminated wheat. The results showed that the addition of these extracts was efficient with OTA content decreasing. On treatment with these extracts the loss of OTA content after 14 days was in the range 7.8–28.3% relative to the control sample, but increased up to 26.48–37% after 28 days. The highest loss in OTA content was recorded for treatment with GPE at the 500 ppm level. Regarding the fungal development, the obtained results show that the total fungal populations were significantly reduced by using natural extracts. The most efficient extract was GPE. Both BHT and GPE inhibit the growth of Penicillium verrrucosum, for all doses, but less with Aspergillus genera. GPE affects the growth of other types of moulds such as Rhizopus microsporus, Fusarium graminearum, Alternaria infectoria and Cladosporium herbarum. Our data suggested that GPE and GSE are able to provide fungicidal and fungistatic protection and to control the OTA accumulation in wheat, at least in a similar manner to BHT

End-digits preference for self-reported height depends on language

BACKGROUND: When individuals report figures, they often prefer to round to specific end-digits (e.g. zero). Such preference has been found in reports of body weight, cigarette consumption or blood pressure measurements. Very little is known about self-reported body height. End-digit preference can distort estimates of prevalence and other statistical parameters. This study examines end-digit preference for self-reported height and how it relates with sex, age, educational level or cultural affiliation. METHODS: We analysed reports of height of 47,192 individuals (aged 15 years or older) living in Switzerland and participating in one of the three population-based Swiss Health Surveys carried out in 1992/93, 1997 and 2002 respectively. Digit preferences were analysed by sex, age group, educational level, survey, smoking status, interview language (only for Swiss nationals) and nationality. Adjusted odds ratios (OR) with 95% confidence interval were calculated by using multivariate logistic regression. RESULTS: Italian and French nationals (44.1% and 40.6%) and Italian and French Swiss (39.6% and 35.3%) more strongly preferred zero and five than Germans and German Swiss (29.2% and 30.3%). Two, four, six and eight were more popular in Germans and German Swiss (both 44.4%). Compared to German Swiss (OR = 1), for the end-digits zero and five, the OR were 1.50 (1.38-1.63) for Italian Swiss and 1.24 (1.18-1.30) for French Swiss; 1.73 (1.58-1.89) for Italian nationals and 1.61 (1.33-1.95) for French nationals. The end-digits two, four, six and eight showed an opposite pattern. CONCLUSION: Different preferences for end-digits depending on language and nationality could be observed consistently in all three national health surveys. The patterns were strikingly similar in Swiss and foreign nationals speaking the same language, suggesting that preferences were specific to language rather than to nationality. Taking into account rounding preferences could allow more valid comparisons in analyses of self-reported data originating from different cultures

Half-Time Strategies to Enhance Second-Half Performance in Team-Sports Players: A Review and Recommendations

The competitive demands of numerous intermittent team sports require that two consecutive periods of play are separated by a half-time break. Typically, half-time allows players to: return to the changing rooms, temporarily relax from the cognitive demands of the first half of match-play, rehydrate, re-fuel, attend to injury or equipment concerns, and to receive tactical instruction and coach feedback in preparation for the second half. These passive practices have been associated with physiological changes which impair physical and cognitive performance in the initial stages of the second half. An increased risk of injury has also been observed following half-time. On the day of competition, modification of half-time practices may therefore provide Sports Scientists and Strength and Conditioning Coaches with an opportunity to optimise second half performance. An overview of strategies that may benefit team sports athletes is presented; specifically, the efficacy of: heat maintenance strategies (including passive and active methods), hormonal priming (through video feedback), post-activation potentiation, and modified hydro-nutritional practices are discussed. A theoretical model of applying these strategies in a manner that compliments current practice is also presented