Mycobacterium avium subsp. paratuberculosis (MAP), the agent of paratuberculosis in ruminants, is suspected to be involved in the aetiology of some human diseases. Notably, the consumption of milk ...and dairy products is considered to be the main route of human exposure to MAP because of its ability to survive during pasteurization and manufacturing processes. The aim of this study was to investigate, through a microbiological challenge test, the survival of MAP during the manufacturing and ripening period of two Italian hard cheeses, Parmigiano Reggiano and Grana Padano, made from raw bovine milk. The challenge test was performed in two different phases: the creaming phase and the manufacturing phase.
The creaming phase, which is the first step of cheese production, was reproduced in the laboratory employing raw cow's milk spiked with a MAP reference strain at a final concentration of 5.58 log10 CFU/mL. After the creaming at 18 °C and 27 °C for 12 h, a decrease of 0.80 log10 and 0.77 log10 was observed in partially skimmed milk, respectively.
In the second phase, two batches of raw cow's milk (1000 L each) were inoculated with MAP reference and wild strains, respectively. Then, the entire manufacturing process for Parmigiano Reggiano and Grana Padano, both of Protective Designation of Origin (PDO), was reproduced in an experimental cheese factory, starting from a concentration in milk of 5.19 ± 0.01 and 5.28 ± 0.08 log10 CFU/mL of MAP reference and wild strains, respectively. Heating the curd at 53 °C for 20 min did not affect MAP survival, however a significant decrease (p < 0.05) in MAP viability was observed during the moulding phase and after salting in brine, regarding the wild strains and the reference strain, respectively. In addition, a significant decrease was observed during the ripening period, at which time the MAP concentration dropped below the limit of detection from the second and the third month of ripening, for the wild and reference strains, respectively.
Taking into account the poor data availability about MAP survival in hard cheeses, this study may improve the knowledge regarding the effect of the cheese manufacturing process on the MAP dynamics, supporting also the safety of traditional raw milk hard cheeses.
•MAP survival in Parmigiano Reggiano and Grana Padano was investigated by cultural assay.•The creaming phase caused a decrease in the MAP load of 0.77–0.8 log10/mL in milk.•No viable MAP cells were detectable after two (wild strains) to three (reference strain) months of ripening.•The results support the safety of these dairy products with respect to MAP.
A 4πγ integral counting system with a NaI(Tl) well-type detector and a digital interface to acquire measurement data was implemented at LMR-CNEA. The detection efficiency as a function of the energy ...was computed by Monte Carlo simulations and the total efficiencies for ampoules and point sources were calculated considering all the decay branches. A computer code was developed to analyse data. This program reads the files generated by the digitizer module, corrects for dead time and calculates source activities and their uncertainties. Ampoules with solutions of 113Sn, 192Ir and 131I were measured.
•Implementation of a new measurement system at LMR-CNEA (Argentina).•Based on a well-type NaI(Tl) detector.•Instruments, methods and uncertainty budget are detailed.
Vitamin A has been linked to the development of allergic diseases although its role is not fully understood, Retinoic acid (RA), a metabolite of Vitamin A, has been previously associated with the ...prostaglandin pathway, and PTGDR, a receptor of PGD2, has been proposed as a candidate gene in allergy and asthma. Considering the role of PTGDR in allergy, the goal of this study was to analyze the effect of RA on the activation of the promoter region of the PTGDR gene.
A549 lung epithelial cells were transfected with 4 combinations of genetic variants of the PTGDR promoter and stimulated with all-trans RA (ATRA); luciferase assays were performed using the Dual Luciferase Reporter System, and real-time quantitative polymerase chain reaction was used to measure the expression of PTGDR, CYP26A1, RARA, RARB, RARG, and RXRA in basal A549 cell cultures and after ATRA treatment. We also performed an in silico analysis.
After ATRA treatment increased expression of CYP26A1 (12-fold) and RARB (4-fold) was detected. ATRA activated PTGDR promoter activity in transfected cells (P<.001) and RA response element sequences were identified in silico in this promoter region.
RA modulated PTGDR promoter activity. Differential response to RA and to new treatments based on PTGDR modulation could depend on genetic background in allergic asthmatic patients.
The International Atomic Energy Agency (IAEA) Network of Analytical Laboratories for the Measurement of Environmental Radioactivity (ALMERA) is a worldwide network of laboratories capable of ...providing reliable and timely analysis of radionuclides in environmental samples during normal or accidental/intentional events that result in the release of radioactivity in the environment. ALMERA is coordinated by the Environment Laboratories of the IAEA and organized into five regional groups, including the North and Latin America region (NLAR), led by the Canadian Nuclear Safety Commission. Capacity building in the NLAR for the measurements of environmental radioactivity is achieved through qualitative study that consists of experiences of ALMERA core activities and perspectives of the regional initiatives, respectively. Outcomes' analysis showed that the ALMERA core activities have contributed to the improvements of the analytical capacity and capability, and the regional initiatives assisted in the strengthening of the collaboration and networking, in the NLAR region. The immediate impact of these achievements consisted of gaining technical competence in measuring environmental radioactivity and establishing a network of laboratories. In the long term, sustained efforts between ALMERA and the regional coordination will continue upgrading national and regional competence in the radio analytical services.
Alkaline phosphatase (ALP) in lung lavage fluids is a marker of tissue damage and type II cell proliferation. Type II pneumocytes are extensively involved in fibrosis. The aim of our study was to ...verify whether local ALP activity can be held as a marker of fibrosis in chronic interstitial lung disorders. ALP was determined in the bronchoalveolar lavage of 140 subjects: 25 control subjects, and 33 patients with silicosis, 38 with idiopathic pulmonary fibrosis (IPF), and 18 with Stage I, 16 with Stage II, and 10 with Stage III/IV sarcoidosis. We observed a significant augmentation of ALP activity in patients with IPF, as well as in those with Stages II and III/IV sarcoidosis. Albumin was higher in those with Stages I and II sarcoidosis, but it was normal in the other groups. Therefore, a significant increase in ALP:albumin ratio characterized IPF (median, 10.3; range, 2.4 to 83.2) and Stage III/IV sarcoidosis, (8.9, 4.4 to 103.1) as compared with Stage I (2, 1.1 to 18) and Stage II (4.7, 0.7 to 26.6) sarcoidosis, silicosis (6.5, 0.5 to 42.7), and the nonsmoker (6.1, 2.9 to 12.9) and smoker (6.4, 2 to 15.2) control groups. We conclude that an increase of the ALP: albumin ratio in bronchoalveolar lavage fluids from patients with chronic interstitial disorders may reflect a fibrosing progression.
Background: Elicitation of broadly neutralizing antibodies remains a challenge to the development of an effective HIV-1 vaccine. While most neutralizing epitopes identified are present on the ...prefusion closed form of HIV-1 envelope trimer (Env), only limited neutralization breadth has been achieved with previous HIV-1 Env immunizations. Methods: Here we characterize the antibody response in non-human primates (NHP) following sequential immunization with HIV-1 Env variants, priming by Env with glycans removed around the CD4-binding site (CD4BS), boosting with Envs with gradually restored glycans and eventually with natively glycosylated Envs. Sera were evaluated with a sentinel panel of viruses. PBMCs of a selected group of NHPs with broad sera neutralization activity were analyzed using probe-based B cell sorting and monoclonal antibody cloning. Antibodies were selected by binding affinity to gp120 core and Env trimer. Large panel neutralization and structural studies were conducted to map major sites of antibody targeting. Results: The sequential Env-only immunization scheme elicited substantial sera neutralization activity in a subgroup of NHP and NHPs primed/boosted with deglycosylated 4501dG5 trimers elicited higher titers against the fusion peptide (FP). Isolated antibodies with gp120-binding activity had limited breadth, only neutralizing HIV-1 strains lacking glycan on Asn276. Structural studies revealed that these antibodies target the CD4BS, however, in an orientation that fails to accommodate glycan 276. In contrast, one of the Env-interactive antibodies, VRC7324, showed 34% breadth on a 208-virus panel and a median IC.sub.50 of 0.98 microg/ml. VRC7324 bound to diverse FP and competed with FP-directed antibodies. Cryo-electron microscopy was carried out to determine the structure of VRC7324-Env complex and elucidate how a single antibody can recognize different FPs to achieve broad neutralization. Conclusions: Immunization with glycan-removed Env increases immune responses. However, optimal glycan shield around the CD4BS may be essential to train developing antibodies to accommodate glycans on Env. In contrast, Env-only immunizations can elicit broadly neutralizing FP-targeting antibodies and we identified that the 4501dG5 Env can be a promising priming immunogen for eliciting the FP-targeting antibodies.
Within the framework of a smartphone-based application, helping people to identify plant species in the wild, a sub-classifier strategy has been introduced. It aims at recognizing the botanical ...properties of a leaf, relatively to various global and local shape criteria used in flora books. A decision function is applied on these classified shape categories to produce a final decision on the species of the leaf. In this paper, the fusion strategy and its corresponding Random-Forest-based sub-classifiers are described. The results of these algorithms for botanical leaf shape recognition demonstrate that our classification algorithm can achieve good performance on leaf species identification while providing the user with relevant information for educational purposes.
This work presents the experience developed by the Radioisotope Metrology Laboratory (LMR), of the Argentine National Atomic Energy Commission (CNEA), as result of the accreditation process of the ...Quality System by ISO 17025 Standard. Considering the LMR as a calibration laboratory, services of secondary activity determinations and calibration of activimeters used in Nuclear Medicine were accredited. A peer review of the
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coincidence system was also carried out. This work shows in detail the structure of the quality system, the results of the accrediting audit and gives the number of non-conformities detected and of observations made which have all been resolved.
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