In vitro and in vivo expression of foreign genes by transmissible gastroenteritis coronavirus-derived minigenomes

A helper-dependent expression system based on transmissible gastroenteritis coronavirus (TGEV) has been developed using a minigenome of 3·9 kb (M39). Expression of the reporter gene {beta}-glucuronidase (GUS) (2–8 µg per 106 cells) and the porcine respiratory and reproductive syndrome virus (PRRSV) ORF5 (1–2 µg per 106 cells) has been shown using a TGEV-derived minigenome. GUS expression levels increased about eightfold with the m.o.i. and were maintained for more than eight passages in cell culture. Nevertheless, instability of the GUS and ORF5 subgenomic mRNAs was observed from passages five and four, respectively. About a quarter of the cells in culture expressing the helper virus also produced the reporter gene as determined by studying GUS mRNA production by in situ hybridization or immunodetection to visualize the protein synthesized. Expression of GUS was detected in the lungs, but not in the gut, of swine immunized with the virus vector. Around a quarter of lung cells showing replication of the helper virus were also positive for the reporter gene. Interestingly, strong humoral immune responses to both GUS and PRRSV ORF5 were induced in swine with this virus vector. The large cloning capacity and the tissue specificity of the TGEV-derived minigenomes suggest that these virus vectors are very promising for vaccine development

Feasibility of using teleradiology to improve tuberculosis screening and case management in a district hospital in Malawi.

Malawi has one of the world's highest rates of human immunodeficiency virus (HIV) infection (10.6%), and southern Malawi, where Thyolo district is located, bears the highest burden in the country (14.5%). Tuberculosis, common among HIV-infected people, requires radiologic diagnosis, yet Malawi has no radiologists in public service. This hinders rapid and accurate diagnosis and increases morbidity and mortality

Component-resolved diagnosis of pollen allergy based on skin testing with profilin, polcalcin and lipid transfer protein pan-allergens

BACKGROUND Allergy diagnosis needs to be improved in patients suffering from pollen polysensitization due to the existence of possible confounding factors in this type of patients. OBJECTIVE To evaluate new diagnostic strategies by comparing skin responses to pan-allergens and conventional allergenic extracts with specific IgE (sIgE) to purified allergen molecules. METHODS One thousand three hundred and twenty-nine pollen-allergic patients were diagnosed by a combination of an in vitro method with a panel of 13 purified allergens, including major allergens and pan-allergens, using a high-capacity screening technology (ADVIA-Centaur®) and skin prick test (SPT) to pan-allergens and conventional extracts. RESULTS There was a high concordance (κ index) between in vitro (sIgE to major allergens) and in vivo (SPT to conventional extracts) methods in patients who were not sensitized to pan-allergens, but SPT with conventional extracts failed to diagnose patients with sensitization to pan-allergens. In patients who were simultaneously sensitized to polcalcins and profilins, there was a duplication both in the number of sensitizations to major allergens and in the years of disease evolution. There was a statistical association between sensitization to profilins and/or lipid transfer proteins and food allergy (P<0.0001). CONCLUSION The novel diagnostic strategy has proven to be a valuable tool in daily clinical practice. Introduction of routine SPT to pan-allergens is a simple and feasible way of improving diagnostic efficacy. Patients sensitized to pan-allergens should be tested by an adequate panel of allergenic molecules in order to identify the allergens that are responsible for the allergic disease

Component-Resolved in Vitro Diagnosis in Peach-Allergic Patients

BACKGROUND: The in vitro diagnosis of pollen-related food allergy presents low specifi city and reproducibility with many conventional extracts. This can be improved using natural purifi ed allergens, recombinant purifi ed allergens, or both. OBJECTIVE: We compared specifi c immunoglobulin (Ig) E determination (sIgE), the basophil activation test (BAT), the histamine release test (HRT), and the cellular allergen stimulation test (CAST) using natural and recombinant allergens in the diagnosis of peach allergy. METHODS: Thirty-two peach allergic patients were studied. Skin prick tests were performed with commercial peach and extract with Mal d 1, nPru p 3, and profi lin (nPho d 2). sIgE, BAT, CAST, and HRT were determined using rPru p 3, rMal d 3, rBet v 1, rMal d 1, and rMal d 4. RESULTS: Agreement between the techniques was good with all the allergens, except HRT with rMal d 1 and rMal d 4. With rPru p 3, sIgE, CAST, BAT, and HRT showed sensitivity values of 88%, 81%, 72%, and 69% and specifi city values of 100%, 93%, 97%, and 83%, respectively. In patients with systemic symptoms or contact urticaria, the values were 100%, 85%, 81%, and 81%. In patients with oral allergy syndrome, sensitivity to profi lins or homologues of Bet v 1 was detected in 100% of the cases by all the techniques, except by HRT with rMal d 1, which detected 66% of the cases. CONCLUSIONS: The use of single allergens in the in vitro diagnosis of peach allergy by specifi c IgE determination, BAT, and CAST offers high specifi city and sensitivity, with better results than the HRT

Introducció

Dins la secció: "Dossier : Món rural i paisatge a l'època roman

Subdiffusive axial transport of granular materials in a long drum mixer

Granular mixtures rapidly segregate radially by size when tumbled in a partially filled horizontal drum. The smaller component moves toward the axis of rotation and forms a buried core, which then splits into axial bands. Models have generally assumed that the axial segregation is opposed by diffusion. Using narrow pulses of the smaller component as initial conditions, we have characterized axial transport in the core. We find that the axial advance of the segregated core is well described by a self-similar concentration profile whose width scales as $t^\alpha$, with $\alpha \sim 0.3 < 1/2$. Thus, the process is subdiffusive rather than diffusive as previously assumed. We find that $\alpha$ is nearly independent of the grain type and drum rotation rate within the smoothly streaming regime. We compare our results to two one-dimensional PDE models which contain self-similarity and subdiffusion; a linear fractional diffusion model and the nonlinear porous medium equation.Comment: 4 pages, 4 figures, 1 table. Submitted to Phys Rev Lett. For more info, see http://www.physics.utoronto.ca/nonlinear