PCR-based procedures in detection and DNA-fingerprinting of Salmonella from samples of animal origin

The usefulness of selected PCR-protocols for the detection of Salmonella in 117 samples of animal origin (17 raw minced meat, 27 raw chicken meat, 8 raw sausages, and 25 egg samples, as well as 18 poultry faecal, and caecal swabs samples) and DNA-fingerprinting typing is shown. To establish an accurate PCR-procedure for Salmonella detection the following parameters were evaluated: two pre-PCR concentration procedures, centrifugation and immunomagnetic separation (IMS) using Dynabeads anti- Salmonella; the specificity and sensitivity of 10 sets of primers; and different conditions of the amplification reaction. In light of the results obtained from the use of PCR-based procedures alone or in combination with conventional methods, the following findings can be underlined: First, IMS is more efficient than centrifugation in the recovery of Salmonella. Second, the selected IMS/PCR-detection protocol is less time-consuming (45 h) than the IMS/culture procedure (90 h), and a good concordance between them was found when the Kappa coefficient was calculated (0·87). Third, PCR-ribotyping technique showed a very low discrimination power, being able to differentiate only three profiles. Fourth, RAPD technique using specific primers supports previous works in which it was proposed as a simple and useful tool for discriminating isolates between and within serotypes. Fifth, The efficiency, rapidity, and flexibility of the PCR-protocols applied were high, and they can be performed using two PCR-programs and the same basic equipment.     

Minors held by majors: the H13 minor histocompatibility locus defined as a peptide/MHC class I complex

The products of minor histocompatibility (H) loci are serious barriers to tissue transplantation even among major histocompatibility complex (MHC) identical individuals, frequently causing chronic graft rejection and graft versus host disease. Over 50 minor H loci map to mouse autosomal chromosomes but none are known at the molecular level. By expression cloning, we identified the H13 locus, a classical minor H locus first detected 30 years ago by the trait of graft rejection. The H13a allele is located on chromosome 2 and encodes a novel protein that yields the rare naturally processed nonapeptide SSVVGVWYL (SVL9) for presentation by the Db MHC class I molecule. The SVL9 peptide binds Db MHC despite the absence of the consensus binding motif, and a conservative methyl group substitution (Valine 4 <--> Isoleucine) explains why reciprocal T cell responses are elicited in H13a and H13b congenic strains.     

Mechanisms of resistance in a human cell line exposed to sequential topoisomerase poisoning

Camptothecins are a new class of anticancer drugs that target DNA topoisomerase I; current efforts are directed toward elucidating optimal combinations of these drugs with other antineoplastic agents. A rationale for the use of sequential therapy involving the combination of camptothecins with topoisomerase II-targeting drugs, such as etoposide, has arisen from observations of increased topoisomerase II protein levels in cell lines resistant to camptothecin. In an effort to understand potential mechanisms of resistance to this strategy, we developed a U-937 cell subline, denoted RERC, that is capable of surviving exposure to sequential topoisomerase poisoning. The RERC cells are 200-fold resistant to camptothecin, 8-fold resistant to etoposide, and 10-fold hypersensitive to cisplatin compared to the parental U-937 cells. Biochemical analyses indicate that the resistant phenotype involves alterations in both topoisomerase I and topoisomerase IIalpha. Topoisomerase I catalytic activity in the resistant cells is similar to that of the parental line but is resistant to camptothecin. Moreover, the resistant cells express a single mRNA species of topoisomerase I that codes for a mutation in codon 533. In addition, topoisomerase IIalpha protein levels are decreased 10-fold in the resistant line, coincident with a two-fold decrease in the expression of topoisomerase IIalpha mRNA. Collectively, these results indicate that resistance to sequential topoisomerase poisoning may involve a reduction in total cellular topoisomerase activity.     

Adequacy of medical referrals to hospital emergency service

OBJECTIVE: To assess the correctness and diagnostic concordance in referrals of patients to hospital Casualty Departments (CD); and the possible differences between rural and urban health centres (HC) as well as between general practitioners (GP) and family and community medicine (FCM) specialists. DESIGN: Observational study of a crossover nature. SETTING: Health area 1 in the province of Badajoz. PATIENTS AND OTHER PARTICIPANTS: 800 patients, referred to their local hospital Casualty Department. MEASUREMENTS AND MAIN RESULTS: 50.5% of all referrals were considered justified. 15.1% of the patients referred were admitted to hospital. 50.7% of referrals from GPs were found to be correct, against 47% from FCM specialists; and 49% of those referred from rural HCs were correct, as against 45.9% of those referred from urban HCs (no significant differences). Only 54% of referrals had a diagnostic hypothesis, with diagnostic concordance at 50%. There was diagnostic concordance in 28.1% of GP referrals, against 33.3% of FCM specialists; and in 27.5% of those from rural HCs against 26.1% from urban HCs (no significant differences). CONCLUSIONS: We found no differences in the correctness of referrals or diagnostic concordance of patients seen in the CD of our city's health area, neither between patients referred from rural and urban HCs, nor between patients referred by GPs and FCM specialists.     

The effect of nanoclays on the nucleation,crystallization, and melting mechanisms of isotactic polypropylene

The influence of organomodified nanoclay (montmorillonite) on the crystallization and melting mechanisms of isotactic polypropylene (iPP) was studied. Films of both neat polymer and clay nanocomposites were prepared after molecular weight optimization through melt extrusion. Products exhibited the tactoidlike morphology since no compatibilizers were used. The effect of introduction of nanoclay within the polymer was studied through isothermal crystallization and linear heating. The results indicated that low nanoclay concentrations induce the formation of the β‐crystalline structure, its formation being inhibited with high nanoclay contents. Dynamic nonisothermal bulk crystallization experiments indicated that nanoclays act as nucleating agents of iPP. Isothermal, bulk crystallization studies indicated that the crystallization process (induction time and crystallization kinetics) is proportionally accelerated by the nanoclay presence. There was also an accelerated formation of secondary crystallization when nanoclays were present in high concentrations. POLYM. ENG. SCI., 47:1889–1897, 2007. © 2007 Society of Plastics Engineers     

A memetic algorithm for the multi-compartment vehicle routing problem with stochastic demands

The multi-compartment vehicle routing problem (MC-VRP) consists of designing transportation routes to satisfy the demands of a set of customers for several products that, because of incompatibility constraints, must be loaded in independent vehicle compartments. Despite its wide practical applicability the MC-VRP has not received much attention in the literature, and the few existing methods assume perfect knowledge of the customer demands, regardless of their stochastic nature. This paper extends the MC-VRP by introducing uncertainty on what it is known as the MC-VRP with stochastic demands (MC-VRPSD). The MC-VRPSD is modeled as a stochastic program with recourse and solved by means of a memetic algorithm. The proposed memetic algorithm couples genetic operators and local search procedures proven to be effective on deterministic routing problems with a novel individual evaluation and reparation strategy that accounts for the stochastic nature of the problem. The algorithm was tested on instances of up to 484 customers, and its results were compared to those obtained by a savings-based heuristic and a memetic algorithm (MA/SCS) for the MC-VRP that uses a spare capacity strategy to handle demand fluctuations. In addition to effectively solve the MC-VRPSD, the proposed MA/SCS also improved 14 best known solutions in a 40-problem testbed for the MC-VRP.     

Temporarily Expanding the Boundaries of the Self: Motivations for Entering the Story World and Implications for Narrative Effects

A wide variety of motivations for engaging with narratives have been proposed and studied. We propose that underlying these motivations is another, more fundamental motivation. Our premise is that maintenance, defense, and regulation of the personal and social self in daily life are demanding both emotionally and cognitively. Moreover, any individual self is constrained by capability, situation, and social role. Stories and identification with story characters provide a means individuals may use for temporary relief from the task of self‐regulation and from the limitations of individual personal and social identities. Existing supportive research is acknowledged and implications explored, concerning contexts in which story involvement will be particularly attractive and possible impacts on attitudes and acceptance of out‐groups including stigmatized others.     

Virulence-resistance plasmids (pUO-StVR2-like) in meat isolates of Salmonella enterica serovar Typhimurium

During a screening of Salmonella enterica in foods of animal origin four isolates of serovar Typhimurium carrying hybrid virulence-resistance plasmids were detected. Three of them, one from pork and two from chicken meat, contained pUO-StVR2, a derivative of the pSLT virulence plasmid with the bla_OXA-1, catA1, aadA1, sulI and tet(B) genes, which confer resistance to ampicillin, chloramphenicol, streptomycin-spectinomycin, sulfonamides and tetracycline, respectively. The fourth isolate, from pork, harbored a pUO-StVR2 variant (termed ν8) with an additional dfrA10 gene responsible for resistance to trimethoprim. This gene is part of the orf513-dfrA10-qacEΔ1-sul1 element characteristically found in complex class 1 integrons. Pulsed-field gel electrophoresis identified two XbaI-BlnI combined profiles: X2-B2 generated from the three pUO-StVR2 isolates, and X12-B17 shown by the pUO-StVR2-ν8 isolate. The same profiles have also been found in clinical and ill pig isolates, supporting chicken and pork meat as vehicles for transmission of Typhimurium carrying pUO-StVR2-like plasmids.     

Anthocyanin composition in aged Chilean Cabernet Sauvignon red wines

Anthocyanins in aged Cabernet Sauvignon red wines were studied by HPLC–MS. The major anthocyanin in 6, 7, and 8 year old red wine extracts was the dimer vitisin A produced by condensation of malvidin-3-O-glucoside mediated by pyruvic acid. In aged wine, the content of malvidin-3-O-glucoside decreased with age with a concomitant increase of vitisin A. The latter is accompanied by several condensation products with molecular weight up to 1500 m/z. Differential pulse voltammetry indicated that aged wines have lower antioxidant capacity than young wine (400 mV), but higher than malvidin-3-O-glucoside (483 mV). Tafel’s plots showed that the electrochemical process occurring in aged wines is different from young wines. Six, seven and eight year old wines show similar behaviour with plots of 234, 177 and 188 mV/dec, respectively. These values are higher than the expected 120 mV/dec corresponding to a first electronic transfer but smaller compared to the 523 mV/dec corresponding to young wine.