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.     

Burner Performance Evaluation Using Test Code

Development of new burner technology is common practice nowadays. Standardized test codes are available to conduct tests to determine operating characteristics and thermal performance of different parts of steam generating units. This paper describes the results of testing performed to evaluate thermal efficiency of a 74.5?kW fire-tube steam generator fired with a new premixed flame burner in comparison to a typical standard-retrofit diffusion flame burner system. Two premixed flame burners sized at 7.6 and 10.1?cm were tested. Testing was performed in accordance with the ASME PTC 4.1 Power Test Code. The evaluation was conducted in the range of small- to mid-sized, natural gas-fired applications, from 1.06?to?2.64×106?kJ/h. The environmental nitrogen oxide (NOx) emissions performance was also compared for both burner technologies. The results of this evaluation indicate that firing the test boiler with the 7.6?cm new burner slightly improves unit thermal efficiency by 4.0–3.3% for a unit load ranging from approximately 1.06?to?1.79×106?kJ/h, respectively. Operation with the 10.1?cm new burner improves the thermal efficiency by approximately 1.6% at 2.32×106?kJ/h. The uncertainty in the measurements used in the efficiency calculations should account for approximately ±1.5% uncertainty in the reported gross efficiency. Reductions in NOx emissions of the order of 10% resulted from operation with the new premixed flame burners.     

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.     

Long-term energy scenarios for Mexico: Policy options for carbon savings and main barriers

This paper shows that possibilities exist for maintaining current energy-related carbon emissions per capita in Mexico at almost constant levels in the long term. It is argued, however, that the identified carbon emissions reductions will not be achieved easily or rapidly, as they require a restructuring of Mexico's current technological base and an unusual degree of coordination among the government, lending institutions, equipment manufacturers, and final users. Such changes also will entail surmounting major institutional and financial barriers and creating a better international environment for technology transfer and capital lending.     

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.     

Altered hippocampal kainate-receptor mRNA levels in temporal lobe epilepsy patients

This study determined whether hippocampal kainate (KA) receptor mRNA levels were increased or decreased in temporal lobe epilepsy patients compared with nonseizure autopsies. Hippocampal sclerosis (HS; n = 17), nonsclerosis (non-HS; n = 11), and autopsy hippocampi (n = 9) were studied for KA1-2 and GluR5-7 mRNA levels using semiquantitative in situ hybridization techniques, along with neuron densities. Compared with autopsy hippocampi, HS and non-HS cases showed decreased GluR5 and GluR6 hybridization densities per CA2 and/or CA3 pyramid. Furthermore, HS patients demonstrated increased KA2 and GluR5 hybridization densities per granule cell compared with autopsy hippocampi. These findings indicate that chronic temporal lobe seizures were associated with differential changes in hippocampal KA1-2 and GluR5-7 hybridization densities that vary by subfield and pathology group. In temporal lobe epilepsy patients, these results support the hypothesis that pyramidal cell GluR5 and GluR6 mRNA levels are decreased as a consequence of seizures, and in HS patients granule cell KA2 and GluR5 mRNA levels are increased in association with aberrant fascia dentata mossy fiber sprouting and/or hippocampal neuronal loss.