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.     

From dust till drawn

We present a system for drawing pastel media in real-time as an effective alternative to most existing digital solutions that basically allow for drawing arbitrary strokes in a particular style. Our approach is focused on the simulation of the natural material itself and on its interaction with the drawing surface and the drawing tool. Upon free-form drawing, a bidirectional transfer of pigment takes place. In one direction, the paper surface is dusted with new pigment particles broken off the tip (i.e., the end of the drawing tool). A large part of these particles will be deposited or blended together with previously deposited ones whereas the remainder does not contribute to the drawing and is blown off. On the other hand, a certain amount of previously deposited pigment is scraped off and picked up again soiling the tip. This is noticeable in the next strokes to be drawn. Furthermore, both the tip and the paper surface are subject to weathering depending on the exerted pressure and friction of the drawing tool, and the bumpiness of the paper. As a result, the paper surface becomes slightly damaged, limiting the deposition of new pigment. The tip, on the other hand, becomes blunt making new strokes wider. From a stylistic point of view, similarly to traditional drawings our results convey the artists’ characteristics (e.g., the way of wielding the brush, skillfulness, feeling for the medium). Therefore, we believe that our system allows an artist to create realistically looking pastel images without losing his/her personal touch.     

Out-of-Field Hippocampus from Partial-Body Irradiated Mice Displays Changes in Multi-Omics Profile and Defects in Neurogenesis

The brain undergoes ionizing radiation exposure in many clinical situations, particularly during radiotherapy for brain tumors. The critical role of the hippocampus in the pathogenesis of radiation-induced neurocognitive dysfunction is well recognized. The goal of this study is to test the potential contribution of non-targeted effects in the detrimental response of the hippocampus to irradiation and to elucidate the mechanisms involved. C57Bl/6 mice were whole body (WBI) or partial body (PBI) irradiated with 0.1 or 2.0 Gy of X-rays or sham irradiated. PBI consisted of the exposure of the lower third of the mouse body, whilst the upper two thirds were shielded. Hippocampi were collected 15 days or 6 months post-irradiation and a multi-omics approach was adopted to assess the molecular changes in non-coding RNAs, proteins and metabolic levels, as well as histological changes in the rate of hippocampal neurogenesis. Notably, at 2.0 Gy the pattern of early molecular and histopathological changes induced in the hippocampus at 15 days following PBI were similar in quality and quantity to the effects induced by WBI, thus providing a proof of principle of the existence of out-of-target radiation response in the hippocampus of conventional mice. We detected major alterations in DAG/IP3 and TGF-β signaling pathways as well as in the expression of proteins involved in the regulation of long-term neuronal synaptic plasticity and synapse organization, coupled with defects in neural stem cells self-renewal in the hippocampal dentate gyrus. However, compared to the persistence of the WBI effects, most of the PBI effects were only transient and tended to decrease at 6 months post-irradiation, indicating important mechanistic difference. On the contrary, at low dose we identified a progressive accumulation of molecular defects that tended to manifest at later post-irradiation times. These data, indicating that both targeted and non-targeted radiation effects might contribute to the pathogenesis of hippocampal radiation-damage, have general implications for human health.     

Solubility and Aggregation of Selected Proteins Interpreted on the Basis of Hydrophobicity Distribution

Protein solubility is based on the compatibility of the specific protein surface with the polar aquatic environment. The exposure of polar residues to the protein surface promotes the protein’s solubility in the polar environment. The aquatic environment also influences the folding process by favoring the centralization of hydrophobic residues with the simultaneous exposure to polar residues. The degree of compatibility of the residue distribution, with the model of the concentration of hydrophobic residues in the center of the molecule, with the simultaneous exposure of polar residues is determined by the sequence of amino acids in the chain. The fuzzy oil drop model enables the quantification of the degree of compatibility of the hydrophobicity distribution observed in the protein to a form fully consistent with the Gaussian 3D function, which expresses an idealized distribution that meets the preferences of the polar water environment. The varied degrees of compatibility of the distribution observed with the idealized one allow the prediction of preferences to interactions with molecules of different polarity, including water molecules in particular. This paper analyzes a set of proteins with different levels of hydrophobicity distribution in the context of the solubility of a given protein and the possibility of complex formation.     

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     

An empirical study of control logic specifications for programmable logic controllers

This paper presents an empirical study of control logic specifications used to document industrial control logic code in manufacturing applications. More than one hundred input/output related property specifications from ten different reusable function blocks were investigated. The main purpose of the study was to provide understanding of how the specifications are expressed by industrial practitioners, in order to develop new tools and methods for specifying control logic software, as well as for evaluating existing ones. In this paper, the studied specifications are used to evaluate linear temporal logic in general and the specification language ST-LTL, tailored for functions blocks, in particular. The study shows that most specifications are expressed as implications, that should always be fulfilled, between input and output conditions. Many of these implications are complex since the input and output conditions may be mixed and involve sequences, timer issues and non-boolean variables. Using ST-LTL it was possible to represent all implications of this study. The few non-implication specifications could be specified in ST-LTL as well after being altered to suit the specification language. The paper demonstrates some advantages of ST-LTL compared to standard linear temporal logic and discusses possible improvements such as support for automatic rewrite of complex specifications.