Effect of acid adaptation on inactivation of Salmonella during drying and storage of beef jerky treated with marinades

This study evaluated the influence of pre-drying marinade treatments on inactivation of acid-adapted or nonadapted Salmonella on beef jerky during preparation, drying and storage. The inoculated (five-strain composite, 6.0 log CFU/cm2) slices were subjected to the following marinades (24 h, 4 degrees C) prior to drying at 60 degrees C for 10 h and aerobic storage at 25 degrees C for 60 days: (1) no marinade, control (C), (2) traditional marinade (TM), (3) double amount of TM modified with added 1.2% sodium lactate, 9% acetic acid, and 68% soy sauce with 5% ethanol (MM), (4) dipping into 5% acetic acid and then TM (AATM), and (5) dipping into 1% Tween 20 and then into 5% acetic acid, followed by TM (TWTM). Bacterial survivors were determined on tryptic soy agar with 0.1% pyruvate and xylose-lysine-tergitol 4 (XLT4) agar. Results indicated that drying reduced bacterial populations in the order of pre-drying treatments TWTM (4.8-6.0 log CFU/cm2)> or =AATM> or =MM>TM> or =C (2.6-5.0 log CFU/cm2). Nonadapted Salmonella were significantly (P<0.05) more resistant to inactivation during drying than acid-adapted Salmonella in all treatments. Bacterial populations decreased below the detection limit (-0.4 log CFU/cm2) as early as 7 h during drying or remained detectable even after 60 days of storage, depending on acid adaptation, pre-drying treatment, and agar media. The results indicated that acid adaptation may not cause increased resistance of Salmonella to the microbial hurdles involved in jerky processing and that use of modified marinades in manufacturing jerky may improve the effectiveness of drying in inactivating Salmonella.     

An Empirically-Based Process for Software Architecture Evaluation

Software systems undergo constant change causing the architecture of the system to degenerate over time. Reversing system degeneration takes extra effort and delays the release of the next version. Improved architecture is intangible and does not translate into visible user features that can be marketed. Due to a lack of representative metrics, technical staff has problems arguing that stopping degeneration is indeed necessary and that the effort will result in an improved architecture that will pay off. We believe that architectural metrics would give technical staff better tools to demonstrate that the architecture has improved. This paper defines and uses a set of architectural metrics and outlines a process for analyzing architecture to support such an argument. The paper reports on a case study from a project where we restructured the architecture of an existing client-server system written in Java while adding new functionality. The modules of the existing version of the system were library-oriented and had a disorganized communication structure. The new architecture is based on components and utilizes the mediator design pattern. The goal of the study is to evaluate the new architecture from a maintainability perspective. The paper describes our evaluation process, the metrics used, and provides some preliminary results. The architectural evaluation shows that the components of the system are only loosely coupled to each other and that an architectural improvement has occurred from a maintenance perspective. The process used to evaluate the architecture is general and can be reused in other contexts.     

Development of two multiplex polymerase chain reactions for the detection of enterotoxigenic strains of Staphylococcus aureus isolated from foods

Two multiplex polymerase chain reactions were developed for the detection of enterotoxigenic strains of Staphylococcus aureus: one multiplex reaction for the simultaneous detection of enterotoxigenic strains type A (entA), type B (entB), and type E (entE) and another for the simultaneous detection of enterotoxigenic strains type C (entC) and type D (entD). Both reactions were standardized with the use of the reference enterotoxigenic strains of S. aureus: FRI 722, producer of staphylococcal enterotoxin (SE) type A (SEA); FRI 1007, producer of SEB; FRI 137, producer of SEC1; FRI 472, producer of SED; and FRI 326, producer of SEE. Optimized methods were used to determine the presence of enterotoxigenic types for 51 S. aureus strains isolated from meat (sausage, ham, and chorizo) and dairy (powdered milk and cheese) products by the Baird-Parker technique. The enterotoxigenic capacities of the strains were determined by the indirect enzyme-linked immunosorbent assay (ELISA) with the use of reference staphylococcal toxins and antitoxins. Fifty of the 51 strains isolated were enterotoxigenic and produced one to four enterotoxin types, with the most frequently produced types being SEA and SED. Levels of correlation between the presence of genes that code for the production of SE (as determined by polymerase chain reaction) and the expression of these genes (as determined by the indirect ELISA) were 100% for SEA and SEE, 86% for SEC, 89% for SED, and 47% for SEB.     

Detection of attomole quantities [correction of quantitites] of DNA targets on gold microelectrodes by electrocatalytic nucleobase oxidation

The electrochemical detection of nucleic acid targets at low concentrations has a number of applications in diagnostics and pharmaceutical research. Self-assembled monolayers of alkanethiol-derivatized oligonucleotides on gold electrodes provide a useful platform for such detectors, and the electrocatalytic oxidation of nucleobases included in the DNA targets is a particularly sensitive method of electrochemical detection. A strategy has been developed for combining these two aspects by substituting either 7,8-dihydro-8-oxoguanine (8G) or 5-aminouridine (5U) into DNA targets. Upon hybridization of targets containing these modified nucleobases, electrocatalytic signals at probe-modified gold electrodes are observed in the presence of Os(bpy)(3)(2+), which oxidizes both 8G and 5U upon oxidation to the Os(III) state. Self-assembled monolayers were prepared on both macro (1.6 mm) and micro (25 microm) gold electrodes using published procedures involving C6-terminated alkanethiol oligonucleotides and mercaptohexanol as the diluent. The extent of electrode modification by the modified probe was assessed using radiolabeling and a standard chronocoulometry method; both approaches gave loading levels within expected ranges ((1-6) x 10(12) molecules/cm(2)). Hybridization of the modified targets where the non-native nucleobase was incorporated by solid-phase synthesis produced electrocatalytic signals from strands that were independently detected using radiolabeling and chronocoulometry. This result was used as a basis to develop an on-electrode amplification scheme where Taq polymerase was used to extend the immobilized DNA probes from solution-phase polymeric templates using modified nucleotriphosphates. This reaction produced an electrode that was modified with extended DNA containing the appropriate modified nucleotide. Radiolabeled nucleotide triphosphates were used to confirm the desired on-electrode DNA synthesis. When these electrodes were cycled in the presence of Os(bpy)(3)(2+), electrocatalytic signals were observed when as little as 40 amol (400 fM) of the desired target was present in the hybridization solution.     

Effects of high-γ-linolenic acid canola oil compared with borage oil on reproduction,growth, and brain and behavioral development in mice

Previous research in rats and mice has suggested that γ-linolenic acid (GLA) derived from borage oil (BO: 23% GLA) may be an appropriate source for increasing levels of long-chain n−6 FA in the developing brain. Recently, transgenic technology has made available a highly enriched GLA seed oil from the canola plant (HGCO: 36% GLA). The first objective of this study was to compare the effects of diets containing equal levels of GLA (23%) from either BO or HGCO on reproduction, pup development, and pup brain FA composition in mice. The second objective was to compare the effects of the HGCO diluted to 23% GLA (GLA-23) with those of undiluted HGCO containing 36% GLA (GLA-36). The diets were fed to the dams prior to conception and throughout pregnancy and lactation, as well as to the pups after weaning. The behavioral development of the pups was measured 12 d after birth, and anxiety in the adult male offspring was assessed using the plus maze. The findings show that despite equivalent levels of GLA, GLA-23 differed from BO in that it reduced pup body weight and was associated with a slight increase in neonatal pup attrition. However, there were no significant effects on pup behavioral development or on performance in the plus maze. An increase in dietary GLA resulted in an increase in brain 20∶4n−6 and 22∶4n−6, with a corresponding decrease in 22∶6n−3. Again, despite their similar levels of GLA, these effects tended to be larger in GLA-23 than in BO. In comparison with GLA-23, GLA-36 had larger effects on growth and brain FA composition but no differences with respect to effects on reproduction and behavioral development. These findings suggest that the HGCO can be used as an alternative source of GLA.     

Application of a breeder genetic algorithm for filter optimization

In this work we are optimizing an adaptive finite impulse response filter applied to the problem of system identification. We are proposing a breeder genetic algorithm (BGA) for performing the parametric search in highly multimoldal landscapes since in this kind of filters there exits epistiasis. The results of BGA were compared to the traditional genetic algorithm, and we found that the BGA was clearly superior (in accuracy) in most of the cases. We used the statistical least mean squared for validating the results. We suggest to hybridized both methods for real world applications.     

Contamination-free continuous flow microfluidic polymerase chain reaction for quantitative and clinical applications

We present a method for performing polymerase chain reaction (PCR) using isolated droplets flowing in an immiscible fluorinated solvent system. Thanks to an optimized control of interfacial properties, we could achieve in this capillary-based system reproducible amplification factors, without any detectable contamination between neighboring droplets. The system is readily amenable to further miniaturization and automation and serves as the first step toward a clinically viable, high-throughput, quantitative continuous flow PCR apparatus.     

Substrate interactions with the nitrogenase active site

The chemical mechanism for biological cleavage of the N(2) triple bond at ambient pressure and temperature has been the subject of intense study for many years. The site of substrate activation and reduction has been localized to a complex cofactor, called FeMo cofactor, yet until now the complexity of the system has denied information concerning exactly where and how substrates interact with the metal-sulfur framework of the active site. In this Account, we describe a combined genetic, biophysical, and biochemical approach that was used to provide direct and detailed information concerning where alternative alkyne substrates interact with FeMo cofactor during catalysis. The relevance and limitations of this work with respect to N(2) binding and reduction also are discussed.     

Comparative Investigation of the Volatile Urinary Profiles in Different <Emphasis Type="Italic">Phodopus</Emphasis> Hamster Species

Stir bar sorptive extraction method was used for investigation of the urinary volatile profiles in male and female Phodopus campbelli and Phodopus sungorus hamsters. Additionally, female Phodopus roborowsky urinary profiles were characterized. A quantitative analytical approach allowed comparisons of 17 selected compounds in urine. Results showed that campbelli and sungorus species show similar urinary volatile profiles for males and females. Differences appeared only in concentrations. Several unique compounds, such as pyrazine derivatives, were found to be gender- and age-specific. P. roborowsky females exhibited a completely different urinary volatile profile from campbelli and sungorus females, featuring a unique set of substituted quinoxalines.