A multiplex RTi-PCR reaction for simultaneous detection of Escherichia coli O157:H7, Salmonella spp. and Staphylococcus aureus on fresh, minimally processed vegetables

In this work, a new multiplex single-tube real-time PCR approach is presented for the detection of Escherichia coli O157:H7, Salmonella spp. and Staphylococcus aureus, three of the more frequent food-borne bacterial pathogens that are usually investigated in a variety of food matrices. The study includes the design and specificity testing, of a new primer and probe specific for Salmonella spp. Reaction conditions were adjusted for the simultaneous amplification and detection of specific fragments in the beta-glucuronidase (uidA, E. coli) and Thermonulease (nuc, Sta. aureus) genes, and in the replication origin sequence (oriC, Salmonella spp.). Melting-curve analysis using a SYBR Green I RTi-PCR approach showed characteristic T(m) values demonstrating the specific and efficient amplification of the three fragments. Subsequently, a TaqMan RTi-PCR approach was settled, using FAM, NED and VIC fluorescently labelled specific probes for an automated detection. It was equally sensitive than uniplex RTi-PCR reactions in Sta. aureus and E. coli O157:H7, using same amounts of purified DNA, and allowed detection of 10 genome equivalents in the presence of 10(2) or 10(4) genome equivalents of the other two pathogens. Finally, it was tested in artificially inoculated fresh, minimally processed vegetables, revealing a sensitivity of 10(3)CFUg(-1) each of these pathogens in direct detection, following DNA extraction with DNeasy Tissue Kit (Qiagen). The multiplex RTi-PCR developed scored the sensitivity recognised for PCR in food and it allows a high-throughput and automation, thus it is promising as a rapid and cost-effective test for the food industry.     

Short communication: Addition of sodium bicarbonate to maternal colostrum: Effects on immunoglobuling absorption and hematocrit in neonatal calves

Twenty-six Holstein bull calves born from primiparous and multiparous cows without dystocia were assigned in a randomized complete block design to 1 of 2 treatments: pooled maternal colostrum (PMC) or PMC supplemented with 30g of sodium bicarbonate (NaHCO(3)). Calves were fed PMC from 9 different batches containing (mean ± SD) 82.05±8.45g/L of IgG. Calves were fed 2.68L of PMC at birth (referred to as 0h) and 1.32L of PMC 6h later. The total amount of IgG fed was 329.89±34.56g. Calves were fed 2L of milk replacer at 24, 36, and 48h postpartum. The addition of NaHCO(3) had no effect on IgG absorption. Serum IgG concentrations at 0, 6, 12, 24, and 48h postpartum were not different between calves supplemented with or without 30g of NaHCO(3) to colostrum. Area under the curve, apparent efficiency of absorption, and hematocrit were not affected by the NaHCO(3) treatment.     

Local Notions of Healthy Eating and National Dietary Guidelines: A Comparison in Vulnerable Salvadoran Communities

This study presents an assessment of local definitions and perceptions concerning healthy eating through a study in four resource-poor border communities in El Salvador. The study included focus groups, key-informant interviews, and observations of the food environment. Local definitions of healthy eating elicited through focus groups were compared to the national Salvadoran dietary guidelines recommendations. The comparison revealed several areas of overlap (including the importance of dietary variety, fruits, and vegetables, among others) and omissions (mention of limiting sweets/candy, salt, sugar, and alcohol). Focus group participants expressed concerns over the origin of their foods and whether food contained harmful chemicals. These conversations also revealed the contradictions between nutrition knowledge and preferences for foods classified as unhealthy. This article concludes with a discussion about barriers to healthy eating identified in the focus groups and through the food environment assessment.     

重组华根霉脂肪酶协同转谷氨酰胺酶提高冷冻面团抗冻发酵特性的研究

采用差示扫描量热仪和F3流变发酵仪研究重组华根霉脂肪酶(RCL)和转谷氨酰胺酶(TG)共同作用对冷冻面团抗冻发酵特性的影响。将面团于-18℃冻藏0、7、21、35d,结果发现:随着冻藏时间的延长,甘油含量有所降低,可冻结水含量增加;引入RCL和TG到冷冻面团中可以显著增加面团中的甘油含量,显著降低面团中可冻结水的含量,减少冰晶体的形成,并且可以提高酵母的存活数。F3流变发酵仪测定面团的发酵流变学特性,结果表明:RCL和TG同时作用可以显著降低冻藏对面团发酵高度的削弱作用,改善酵母的发酵性能和增加面团的持气率。     

Genome-wide bacterial toxicity screening uncovers the mechanisms of toxicity of a cationic polystyrene nanomaterial

By exploiting a genome-wide collection of bacterial single-gene deletion mutants, we have studied the toxicological pathways of a 60-nm cationic (amino-functionalized) polystyrene nanomaterial (PS-NH(2)) in bacterial cells. The IC(50) of commercially available 60 nm PS-NH(2) was determined to be 158 μg/mL, the IC(5) is 108 μg/mL, and the IC(90) is 190 μg/mL for the parent E. coli strain of the gene deletion library. Over 4000 single nonessential gene deletion mutants of Escherichia coli were screened for the growth phenotype of each strain in the presence and absence of PS-NH(2). This revealed that genes clusters in the lipopolysaccharide biosynthetic pathway, outer membrane transport channels, ubiquinone biosynthetic pathways, flagellar movement, and DNA repair systems are all important to how this organism responds to cationic nanomaterials. These results, coupled with those from confirmatory assays described herein, suggest that the primary mechanisms of toxicity of the 60-nm PS-NH(2) nanomaterial in E. coli are destabilization of the outer membrane and production of reactive oxygen species. The methodology reported herein should prove generally useful for identifying pathways that are involved in how cells respond to a broad range of nanomaterials and for determining the mechanisms of cellular toxicity of different types of nanomaterials.     

Altered Distribution and Expression of Syndecan-1 and -4 as an Additional Hallmark in Psoriasis

Syndecans act as independent co-receptors to exert biological activities and their altered function is associated with many pathophysiological conditions. Here, syndecan-1 and -4 were examined in lesional skin of patients with psoriasis. Immunohistochemical staining confirmed altered syndecan-1 distribution and revealed absence of syndecan-4 expression in the epidermis. Fibronectin (FN)—known to influence inflammation and keratinocyte hyperproliferation via α5β1 integrin in psoriasis—was also decreased. Syndecan-1 and -4 expression was analyzed in freshly isolated lesional psoriatic human keratinocytes (PHK) characterized based on their proliferation and differentiation properties. mRNA levels of syndecan-1 were similar between healthy and PHK, while syndecan-4 was significantly decreased. Cell growth and release of the pro-inflammatory Tumor Necrosis Factor-alpha (TNFα) were selectively and significantly induced in PHKs plated on FN. Results from co-culture of healthy keratinocytes and psoriatic fibroblasts led to the speculation that at least one factor released by fibroblasts down-regulate syndecan-1 expression in PHK plated on FN. To assay if biological treatments for psoriasis target keratinocyte proliferation, gelatin-based patches enriched with inteleukin (IL)-17α or TNFα blockers were prepared and tested using a full-thickness healthy epidermal model (Phenion^®). Immunohistochemistry analysis showed that both blockers impacted the localisation of syndecan-1 within the refined epidermis. These results provide evidence that syndecans expression are modified in psoriasis, suggesting that they may represent markers of interest in this pathology.     

Real-Time Differential Pricing Scheme for Active Consumers with Electric Vehicles

Electric vehicles (EVs) are becoming increasingly popular because of their low operating costs. However, the anticipated increase in EV usage presents opportunities as well as challenges for utilities. This paper models EV charging behavior of active consumers who strategize their EV charging schedule to minimize the charging cost. The electric utility employs a temporal and spatially varying real-time pricing scheme that minimizes peak-to-average load ratio by influencing charging behavior of multiple active consumers. These active consumers decide their optimal charging strategy based on real-time electricity prices in order to minimize their EV charging cost. A model predictive control (MPC)-based optimal pricing scheme for electric utility and optimal charging strategy for active consumers is proposed. Effects of the proposed approach on load profile is verified using simulations.     

Structural performance of a climbing cactus: making the most of softness

Climbing plants must reach supports and navigate gaps to colonize trees. This requires a structural organization ensuring the rigidity of so-called ‘searcher’ stems. Cacti have succulent stems adapted for water storage in dry habitats. We investigate how a climbing cactus Selenicereus setaceus develops its stem structure and succulent tissues for climbing. We applied a ‘wide scale’ approach combining field-based bending, tensile and swellability tests with fine-scale rheological, compression and anatomical analyses in laboratory conditions. Gap-spanning ‘searcher’ stems rely significantly on the soft cortex and outer skin of the stem for rigidity in bending (60–94%). A woody core contributes significantly to axial and radial compressive strength (80%). Rheological tests indicated that storage moduli were consistently higher than loss moduli indicating that the mucilaginous cortical tissue behaved like a viscoelastic solid with properties similar to physical or chemical hydrogels. Rheological and compression properties of the soft tissue changed from young to old stages. The hydrogel–skin composite is a multi-functional structure contributing to rigidity in searcher stems but also imparting compliance and benign failure in environmental situations when stems must fail. Soft tissue composites changing in function via changes in development and turgescence have a great potential for exploring candidate materials for technical applications.     

Radiological Prediction and Monitoring of Tests at the Nuclear Rocket Development Station

The procedure and formula used in the radiological analysis of reactor testing at the Nuclear Rocket Development Station are described. Monitoring methods are discussed and comparison is made between measured dose levels and guide values.