Detection of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in kimchi by multiplex polymerase chain reaction (mPCR)

We developed an mPCR assay for the simultaneous detection, in one tube, of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes using species-specific primers. The mPCR employed the E. coli O157:H7 specific primer Stx2A, Salmonella spp. specific primer Its, S. aureus specific primer Cap8A-B and L. monocytogenes specific primer Hly. Amplification with these primers produced products of 553, 312, 405 and 210 bp, respectively. All PCR products were easily detected by agarose gel electrophoresis, and the sequences of the specific amplicons assessed. Potential pathogenic bacteria, in laboratory-prepared and four commercially available kimchi products, were using this mPCR assay, and the amplicons cloned and sequenced. The results correlated exactly with sequences derived for amplicons obtained during preliminry tests with known organisms. The sensitivity of the assay was determined for the purified pathogen DNAs from four strains. The mPCR detected pathogen DNA at concentrations ranging from approximately 0.45 to 0.05 pM/microl. Thus, this mPCR assay may allow for the rapid, reliable and cost-effective identification of four potentially pathogens present in the mixed bacterial communities of commercially available kimchi.     

Localization of hydrogen peroxide in pumpkin (Cucurbita ficifolia bouché) seedlings exposed to high-dose gamma ray

Hydrogen peroxide (H₂O₂) was detected cytochemically, via transmission electron microscopy (TEM), in pumpkin tissues exposed to high-dose gamma ray. Its reaction with cerium chloride produced electron-dense precipitates of cerium perhydroxides. Their patterns of deposition in the tissues of both control plants and those irradiated with gamma ray (PIG) were typically found in the plasma membranes and cell walls. However, gamma irradiation remarkably increased the intensities of cerium perhydroxide deposits (CPDs) in the plasma membranes and cell walls for all tissue types, but especially the leaves. The only exception was for vessels in the cotyledons. After gamma irradiation, the (H₂O₂) content in all tissues was higher than in the control samples, except for the cotyledons of PIG, where the (H₂O₂) content was lower than for all others. The increased appearance of CPDs may have been due to the enhancement of (H₂O₂) accumulation by gamma radiation. This accumulation also varied according to the cell or tissue type examined.     

Sclerophylly in Qualea Parviflora (Vochysiaceae): Influence of Herbivory, Mineral Nutrients, and Water Status

Qualea parviflora Mart. (Vochysiaceae) is a deciduous tree, commonly observed in campo sujo, cerrado sensu stricto and cerradão vegetation types in Brazilian cerrado (savannas). In this study we investigated herbivory, nutritional, and water status effects on leaf sclerophylly of Q. parviflora. Twenty fully expanded leaves were taken from 10 plants in each vegetation type four times a year. Mean leaf concentration of N, P, K, Ca, C, Al, Si, and percentage of total phenols, herbivory and tannins were measured on a plant basis. Leaf specific mass (LSM) (g m^?2), a sclerophylly index, and pre-dawn leaf water potential (MPa) were also recorded. Soil samples below each tree were collected to quantify N–NO₃, N–NH₄, P, K, Mn, soil moisture, organic matter, Si, and Al. Qualea parviflora showed a LSM from 69 to 202 g m^?2 and leaves were younger and less sclerophyllous in November (beginning of rainy season). Q. parviflora inhabiting the cerradão had leaves with higher concentration of nutrients and lower sclerophylly while trees in campo sujo and cerrado sensu stricto did not show significant differences in leaf sclerophylly. The concentrations of N, P, K and tannins had an inverse relationship with leaf age. Concentration of phenols, Al, C, Ca, Si, C/N and Ca/K increased with leaf age. The concentrations of P and Ca/K ratio in leaves explained 60% of variation observed in leaf sclerophylly. We did not find any significant relationship between the level of sclerophylly and water potential or herbivory. Our results corroborate the hypothesis that predicts lower concentrations of essential macronutrients would be the main factors influencing higher sclerophylly in leaves of Q. parviflora plants in Cerrado.     

Upscaling Regional Emissions of Greenhouse Gases from Rice Cultivation: Methods and Sources of Uncertainty

One of the important sources of greenhouse gases is the emission of methane from rice fields. Methane emission from rice fields is the result of a complex array of soil processes involving plant-microbe interactions. The cumulative effects of these processes at the level of individual plants influence the global atmospheric composition and make it necessary to expand our research focus from small plots to large landscapes and regions. However, present extrapolations (‘upscaling’) are tenuous at best because of methodological and practical problems. The different steps taken to calculate regional emission strengths are discussed and illustrated by calculations for a case-study in the Philippines. The applicability of high quality, process-based, models of methane emission at the level of individual plants is limited for regional analysis by their large data requirements. Simplified models can be used at the regional level but are not able to capture the complex emission situation. Data availability and model accuracy are therefore often difficult to match. Other common sources of uncertainty are the quality of input data. A critical evaluation of input data should be made in every upscaling study to assess the suitability for calculating regional emissions. For the case-study we show effects of differences in input data caused by data source and interpolation technique. The results from the case-study and similar studies in literature indicate that upscaling techniques are still troublesome and a cause of large uncertainties in regional estimates. The results suggest that some of the stumbling blocks in the conventional upscaling procedure are almost impassable in the near future. Based on these results, a plea is made for meso-level measurements to calibrate and validate upscaling methods in order to be better able to quantify and reduce uncertainties in regional emission estimates.     

Targeting of solid tumors and blood malignancies by antibody-based therapies — EGFR-pathway as an example

A well-coordinated interaction between extracellular signals and intracellular response forms the basis of life within multicellular organisms, with growth factors playing a crucial role in these interactions. Discoveries in recent years have shown that components of the Epidermal Growth Factor (EGF) signaling system have frequently been used by cancer cells to autonomously provide survival and proliferation signals. The main focus of this review is the ErbB epidermal growth factor receptor (EGFR) family of receptor tyrosine kinases including ErbB1/EGFR, ErbB2/HER2/neu, ErbB3/HER3, and ErbB4/HER4 as therapeutic targets. Since the ErbB receptor family regulates cell proliferation through the Ras-mitogen-activated protein kinase (RAS/MAPK) pathway, and cell survival and transformation through the phosphatidylinositol 3-kinase (PI3K/AKT) pathway, pharmacological targeting of these pathways is also discussed. We will also address the clinical studies that have been conducted to evaluate antibody-based therapies mostly on solid tumors and hematologic malignancies.     

Heritability of cardiovascular and personality traits in 6,148 Sardinians

In family studies, phenotypic similarities between relatives yield information on the overall contribution of genes to trait variation. Large samples are important for these family studies, especially when comparing heritability between subgroups such as young and old, or males and females. We recruited a cohort of 6,148 participants, aged 14–102 y, from four clustered towns in Sardinia. The cohort includes 34,469 relative pairs. To extract genetic information, we implemented software for variance components heritability analysis, designed to handle large pedigrees, analyze multiple traits simultaneously, and model heterogeneity. Here, we report heritability analyses for 98 quantitative traits, focusing on facets of personality and cardiovascular function. We also summarize results of bivariate analyses for all pairs of traits and of heterogeneity analyses for each trait. We found a significant genetic component for every trait. On average, genetic effects explained 40% of the variance for 38 blood tests, 51% for five anthropometric measures, 25% for 20 measures of cardiovascular function, and 19% for 35 personality traits. Four traits showed significant evidence for an X-linked component. Bivariate analyses suggested overlapping genetic determinants for many traits, including multiple personality facets and several traits related to the metabolic syndrome; but we found no evidence for shared genetic determinants that might underlie the reported association of some personality traits and cardiovascular risk factors. Models allowing for heterogeneity suggested that, in this cohort, the genetic variance was typically larger in females and in younger individuals, but interesting exceptions were observed. For example, narrow heritability of blood pressure was approximately 26% in individuals more than 42 y old, but only approximately 8% in younger individuals. Despite the heterogeneity in effect sizes, the same loci appear to contribute to variance in young and old, and in males and females. In summary, we find significant evidence for heritability of many medically important traits, including cardiovascular function and personality. Evidence for heterogeneity by age and sex suggests that models allowing for these differences will be important in mapping quantitative traits.     

Quantitative detection of doping substances by a localised surface plasmon sensor

Within this communication, consistent evidence of a quantitative biosensing principle for steroidal residue analysis is presented. Our approach uses a simple method for the quantitative determination of an anabolic agent called stanozolol (Sz). Sz (Mw 328) is widely used in sports, horse racing and as a growth promoter in animals for human consumption. Through the use of localised surface plasmons (LSPs), sustained by three-dimensional noble metal nanostructures, we have developed a highly specific, label-less immunosensor for the detection of this small organic molecule to low levels (nM range). A main practical advantage over conventional flat extended film surface plasmon resonance (SPR) systems is the simplicity of the optical configuration, since there is no need for cumbersome total internal reflection illumination, thus making integration easier. In addition, the active area of the LSP-based sensor is smaller, decreasing the minimum detectable number of molecules involved in the binding event. Assay times are short and the set-up is comprised of relatively cheap instrumentation. Detection levels found here are comparable with SPR, even at this early stage of development and with further modifications, we envisage sensing down to pM (10(-12)) levels.     

Karyotypic and molecular polymorphisms in <Emphasis Type="Italic">Ctenomys</Emphasis><Emphasis Type="Italic">torquatus</Emphasis> (Rodentia: Ctenomyidae): taxonomic considerations

The rodent genus Ctenomys (tuco-tucos) comprises more than 60 described species, and shows extraordinary inter- and intraspecific karyotypic variation. The most widely distributed species of Ctenomys in Brazil is C. torquatus. Although several cytogenetic studies have been done, the karyotypic variability of this species is still poorly known. In this paper we report two new diploid numbers for C. torquatus: 2n = 40 and 2n = 42, both showing AN = 72. The new distribution limits of C. torquatus here reported include localities in the southern, central and western parts of Rio Grande do Sul (RS) State in southern Brazil. The phylogenetic relationship between C. torquatus from Alegrete, RS, and Ctenomys sp. from Corrientes, Argentina, is described by means of mtDNA cytochrome b analysis. Although both entities share similar karyotypes and sperm morphology, these two species are not phylogenetically close.     

Homologous recombination and non-homologous end-joining repair pathways in bovine embryos with different developmental competence

This study investigated the expression of genes controlling homologous recombination (HR), and non-homologous end-joining (NHEJ) DNA-repair pathways in bovine embryos of different developmental potential. It also evaluated whether bovine embryos can respond to DNA double-strand breaks (DSBs) induced with ultraviolet irradiation by regulating expression of genes involved in HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro insemination and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation. All studied genes were expressed before, during and after the genome activation period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before genome activation in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-insemination in response to DSBs induced by ultraviolet irradiation. In conclusion, important genes controlling HR and NHEJ DNA-repair pathways are expressed in bovine embryos, however genes participating in these pathways are only regulated after the period of embryo genome activation in response to ultraviolet-induced DSBs.     

Influence of gender and estrous cycle on plasma and renal catecholamine levels in rats

Several studies have demonstrated that gonadal hormones show significant effects on the brain and signaling pathways of effector organs/cells that respond to neurotransmitters. Since little information is available concerning the impact of male and female gonadal hormones on the renal and peripheral sympathetic system, the objective of this study was to further assess whether and how the renal content and plasma concentration of catecholamines are influenced by gender and the estrous cycle in rats. To achieve this, males Wistar rats were divided into 4 groups: (i) sham (i.e., control), (ii) gonadectomized, (iii) gonadectomized and nandrolone decanoate replacement at physiological levels or (iv) gonadectomized and nandrolone decanoate replacement at high levels. Female Wistar rats were divided into 6 groups: (i) ovariectomized (OVX), (ii) estrogen replacement at physiological levels and (iii) estrogen replacement at at high levels, (iv) progesterone replacement at physiological levels and (v) progesterone replacement at at high levels, and (vi) sham. The sham group was subdivided into four subgroups: (i) proestrus, (ii) estrus, (iii) metaestrus, and (iv) diestrus. Ten days after surgery, the animals were sacrificed and their plasma and renal catecholamine levels measured for intergroup comparisons. Gonadectomy led to an increase in the plasma catecholamine concentration in females, as well as in the renal catecholamine content of both male and female rats. Gonadectomized males also showed a lower level of plasma catecholamine than the controls. The urinary flow, and the fractional excretion of sodium and chloride were significantly increased in gonadectomized males and in the OVX group when compared with their respective sham groups.