New PCR Assays for the Identification of Fusarium verticillioides, Fusarium subglutinans, and Other Species of the Gibberella fujikuroi Complex

Fusarium verticillioides and Fusarium subglutinans are important fungal pathogens of maize and other cereals worldwide. In this study, we developed PCR-based protocols for the identification of these pathogens targeting the gaoB gene, which codes for galactose oxidase. The designed primers recognized isolates of F. verticillioides and F. subglutinans that were obtained from maize seeds from several producing regions of Brazil but did not recognize other Fusarium spp. or other fungal genera that were either obtained from fungal collections or isolated from maize seeds. A multiplex PCR protocol was established to simultaneously detect the genomic DNA from F. verticillioides and F. subglutinans. This protocol could detect the DNA from these fungi growing in artificially or naturally infected maize seeds. Another multiplex reaction with a pair of primers developed in this work combined with a pre-existing pair of primers has allowed identifying F. subglutinans, F. konzum, and F. thapsinum. In addition, the identification of F. nygamai was also possible using a combination of two PCR reactions described in this work, and another described in the literature.     

Mechanical and dynamical mechanical properties of chloroprene rubber and cellulose II composites

Vulcanized composites of chloroprene rubber (CR) with cellulose II (Cel II) as a filler were investigated. Cel II, obtained by the coagulation of cellulose xanthate, was incorporated in the rubber by the traditional method. The filler content varied from 0 to 30 phr. For comparison purposes, carbon black (CB)–CR composites were also studied. The CB amount varied from 0 to 45 phr. The mechanical and dynamic mechanical properties were determined, and the CR composite containing 20 phr of Cel II showed the best set of properties. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 2425–2430, 2004     

A novel electron beam-based method for the immobilization of trypsin on poly(ethersulfone) and poly(vinylidene fluoride) membranes

A novel technique for the covalent immobilization of trypsin in a one-step reaction using low-energy electron beam is described. The enzyme immobilization was applied on poly(ethersulfone) and poly(vinylidene fluoride) microfiltration membranes. For this purpose, the membranes were dipped in an aqueous solution of trypsin followed by electron beam treatment.The effect of irradiation conditions on the immobilization was investigated, as well as the resulting membrane properties with respect to enzymatic activity, immobilized enzyme concentration, pure water flux, scanning electron microscopy, and porosimetry. This technique shall provide a simple, inexpensive method for enzyme immobilization on various polymer membranes and offer a tool for the application in enzymatic membrane reactors.     

Genome-Wide Investigation of Multifocal and Unifocal Prostate Cancer—Are They Genetically Different?

Prostate cancer is widely observed to be biologically heterogeneous. Its heterogeneity is manifested histologically as multifocal prostate cancer, which is observed more frequently than unifocal prostate cancer. The clinical and prognostic significance of either focal cancer type is not fully established. To investigate prostate cancer heterogeneity, the genetic profiles of multifocal and unifocal prostate cancers were compared. Here, we report observations deduced from tumor-tumor comparison of copy number alteration data of both focal categories. Forty-one fresh frozen prostate cancer foci from 14 multifocal prostate cancers and eight unifocal prostate cancers were subjected to copy number variation analysis with the Affymetrix SNP 6.0 microarray tool. With the investigated cases, tumors obtained from a single prostate exhibited different genetic profiles of variable degrees. Further comparison identified no distinct genetic pattern or signatures specific to multifocal or unifocal prostate cancer. Our findings suggest that samples obtained from multiple sites of a single unifocal prostate cancer show as much genetic heterogeneity and variability as separate tumors obtained from a single multifocal prostate cancer.     

Scanning droplet cell investigations on single grains of a FeAlCr light weight ferritic steel

Polycrystalline Fe7.5Al7Cr is a ferritic light weight steel that shows a strong anisotropic dissolution behaviour. This electrochemical phenomenon was investigated by use of a scanning droplet cell (SDC). The crystallographic orientation of the single grains was determined by electron back scatter diffraction (EBSD). The fraction of grains with a near 〈1 1 1〉 orientation (<10°) was 1.88 times higher than that of the near 〈0 0 1〉 orientation prior to annealing and 3.58 times higher after annealing. Voltammograms were performed on single grains with low index planes. The critical current density was 53% higher on 〈0 0 1〉 grains as compared to 〈1 1 1〉 grains. These results are used to discuss the electrochemical behaviour of the polycrystalline FeAlCr.     

Effect of Bacterial Infection on Ghrelin Receptor Regulation in Periodontal Cells and Tissues

The effect of bacterial infection on the expression of growth hormone secretagogue receptor (GHS-R) was investigated in periodontal cells and tissues, and the actions of ghrelin were evaluated. GHS-R was assessed in periodontal tissues of rats with and without periodontitis. Human gingival fibroblasts (HGFs) were exposed to Fusobacterium nucleatum in the presence and absence of ghrelin. GHS-R expression was determined by real-time PCR and immunocytochemistry. Furthermore, wound healing, cell viability, proliferation, and migration were evaluated. GHS-R expression was significantly higher at periodontitis sites as compared to healthy sites in rat tissues. F. nucleatum significantly increased the GHS-R expression and protein level in HGFs. Moreover, ghrelin significantly abrogated the stimulatory effects of F. nucleatum on CCL2 and IL-6 expressions in HGFs and did not affect cell viability and proliferation significantly. Ghrelin stimulated while F. nucleatum decreased wound closure, probably due to reduced cell migration. Our results show original evidence that bacterial infection upregulates GHS-R in rat periodontal tissues and HGFs. Moreover, our study shows that ghrelin inhibited the proinflammatory actions of F. nucleatum on HGFs without interfering with cell viability and proliferation, suggesting that ghrelin and its receptor may act as a protective molecule during bacterial infection on periodontal cells.     

Inflammation in the Human Periodontium Induces Downregulation of the α1- and β1-Subunits of the sGC in Cementoclasts

Nitric oxide (NO) binds to soluble guanylyl cyclase (sGC), activates it in a reduced oxidized heme iron state, and generates cyclic Guanosine Monophosphate (cGMP), which results in vasodilatation and inhibition of osteoclast activity. In inflammation, sGC is oxidized and becomes insensitive to NO. NO- and heme-independent activation of sGC requires protein expression of the α₁- and β₁-subunits. Inflammation of the periodontium induces the resorption of cementum by cementoclasts and the resorption of the alveolar bone by osteoclasts, which can lead to tooth loss. As the presence of sGC in cementoclasts is unknown, we investigated the α₁- and β₁-subunits of sGC in cementoclasts of healthy and inflamed human periodontium using double immunostaining for CD68 and cathepsin K and compared the findings with those of osteoclasts from the same sections. In comparison to cementoclasts in the healthy periodontium, cementoclasts under inflammatory conditions showed a decreased staining intensity for both α₁- and β₁-subunits of sGC, indicating reduced protein expression of these subunits. Therefore, pharmacological activation of sGC in inflamed periodontal tissues in an NO- and heme-independent manner could be considered as a new treatment strategy to inhibit cementum resorption.     

Occurrence of Listeria and Enterobacteriaceae in domestic refrigerators

Consumers' refrigeration practices have a significant impact on the safety and quality of foods. To determine the prevalence and the identity of microorganisms in domestic refrigerators, swab samples were taken from various locations in the refrigerators from 137 households in middle Tennessee. The swabs were inoculated into different media, and standard procedures were used to characterize the isolates. API 20E and API Listeria were used for identification of Enterobacteriaceae and Listeria spp., respectively. The Kirby-Bauer technique was used to test resistance of the isolates. Actual counts for aerobic and Enterobacteriaceae ranged from not detected to 8.53 and 8.39 log CFU per sample, respectively. Klebsiella pneumoniae (23.4%), Klebsiella oxytoca (6.8%), Klebsiella terrigena (4.0%), Enterobacter sakazakii (2.2%), and Yersinia enterocolitica (0.7%) were some of the bacteria of concern that were isolated from domestic refrigerators. Resistance to antibiotics was most common in erythromycin (39.9%), followed by ampicillin (33.8%), cefoxitin (12.8%), tetracycline (5%), streptomycin (4.0%), nalidixic acid (2.1%), kanamycin (1.4%), and colistin (0.7%). None of the isolates tested was resistant to ciprofloxacin or gentamycin. Listeria spp. were also detected in six refrigerators. These findings underline the need for greater consumer education regarding proper refrigerator cleaning and safe food handling practices.     

The yield,fatty acid composition and physical properties of milk fat obtained by feeding soya oil to dairy cows

The presence in the bovine mammary gland of a desaturase specific for the conversion of stearic to oleic acid allows the manipulation of the physical properties of milk fat by varying the dietary intake of C₁₆ and C₁₈ fatty acids. In particular, feeding the dairy cow an oil with a high C₁₈: C₁₆ ratio should allow the proportion of milk fat that is liquid at 5°C to be increased, with a consequent improvement in the low-temperature spreadability of butter. In this paper, the effect of feeding concentrate rations containing (a) soya oil as the free oil and as cracked soya beans; (b) various levels of soya oil, the ration being offered twice or 24-times daily, on the thermal properties of the milk fat and on the yield of milk constituents is examined in detail. Feeding ratios containing free soya oil greatly increased the proportion of milk fat liquid at 5°C, whereas cracked soya oil gave only slight increases. Proportion and yield of milk fat were decreased by feeding soya oil twice daily but were increased by continuous feeding.     

Quantitative electroencephalographic profiles for children with autistic spectrum disorder.

The present study examined quantitative electroencephalographic (QEEG) profile for children with autistic spectrum disorder (ASD). Five-minute QEEG data were obtained from 90 normal controls (NCs) and 66 children with ASD. Spectrum analyses revealed that ASD children showed significantly less relative alpha and more relative delta than NC. Specifically, 26% of ASD children and 2% of NCs showed 1.5 SDs of relative alpha below the normative mean. Children with this QEEG profile had 17 times the risk of having ASD than those without such a profile. Sensitivity and specificity of relative alpha were 91% and 73%, respectively. Split-half cross-validation yielded a sensitivity of 76%. (PsycINFO Database Record (c) 2010 APA, all rights reserved)