Evaluation of the effect of probiotic cultures added to commercial yogurt over a known population of Listeria monocytogenes and Escherichia coli O157:H7

The effect of probiotic cultures over Listeria monocytogenes and Escherichia coli O157:H7 during yogurt storage was evaluated. Two different yogurt brands, one with additional probiotic cultures (Lactobacillus casei and L. acidophilus) were inoculated with known populations (106 UFC/g) of either L. monocytogenes or E. coli O157:H7 in three different times and stored for 32 days at 5 degrees C. Every four days the count of lactic bacteria, the added pathogens and pH was evaluated, according to the methodology described in the Bacteriological Analytical Manual. The pH and lactic bacteria population were constant during the testing period. Yogurt with additional probiotic cultures reduced the population of L. monocytogenes in 8 days, the population of E. coli O157:H7 in 16; yogurt with no additional probiotics took 20 days to reduce L. monocytogenes to non-detectable levels and even after 28 days of storage, E. coli O157:H7 was cultured. In this work, the beneficial effects of additional probiotic cultures in yogurt is confirmed again.     

Mapping the Nicotinic Acetylcholine Receptor Nanocluster Topography at the Cell Membrane with STED and STORM Nanoscopies

The cell-surface topography and density of nicotinic acetylcholine receptors (nAChRs) play a key functional role in the synapse. Here we employ in parallel two labeling and two super-resolution microscopy strategies to characterize the distribution of this receptor at the plasma membrane of the mammalian clonal cell line CHO-K1/A5. Cells were interrogated with two targeted techniques (confocal microscopy and stimulated emission depletion (STED) nanoscopy) and single-molecule nanoscopy (stochastic optical reconstruction microscopy, STORM) using the same fluorophore, Alexa Fluor 647, tagged onto either α-bungarotoxin (BTX) or the monoclonal antibody mAb35. Analysis of the topography of nanometer-sized aggregates (“nanoclusters”) was carried out using STORMGraph, a quantitative clustering analysis for single-molecule localization microscopy based on graph theory and community detection, and ASTRICS, an inter-cluster similarity algorithm based on computational geometry. Antibody-induced crosslinking of receptors resulted in nanoclusters with a larger number of receptor molecules and higher densities than those observed in BTX-labeled samples. STORM and STED provided complementary information, STED rendering a direct map of the mesoscale nAChR distribution at distances ~10-times larger than the nanocluster centroid distances measured in STORM samples. By applying photon threshold filtering analysis, we show that it is also possible to detect the mesoscale organization in STORM images.     

Down-regulation of brain muscarinic cholinergic receptor promoted by diacylglycerols and phorbol ester

Sustained agonist stimulation induces an asymmetric down-regulation of brain muscarinic acetylcholine receptor (mAChR): 43 +/- 2% in the right and 26 +/- 2% in the left cerebral hemisphere, respectively (Ref. 1). In order to determine the possible involvement of endogenous diacylglycerols produced under muscarinic stimulation in the down-regulation phenomenon, here we have studied the effects of synthetic diacylglycerols and a phorbol ester on cells dissociated from rat cerebral cortex. Oleylacetylglycerol decreased the amount of cell-surface mAChR by 37 +/- 2% and 25 +/- 2% in right and left cerebral cortex, respectively. Long-term treatment with phorbol dibutyrate also produced internalization of the mAChR (25 +/- 1.5% and 33 +/- 2% in right and left cortical cells, respectively). These changes occurred without modification of the Kdapp for the selective antagonist pirenzepine. The action of calcium ions was also studied using incubation of cells with the ionophore A23187. No changes were observed in the amount of mAChR detected at the plasma membrane with the ionophore alone, but when used in combination with phorbol dibutyrate and the agonist carbamylcholine a sinergistic decrease in mAChR was apparent. It is concluded that long-term exposure to exogenously added diacyglycerols and phorbol ester significantly reduces the amount of mAChR detected at the plasma membrane and abolishes the asymmetry of the down-regulation phenomenon observed under specific muscarinic stimulation, suggesting that diacylglycerols may be one of the factors responsible for such asymmetry.     

Crosslinking of unfilled carboxylated nitrile rubber with different systems: Influence on properties

The presence of two functional groups, nitrile and carboxyl, in carboxylated nitrile rubber allows it to be crosslinked with different agents. In this study, we examined the influence of different crosslinking agents on the properties of unfilled carboxylated nitrile rubber. Significant differences were found when different crosslinking agents were used, as shown in the vulcanization curves, especially in the variation of the viscous component with the reaction time. The reaction rate was highest when organic peroxide was used, and it was lowest when metallic peroxide or copper sulfate was used. When the crosslinking agents led to the formation of ionic bonds (metallic oxide and metallic peroxide), the carboxylic groups ? COOH had a greater participation in the crosslinking. However, when copper sulfate was used, coordination bonds were formed, and the main contribution was due to the nitrile groups. Tensile strength, tear strength, and abrasion properties were superior when the crosslinking systems used led to the formation of ionic bonds. On the contrary, the compression set was optimum when covalent bonds were formed. Copper sulfate behaved as an intermediate between the two previous situations. The mechanodynamic response of the compounds also depended on the crosslinking agent used. The correlation between crosslink density by swelling in dichloromethane, maximum damping temperature by dynamic measurements, and glass‐transition temperature by differential scanning calorimetry had to be explained in terms of the crosslink type. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Crosslinking of carboxylated nitrile rubber (XNBR) induced by coordination with anhydrous copper sulfate

BACKGROUND: Crosslinking induced by non‐covalent interactions in rubber materials is currently attracting a great deal of interest because of its perceived reversibility. Some thermoplastic elastomers and ionic elastomers, which are crosslinked by physical entanglements and ionic bonds, respectively, have already been investigated. The present work is centred on the study of the influence of coordination bonds on the physical properties of crosslinked materials. These bonds are formed on addition of copper sulfate. RESULTS: In this context, the effect of crosslinking of carboxylated nitrile rubber (XNBR) by anhydrous copper sulfate (CuSO₄) on crosslink reaction and mechanical and mechano‐dynamic properties is studied. The influence of different levels of CuSO₄ and the addition of conventional filler, carbon black, on the network is studied. The presence of nitrile (? CN) in the elastomeric chains leads to the formation of coordination bonds with the nitrile groups. The formation of ionic bonds between copper ions and carboxylic groups can be deduced from X‐ray photoelectron, attenuated total reflectance Fourier transform infrared and NMR analyses and vulcanization measurements. The dynamic response of the material does not show a secondary transition, the so‐called ionic transition typical of ionic elastomers, since ionic bonds do not lead to a separated ionic microphase, immersed in the elastomeric matrix. CONCLUSION: As expected, the crosslink density of the polymer network increases with the amount of crosslinking agent. Because of the presence of polymer–filler interactions, an increase in crosslink density is observed. However, these interactions seem not to affect the formation of coordination or ionic bonds. Copyright © 2008 Society of Chemical Industry     

High cancer susceptibility and embryonic lethality associated with mutation of the PTEN tumor suppressor gene in mice

BACKGROUND: Germ-line and sporadic mutations in the tumor suppressor gene PTEN (also known as MMAC or TEP1), which encodes a dual-specificity phosphatase, cause a variety of cancers such as Cowden disease, glioblastoma, endometrial carcinoma and prostatic cancer. PTEN is widely expressed, and Cowden disease consistently affects various organ systems, suggesting that the PTEN protein must have an important, although as yet poorly understood, function in cellular physiology. RESULTS: Homozygous mutant mice lacking exons 3-5 of the PTEN gene (mPTEN3-5) had severely expanded and abnormally patterned cephalic and caudal regions at day 8.5 of gestation. Embryonic death occurred by day 9.5 and was associated with defective chorio-allantoic development. Heterozygous mPTEN3-5 mice had an increased incidence of tumors, especially T-cell lymphomas; gamma-irradiation reduced the time lapse of tumor formation. DNA analysis of these tumors revealed the deletion of the mPTEN gene due to loss of heterozygosity of the wild-type allele. Tumors associated with loss of heterozygosity in mPTEN showed elevated phosphorylation of protein kinase B (PKB, also known as Akt kinase), thus providing a functional connection between mPTEN and a murine proto-oncogene (c-Akt) involved in the development of lymphomas. CONCLUSIONS: The mPTEN gene is fundamental for embryonic development in mice, as mPTEN3-5 mutant embryos died by day 9.5 of gestation, with patterning defects in cephalic and caudal regions and defective placentation. Heterozygous mice developed lymphomas associated with loss of heterozygosity of the wild-type mPTEN allele, and tumor appearance was accelerated by gamma-irradiation. These lymphomas had high levels of activated Akt/PKB, the protein product of a murine proto-oncogene with anti-apoptotic function, associated with thymic lymphomas. This suggests that tumors associated with mPTEN loss of heterozygosity may arise as a consequence of an acquired survival advantage. We provide direct evidence of the role of mPTEN as a tumor suppressor gene in mice, and establish the mPTEN mutant mouse as an experimental model for investigating the role of PTEN in cancer progression.     

"Cholos de Coclé": determination of their racial mixture and genetic origins]

We have found, in this first genetic study of a supposedly admixed Panamanian population, that the cultural group known as "cholos of Coclé" constitute a trihybrid mixture, whose genetic pool has the following composition: 44% Amerindian, 38% Caucasoid and 18% Negroid. Similarly, we have detected Amerindian genes, such as LDHB--Gua and TFchi, in proportions that relate this population with the extant Ngawbé (Guaymí). Nevertheless, the very high frequency of variant PEPA--KUN seems to indicate the genetic contribution of Amerindian populations from Eastern Panama, possibly from the extinct indigenous group cueva. This variant is frequently found among the present-day Kuna, but has not been detected among Nagawbé and Buglé.