Combined effect of synthetic enterocin CRL35 with cell wall,membrane‐acting antibiotics and muranolytic enzymes against Listeria cells

Aims: To evaluate the inhibition effectiveness of enterocin CRL35 in combination with cell wall, membrane‐acting antibiotics and muranolytic enzymes against the foodborne pathogen Listeria. Methods and Results: Synthetic enterocin CRL35 alone and in combination with monensin, bacitracin, gramicidin, mutanolysin and lysozyme were used in this study. Minimal inhibitory concentration (MIC) and fractional inhibitory concentration (FIC) index assays were performed using Listeria innocua 7 and Listeria monocytogenes FBUNT as sensitive strains. Antibiotics showed positive interactions with the bacteriocin in both strains tested. On the other hand, when mutanolysin and enterocin CRL35 were added to resting cells in a buffer system, the lytic effect of mutanolysin was enhanced. However, the addition of mutanolysin showed no effect on the growth of L. innocua 7 cells in a culture medium. Moreover, mutanolysin allowed the overgrowth of L. innocua 7 cells to an OD similar to control cells in the presence of inhibitory concentration of enterocin CRL35. In contrast, the combination of lysozyme and enterocin CRL35 resulted in a 50% inhibition of the L. innocua 7 growth. Conclusions: Based on our results, we conclude that the combination of synthetic enterocin CRL35 with some antibiotics is effective against L. innocua 7 and L. monocytogenes FBUNT cells, and more importantly the amount of these agents to be used was considerably reduced. The effectiveness of the combination of synthetic enterocin CRL35 with muramidases seems to depend on complex environments, and more detailed studies need to be performed to elucidate this issue. Significance and Impact of the Study: Enterocin CRL35 represents a promising agent that not only can ensure the quality and safety of food but it can also be combined with several antimicrobial agents important in the medical field.     

New differences between the Wistar rat and Octodon degus, a putative laboratory animal resistant to morphine

1. The effects of CNS depressants (methadone and alcohol) and natural neurotransmitters (NA and ACh) are studied in O. degus. 2. O. degus shows resistance to methadone in the formalin algesiometric test and EEG. 3. Ethanol elimination profile suggest the presence of an atypical alcohol dehydrogenase in O. degus 4. O. degus is extremely resistant to the pressor effects of noradrenaline 5. the isolated atrium of this rodent is 40 times more sensitive to the negative chronotropic effect of methadone, than the rat atrium. 6. These effects could be explained in terms of an important catecholamine and endorphin co-secretion from adrenal glands in O. degus.     

Human T cell clone identifies a potentially protective 54-kDa protein antigen of Toxoplasma gondii cloned and expressed in Escherichia coli.

Protective immunity against Toxoplasma gondii is recognized to be cell mediated and IFN-gamma is considered to be the major mediator of resistance. Thus, protective Ag of the parasite must induce IFN-gamma-producing T cells. In order to identify such Ag, we have constructed a T. gondii cDNA library in the cloning/expression vector lambda gt11, screened this library with a pool of sera of immune donors, and further screened the set of selected recombinant Ag using, as probe, a T. gondii-reactive T-cell clone (TCC) derived from an infected/immune individual and producing a high level of IFN-gamma. One recombinant Ag was shown to induce TCC proliferation and was characterized. The corresponding mature T. gondii Ag has an apparent molecular mass of 54 kDa and the sequence of the cDNA clone suggests that it is membrane associated. The epitope defined by the TCC on this Ag was found to be present in three Toxoplasma strains independently of their phenotype (virulent or cyst forming). Recognition of this Ag by the TCC was shown to be restricted by HLA-DPw4, the most frequent allele in the Caucasian population (approximately 40%). The use of this Ag as a vaccine component is proposed.     

Influence of resin cement rigidity on the stress distribution of resin-bonded fixed partial dentures

The mechanical properties of the adhesive cement used in resin-bonded fixed partial dentures (RBFPD) can modify the clinical performance of the rehabilitation. The goal of this study was to evaluate the influence of the elastic modulus of different cements on the stress distribution in RBFPD using finite element analysis. For that an anterior 3-unit prosthesis was modeled based in a stereolithography file. The model was meshed with tetrahedral elements and materials considered isotropic, linearly elastic and homogeneous. The force applied to the palatal area of the lateral incisor (pontic) at 45° was 100?N. The cements used presented 7 different elastic modulus (E): 2, 6, 10, 14, 18, 22 or 26?GPa. The total deformation, von-Mises stress and maximum principal stress criteria were used to calculate the results. The lower tensile stress occurred in the cement layer with E?=?2?GPa [25.6 (canine) and 16.32?MPa (incisor)]. For the prosthesis, the model with the lower tensile stress [287 (canine) and 248?MPa (incisor)] occurred when the cement presented E?=?26?GPa.

In this way, the stress concentration may have its magnitude modified depending on the stiffness of the cement. Since more flexible cements concentrate less tensile stress in its structure, but allow an increased displacement of the prosthesis, which is friable and rigid and ends up concentrating more tensile stress at its connector. In that way the clinician should avoid the use of adhesive cement with lower elastic modulus due to it increases the stress concentration in the ceramic.     

Six Commercially Available Angiotensin II AT1 Receptor Antibodies are Non-specific

Commercially available Angiotensin II AT₁ receptor antibodies are widely employed for receptor localization and quantification, but they have not been adequately validated. In this study, six commercially available AT₁ receptor antibodies were characterized by established criteria: sc-1173 and sc-579 from Santa Cruz Biotechnology, Inc., AAR-011 from Alomone Labs, Ltd., AB15552 from Millipore, and ab18801 and ab9391 from Abcam. The immunostaining patterns observed were different for every antibody tested, and were unrelated to the presence or absence of AT₁ receptors. The antibodies detected a 43?kDa band in western blots, corresponding to the predicted size of the native AT₁ receptor. However, identical bands were observed in wild-type mice and in AT_1A knock-out mice not expressing the target protein. Moreover, immunoreactivity detected in rat hypothalamic 4B cells not expressing AT₁ receptors or transfected with AT_1A receptor construct was identical, as revealed by western blotting and immunocytochemistry in cultured 4B cells. Additional prominent immunoreactive bands above and below 43?kDa were observed by western blotting in extracts from tissues of AT_1A knock-out and wild-type mice and in 4B cells with or without AT₁ receptor expression. In all cases, the patterns of immunoreactivity were independent of the AT₁ receptor expression and different for each antibody studied. We conclude that, in our experimental setup, none of the commercially available AT₁ receptor antibodies tested met the criteria for specificity and that competitive radioligand binding remains the only reliable approach to study AT₁ receptor physiology in the absence of full antibody characterization.     

Expression of the ubiE Gene of Geobacillus stearothermophilus V in Escherichia coli K-12 Mediates the Evolution of Selenium Compounds into the Headspace of Selenite- and Selenate-Amended Cultures

The ubiE gene of Geobacillus stearothermophilus V, with its own promoter, was cloned and introduced into Escherichia coli. The cloned gene complemented the ubiE gene deficiency of E. coli AN70. In addition, the expression of this gene in E. coli JM109 resulted in the evolution of volatile selenium compounds when these cells were grown in selenite- or selenate-amended media. These compounds were dimethyl selenide and dimethyl diselenide.     

Identification of biogenic dimethyl selenodisulfide in the headspace gases above genetically modified Escherichia coli

Escherichia coli JM109 cells were modified to express the genes encoded in a 3.8-kb chromosomal DNA fragment from a metalloid-resistant thermophile, Geobacillus stearothermophilus V. Manual headspace extraction was used to collect the gases for gas chromatography with fluorine-induced sulfur chemiluminescence analysis while solid-phase microextraction was used for sample collection in gas chromatography/mass spectrometry (GC/MS) analysis. When grown in the presence of selenate or selenite, these bacteria produced both organo-sulfur and organo-selenium in the headspace gases above the cultures. Organo-sulfur compounds detected were methanethiol, dimethyl sulfide, dimethyl disulfide, and dimethyl trisulfide. Organo-selenium compounds detected were dimethyl selenide and dimethyl diselenide. Two mixed sulfur-selenium compounds, dimethyl selenenyl sulfide and a chromatographically late-eluting compound, were detected. Dimethyl selenodisulfide, CH(3)SeSSCH(3), and dimethyl bis(thio)selenide, CH(3)SSeSCH(3), were synthesized and analyzed by GC/MS and fluorine-induced chemiluminescence to determine which corresponded to the late-eluting compound that was bacterially produced. CH(3)SeSSCH(3) was positively identified as the compound detected in bacterial headspace above Se-amended cultures. Using GC retention times, the boiling point of CH(3)SeSSCH(3) was estimated to be approximately 192 degrees C. This is the first report of CH(3)SeSSCH(3) produced by bacterial cultures.