Proteomic analyses of Ehrlichia ruminantium highlight differential expression of MAP1-family proteins

The Rickettsiales Ehrlichia ruminantium (ER) is the causative agent of heartwater, a fatal tick-borne disease of livestock in sub-Saharan Africa and in the Caribbean, posing strong economical constraints to livestock production. In an attempt to identify the most prominent proteins expressed by this bacterium, especially those encoded by the major antigenic protein 1 (map1) multigene family, a proteome map of ER cultivated in endothelial cells was constructed by using two dimensional gel electrophoresis combined with mass spectrometry. Among the sixty-four spots detected, we could identify only four proteins from the MAP1-family; the other proteins detected were mainly related to energy, amino acid and general metabolism (26%), to protein turnover, chaperones and survival (21%) and to information processes (14%) or classified as hypothetical proteins (23%). Additional studies on MAP1-family protein using immunochemical labeling also revealed that these proteins are differentially expressed along the bacterium life cycle, presenting different structural organization. Interestingly, when infectious elementary bodies (EBs) are released from host cells, MAP1 appears to be organized in SDS and heat-resistant dimers and trimers stabilized by disulfide bridges. Overall, the results presented herein not only reveal the first partial proteome map of ER but provide new insights on the expression ER MAP1-family proteins in host endothelial cells.     

In situ adherence of Vibrio spp. to cryosections of Atlantic salmon, Salmo salar L., tissue.

Pathogenic and presumed non-pathogenic bacteria isolated from fish were tested for their adhesion to cryosections from different mucosal surfaces of Atlantic salmon, Salmo salar L. Adhered bacteria were detected by immunohistochemistry. Mucus was stained and fixed with Alcian blue after incubation of bacteria. The majority of the bacteria tested, i.e. Vibrio anguillarum serotype O1 , Vibrio salmonicida , Vibrio viscosus, Flexibacter maritimus and 'gut vibrios', i.e. Vibrio iliopiscarius and intestinal isolates of V. salmonicida , all adhered to mucus on all salmon epithelial surfaces tested, including sections from the foregut, hindgut, pyloric caeca, gills and skin. In contrast, V. anguillarum serotype O2, including both serotypes O2a and O2b, did not adhere to mucus, but did adhere to all other components of the tissues. As a positive control for adhesion of bacteria on cryosections, Escherichia coli was bound to piglet ileal mucosal lining, and as a negative control for adhesion, Staphylococcus aureus was found not to bind to any of the tissues tested. The present study shows that adhesion to mucus was not restricted to pathogenic bacteria, and furthermore, that not all pathogenic bacteria studied adhered to mucus. Hence, on the basis of these findings, the present authors suggest that V. anguillarum O2 may have an invasion strategy which does not involve adhesion to mucus, and thus, differs from the other pathogenic bacteria in the present study, which all bound to salmon mucus.     

Heterofucan from Sargassum filipendula induces apoptosis in HeLa cells

Fucan is a term used to denominate a family of sulfated polysaccharides rich in sulfated l-fucose. Heterofucan SF-1.5v was extracted from the brown seaweed Sargassum filipendula by proteolytic digestion followed by sequential acetone precipitation. This fucan showed antiproliferative activity on Hela cells and induced apoptosis. However, SF-1.5v was not able to activate caspases. Moreover, SF-1.5v induced glycogen synthase kinase (GSK) activation, but this protein is not involved in the heterofucan SF-1.5v induced apoptosis mechanism. In addition, ERK, p38, p53, pAKT and NFκB were not affected by the presence of SF-1.5v. We determined that SF-1.5v induces apoptosis in HeLa mainly by mitochondrial release of apoptosis-inducing factor (AIF) into cytosol. In addition, SF-1.5v decreases the expression of anti-apoptotic protein Bcl-2 and increased expression of apoptogenic protein Bax. These results are significant in that they provide a mechanistic framework for further exploring the use of SF-1.5v as a novel chemotherapeutics against human cervical cancer.     

Heterofucans from the brown seaweed Canistrocarpus cervicornis with anticoagulant and antioxidant activities

Fucan is a term used to denominate a family of sulfated polysaccharides rich in sulfated l-fucose. We extracted six fucans from Canistrocarpus cervicornis by proteolytic digestion followed by sequential acetone precipitation. These heterofucans are composed mainly of fucose, glucuronic acid, galactose and sulfate. No polysaccharide was capable of prolonging prothrombin time (PT) at the concentration assayed. However, all polysaccharides prolonged activated partial thromboplastin time (aPTT). Four sulfated polysaccharides (CC-0.3/CC-0.5/CC-0.7/CC-1.0) doubled aPTT with only 0.1 mg/mL of plasma, only 1.25-fold less than Clexane, a commercial low molecular weight heparin. Heterofucans exhibited total antioxidant capacity, low hydroxyl radical scavenging activity, good superoxide radical scavenging efficiency (except CC-1.0), and excellent ferrous chelating ability (except CC-0.3). These results clearly indicate the beneficial effect of C. cervicornis polysaccharides as anticoagulants and antioxidants. Further purification steps and additional studies on structural features as well as in vivo experiments are needed to test the viability of their use as therapeutic agents.     

Replacement of fish oil with vegetable oil blends in feeds for greater amberjack (Seriola dumerili) juveniles: Effect on growth performance,feed efficiency,tissue fatty acid composition and flesh nutritional value

This study was undertaken to assess the effects of fish oil (FO) substitution by a mixture of alternative vegetable oils (VO) on Seriola dumerili culture performance. A 154‐day feeding experiment was conducted using juveniles (39.2 ± 1.6 g average weight). Three isolipidic and isoenergetic meal‐based diets were formulated varying their lipid component. The control diet contained 100% FO (FO100), whereas diets VO50 and VO100 included 1/2 of oil blend and all the oil from blend of palm oil (PO) and linseed oil (LO) as substitute for FO, respectively. Dietary regime did not significantly affect growth performance, biometric indices, feed efficiency, plasma chemistry and liver and muscle lipid contents. Nonetheless, dietary VO inclusion impacted on the fatty acid profile of target tissues, especially in the liver. Fatty acid profiles of the fillets reflected those of the dietary oils except that there was apparent selective utilization of palmitic acid (C16:0) and oleic acid (C18:1n‐9) and apparent selective retention of long‐chain polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA, C20:5n‐3) and docosahexaenoic acid (DHA, C22:6n‐3). The nutritional value and the potential ability to prevent the development of coronary heart diseases of the flesh lipid fraction decreased with gradual FO substitution.     

Flow cytometry in Spermatology: A bright future ahead

Techniques such as mass spectrometry have led to unprecedented knowledge of the proteins that are present in the spermatozoa of humans and other mammals. However, in spite of their high‐throughput and fractioning techniques, most of the techniques in use only offer average values for the entire sperm population. Yet, ejaculate is very heterogeneous, and average values may mask relevant biological information.The application of flow cytometry may overcome this disadvantage, allowing proteomic analysis at the single‐cell level. Moreover, recent advances in cytometry, allowing multiple analyses within a single cell combined with powerful statistical tools, as an expanding subfield in spermatology, are described. The increased use of advanced flow cytometers in andrology laboratories will allow the rapid development of multiparametric, multicolour flow cytometry in andrology that will expand the clinical applications and research possibilities of flow cytometry‐based proteomic approaches, especially in the subfields of clinical andrology and sperm biotechnology.     

Maya Blue Paint: An Ancient Nanostructured Material

Maya blue paint was often used in Mesoamerica. The origin of its color and its resistance to acids and biocorrosion have not been fully understood. High-resolution transmission electron microscopy, electron energy loss spectroscopy, and x-ray microanalysis studies of authentic samples show that palygorskite crystals in the paint form a superlattice that probably occurs as a result of mixing with indigo molecules. An amorphous silicate substrate contains inclusions of metal nanoparticles encapsulated in the substrate and oxide nanoparticles on the surface. The beautiful tone of the color is obtained only when both the particles and the superlattice are present.     

The reversal of xylazine/ketamine immobilisation of fallow deer with yohimbine

Fallow deer were immobilised using a combination of xylazine and ketamine. Adult males (n = 10) and adult females (n = 10) received 4 mg/kg of each drug intramuscularly. Juveniles (n = 11) received 2 mg/kg of each drug, intravenously. Times to recumbency were as follows: adult males 4.9 +/- 2.9 min, adult females 4.1 +/- 1.9 min, juveniles 2.3 +/- 1.1 min. After 30 min each deer received 0.2 mg/kg of yohimbine, or an equal volume of sterile diluent intravenously. Yohimbine substantially reduced the recovery times of treated deer. Adults males were releasable 7.2 +/- 4.3 min after yohimbine administration, whereas control males were not releasable until 165 +/- 18 min. Treated adult females were releasable after 6.6 +/- 4.3 min, while control females were not releasable until 84 +/- 29 min. Juveniles were releasable 2.1 +/- 0.8 min after administration of yohimbine but control juveniles were not releasable until 62 +/- 16 min. Xylazine/ketamine administration produced statistically significant changes in packed cell volume, total plasma protein, albumin, sodium, glucose, creatine phosphokinase and inorganic phosphate values after 30 min. Yohimbine administration had no effect on these changes.     

Secondary Corpora Lutea Induced by hCG Treatment Enhanced Demi‐Embryo Survival in Lactating High‐Yielding Dairy Cows

Using a novel in vivo model considering a low developmental competence embryo (demi‐embryo) and a subnormal fertility recipient (lactating high‐yielding dairy cow), this experiment evaluated the effect of human chorionic gonadotrophin (hCG) treatment at embryo transfer (ET) on embryonic size at implantation, embryonic survival and recipient plasma progesterone (P₄) and bovine pregnancy‐specific protein B (PSPB) concentrations until day 63 of pregnancy. Embryos were bisected and each pair of demi‐embryos was bilaterally transferred to recipients (n = 61) on day 7 of the oestrous cycle. At ET recipients were randomly assigned to treatment with 1500 IU hCG or to untreated controls. Higher (p < 0.01) pregnancy rates on days 25, 42 and 63, and embryo survival rate on day 63 were observed in hCG‐treated cows with secondary CL than in hCG‐treated cows without secondary CL and in untreated cows. Pregnancy rates and embryo survival rate were similar in hCG‐treated cows without secondary CL and untreated cows. Embryonic size on day 42 was not affected by treatment with hCG, presence of secondary CL and type of pregnancy (single vs twin). Presence of secondary CL increased (p < 0.05) plasma P₄ concentrations of pregnant cows on days 14, 19 and 25 but not thereafter and of non‐pregnant cows on days 14–21. Treatment with hCG and presence of secondary CL had no effect on plasma PSPB concentrations, which were higher (p < 0.05) in twin than in single pregnancies. In conclusion, secondary CL induced by hCG treatment at ET significantly increased plasma P₄ concentrations, the survival rate of demi‐embryos and the pregnancy rate of high‐yielding lactating dairy cows. Embryos were rescued beyond maternal recognition of pregnancy, but later embryonic survival, growth until implantation and placental PSPB secretion until day 63 of pregnancy were not affected by treatment or presence of secondary CL.