Étude du mécanisme de protection des spermatozoïdes de mammifères par le lait

Le lait écrémé est utilisé depuis plus d’un demi-siècle comme diluant protecteur des spermatozoïdes de mammifères. Depuis quelques années, il existe une demande grandissante pour des diluants exempts de produits d’origine animale. Toutefois, le mécanisme par lequel le lait protège les spermatozoïdes n’est pas connu, ce qui rend difficile de lui trouver un substitut. Les protéines majeures du plasma séminal de taureau, les protéines « Binder of SPerm » (BSP), sont néfastes lors de la conservation de la semence. Les spermatozoïdes sont en contact avec une grande concentration de protéines BSP qui stimulent une extraction continuelle de cholestérol/phospholipides de leur membrane plasmique. Les lipoprotéines de faible densité (LDL) du jaune d’oeuf, un autre composé utilisé dans les diluants, empêcheraient les protéines BSP de se lier à la membrane des spermatozoïdes de taureaux et de stimuler un efflux des lipides membranaires, ce qui les protégerait durant la conservation. Notre hypothèse était que les protéines du lait protègent les spermatozoïdes durant la conservation en séquestrant les protéines BSP. Premièrement, nous avons démontré par filtration sur gel qu’il y a une interaction entre les protéines BSP bovines et les protéines du lait. Le lait écrémé a été fractionné en trois fractions : F1 (alpha-lactalbumine, bêta-lactoglobuline et caséine kappa), F2 (toutes les protéines du lait) et F3 (sels, sucres et petits peptides). Les protéines BSP1 et BSP5 ont une affinité plus grande pour F1 que BSP3, tandis que toutes les protéines BSP ont une affinité pour F2. Le titrage calorimétrique isotherme a permis de confirmer l’interaction entre les protéines BSP et les protéines du lait. L’association entre la protéine BSP1 bovine et les micelles de caséines est caractérisée par une constante d’affinité (Ka) de 3.5 × 10^5 M-1 et un paramètre stoichiométrique (n) de 4,5 BSP1 pour une caséine. L’association entre la protéine BSP1 bovine et l’alpha-lactalbumine (une protéine du sérum principale), est caractérisée par un Ka de 2.4 × 10^5 M-1 et une valeur “n” de 0,8. Ces résultats indiquent que le lait protège les spermatozoïdes bovins en séquestrant les protéines BSP grâce à une interaction protéine : protéine, tandis que le jaune d’oeuf les protège grâce à une interaction protéine : lipoprotéine. Deuxièmement, nous avons démontré par filtration sur gel que les protéines homologues aux BSP bovines retrouvées dans le plasma séminal de porc, d’étalon et de bélier ont une affinité avec les protéines du lait, ce qui suggère que le mécanisme de protection des spermatozoïdes par le lait pourrait être le même chez ces espèces. Troisièmement, nous avons caractérisé l’interaction entre BSP1 bovine et les LDL du jaune d’oeuf qui a un Ka de 3.4 ± 0.4 × 10^6 M-1 et une valeur de « n » de 104 BSP1 pour une particule de LDL, indiquant qu’il existe des différences entre le mécanisme de protection des spermatozoïdes par le lait et le jaune d’oeuf. Nous croyons que les résultats présentés dans cette thèse aideront à créer de nouveaux diluants ne contenant pas de produits d’origine animale afin de cryoconserver les spermatozoïdes des mammifères.

Chemical and Biochemical Studies on Natural Antioxidants from sesamum Species

In the present study on natural antioxidants, the focus has been kept mainly on oil seeds, especially sesame and its by-products. Sesame, which has been under cultivation in India for centuries is called the 'Queen of oil seed crops' because of the high yield of oil obtained and the nutritional qualities of the seed, oil, and meal. Though India is the largest producer of sesame in the world, research on the various health benefits of sesame has been carried out by Japanese Sesame has an important place in the foods and tradit..ional medicine of India from time immemorial. Foreseeing the potential of sesame and its byproducts as an important antioxidant source and its availability in bulk, the present study was focussed on Sesamum species. There are not many reports on the wild species of Sesamum in India, especially of the Kerala region. Hence, in the present study we also included antioxidants of Sesamurnrnalabaricumdistributed throughout the coastal region.The important characteristics of sesame are attributed to the presence of the umquc compounds lignans. Lignans arc a group of natural products of phenyl propanoid ongm, whieh are widely distributed in nature. They display important physiological functions in plants, in human nutrition and medicine, given their extensive health promotive and curative properties. Much interest has been focussed on their effectiveness as antineoplastic agents and research in this area has revealed several modes of action by which they can regulate the growth of mammalian cells. Sesame is an important source of furofuran lignans, of which sesamin and the rare oxygenated derivative sesamoIin are the most abundant. Others include sesamol and glucosides of lignans. Sesarnin and episesamin are reported to have hypocholesterolemic effect, suppressive effect on chemically induced cancer, alleviation of allergy symptoms etc. Sesamol, sesamolin and the lignan glycosides are reported to inhibit lipid peroxidation. Present investigation on sesame and its byproducts have been carried out to explore the possibility of developing a natural antioxidant extract from available resources to be used as a substitute to synthetic ones in vegetable oils and foods. Preliminary analysis showed that sesame cake, a byproduct could still be utilized as a major source of lignans. Sesame cake, which is now used only as a cattlefeed, can be better utilized in the form of a valuable antioxidant source. The present study explains the development of a feasible process for the extraction of antioxidant compounds from sesame cake. The antioxidant extract so prepared from sesame cake has been tested for vegetable oil protection and is found to be effective at low concentration. In addition, studies also include the antioxidant, radical scavenging, anticancer, mosquitocidal and pesticidal activities of extract and individual compounds.

Population Trends of a Mixed-Species Colony of Humboldt and Magellanic Penguins in Southern Chile after Establishing a Protected Area

Worldwide marine protected areas (MPAs) have been designated to protect marine resources, including top predators such as seabirds. There is no conclusive information on whether protected areas can improve population trends of seabirds when these are further exploited as tourist attractions, an activity that has increased in past decades. Humboldt Penguins (Spheniscus humboldti) and Magellanic Penguins (S. magellanicus) breed sympatrically on Puñihuil Islets, two small coastal islands off the west coast of Chiloé Island (41° S) in southern Chile that are subject to exploitation for tourism. Our goal was to compare the population size of the mixed colony of Humboldt and Magellanic Penguins before and after protection from unregulated tourism and freely roaming goats in 1997. For this purpose, two censuses were conducted in 2004 and 2008, and the numbers compared with those obtained in 1997 by other authors. The proportion of occupied, unoccupied, and collapsed/flooded burrows changed between years; there were 68% and 34% fewer collapsed burrows in 2004 and 2008, respectively, than in 1997. For the total number of burrows of both species, we counted 48% and 63% more burrows in 2004 and 2008, respectively, than in 1997. We counted 13% more burrows of Humboldt Penguins in 2008 than in 1997, and for Magellanic Penguins, we estimated a 64% increase in burrows in 2008. Presumably, this was as a result of habitat improvement attributable to the exclusion of tourists and the removal of goats from the islets. Although tourist visits to the islets are prohibited, tourism activities around the colonies are prevalent and need to be taken into account to promote appropriate management.

Macrophage antioxidant protection within atherosclerotic plaques

Macrophage cells within inflammatory lesions are exposed to a wide range of degrading and cytotoxic molecules including reactive oxygen species. Unlike neutrophils, macrophages do not normally die in this environment but continue to generate oxidants, phagocytose cellular remains, and release a range of cytoactive agents which modulate the immune response. It is this potential of the macrophage cell to survive in an oxidative environment that allows the growth and complexity of advanced atherosclerotic plaques. This review will examine the oxidants encountered by macrophages within an atherosclerotic plaque and describe some of the potential antioxidant mechanisms which enable macrophages to function within inflammatory lesions. Ascorbate, alpha-tocopherol, and glutathione appear to be central to the protection of macrophages yet additional antioxidant mechanisms appear to be involved. gamma-Interferon causes macrophages to generate 7,8dihydroneopterin/neopterin and 3-hydroxyanthranilic acid both of which have antioxidant properties. Manganese superoxide dismutase is also upregulated in macrophages. The evidence that these antioxidants provide further protection, so allowing the macrophage cells to survive within sites of chronic inflammation such as atherosclerotic plaques, will be described.

Peroxynitrite induced formation of the neurotoxins 5-S-cysteinyl-dopamine and DHBT-1: implications for Parkinson's disease and protection by polyphenols

Mechanisms of nigral cell injury in Parkinson's disease remain unclear, although a combination of increased oxidative stress, the formation of catecholamine-quinones and the subsequent formation of neurotoxic cysteinyl-catecholamine conjugates may contribute. In the present study, peroxynitrite was observed to generate both 2-S- and 5-S-cysteinyl-dopamine and a dihydrobenzothiazine species, DHBT-1, following the reaction of dopamine with L-cysteine. The formation of 5-S-cysteinyl-dopamine and DHBT-1 in the presence of peroxynitrite induced significant neuronal injury. Pre-treatment of cortical neurons with pelargonidin, quercetin, hesperetin, caffeic acid, the 4'-O-Me derivatives of catechin and epicatechin (0.1-3.0 mu M) resulted in concentration dependant protection against 5-S-cysteinyl-dopamine-induced neurotoxicity. These data suggest that polyphenols may protect against neuronal injury induced by endogenous neurotoxins relevant to the aetiology of the Parkinson disease. (C) 2008 Elsevier Inc. All rights reserved.

Leishmania amazonensis META2 protein confers protection against heat shock and oxidative stress

The META cluster of Leishmania amazonensis contains both META1 and META2 genes, which are upregulated in metacyclic promastigotes and encode proteins containing the META domain. Previous studies defined META2 as a 48.0-kDa protein, which is conserved in other Leishmania species and in Trypanosoma brucei. In this work, we demonstrate that META2 protein expression is regulated during the Leishmania life cycle but constitutive in T. brucei. META2 protein is present in the cytoplasm and flagellum of L amazonensis promastigotes. Leishmania META2-null replacement mutants are more sensitive to oxidative stress and, upon heat shock, assume rounded morphology with shortened flagella. The increased susceptibility of null parasites to heat shock is reversed by extra-chromosomal expression of the META2 gene. Defective Leishmania promastigotes exhibit decreased ability to survive in macrophages. By contrast, META2 expression is decreased by 80% in RNAi-induced T. brucei bloodstream forms with no measurable effect on survival or resistance to heat shock. (C) 2010 Elsevier Inc. All rights reserved.

Evaluation of the Conservation Lands of Maine Based on Presence of Habitat Suitable for Listed Species

“Conservation Lands” include both public and private lands that are devoted to the protection of wildlife and natural resources. “Listed Species” include federally and state endangered and threatened animals that are in danger of extermination in the state of Maine. The purpose of this study is to determine how well conservation lands are protecting the habitats of listed species. GAP data was used for 13 terrestrial vertebrate species indicating the presence or absence of suitable habitats. This data was compiled in GIS, generating a layer showing the number of listed species an area is suitable for. The areas that were suitable for at least one habitat were compared to answer three questions: (1) Is there is a difference between the presence and absence of listed species on protected lands and lands that are not protected? (2) Is there is a difference between the presence and absence of listed species on public lands managed by the state and the federal government and private land? (3) Is there is a difference between the presence and absence of listed species on lands protected under easements and lands that are protected fee simple? We found significant differences between all three categories. Conservation lands, private lands, and lands held under easement protect the habitat of listed species most effectively. We believe that this is due to the large number of private land trusts in the state of Maine and the effective management strategies of state lands.

Protection of insulin-producing cells against toxicity of dexamethasone by catalase overexpression

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detection of hybrids and genetic introgression in wild stocks of two catfish species (Siluriformes: Pimelodidae): The impact of hatcheries in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Protection of calves against Haemonchus placei and Haemonchus contortus after immunization with gut membrane proteins from H. contortus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bird species of black water inundation forests in the Jau National Park (Amazonas state, Brazil): their contribution to regional species richness

We compared the relative contribution of blackwater inundation forest (igapo) and unflooded (terra firme) forest for regional bird species diversity in the lower Rio Negro region. For analyses of habitat preference we used mist-net samples and an unpublished bird species check-list. The igapo forest had lower bird species diversity than the terra firme forest. However, some 14% of the forest bird species in the region are restricted to igapo forest. Species composition was found to be distinct between the understories of the two forest types. Common forest bird guilds in the Neotropics such as ant-following birds avoid igapo forest. The differential habitat selection among birds in the inundated and unflooded forest is likely to result from vegetation structure and biological interactions such as aggressive behaviour. Since the inundated forest has a typical biota, conservation efforts needs to be directed to protect these habitats. Unfortunately, inundation forests are threatened by several human actions, especially dam construction. The extent of protected areas in the Brazilian Amazon is insufficient for the adequate protection of the inundated habitats, because it includes only 3% of the total area.

XPS study on water corrosion of fluorzirconate glasses and their protection by a layer of surface modified tin dioxide nanoparticles

The surface corrosion process associated with the hydrolysis of fluorozirconate glass, Z-BLAN (53ZrF(4), 20BaF(2), 20NaF, 4LaF(2), 3AlF(3)), and the corrosion protection efficiency of a nanocrystalline transparent SnO2 layer were investigated by X-ray photoelectron spectroscopy. The tin oxide film was deposited by the sol-gel dip-coating process in the presence of Tiron(R) as particle surface modifier agent. The chemical bonding structure and composition of the surface region of coated and non-coated ZBLAN were studied before water contact and after different immersion periods (5-30 min). In contrast to the effects occurring for non-coated glass, where the surface undergoes a rapid selective dissolution of the most soluble species inducing the formation of a new surface phase consisting of stable zirconium oxyfluoride, barium fluoride and lanthanum fluoride species, the results for the SnO2-coated glass showed that the hydrolytic attack induces a filling of the film nanopores by dissolved glass material and the formation of tin oxylluoride and zirconium oxyfluoride species. This process results in a modified film, which acts as a hermetic diffusion barrier protecting efficiently the glass surface. (C) 2006 Elsevier B.V. All rights reserved.

AN IMPROVED ENRICHMENT PROCEDURE FOR THE ISOLATION OF YERSINIA-ENTEROCOLITICA AND RELATED SPECIES FROM MILK

A new enrichment procedure is proposed to improve the isolation of Yersinia enterocolitica and related species from milk. This procedure uses tryptic soy broth plus Polymyxin (5 IU/ml) and Novobiocin (10 mug/ml) - TSPN broth - incubated at 18-degrees-C for 3 d. Using raw milk and pasteurized milk inoculated with Yersinia strains, the efficiency of this procedure was compared to that of SB broth (sorbitol bile salts broth) incubated at 4-degrees-C for up to 21 d. Despite of the presence of antibiotics in TSPN broth, there were difficulties in recovering Yersinia organisms. Nevertheless, TSPN broth incubated at 18-degrees-C for 3 d showed better efficiency than that other method. In pasteurized milk samples, TSPN medium at 18-degrees-C for 3 d gave better results than the SB broth at 4-degrees-C for 7 d, showing that the proposed procedure is the preferable one due to the shorter period of incubation.

ANTI-HERBIVORE PROTECTION BY MUTUALISTIC SPIDERS and THE ROLE of PLANT GLANDULAR TRICHOMES

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aquatic macrophytes of Itaipu Reservoir, Brazil: Survey of species and ecological considerations

In a survey of the aquatic macrophytes of the Itaipu Reservoir, we identified 62 taxa in 25 families and 42 genera. The highest number taxa was observed for the emergent macrophytes (40 taxa). Reduced fluctuation in water level, increased light penetration, and sediment enrichment by nutrients and organic matter following the formation of the reservoir favored the appearance of a species-rich community of submerged macrophytes (23% of the taxa identified). The aquatic macrophytes were found mainly near the mouths of the main tributaries of the reservoir, in shallow area of depth less than 2 meters. In addition to the shallow depth, the greater nutrient input from the tributaries and relative protection from wind explained this distribution. Among the species found, Egeria najas merits mention for its occurrence in all localities sampled, with biomass values varying between 98 and 186 gDW/m2. Some potential nuisance species such as Eichhornia crassipes, Salvinia auriculata, and Pistia stratiotes also deserve attention, since they were also observed to be covering large areas of Itaipu Reservoir. © 1999, Paraná Institute of Technology Publication.