Induction, by adriamycin and mitomycin C, of modifications in lipid composition, size distribution, membrane fluidity and permeability of cultured RDM4 lymphoma cells

Adriamycin and mitomycin C were previously found to modulate the sensitivity of lymphoma cells to lysis by certain effectors of immunity and this modulation was dependent on drug concentration. In the present studies, RDM4 lymphoma cells were treated with different concentrations of the two drugs for 24 h in culture. These treatments resulted in changes in the lipid composition, membrane fluidity, cell size distribution, and permeability to 51CrO4, Trypan blue, Acridine orange and trimethylaminodiphenylhexatriene (TMA-DPH) of the cells. Changes in some of these parameters, as a function of drug concentration, resulted in dose-response curves which were bell-like shaped, hence paradoxical similarities between non-drug-treated cells and cells treated with higher drug concentrations were observed.     

Current knowledge of fungi from Neotropical montane cloud forests: distributional patterns and composition

Montane cloud forests in the Neotropics harbor a great wealth of biological diversity and a large number of endemic species. Here, we present (i) a comprehensive data mining exercise of fungi from Neotropical montane cloud forests (NMCF), (ii) an extensive review of the current knowledge of fungal richness, distribution and composition, and (iii) a preliminary analysis of fungal endemicity in Mexican montane cloud forests. Based on a survey of literature and other sources, we assembled a database of 6349 records representing 2962 fungal species in NMCF. The computed individual-based species rarefaction curve remained non-asymptotic, and the extrapolation curve estimated an expected increment of 42% in the number of species by doubling the sampling effort. Fungal species richness was highest in NMCF from Mesoamerica, particularly from Mexico and Costa Rica. Fungi from Mesoamerica, Caribbean and South America are significantly different at diverse taxonomic levels, and there is a little overlap in the fungal species recorded from these regions. The analyses of endemicity of the Mexican dataset performed with parsimony and Bayesian methods were highly complementary. They showed the following areas of endemicity supported by the congruent distribution of fungal species: (i) two main regions in the Trans-Mexican Volcanic Belt (TMVB); (ii) a region in the southern part of Veracruz; and (iii) a region located in the eastern part of TMVB with affinities with Sierra Madre Oriental and the Chiapan-Guatemala Highlands. This last area was supported by five species of Glomeromycota and is consistent with an area of endemicity previously found in vascular plants. In this study, we provide a perspective on gaps in knowledge regarding the diversity and distribution of fungi in NMCF, and provide a full dataset of fungal records with geographical, bibliographic and taxonomic information.     

Biogeography, current knowledge and conservation of threatened vascular plants characteristic of Mexican temperate forests

The main goal of this study is to document the biogeographical patterns, current status and conservation of 24 species of vascular plants in Mexico, all of them recorded in some risk category in the Mexican official publication named ‘Norma Oficial Mexicana 059’ (NOM-059) and some of them in the IUCN red lists (the World Conservation Union) and CITES (Convention on International Trade in Endangered Species); many of these species are linked to Mexican cloud forests and other temperate vegetation types. Distribution maps of these 24 species were generated with information obtained from specialized literature, herbarium specimens, institutional databases, and field work; with this information, the patterns of richness and endemism of these species were obtained. For this, the Mexican territory was divided using a grid system based on a chart index (scale 1:50,000 system composed by grids of 15' × 20'). Also, we up to date the knowledge of these species and their populations and current habitats, and evaluate their geographic distribution in relation to the current Mexican National Parks and Reserves System and Mexican Priority Regions for Conservation. We also discuss some changes of categories in the NOM-059 and suggest the urgent incorporation of some of the species in the recent IUCN Red Data List of Threatened Species (2003). Most of the species studied require special policies for their conservation due to problems that affect their natural populations; also we consider that these taxa are underrepresented in the current Mexican System of Natural Protected Areas, especially those with restricted distributions.     

Cytoskeletal proteins F-actin and β-dystrobrevin are altered by the cryopreservation process in bull sperm

The cryopreservation process has an important impact on sperm structure and physiology. The negative effects have been mainly observed on the plasma membrane, which is directly stabilized by the cytoskeleton. Since cytoskeleton proteins are osmosensitive and thermosensitive, the aim of this study was to evaluate the damage caused to the bull sperm cytoskeleton by cryopreservation (freezing-thawing). Fresh and frozen-thawed bull semen samples were exposed to a treatment with the neutral detergent Brij 36-T. Electron microscopy evidenced important damages at the sperm perinuclear theca after the protein extraction protocol; the perinuclear theca was partially solubilized, the perinuclear theca substructure disappeared in the cryopreserved samples. Furthermore, the sperm head's shape was significantly altered on the cryopreserved samples. Fluorescence analysis showed a decrease of the intensity of actin and dystrobrevin on the frozen-thawed samples. Western blot assays revealed a stronger signal for actin and β-dystrobrevin in the frozen-thawed sperm samples than in the fresh ones. Our results suggest that the cryopreservation process highly alters the sperm cytoskeleton stability, causing its proteins to become more fragile and therefore more susceptible to be extracted.     

Visualization of different types of hapten-cell membrane interaction by fluorine high-field nuclear magnetic resonance

Using a high field spectrometer (5.9 Tesla) 19F-NMR spectrum of soluble material from 4 trifluoromethyl 2,6 dinitrobenzene sulphonate (CF3-DNBS) treated murine lymphocytes was recorded. CF3-DNBS is a fluorinated analog of 2,4,6 trinitrobenzene sulphonate (TNBS), and these compounds have been found to create cross-reacting antigenic modifications of cell surface. At least 4 distinguishable signals have been detected, and we think that 19F-NMR could be used to study, at a molecular level, some immunochemical problems concerning modification with TNBS.     

Patterns of diversity,endemism and conservation: an example with Mexican species of Ternstroemiaceae Mirb. ex DC. (Tricolpates: Ericales)

The Ternstroemiaceae is a family of vascular plants with many taxonomic problems, basically in the delimitation of some of its species. Little is known about its world distribution and about the real state of conservation of its species. An analysis of the distribution patterns of the Mexican species was undertaken to provide information about the present status of species and possible conservation areas. Also, an analysis of the species richness, endemism centres, extension and location of the areas occupied by these species with a 1:50000 chart index was made. Finally, we propose some of the species of this family to be included in the IUCN Red List of Threatened Species and in the next Mexican Norma Oficial Mexicana 059.     

The association of the rabies glycoprotein with liposome (immunosome) induces an in vitro specific release of interleukin 2

BALB/c mice were primed by receiving a unique intraperitoneal injection of rabies virus antigens presented as complete inactivated virus (P.V. strain) or as purified glycoproteins either in the aggregated form or in physical combination with liposomes (i.e., in the form of "immunosomes"). The splenocytes of these mice were restimulated, 6-15 days after priming, in culture with rabies virus antigens, and antigen-specific IL-2 production was measured. It was found that rabies antigens presented as immunosomes were as active as the inactivated virus, whereas equivalent amounts of purified glycoproteins were inactive. The optimal amounts of rabies immunosomes used for priming was found to be 0.5 to 0.05 micrograms per mouse.     

Role of liposomes in the presentation of HIV envelope glycoprotein and the immune response in mice

The role of antigen presentation in the induction of humoral as well as cell-mediated immune responses has been investigated by anchoring HIV-1 envelope glycoprotein (gp 160/120) into the phospholipidic bilayer of preformed liposomes to produce HIV-Immunosomes. HIV-Immunosomes induced high titres of HIV-specific antibodies when tested by ELISA, IFA and neutralization, whereas equal amounts of purified glycoprotein alone produced lower antibody response. Similarly, HIV-Immunosomes induced antigen-specific Interleukin-2 production and blastogenic response upon restimulation with the same antigen, in animals vaccinated with HIV-Immunosomes, whereas no secondary response was observed in animals vaccinated with equal amounts of purified gp 160/120. Taken together, these results underline the importance of antigen presentation in the establishment of an adequate immune status and show the potential of HIV-Immunosomes as vaccine against AIDS.