The p53 isoforms are differentially modified by Mdm2

The discovery that the single p53 gene encodes several different p53 protein isoforms has initiated a flurry of research into the function and regulation of these novel p53 proteins. Full-length p53 protein level is primarily regulated by the E3-ligase Mdm2, which promotes p53 ubiquitination and degradation. Here, we report that all of the novel p53 isoforms are ubiquitinated and degraded to varying degrees in an Mdm2-dependent and -independent manner, and that high-risk human papillomavirus can degrade some but not all of the novel isoforms, demonstrating that full-length p53 and the p53 isoforms are differentially regulated. In addition, we provide the first evidence that Mdm2 promotes the NEDDylation of p53β. Altogether, our data indicates that Mdm2 can distinguish between the p53 isoforms and modify them differently.     

The effect of sperm preparation and co-incubation time on in vitro fertilization of Bos indicus oocytes.

The objective of the present study was to evaluate the effect of various methods of sperm selection and various sperm-oocyte co-incubation times on in vitro fertilization (IVF) of zebu (Bos indicus) oocytes. Frozen semen from one ejaculate of a single bull was used for all treatments and replicates. After thawed, sperm was subjected to one of the three treatments: 45 and 90% discontinuous Percoll gradient, swim-up and washing by centrifugation. In all treatments, the spermatozoa were incubated with in vitro matured oocytes for 3, 6, 12 and 18h. After co-incubation oocytes were transferred to the culture medium and culture for 44h, when the cleavage was evaluated. The uncleavaged oocytes were fixed and stained to determine penetration, pronucleus formation and polyspermy. The sperm selection method did not influence (P<0.05) polyspermy, pronucleus formation, penetration and cleavage rates. No interaction between method of selection and sperm-oocyte co-incubation time was observed (P>0.05). However, sperm-oocyte co-incubation time affected fertilization. The lower penetration (26.5%) and cleavage rates (13.1%) were obtained at 3-h period. The penetration and cleavage percentages increased (P<0.05) progressively at 6h (63.3 and 54.4%) and 12h (77.6 and 67.6%). No differences (P>0.05) were observed between 12 and 18h of incubation for penetration and cleavage rates. The incidence of polyspermy and pronucleus formation was similar (P>0.05) for all time points. It is concluded that the methods used in this study for sperm selection do not affect fertilization; therefore, they all can be used for bovine IVF. In addition, regardless the method used better fertilization results were obtained when sperm and oocytes were co-incubated for 12h, and the prolongation of that time for up to 18h had no detrimental effect on fertilization.     

Membrane microheterogeneity: Förster resonance energy transfer characterization of lateral membrane domains

Lateral membrane heterogeneity, in the form of lipid rafts and microdomains, is currently implicated in cell processes including signal transduction, endocytosis, and cholesterol trafficking. Various biophysical techniques have been used to detect and characterize lateral membrane domains. Among these, Förster resonance energy transfer (FRET) has the crucial advantage of being sensitive to domain sizes smaller than 50-100 nm, below the resolution of optical microscopy but, apparently, similar to those of rafts in cell membranes. In the last decade, several formalisms for the analysis of FRET in heterogeneous membrane systems have been derived and applied to the study of microdomains. They are critically described and illustrated here.     

Sex differences in the electrocommunication signals of the electric fish Apteronotus bonapartii

The South American weakly-electric knifefish (Apteronotidae) produce highly diverse and readily quantifiable electrocommunication signals. The electric organ discharge frequency (EODf), and EOD modulations (chirps and gradual frequency rises (GFRs)), vary dramatically across sexes and species, presenting an ideal opportunity to examine the proximate and ultimate bases of sexually dimorphic behavior. We complemented previous studies on the sexual dimorphism of apteronotid communication signals by investigating electric signal features and their hormonal correlates in Apteronotus bonapartii, a species which exhibits strong sexual dimorphism in snout morphology. Electrocommunication signals were evoked and recorded using a playback paradigm, and were analyzed for signal features including EOD frequency and the structure of EOD modulations. To investigate the androgenic correlates of sexually dimorphic EOD signals, we measured plasma concentrations of testosterone and 11-ketotestosterone. A. bonapartii responded robustly to stimulus playbacks. EODf was sexually monomorphic, and males and females produced chirps with similar durations and amounts of frequency modulation. However, males were more likely than females to produce chirps with multiple frequency peaks. Sexual dimorphism in apteronotid electrocommunication signals appears to be highly evolutionarily labile. Extensive interspecific variation in the magnitude and direction of sex differences in EODf and in different aspects of chirp structure suggest that chirp signals may be an important locus of evolutionary change within the clade. The weakly-electric fish represent a rich source of data for understanding the selective pressures that shape, and the neuroendocrine mechanisms that underlie, diversity in the sexual dimorphism of behavior.     

Familial breast/ovarian cancer and BRCA1/2 genetic screening: the role of immunohistochemistry as an additional method in the selection of patients.

Only 20-25% of families screened for BRCA1/2 mutations are found positive. Because only a positive result is informative, we studied the role of BRCA1/2 immunohistochemistry as an additional method for patient selection. From 53 high-risk-affected probands, 18 (34%) had available paraffin blocks of their tumors and were selected for this study. Mutation screening was done by conformation-sensitive gel electrophoresis and multiplex ligation-dependent probe amplification. For immunohistochemistry, 21 neoplastic specimens (15 breast carcinomas, 5 ovary neoplasms, and 1 rectal adenocarcinoma) were analyzed with BRCA1 (monoclonal antibody, Ab-1, oncogene) and BRCA2 (polyclonal antibody, Ab-2, oncogene) antibodies. Absence of the BRCA1 protein was confirmed in negative tumors by Western blotting. Seven patients were positive for BRCA1/2 mutations: 5 for BRCA1 and 2 for BRCA2. Four out of five positive patients had tumors negative for BRCA1 immunostaining, and the remaining 13 BRCA1-negative patients had positive BRCA1 immunostaining in all tumor samples. Sensitivity to predict for BRCA1 mutation carriers was 80%, and specificity was 100%, with a positive predictive value of 100% and a negative predictive value of 93%. This correlation was statistically significant (p=0.001). No correlation was observed for BRCA2. If larger studies confirm these results, high-risk patients with BRCA1-negative tumors should be screened first for this gene.     

Comparative study of helminth parasites of fishes from the Mogi Guassu river, collected during expeditions between 1927 and 1985

This paper concerns the parasitism of Nematoda, Trematoda, Acanthocephala and Cestoda from 1027 fishes of 45 species from Mogi Guassu river, examined in the years 1927, 1946, 1947, 1962, 1983 and 1985 as a contribution to the knowledge of the parasitological fauna in 58 years. Thirty two species of nematodes, 21 of trematodes, 3 of acanthocephalans and 2 of cestodes are reported. New hosts are presented for the nematodes Cucullanus pinnai, Spirocamallanus inopinatus and Travnema travnema, for the trematode Pararhipidocotyle jeffersoni and for the acanthocephalan Gorytocephalus spectabilis. New measurements for the trematode Creptotrema lynchy, tables, graphics and a list of the examined hosts with their parasites are presented.     

Revision of the Brazilian species of the genus Halipegus Looss, 1899 (Trematoda: Derogenidae)

Halipegus dubius Klein, 1905 is redescribed and illustrated from a Brazilian frog Leptodactylus ocellatus. The type-material of Halipegus genarchella (Travassos, Artigas & Pereira, 1928) and Halipegus parva (Travassos, Artigas & Pereira, 1928) are redescribed with original figures. New synonyms are proposed: Halipegus salmini Yamaguti, 1958 is considered a synonym of H. genarchella, H. tropicus (Manter, 1936) sensu Kloss (1966) of H. parva and Genarchella lenti Fábio, 1984 of H. overstreeti (Brooks, Mayes & Thorson, 1979) n. comb.     

Behavioral changes in pottos (Perodicticus potto): Effects of naturalizing an exhibit

Observations of the potto (Perodicticus potto),a nocturnal prosimian primate,indicated a limited behavioral repertoire and prompted renovation of their exhibit at the Frankin Park Zoo (Boston, MA). We used the natural history of this species to direct the exhibit modifications, which used only nonsynthetic items. We added live plants, soil, bamboo, shelf fungi, grapevines, a hollow tree, and a log containing an insect dispenser to the exhibit at little expense. They provided new textures, odors, pathways, stimulation and cover for this secretive species. The changes produced a heightened aesthetic appeal to the nocturnal exhibit and improved its educational value by representing the animals’ habitat more accurately. The exhibit modifications also resulted in a significant increase in activity, an expansion of the pottos’ behavioral repertoire,the emergence of sexual behaviors, and an increased visibility to the public.     

The management of European estuaries: A comparison of the features,controls and management framework of the Tagus (Portugal) and Humber (England)

The paper describes the local, national, European and wider-area framework, statutes, and formal and voluntary mechanisms for managing European estuaries. These aspects are discussed in relation to two large and representative estuarine systems, the Tagus, Portugal, and the Humber, on the English North Sea coast As estuaries are sites of many activities and uses, most of which are encouraged or at least condoned, management has the role of preventing and resolving conflicts between those uses and users. Accepted uses of estuaries include the discharge and dumping of waste materials, fin and shell-fisheries, conservation, land reclamation, natural usage, abstraction by industry, and recreation. Estuarine management is now being carried out within the constraints of local and regional government planning, planning and activities of water pollution control bodies, fisheries control bodies, and navigation and port authorities The Tagus and Humber estuaries support all of the above activities and uses, and have controls within a European legislative framework but have differing histories of management and planning in order to resolve conflicts. In addition the Humber is subject to controls placed on North Sea areas. The paper discusses the relevant national and European legislation (Directives) and accepted practices for management. Furthermore, the paper discusses the formulation and practice of estuarine management plans as used by various bodies (nature conservation, water quality and regional authority). It is of particular note that the lessons from these two estuaries are relevant to many other European estuaries.