Discriminating analysis of “in vitro” prostaglandin release by myometrial and luminal sides of the ewe endometrium

An original perifusion device which allows a discrimination between the 30 mn releases of prostaglandins F₂α and E₂ by the luminal and the myometrial faces of sheep endometrium is described. Tissue was sampled on day 4, 14, 16 or 17 of the cycle and on day 14 or 17 of pregnancy. Total prostaglandin (PG) release measured with this device was in good agreement with PG's concentrations in media of in vitro endometrium incubations already described.Discrimination analysis of the PGs release by each side of the endometrial tissue during the 30 mn perifusion time releaved that PGF₂α concentrations of the perifusion medium issued from the lumen compartment were higher than those of the myometrial compartment in all physiological status where corpus luteum is active (including early pregnancy). Therefore in the ewe, it seems that luteal structure maintenance during early pregnancy is not due, as in the giltm to a shift in PGF₂α secretion towards the uterine lumen.     

Identification and genetic diversity analysis of strain Pseudomonas syringae S5: A pathogen responsible for bacteriosis in Avena sativa plants

The genus Pseudomonas includes pathogenic species P. syringae, which can be found in various agricultural environments and which can affect a wide variety of plants, causing significant economic losses when the environmental conditions for its proliferation are optimal. Comprehensive characterizations of phytopathogenic bacteria belonging to the genus Pseudomonas are scarce in Argentina. In this work, the tabtoxin‐producing strain Pseudomonas S5, isolated from oat, was identi?ed as a P. syringae through biochemical tests such as the LOPAT test, and genetic tests such as the analysis of the small subunit ribosomal RNA gene (16S rRNA) sequence and repetitive elements, using BOX and ERIC primers. It was also determined that this phytopathogen is potentially capable of infecting other crops of agricultural importance for our region, such as soybean. This ability to infect different hosts gives it an adaptive advantage that allows it to endure seasonal changes in the environment where it lives. Our work contributes to the physiological classification of the phytopathogen P. syringae S5 isolated from our region, as well as to the knowledge about its range of potential hosts.     

Novel,potent, selective,and brain penetrant phosphodiesterase 10A inhibitors

Herein we report the discovery of a novel series of phosphodiesterase 10A inhibitors. Optimization of a HTS hit (17) resulted in potent, selective, and brain penetrant 23 and 26; both exhibited much lower clearance in vivo and decreased volume of distribution (rat PK) and have thus the potential to inhibit the PDE10A target in vivo at a lower efficacious dose than the reference compound WEB-3.     

Protein interactions and disease: computational approaches to uncover the etiology of diseases

The genomic era has been characterised by vast amounts of data from diverse sources, creating a need for new tools to extract biologically meaningful information. Bioinformatics is, for the most part, responding to that need. The sparseness of the genomic data associated with diseases, however, creates a new challenge. Understanding the complex interplay between genes and proteins requires integration of data from a wide variety of sources, i.e. gene expression, genetic linkage, protein interaction, and protein structure among others. Thus, computational tools have become critical for the integration, representation and visualization of heterogeneous biomedical data. Furthermore, several bioinformatics methods have been developed to formulate predictions about the functional role of genes and proteins, including their role in diseases. After an introduction to the complex interplay between proteins and genetic diseases, this review explores recent approaches to the understanding of the mechanisms of disease at the molecular level. Finally, because most known mechanisms leading to disease involve some form of protein interaction, this review focuses on the recent methodologies for understanding diseases through their underlying protein interactions. Recent contributions from genetics, protein structure and protein interaction network analyses to the understanding of diseases are discussed here.     

Trophic interactions between brown and south polar skuas at Deception Island,Antarctica

It is broadly accepted that the brown skua (Stercorarius antarcticus lonnbergi) competitively excludes the south polar skua (S. maccormicki) from penguin colonies when breeding sympatrically, forcing the latter to feed on marine resources. The purpose of this work was to examine the diets and trophic niche breadths of each species where they co-occur and to determine the degree of overlap. To this end, we analyzed 169 pellets of brown skuas, collected in two different areas (20 individuals), and 152 of south polar skuas, collected in three different areas (18 individuals), on Deception Island, South Shetland Islands, Antarctica, during the austral summer 2000. Pellet analysis often underestimates the amount of easily digestible prey, but allows for comparisons of the relative contributions of different items in the diet. South polar skuas at our study locations consumed seven different food items and had a trophic niche breadth of 0.133 compared to brown skuas that fed on 10 different items and had a trophic niche breadth of 0.078. The niche overlap between the species was 82.1%. Penguins were the principal food source of both species, however, brown skuas fed mostly on chicks, while south polar skuas fed on adults (carcasses). The use of different age classes of penguins as a food source offers an alternative to competitive exclusion, allowing the coexistence of these species on Deception Island.     

Nuclear Entry of Hepatitis B Virus Capsids Involves Disintegration to Protein Dimers followed by Nuclear Reassociation to Capsids

Assembly and disassembly of viral capsids are essential steps in the viral life cycle. Studies on their kinetics are mostly performed in vitro, allowing application of biochemical, biophysical and visualizing techniques. In vivo kinetics are poorly understood and the transferability of the in vitro models to the cellular environment remains speculative. We analyzed capsid disassembly of the hepatitis B virus in digitonin-permeabilized cells which support nuclear capsid entry and subsequent genome release. Using gradient centrifugation, size exclusion chromatography and immune fluorescence microscopy of digitonin-permeabilized cells, we showed that capsids open and close reversibly. In the absence of RNA, capsid re-assembly slows down; the capsids remain disintegrated and enter the nucleus as protein dimers or irregular polymers. Upon the presence of cellular RNA, capsids re-assemble in the nucleus. We conclude that reversible genome release from hepatitis B virus capsids is a unique strategy different from that of other viruses, which employs irreversible capsid destruction for genome release. The results allowed us to propose a model of HBV genome release in which the unique environment of the nuclear pore favors HBV capsid disassembly reaction, while both cytoplasm and nucleus favor capsid assembly.     

Human RBM28 protein is a specific nucleolar component of the spliceosomal snRNPs

The biogenesis of spliceosomal small nuclear RNAs (snRNAs) involves organized translocations between the cytoplasm and certain nuclear domains, such as Cajal bodies and nucleoli. Here we identify human RBM28 protein as a novel snRNP component, based on affinity selection of U6 small nuclear ribonucleoprotein (snRNP). As shown by immunofluorescence, RBM28 is a nucleolar protein. Anti-RBM28 immunoprecipitation from HeLa cell lysates revealed that this protein specifically associates with U1, U2, U4, U5, and U6 snRNAs. Our data provide the first evidence that RBM28 is a common nucleolar component of the spliceosomal ribonucleoprotein complexes, possibly coordinating their transition through the nucleolus.     

Serum insulin-like growth factor I reference values for an automated chemiluminescence immunoassay system: results from a multicenter study

BACKGROUND: Analysis of insulin-like growth factor I in serum (S-IGF-I) is an integral component in the diagnosis of GH-related disorders and is going to be of interest in the diagnosis and follow-up of many disorders. The objective of the present study was to develop cross-sectional reference values for S-IGF-I measured by an automated chemiluminescence immunoassay (Nichols Advantage). METHODS: The study included samples from 3,961 healthy subjects (2,201 males, 1,760 females) aged 1 month to 88 years. Six laboratories were involved in this study and the samples were analyzed by one of seven automated immunoassay systems run in these laboratories. For data analysis, polynomial age and sex-specific models were fitted after transformation of S-IGF-I values. RESULTS: The results show the well-known age dependency of S-IGF-I levels. At ages <20, higher S-IGF-I levels were seen in girls with an estimated mean peak of 410 microg/l at age 14 and an estimated mean peak of 382 microg/l at age 16 in boys. Thereafter, a rapid decrease was seen to approximately 25 years of age, followed by a slow age-dependent decrease. In adulthood, S-IGF-I in males were slightly, but significantly higher than in females. It could be shown that the mean values of some reference sample subgroups differed significantly from the total mean. However, the multicenter approach used in this study reduces the impact of systematic population, sample handling and laboratory differences on the calculated reference mean. CONCLUSION: The present study establishes age- and sex-specific reference values for a fully automated immunoassay system based on a large population of healthy subjects. The established reference values may be used for this immunoassay system in different laboratories provided that the systematic difference between systems is low.