Spatial distribution and diet of the Neotropical otter<Emphasis Type="Italic">Lontra longicaudis</Emphasis> in the Ibera Lake (northern Argentina)

The seasonal spatial distribution and diet of the Neotropical otterLontra longicaudis (Olfers, 1818) were studied on Ibera Lake within the Esteros del Ibera, an important wetland of NE Argentina (Ramsar site no. 1162). Twelve lake perimeter sites were regularly checked for otter tracks and signs. The amount of signs was compared to environmental parameters to test their influence on otter presence and movements. A positive correlation was found between the amount of signs and the physical structure of the littoral areas, as well as vegetation structure. Lakeside otter presence decreased during the summer sampling throughout all sites. In order to determine otter diet, 205 spraints were examined. Otters fed mainly on fish (mostly Cichlidae), but also on crustaceans and molluscs. Other fish consumed were Characidae, Synbranchidae, Loricariidae and Erythrinidae. Seasonal variation was observed in diet composition: in summer, diet crustaceans and vertebrates other than fish increased. A higher percentage of benthic fish species was also observed in summer, while pelagic and benthopelagic species increased in winter. Such dietary changes may be explained by the different habitat use of otters in different seasons, from the lake coast (winter) towards more internal marshy areas of the wetland (summer).     

High-Throughput Multiplex Single-Nucleotide Polymorphism (SNP) Analysis in Genes Involved in Methionine Metabolism

Hyperhomocysteinemia is a well-known independent marker factor for atherothrombotic diseases and may result from acquired and genetic influences. Several polymorphisms are suspected to be associated with hyperhomocysteinemia, but data are limited and inconsistent. High-throughput genotyping technologies, such as GenomeLab SNPStream, are now available. Moreover, an appropriate selection of SNPs to be analyzed could represent a strong resource to define the role of genetic risk factors. We developed a multiplex PCR-oligonucleotide extension approach by GenomeLab platform. We selected 72 SNPs based on their putative function and frequency in the candidate genes AHCY, BHMT, BHMT2, CBS, ENOSF1, FOLH1, MTHFD1, MTHFR, MTR, MTRR, NNMT, PON1, PON2, SLC19A1, SHMT1, TCN2, and TYMS. We were able to analyze 57 of the SNPs (79%). For MTHFR C677T and A1298C and MTR A2756G SNPs, we compared data obtained with an electronic microchip technology and found 99.2% concordance. We also performed a haplotype analysis. This approach could represent a useful tool to investigate the genotype-phenotype correlation and the association of these genes with hyperhomocysteinemia and correlated diseases.     

Cloning and characterization of a novel WD-40 repeat protein that dramatically accelerates osteoblastic differentiation

The bone morphogenetic proteins (BMPs) play a pivotal role in endochondral bone formation. Using differential display polymerase chain reaction, we have identified a novel gene, named BIG-3 (BMP-2-induced gene 3 kb), that is induced as a murine prechondroblastic cell line, MLB13MYC clone 17, acquires osteoblastic features in response to BMP-2 treatment. The 3-kilobase mRNA encodes a 34-kDa protein containing seven WD-40 repeats. Northern and Western analyses demonstrated that BIG-3 mRNA and protein were induced after 24 h of BMP-2 treatment. BIG-3 mRNA was expressed in conditionally immortalized murine bone marrow stromal cells, osteoblasts, osteocytes, and growth plate chondrocytes, as well as in primary calvarial osteoblasts. Immunohistochemistry demonstrated that BIG-3 was expressed in the osteoblasts of calvariae isolated from mouse embryos. To identify a role for BIG-3 in osteoblast differentiation, MC3T3-E1 cells were stably transfected with the full-length coding region of BIG-3 (MC3T3E1-BIG-3) cloned downstream of a cytomegalovirus promoter in pcDNA3.1. Pooled MC3T3E1-BIG-3 clones expressed alkaline phosphatase activity earlier and achieved a peak level of activity 10-fold higher than cells transfected with the empty vector (MC3T3E1-EV) at 14 days. Cyclic AMP production in response to parathyroid hormone was increased 10- and 14-fold at 7 and 14 days, respectively, in MC3T3E1-BIG-3 clones, relative to MC3T3E1-EV clones. This increase in cAMP production was associated with an increase in PTH binding. Expression of BIG-3 increased mRNA levels encoding Cbfa1, type I collagen, and osteocalcin and accelerated formation of mineralized nodules. In conclusion, we have identified a novel WD-40 protein, induced by BMP-2 treatment, that dramatically accelerates the program of osteoblastic differentiation in stably transfected MC3T3E1 cells.     

Immune-reconstituted influenza virosome containing CD40L gene enhances the immunological and protective activity of a carcinoembryonic antigen anticancer vaccine

The correct interaction of a costimulatory molecule such as CD40L with its contrareceptor CD40 expressed on the membrane of professional APCs, provides transmembrane signaling that leads to APC activation. This process can be exploited to significantly improve the efficacy of cancer vaccines and the outcome of a possible cancer vaccine-induced, Ag-specific CTL response. Therefore, we investigated whether a novel intranasal delivery of immune-reconstituted influenza virosomes (IRIV), assembled with the CD40L gene (CD40L/IRIV), could be used to improve protective immunity and the Ag-specific CTL response against carcinoembryonic Ag (CEA) generated with a novel vaccine constituted of IRIV assembled with the CEA gene (CEA/IRIV). Our results suggest that CD40L/IRIV was able to augment CEA-specific CTL activity and CEA-specific protective immunity induced by CEA/IRIV most likely through the induction of a CTL response associated with a Th1 phenotype. In conclusion, we provide evidence that CD40L/IRIV, by acting through the CD40L/CD40 signaling pathway, acts as an immune-adjuvant that could increase the efficacy of a CEA-specific cancer vaccine, which could provide an efficacious new strategy for cancer therapy.     

Sequence- and Structure-Based Immunoreactive Epitope Discovery for Burkholderia pseudomallei Flagellin

Burkholderia pseudomallei is a Gram-negative bacterium responsible for melioidosis, a serious and often fatal infectious disease that is poorly controlled by existing treatments. Due to its inherent resistance to the major antibiotic classes and its facultative intracellular pathogenicity, an effective vaccine would be extremely desirable, along with appropriate prevention and therapeutic management. One of the main subunit vaccine candidates is flagellin of Burkholderia pseudomallei (FliC_Bp). Here, we present the high resolution crystal structure of FliC_Bp and report the synthesis and characterization of three peptides predicted to be both B and T cell FliC_Bp epitopes, by both structure-based in silico methods, and sequence-based epitope prediction tools. All three epitopes were shown to be immunoreactive against human IgG antibodies and to elicit cytokine production from human peripheral blood mononuclear cells. Furthermore, two of the peptides (F51-69 and F270-288) were found to be dominant immunoreactive epitopes, and their antibodies enhanced the bactericidal activities of purified human neutrophils. The epitopes derived from this study may represent potential melioidosis vaccine components.     

Changes of glycogen metabolism in phosphorylcreatine-depleted muscles taken from rats fed with beta-guanidine propionate

Changes in glycogen metabolism were explored in fast and slow muscles taken from rats fed with a diet containing 1% beta-guanidine propionate (GPA), a synthetic analog that inhibits the entry of creatine into muscle cells competitively and causes phosphorylcreatine depletion. Feeding with the GPA-containing diet increased glycogen levels in the two types of muscles to a different extent and with different temporal patterns; it did not change significantly the rate of glycogen turnover both at rest and during exercise; it did not affect the net degradation of glycogen during exercise. Diet could affect the activity of several enzymes of sugar metabolism. These latter changes too were different in fast-twitch and in slow-twitch muscles.     

An Unusual Cytoplasmic Component in Euphorbia peplus L. Endosperm

The coenocytic endosperm of Euphorbia peplus contains a complexsystem of flat, tightly-stacked membrane saccules which havea sinuous profile and a smooth or smooth-rough surface. Theyare joined by membrane-to-membrane links. The usual ergastoplasmiccisternae are an integral component of this system which canessentially be regarded as a sui generis from of endoplasmicreticulum.Copyright 1995, 1999 Academic Press Euphorbia peplus, TEM, ultrastructure, endosperm, endoplasmic reticulum, cell organelles