Calcination Method of 45Ca Samples for Isotope Ratio Analysis via Liquid Scintillation

This study was designed to optimize calcium-45 (⁴⁵Ca) sample preparation method in a Liquid Scintillation Counter (LSC) for use with vegetables and soil samples. Different tests were conducted on fruit and soil samples to determine the best conditions related to drying, time and centrifuge speed (rpm) after calcination at 450°C and 550°C for vegetable and soil samples, respectively. The homogeneity of samples, linearity, detection and quantification limits were also determined. The quench curve was calculated to establish the parameters to program the LSC. Six ⁴⁵Ca sample replicates were measured with a coefficient of variation (CV) of 1.74%, which indicates good sample homogeneity. The method is linearity over the range of activity studied for 0–30,000 disintegrations per minute (dpm). Our results suggest that the optimum homogenized sample preparation for calcination purposes was lyophilization. Techniques using ⁴⁵Ca applied to soil and plant tissues are promising tracers of Ca in plants, which can be very useful for the study of Ca kinetics in plants and can serve to guide efficient fertilization programs.     

Assessment of the bacterial community diversity associated with the queen conch Strombus gigas (Linnaeus, 1758) from the Caribbean coast of Colombia using denaturing gradient gel electrophoresis and culturing

The biotic diversity of Strombus gigas has not been thoroughly studied despite the status of the queen conch as an important ecological resource. The bacteria associated with the conch influence their host in several ways, including through the metabolism of nutrients, protection against invasive bacteria and the regulation of the physical conditions. In this study, conventional microbiological methods and molecular tools such as denaturing gradient gel electrophoresis (DGGE) were used to assess the composition of the bacterial communities associated with the queen conch (S. gigas) in wild and captive habitats in Colombia. A genetic analysis of the bacterial communities revealed a high level of diversity based on the large number of bands detected using DGGE. In addition, differences in bacterial community structure were found between the conchs in captivity and the wild populations. The dominant phylogenetic affiliations of the bacteria, as determined using 16S rRNA gene sequencing, were grouped into four classes, namely, Betaproteobacteria (16%), Gammaproteobacteria (70%), Firmicutes (7%) and Actinobacteria (2%). These groups are related to host defence processes and the decomposition of organic matter. The 16S rDNA sequence analysis of the cultured bacteria and the resulting DGGE profiles are useful tools for characterizing the diversity of the bacteria associated with the analyzed conchs.     

Cloning and nucleotide sequence of the equine and elk pituitary pre-prolactin cDNA

We report the equine (Equs equs) and elk (Cervus elaphus) pituitary pre-prolactin (PRL) cDNA cloning, and their nucleotide and deduced amino acid sequences. Pre-PRL cDNA was obtained by RNA ligation mediated-rapid amplification of cDNA ends (RLM-RACE) and polymerase chain reaction (PCR). The elk pre-PRL cDNA exhibits two polymorphisms at positions 96 and 672, which are silent since they encode for the same amino acids, proline and isoleucine, respectively. We found no polymorphisms in the equine pre-PRL cDNA. The deduced amino acid sequence of the equine pre-PRL is 99% identical to the previously reported protein sequence. Pre-PRL mRNA is <1 kb in length and is highly expressed in the anterior pituitary gland, as demonstrated by Northern hybridization analysis. In summary, we cloned and sequenced the equine and elk pre-PRL cDNAs. The deduced amino acid sequence of elk and equine pre-PRL appears to be moderately conserved among other mammalian species. The polymorphic sites found in the elk cDNA could potentially be used in parentage testing and gene mapping.     

Acid phosphatase and cathepsin D are active expressed enzymes in the placenta of the cat

Enzymes are crucial for the metabolism of macromolecular substrates. In the great majority of cells, most enzymes are constitutive. Nevertheless, inducible enzymes can predominate, determining specialized cell functions. Within this context, histochemistry/immunohistochemistry and biochemistry were used to investigate expression of peroxidase and reduced nicotinamide-adenine dinucleotide phosphate (NADPH)-oxidase, as well as the expression and activity of cathepsin D and acid phosphatase, in trophoblast cells within the endotheliochorial labyrinth and marginal hematoma of the term cat placenta. In the marginal hematoma, elevated Cathepsin D expression and activity was accompanied by erythrophagocytosis. In contrast, acid phosphatase activity was much more intense in the labyrinth, where metabolic exchanges occur. Peroxidase and NAD(P)H-oxidase were predominantly active in trophoblast cells within endosomal vesicles of different placental compartments, indicating that, although reactive oxygen species might participate in endosomal/lysosomal processes, they are not territorially specific or functional markers. These findings highlight differential characteristics of cathepsin D and acid phosphatase activity within each placental compartment, thereby contributing to the comprehension of the territorial role played by the placenta and facilitating future metabolic studies.     

Drought responses of an exotic tree (Eriobotrya japonica) in a tropical cloud forest suggest the potential to become an invasive species

New Forests - Eriobotrya japonica is a non-native tree expanding in secondary forests and threatening the tropical montane cloud forest of central Veracruz, Mexico. Our objective was to investigate...     

Requirement for caspase-2 in stress-induced apoptosis before mitochondrial permeabilization

A current view is that cytotoxic stress, such as DNA damage, induces apoptosis by regulating the permeability of mitochondria. Mitochondria sequester several proteins that, if released, kill by activating caspases, the proteases that disassemble the cell. Cytokines activate caspases in a different way, by assembling receptor complexes that activate caspases directly; in this case, the subsequent mitochondrial permeabilization accelerates cell disassembly by amplifying caspase activity. We found that cytotoxic stress causes activation of caspase-2, and that this caspase is required for the permeabilization of mitochondria. Therefore, we argue that cytokine-induced and stress-induced apoptosis act through conceptually similar pathways in which mitochondria are amplifiers of caspase activity rather than initiators of caspase activation.