Immunoglobulin light chain class multiplicity and alternative organizational forms in early vertebrate phylogeny

The prototypic chondrichthyan immunoglobulin (Ig) light chain type (type I) isolated from Heterodontus francisci (horned shark) has a clustered organization in which variable (V), joining (J), and constant (C) elements are in relatively close linkage (V-J-C). Using a polymerase chain reaction-based approach on a light chain peptide sequence from the holocephalan, Hydrolagus colliei (spotted ratfish), it was possible to isolate members of a second light chain gene family. A probe to this light chain (type II) detects homologs in two orders of elasmobranchs, Heterodontus, a galeomorph and Raja erinacea (little skate), a batoid, suggesting that this light chain type may be present throughout the cartilaginous fishes. In all cases, V, J, and C regions of the type II gene are arranged in closely linked clusters typical of all known Ig genes in cartilaginous fishes. All representatives of this type II gene family are joined in the germline. A third (kappa-like) light chain type from Heterodontus is described. These findings establish that a degree of light chain class complexity comparable to that of the mammals is present in the most phylogenetically distant extant jawed vertebrates and that the phenomenon of germline-joined (pre-rearranged) genes, described originally in the heavy chain genes of cartilaginous fishes, extends to light chain genes.     

Intracellular aminopeptidases inStreptomyces lividans 66

Summary We have investigated the aminopeptidase activities present inStreptomyces lividans strains. The majority of these activities proved to be intracellular with multiple active species. Two aminopeptidase P genes were identified to be responsible for the ability to hydrolyze amino terminal peptide bonds adjacent to proline residues. Two other broad spectrum aminopeptidases were found to display homology at both the DNA and protein levels. One showed significant homology to PepN proteins, particularly around the putative zinc-binding residues which are important for catalysis. The second broad spectrum activity was not analyzed in detail but showed a different spectrum of substrate specificity to that of PepN.     

Identification of a nucleo-cytoplasmic ionic pathway by osmotic shock in isolated mouse liver nuclei

Summary The observation that the nuclear envelope outer mem brane contains ion channels raises the question of whether these conductances communicate between the cytosol and the nuclear envelope cisternae or between the cytosol and the cytoplasm. Failure to detect large, nonselective holes using the patch-clamp technique has led to the speculation that ion channels and nuclear pores are in fact the same. In this paper we present evidence that the ionic channel, recorded in isolated liver nuclei with the patch-clamp configura tion of “nucleus-attached,” spans the double membrane of the envelope, providing a direct contact between nucleoplasm and cytoplasm.     

Nitrogen fertilizer enhances vegetation establishment of a high-altitude machine-graded ski slope

Machine grading is frequently required to prepare the terrain when building high-altitude ski slopes in the Alps. However, this kind of disturbance alters the natural environment, destroying the vegetation and hampering its reestablishment. Thus, specific restoration plans are necessary to encourage the recovery of vegetation, which is already affected by different natural constraints in this harsh environment. One of the main critical factors affecting plant growth in high-altitude areas is the lack of available nitrogen (N) in the soil. In this context, the addition of a slow-release N fertilizer was carried out in an experimental revegetated ski slope between 2,800 and 2,900 m above sea level in the western Italian Alps. Both vegetation and soil were monitored during a 5-year period in order to test the effectiveness of N addition on the restoration process. Even if effects on soil carbon and N contents were negligible, vegetation was remarkably affected by the fertilization, since the total vegetation cover and the species richness significantly increased. Against the expectations, there was a remarkable increase in spontaneous forbs, rather than in most of the sown graminoids, which slightly varied during the experimental period. Actually, graminoids responded in different ways, mostly increasing (likewise forbs), but the slight decrease of the dominant Festuca nigrescens (Chewing's Fescue) masked their spread. This study confirms the noteworthy role of N in high-altitude alpine soils and consequently its importance to improve the restoration process of degraded ecosystems.     

Evaluation of microbial surfactants for recovery of hydrophobic pollutants from soil

Summary Several microbially produced biosurfactants were evaluated for their ability to remove hydrophobic compounds from soil. The biosurfactants produced byPseudomonas aeruginosa UG2 andAcinetobacter calcoaceticus RAG-1 displayed the best results, with recovery of [¹⁴C]hexachlorobiphenyl from soil slurries of 48.0 and 41.9%, respectively.P. aeruginosa UG2 produced higher levels of extracellular biosurfactants than four otherP. aeruginosa strains.P. aeruginosa UG2 culture filtrate containing biosurfactants enhanced the recovery of several other individual hydrocarbons and polychlorinated biphenyl compounds, as well as several hydrocarbons in a mixture, from soil. The results, suggest that biosurfactants produced byP. aeruginosa UG2 have the potential for remediation of hydrophobic pollutants in soil environments.     

Cumulus expansion is impaired with advanced reproductive age due to loss of matrix integrity and reduced hyaluronan

Reproductive aging is associated with ovulatory defects. Age-related ovarian fibrosis partially contributes to this phenotype as short-term treatment with anti-fibrotic compounds improves ovulation in reproductively old mice. However, age-dependent changes that are intrinsic to the follicle may also be relevant. In this study, we used a mouse model to demonstrate that reproductive aging is associated with impaired cumulus expansion which is accompanied by altered morphokinetic behavior of cumulus cells as assessed by time-lapse microscopy. The extracellular matrix integrity of expanded cumulus–oocyte complexes is compromised with advanced age as evidenced by increased penetration of fluorescent nanoparticles in a particle exclusion assay and larger open spaces on scanning electron microscopy. Reduced hyaluronan (HA) levels, decreased expression of genes encoding HA-associated proteins (e.g., Ptx3 and Tnfaip6), and increased expression of inflammatory genes and matrix metalloproteinases underlie this loss of matrix integrity. Importantly, HA levels are decreased with age in follicular fluid of women, indicative of conserved reproductive aging mechanisms. These findings provide novel mechanistic insights into how defects in cumulus expansion contribute to age-related infertility and may serve as a target to extend reproductive longevity.     

Review on the sublethal effects of pure and formulated glyphosate on bees: Emphasis on social bees

Bees are declining worldwide, and the use of pesticides has been linked to this problem. Studies show that even herbicides can negatively impact bees, causing death or compromising health. As a result, concern about the use of glyphosate (GLY) has increased, as it is the most sold pesticide. Studies have shown that exposure of bees to GLY can trigger sublethal effects. Considering the speed with which information is published, reviews become important for the integration of knowledge, aiding understanding of the topic. Therefore, the present study aimed to review the literature on the sublethal effects of GLY and the different commercial formulations on bees. After the literary review, it was observed that the exposition, acute and chronic, of larvae and adults of social and solitary bees, to GLY and its formulations, can trigger alterations in gene expression, enzyme functioning, oxidative metabolism, cell/tissue structure, intestinal microbiota diversity, learning, food consumption, flight and vertical displacement capacity, circadian cycle and body development of these insects. The most used species in the studies was Apis mellifera L. Studies are still necessary to understand the sublethal effects of GLY on bees, in the medium and long term, on colony homeostasis, especially about the information on the toxicity of some surfactants present in the different commercial formulations.     

iBench: A ground truth approach for advanced validation of mass spectrometry identification method

The discovery of many noncanonical peptides detectable with sensitive mass spectrometry inside, outside, and on cells shepherded the development of novel methods for their identification, often not supported by a systematic benchmarking with other methods. We here propose iBench, a bioinformatic tool that can construct ground truth proteomics datasets and cognate databases, thereby generating a training court wherein methods, search engines, and proteomics strategies can be tested, and their performances estimated by the same tool. iBench can be coupled to the main database search engines, allows the selection of customized features of mass spectrometry spectra and peptides, provides standard benchmarking outputs, and is open source. The proof-of-concept application to tryptic proteome digestions, immunopeptidomes, and synthetic peptide libraries dissected the impact that noncanonical peptides could have on the identification of canonical peptides by Mascot search with rescoring via Percolator (Mascot+Percolator).     

MULTIPLE PATERNITY AND SELF-FERTILIZATION IN RELATION TO FLORAL AGE IN MIMULUS GUTTATUS (SCROPHULARIACEAE)

To gain some understanding of the mechanisms responsible for the intermediate levels of both multiple paternity and selfing observed previously in the common monkeyflower, Mimulus guttatus, we performed a field experiment to assess the role of 1) multiple paternity through sequential pollinator visits and 2) selfing through corolla abscission. In M. guttatus, flowers remain open for several days; then, in the process of corolla abscission anthers drag past the stigma. We predicted that multiple visitation by pollinators over this period should increase the degree of multiple paternity of sibling arrays, and that corolla dragging may be the primary cause of the observed selfing. In three northern California populations corollas were removed after being open for 1 day (preventing further mating), and the type of matings resulting was compared to matings from adjacent flowers with undisturbed corollas. On average, seed set was twice as great in flowers with unmanipulated corollas, indicating that about half of all matings in the lifespan of a M. guttatus flower occur after 1 day. Pollen supplementation revealed that pollen was limiting in one population. Electrophoretic assay of progeny showed multiple paternity was greater in unmanipulated flowers in the two populations that were not pollen limited, suggesting that sequential pollination may be an important source of multiple paternity. However, no difference was detected in the selfing rate between the two treatments, suggesting that corolla dragging was not a source of selfing even under pollen-limited conditions.