Heterogeneity in high performance liquid chromatography of a variant surface glycoprotein of Trypanosoma brucei

High performance liquid chromatography (HPLC) procedures have been used to analyze a preparation of the variant surface glycoprotein AnTat 1.1A of Trypanosoma brucei. The native preparation gives several peaks with a high reproducibility both by reverse-phase (RP-) and gel permeation (GP-) HPLC. Under RP-HPLC conditions, nine fractions are fully resolved. The RP-HPLC fractions migrate with the same molecular weight VSG band on polyacrylamide slab gel electrophoresis and no significant differences are observed in amino acid composition among these fractions. The RP-HPLC resolution is found to be related to the ability of the VSG to polymerize as shown using GP-HPLC. These results suggest the existence of a microheterogeneity of the AnTat 1.1A VSG preparation in relation to post-translational modification of the VSG molecule.     

Immune response in cervical dysplasia induced by human papillomavirus: the influence of human immunodeficiency virus-1 co-infection -- review

Human immunodeficiency virus (HIV-1) has become an important risk factor for human papillomavirus (HPV) infection and the development of HPV associated lesions in the female genital tract. HIV-1 may also increase the oncogenicity of high risk HPV types and the activation of low risk types. The Center for Disease Control and Prevention declared invasive cervical cancer an acquired immunodeficiency virus (AIDS) defining illness in HIV positive women. Furthermore, cervical cancer happens to be the second most common female cancer worldwide. The host's local immune response plays a critical factor in controlling these conditions, as well as in changes in the number of professional antigen-presenting cells, cytokine, and MHC molecules expression. Also, the production of cytokines may determine which arm of the immune response will be stimulated and may influence the magnitude of immune protection. Although there are many studies describing the inflammatory response in HPV infection, few data are available to demonstrate the influence of the HIV infection and several questions regarding the cervical immune response are still unknown. In this review we present a brief account of the current understanding of HIV/HPV co-infection, emphasizing cervical immune response.     

Boat anchoring on Posidonia oceanica beds in a marine protected area (Italy, western Mediterranean): effect of anchor types in different anchoring stages

Seagrasses worldwide are noted for suffering from mechanical damage caused by boat anchoring. This is particularly so in sites highly frequented by boaters (marine protected areas or coastal urbanised areas). In the last decades, different strategies have been put into practice to reduce such impacts on seagrasses (i.e. by anchoring bans or by deploying boat moorings). More recently, in consideration that few marine protected area (MPA) management bodies or local administrations have the resources to enforce their anchorage regulations, the self-regulatory approach based on education and information of boaters has been preferred in several cases. At present, however, very little is known on the correct anchoring practices to ensure the safeguarding of seagrasses. The aim of the present study was to experimentally quantify in the field the damage caused to Posidonia oceanica shoot density by anchoring. A multifactorial experiment was designed to test whether the damage is dependent on (1) different anchor types (Hall, Danforth and Folding grapnel), (2) the use of a chain vs. a rope, (3) the three anchoring stages (anchor fall, dragging/lock-in and weighing), and finally (4) whether the pattern is consistent among different locations of the meadow.As expected, the three anchor types employed in the present study differed in the levels of damage inflicted on the P. oceanica meadows of the Ustica Island MPA. In particular, the use of the Hall type anchor seems to be preferable to minimise this impact in comparison with the other two anchor types. Moreover, the effect on the meadow of the three anchor types is greatly dependent on the anchoring stage. These results confirm that the weighing stage is the critical stage of the anchoring process. The number of damaged shoots of P. oceanica was not affected by the presence of the chain. These patterns were consistent between locations.In the long term, even anchoring on P. oceanica by small boats using low-impact anchors may potentially have detrimental consequences. For this reason, we suggest that in vulnerable sites, it is preferable to implement an educational program based on information of boaters on correct anchoring practices and anchor typology to use, rather than adopting strong restrictions to boat anchoring or deploying mooring buoys. Although the use of these management strategies is still recommended in the case of anchorage frequented by bigger vessels using heavier anchors and chains.     

Magnetic Resonance Imaging Allows the Evaluation of Tissue Damage and Regeneration in a Mouse Model of Critical Limb Ischemia

Magnetic resonance imaging (MRI) provides non-invasive, repetitive measures in the same individual, allowing the study of a physio-pathological event over time. In this study, we tested the performance of 7 Tesla multi-parametric MRI to monitor the dynamic changes of mouse skeletal muscle injury and regeneration upon acute ischemia induced by femoral artery dissection. T2-mapping (T2 relaxation time), diffusion-tensor imaging (Fractional Anisotropy) and perfusion by Dynamic Contrast-Enhanced MRI (K-trans) were measured and imaging results were correlated with histological morphometric analysis in both Gastrocnemius and Tibialis anterior muscles. We found that tissue damage positively correlated with T2-relaxation time, while myofiber regeneration and capillary density positively correlated with Fractional Anisotropy. Interestingly, K-trans positively correlated with capillary density. Accordingly, repeated MRI measurements between day 1 and day 28 after surgery in ischemic muscles showed that: 1) T2-relaxation time rapidly increased upon ischemia and then gradually declined, returning almost to basal level in the last phases of the regeneration process; 2) Fractional Anisotropy dropped upon ischemic damage induction and then recovered along with muscle regeneration and neoangiogenesis; 3) K-trans reached a minimum upon ischemia, then progressively recovered. Overall, Gastrocnemius and Tibialis anterior muscles displayed similar patterns of MRI parameters dynamic, with more marked responses and less variability in Tibialis anterior. We conclude that MRI provides quantitative information about both tissue damage after ischemia and the subsequent vascular and muscle regeneration, accounting for the differences between subjects and, within the same individual, between different muscles.     

Three-dimensional soft tissue preservation revealed in the skin of a non-avian dinosaur

The most commonly preserved soft tissues associated with ornithischian dinosaurs are skin remains. The apparent resistance of hadrosaur skin to decay, and its abundance in the fossil record relative to that of other tetrapods, has been attributed to factors such as thickness and composition. Here we report additional intrinsic factors within hadrosaur skin: 3D-preserved eumelanin-bearing bodies, dermal cells and blood vessel fragments in an organic matrix composed of protein fossilization products. The skin is much thinner than that of living mammals of similar size. It is likely that the preservation of hadrosaur skin is related to the arrangement of the layers composing it.     

Two modes of FEN1 binding to PCNA regulated by DNA

The FEN1 nuclease functions during Okazaki fragment maturation in the eukaryotic cell. Like many other proliferating cell nuclear antigen (PCNA)-binding proteins, FEN1 interacts with the interdomain connector loop (IDCL) of PCNA, and PCNA greatly stimulates FEN1 activity. A yeast IDCL mutant pcna-79 (IL126,128AA) failed to interact with FEN-1, but, surprisingly, pcna-79 was still very active in stimulating FEN1 activity. In contrast, a C-terminal mutant pcna-90 (PK252,253AA) showed wild-type binding to FEN1 in solution, but poorly stimulated FEN1 activity. When PCNA was loaded onto a DNA substrate coupled to magnetic beads, it stabilized retention of FEN1 on the DNA. In this DNA-dependent binding assay, pcna-79 also stabilized retention of FEN1, but pcna-90 was inactive. Therefore, in the absence of DNA, FEN1 interacts with PCNA mainly through the IDCL. However, when PCNA encircles the DNA, the C-terminal domain of PCNA rather than its IDCL is important for binding FEN1. An FF-->GA mutation in the PCNA-interaction domain of FEN1 severely decreased both modes of interaction with PCNA and resulted in replication and repair defects in vivo.     

Pseudomonas community structure and antagonistic potential in the rhizosphere: insights gained by combining phylogenetic and functional gene-based analyses

The Pseudomonas community structure and antagonistic potential in the rhizospheres of strawberry and oilseed rape (host plants of the fungal phytopathogen Verticillium dahliae) were assessed. The use of a new PCR-DGGE system, designed to target Pseudomonas-specific gacA gene fragments in environmental DNA, circumvented common biases of 16S rRNA gene-based DGGE analyses and proved to be a reliable tool to unravel the diversity of uncultured Pseudomonas in bulk and rhizosphere soils. Pseudomonas-specific gacA fingerprints of total-community (TC) rhizosphere DNA were surprisingly diverse, plant-specific and differed markedly from those of the corresponding bulk soils. By combining multiple culture-dependent and independent surveys, a group of Pseudomonas isolates antagonistic towards V. dahliae was shown to be genotypically conserved, to carry the phlD biosynthetic locus (involved in the biosynthesis of 2,4-diacetylphloroglucinol - 2,4-DAPG), and to correspond to a dominant and highly frequent Pseudomonas population in the rhizosphere of field-grown strawberries planted at three sites in Germany which have different land use histories. This population belongs to the Pseudomonas fluorescens phylogenetic lineage and showed closest relatedness to P. fluorescens strain F113 (97% gacA gene sequence identity in 492-bp sequences), a biocontrol agent and 2,4-DAPG producer. Partial gacA gene sequences derived from isolates, clones of the strawberry rhizosphere and DGGE bands retrieved in this study represent previously undescribed Pseudomonas gacA gene clusters as revealed by phylogenetic analysis.     

Genome-wide CRISPR Screens in T Helper Cells Reveal Pervasive Crosstalk between Activation and Differentiation

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Characterization of the receptor for insulin-like growth factor on Leishmania promastigotes

Insulin-like growth factor (IGF)-I constitutively present in the skin is one of the first growth factors that Leishmania parasites encounter after transmission to the vertebrate host. We have previously shown that IGF-I is a potent growth-promoting factor for Leishmania parasites. IGF-I binds specifically to a single-site putative receptor at the parasite membrane, triggering a cascade of phosphorylation reactions. In the present article we characterize the receptor for IGF-I on Leishmania (Leishmania) mexicana promastigotes. The receptor is a monomeric glycoprotein with a molecular mass of 65 kDa and is antigenically related to the alpha chain of human type 1 IGF-I receptor. Upon IGF-I stimulation the receptor undergoes autophosphorylation on tyrosine residues with activation of its signaling pathway. Activation of the IGF-I receptor also leads to phosphorylation of an 185-kDa molecule that is homologous to the substrate of the insulin receptor present in human cells, the insulin receptor substrate 1 (IRS-1).