Development of early erythroid precursors (BFUe) from mouse bone marrow in a serum-free culture media. Effect of hemin

In culture medium containing albumin, iron saturated transferrin, phospholipids, cholesterol and erythropoietin, BFUe growth requires foetal Calf serum. Without serum the BFUe development is advantageously restored by the addition of hemin associated with spleen conditioned medium. In erythropoietin supplemented serum-free medium the growth of primitive erythroid precursor cells is closely dependent on growth factor supply found in spleen conditioned medium. Using this medium allows us to clearly distinguish between the respective effects of erythropoietin and of BPA on erythroid progenitor cells.     

Antisense RNA decreases AP33 gene expression and cytoadherence by <Emphasis Type="Italic">T. vaginalis</Emphasis>

Background  

Host parasitism by Trichomonas vaginalis is complex. Adherence to vaginal epithelial cells (VECs) is mediated by surface proteins. We showed before that antisense down-regulation of expression of adhesin AP65 decreased amounts of protein, which lowered levels of T. vaginalis adherence to VECs. We now perform antisense down-regulation of expression of the ap33 gene to evaluate and confirm a role for AP33 in adherence by T. vaginalis. We also used an established transfection system for heterologous expression of AP33 in T. foetus as an additional confirmatory approach.     

Transmitted Virus Fitness and Host T Cell Responses Collectively Define Divergent Infection Outcomes in Two HIV-1 Recipients

Control of virus replication in HIV-1 infection is critical to delaying disease progression. While cellular immune responses are a key determinant of control, relatively little is known about the contribution of the infecting virus to this process. To gain insight into this interplay between virus and host in viral control, we conducted a detailed analysis of two heterosexual HIV-1 subtype A transmission pairs in which female recipients sharing three HLA class I alleles exhibited contrasting clinical outcomes: R880F controlled virus replication while R463F experienced high viral loads and rapid disease progression. Near full-length single genome amplification defined the infecting transmitted/founder (T/F) virus proteome and subsequent sequence evolution over the first year of infection for both acutely infected recipients. T/F virus replicative capacities were compared in vitro, while the development of the earliest cellular immune response was defined using autologous virus sequence-based peptides. The R880F T/F virus replicated significantly slower in vitro than that transmitted to R463F. While neutralizing antibody responses were similar in both subjects, during acute infection R880F mounted a broad T cell response, the most dominant components of which targeted epitopes from which escape was limited. In contrast, the primary HIV-specific T cell response in R463F was focused on just two epitopes, one of which rapidly escaped. This comprehensive study highlights both the importance of the contribution of the lower replication capacity of the transmitted/founder virus and an associated induction of a broad primary HIV-specific T cell response, which was not undermined by rapid epitope escape, to long-term viral control in HIV-1 infection. It underscores the importance of the earliest CD8 T cell response targeting regions of the virus proteome that cannot mutate without a high fitness cost, further emphasizing the need for vaccines that elicit a breadth of T cell responses to conserved viral epitopes.     

Stand structure influences nekton community composition and provides protection from natural disturbance in Micronesian mangroves

Structurally complex mangrove roots are thought to provide foraging habitat, predation refugia, and typhoon protection for resident fish, shrimp, and crabs. The spatially compact nature of Micronesian mangroves results in model ecosystems to test these ideas. Tidal creek nekton assemblages were compared among mangrove forests impacted by Typhoon Sudal and differing in stand structure. Structurally complex Rhizophora spp. stands were predicted to support higher densities and different communities of nekton and to provide greater protection from typhoons compared to less complex Sonneratia alba/Bruguiera gymnorrhiza stands. Lift net data revealed that structural complexity did not support greater nekton densities, but did support significantly different nekton assemblages. The cardinalfish Apogon ceramensis and goby Oxyurichthys lonchotus had significantly higher densities in S. alba/B. gymnorrhiza mangrove creeks, whereas the silverside Atherinomorus lacunosus and diogenid crabs had significantly higher densities in Rhizophora spp. creeks. Similar nekton densities 17 and 4 months after the typhoon in Rhizophora spp. creeks provided indirect evidence that structural complexity increased protection for resident nekton from disturbances. Findings indicate that studies of structural complexity and nekton densities may be better served when individual species are compared and that diverse mangrove tree assemblages will support diverse nekton assemblages that may be more resilient to disturbance.     

Modulation of lung inflammation by vessel dilator in a mouse model of allergic asthma

Background

Atrial natriuretic peptide (ANP) and its receptor, NPRA, have been extensively studied in terms of cardiovascular effects. We have found that the ANP-NPRA signaling pathway is also involved in airway allergic inflammation and asthma. ANP, a C-terminal peptide (amino acid 99–126) of pro-atrial natriuretic factor (proANF) and a recombinant peptide, NP73-102 (amino acid 73–102 of proANF) have been reported to induce bronchoprotective effects in a mouse model of allergic asthma. In this report, we evaluated the effects of vessel dilator (VD), another N-terminal natriuretic peptide covering amino acids 31–67 of proANF, on acute lung inflammation in a mouse model of allergic asthma.

Methods

A549 cells were transfected with pVD or the pVAX1 control plasmid and cells were collected 24 hrs after transfection to analyze the effect of VD on inactivation of the extracellular-signal regulated receptor kinase (ERK1/2) through western blot. Luciferase assay, western blot and RT-PCR were also performed to analyze the effect of VD on NPRA expression. For determination of VD''s attenuation of lung inflammation, BALB/c mice were sensitized and challenged with ovalbumin and then treated intranasally with chitosan nanoparticles containing pVD. Parameters of airway inflammation, such as airway hyperreactivity, proinflammatory cytokine levels, eosinophil recruitment and lung histopathology were compared with control mice receiving nanoparticles containing pVAX1 control plasmid.

Results

pVD nanoparticles inactivated ERK1/2 and downregulated NPRA expression in vitro, and intranasal treatment with pVD nanoparticles protected mice from airway inflammation.

Conclusion

VD''s modulation of airway inflammation may result from its inactivation of ERK1/2 and downregulation of NPRA expression. Chitosan nanoparticles containing pVD may be therapeutically effective in preventing allergic airway inflammation.     

Causal Characteristics for Ecoepidemiology

We suggest that there are six fundamental characteristics of causation: time order, co-occurrence, preceding causation, sufficiency, interaction, and alteration. The cause precedes the effect (time order). The cause co-occurs with the unaffected entity in space and time (co-occurrence). Causes and their effects are the result of a web of causation (preceding causation). The intensity, frequency, and duration of the cause are adequate and the susceptible entity can exhibit the type and magnitude of the effect (sufficiency). The cause effectively interacts with the entity in a way that induces the effect (interaction). And, the entity is changed by the interactions with the cause (alteration). In contrast to Hill's criteria, the causal characteristics are distinct from the: (1) evidence that is used to document causal characteristics, (2) sources of information used to develop the evidence, and (3) qualities used to evaluate evidence of causal characteristics and body of evidence for the causal relationship. Evidence of causal characteristics can form the basis for assessments of epidemiological studies and can structure an explanatory narrative that is causally relevant and substantive. Six core characteristics may be easier to organize, evaluate, communicate, and for decision-makers to assimilate, remember, and inspire action.     

Causal Assessment of Biological Impairment in the Little Floyd River,Iowa, USA

An assessment of biological impairment in the Little Floyd River (Iowa, USA) was based on evidence of three characteristics of causation: co-occurrence, preceding causation, and sufficiency. Evidence of the physical interaction of the probable causes and the biota, resulting alterations to the biota, as well as the time order of the cause and the effect were consistent within the assessment, but the evidence for these causal characteristics did not discriminate among probable causes or other causes. Deposited sediment, low dissolved oxygen, heat stress, and ammonia toxicity are the probable causes of impaired biological condition in the Little Floyd River compared with other rivers in the ecoregion. Less likely causes are suspended sediment, altered basal food resources, and flow alteration. Very unlikely causes are pH shifts, total dissolved solids, Cyprinus carpio (an invasive species), metal toxicity, and pesticides. Data were insufficient to assess salinity or other toxicants. The assessment was used to develop a recovery plan for the stream. This assessment demonstrates that, even when there are many candidate causes and uncertainties are substantial, the probable causes of biological impairments can be determined with enough certainty to inform decision-making to address environmental problems.