Histamine release factor from Dermanyssus gallinae (De Geer): characterization and in vitro assessment as a protective antigen

A cDNA encoding a 174-amino-acid orthologue of a tick histamine release factor (HRF) was identified from the haematophagous poultry red mite Dermanyssus gallinae. The predicted D. gallinae HRF protein (Dg-HRF-1) sequence is highly conserved with the tick HRFs (identity 52-54%) and to a lesser degree with translationally controlled tumour proteins (TCTP) from mammals and other invertebrates (range 38-47%). Phylogenetically, Dg-HRF-1 partitions with the tick HRF clade suggesting a shared linage and potentially similar function(s). A recombinant Dg-HRF-1 protein (rDg-HRF-1) was produced and shown to induce degranulation of rat peritoneal mast cells in vitro, confirming conservation of the histamine-releasing function in D. gallinae. Polyclonal antibodies were generated in rabbits and hens to rDg-HRF-1. Western blotting demonstrated that native Dg-HRF is a soluble protein and immunohistochemical staining of mite sections revealed that the distribution of Dg-HRF, although ubiquitous, is more common in mite reproductive, digestive and synganglion tissues. A survey of hens housed continuously in a mite-infested commercial poultry unit failed to identify IgY specific for recombinant or native Dg-HRF, indicating that Dg-HRF is not exposed to the host during infestation/feeding and may therefore have potential as a vaccine using the concealed antigen approach. To test the protective capability of rDg-HRF-1, fresh heparinised chicken blood was enriched with yolk-derived anti-Dg-HRF IgY antibodies and fed to semi-starved mites using an in vitro feeding system. A statistically significant increase in mortality was shown (P = 0.004) in mites fed with anti-Dg-HRF IgY after just one blood meal. The work presented here demonstrates, to our knowledge for the first time, the feasibility of vaccinating hens with recombinant D. gallinae antigens to control mite infestation and the potential of rDg-HRF-1 as a vaccine antigen.     

Discovery of the butenyl-spinosyn insecticides: Novel macrolides from the new bacterial strain Saccharopolyspora pogona

A new bacterium, Saccharopolyspora pogona (NRRL30141) was discovered which produced a series of very potent insecticidal compounds structurally related to the ‘classical’ (i.e., C-21-ethyl) spinosyns. A series of fermentations gave sufficient extract to allow the isolation and characterization of a total of 31 new metabolites. The majority of these compounds contained a but-1-enyl group at C-21 of the macrolide in place of the ethyl group in the ‘classical’ spinosyn series, corresponding to an additional acetate group incorporated during their biosynthesis. Additionally a variety of other new functionality was seen including hydroxylations, several novel forosamine sugar replacements, and a novel 14-membered macrolide ring analog.     

Acetylation of PAMAM dendrimers for cellular delivery of siRNA

Background  

The advancement of gene silencing via RNA interference is limited by the lack of effective short interfering RNA (siRNA) delivery vectors. Rational design of polymeric carriers has been complicated by the fact that most chemical modifications affect multiple aspects of the delivery process. In this work, the extent of primary amine acetylation of generation 5 poly(amidoamine) (PAMAM) dendrimers was studied as a modification for the delivery of siRNA to U87 malignant glioma cells.     

QMEANclust: estimation of protein model quality by combining a composite scoring function with structural density information

Background  

The selection of the most accurate protein model from a set of alternatives is a crucial step in protein structure prediction both in template-based and ab initio approaches. Scoring functions have been developed which can either return a quality estimate for a single model or derive a score from the information contained in the ensemble of models for a given sequence. Local structural features occurring more frequently in the ensemble have a greater probability of being correct. Within the context of the CASP experiment, these so called consensus methods have been shown to perform considerably better in selecting good candidate models, but tend to fail if the best models are far from the dominant structural cluster. In this paper we show that model selection can be improved if both approaches are combined by pre-filtering the models used during the calculation of the structural consensus.     

Factors influencing in the response of <i>Schizolobium parahybum</i> (Vell) Blake to <i>Ceratocystis paradoxa</i> and <i>C. moniliformis</i>

In the Ecuadorian coast one of the most destructive diseases of the pachaco is vascular wilt or stem rot caused by Ceratocystis complex, so the aim of this study was to determine the factors that affect the efficiency of the reaction of bark pachaco to this disease. This research was conducted under laboratory conditions, using trees pachaco S38, S41, S98, AE-1, AE-2 and AE-3, and pathogenic species Ceratocystis paradoxa and C. moniliformis. The method utilized was tissue stem bark,with bark sections with 4.5 cm², and a suspension of 3x10⁴ units infection and remained in a humid chamber for 96 hours at 25 ± 5 °C. Were determined grades of resistance/ susceptibility using a scale from 0 to 4, depending on the amount of mycelia and peritecio in each plant sample. Three factors were used: four colonies obtained by several transfers from each fungal specie, four ages of colonies of each fungal specie and four volumes of inoculum applied (units of infection), using for each experiment separately Completely Randomized Design with 4 replications factorial arrangement. For comparison between treatment means was used Tukey test at 5% probability of error. For future trials using this technique, you could use 30-day colonies for C. paradoxa and 40 days for C. moniliformis, and an application volume of 100 μL/cm², it would improve the level of response for the formation of perithecium and mycelia in samples cortex.     

Context alters the ability of clitoral stimulation to induce a sexually-conditioned partner preference in the rat

We have shown previously that clitoral stimulation (CLS) of female rats induces significant conditioned place preference (CPP), indicating that it is rewarding. The present study asked whether CLS could induce a conditioned partner preference. In the first experiment, sexually naïve females received 10 alternating trials of CLS and No-CLS in the presence of a male rat behind a wire-mesh screen. For one group, CLS was made in the presence of the male scented with almond extract. On alternating trials, those females received sham CLS in the presence of an unscented male behind the screen. The order was reversed for the other group. After 5 trials in each condition, females were placed into an open field with two sexually vigorous males, one scented and the other unscented. Contrary to expectation, females displayed a preference for the male associated with sham CLS. The second experiment examined whether a partner preference could be conditioned by associating CLS with the almond odor alone. A new group of sexually naive females received the same CLS-odor, No-CLS-No Odor pairings as above, but with the odor presented on cotton gauze in the chamber. During the final open field test, those females selectively solicited the scented male. We conclude that CLS that induces CPP also induces conditioned partner preference. However, we propose that CLS in the presence of an inaccessible male created a sexual inhibitory state for female rats.     

White-nose syndrome and wing damage index scores among summer bats in the northeastern United States

White-nose syndrome (WNS) adversely affects millions of bats hibernating in caves of the eastern United States. Beginning in 2009, the US Fish and Wildlife Service supported use of a wing damage index (WDI) scoring system (scale of 0 to 3, or no damage to severe) to assess wing damage of bats captured during summer. Based on bat captures at 459 mist net sites in Pennsylvania, New York, Maryland, Virginia, and New Jersey, USA, we questioned whether WDI scores varied by species group, date, and distance to the closest known affected hibernaculum. We also compared relative health (body mass index [BMI] scores) to WDI scores. Of 3,419 bats (nine species), only four individuals (0.1%; little brown [Myotis lucifugus] and northern bats [Myotis septentrionalis]) were scored as a 3 and 47 (1.4%; big brown [Eptesicus fuscus], little brown, and northern bats) as a 2. All tree bats (eastern red [Lasiurus borealis], hoary [Lasiurus cinereus], and silver-haired bats [Lasionycteris noctivagans]) scored a 0 or 1, suggesting that these species were not affected by WNS. The average WDI score decreased as summer progressed, although trends were weak. Average WDI score and number of bats with class 2 and 3 damage increased with proximity to a known WNS-positive hibernaculum. Similarly, the number of bats with severe wing damage (scoring 2 or 3) was greater at sites closer to infected hibernacula, but little variance was explained by the trend. When species-specific BMI was examined, trends were consistent by sex (female BMI scores were higher than those of males), but no relationship was discovered between BMI and WDI scores. We conclude that, at this larger geographic scale, WDI is not a clear indicator of bat health.     

Tropospheric 14CO2 at Wellington, New Zealand: the world??s longest record

Measurements of near-sea-level tropospheric ??¹⁴CO₂ have been made at Wellington, New Zealand since December 1954; these measurements comprise the longest such record available. The ??¹⁴C rose from ?10?? in 1955 peaking at 695?? in 1965 as a result of ??bomb ¹⁴C?? production, before falling thereafter to the present day (2005) value of 73??. The ??¹⁴C peak occurred about 1?year later in the southern hemisphere than in the northern hemisphere. The post-1965 fall is due to the transfer of ¹⁴C-enriched CO₂ to the biospheric and oceanic pools together with ongoing release of ¹⁴C-free CO₂ from fossil fuel combustion, during an era without major atmospheric nuclear-weapon tests. Time series analysis of the data using Loess decomposition and filtering indicates an approximately exponential decline in excess ??¹⁴CO₂ over 1967?C2005 with an e-folding time of 18?years. The seasonal cycle from 1954 until 1980 had a maximum in the late (austral) summer, a minimum in winter, with peak-to-trough amplitude that peaked at 20?? in 1966. For the period 1980?C1989, a new seasonal cycle emerged, with a maximum in winter and a minimum in late summer/early autumn and peak-to-trough amplitude of 3.5??, transitioning to a new seasonal structure after about 1990.     

Subcellular localization of silicon and germanium in grass root and leaf tissues by SIMS: evidence for differential and active transport

Silicon transport and incorporation into plant tissue is important to both plant physiological function and to the influence plants have on ecosystem silica cycling. However, the mechanisms controlling this transport have only begun to be explored. In this study, we used secondary ion mass spectrometry (SIMS) to image concentrations of Si in root and shoot tissues of annual blue grass (Poa annua L.) and orchard grass (Dactylis glomerata L.) with the goal of identifying control points in the plant silica uptake pathway. In addition, we used SIMS to describe the distributions of germanium (Ge); the element used to trace Si in biogeochemical studies. Within root tissue, Si and Ge were localized in the suberized thick-walled region of endodermal cells, i.e. the proximal side of endodermal cells which is in close association to the casparian strip. In leaves, Si was present in the cell walls, but Ge was barely detectable. The selective localization of Si and Ge in the proximal side of endodermal cell walls of roots suggests transport control is exerted upon Si and Ge by the plant. The absence of Si in most root cell walls and its presence in the cell walls of leaves (in areas outside of the transpiration terminus) suggests modifications in the chemical form of Si to a form that favors Si complexation in the cell walls of leaf tissue. The low abundance of Ge in leaf tissue is consistent with previous studies that suggest preferential transport of Si relative to Ge.     

Huh-7 or HepG2 cells: which is the better model for studying human apolipoprotein-B100 assembly and secretion?

Apolipoprotein-B100 (apoB100) is the essential protein for the assembly and secretion of very low density lipoproteins (VLDL) from liver. The hepatoma HepG2 cell line has been the cell line of choice for the study of synthesis and secretion of human apoB-100. Despite the general use of HepG2 cells to study apoB100 metabolism, they secrete relatively dense, lipid-poor particles compared with VLDL secreted in vivo. Recently, Huh-7 cells were adopted as an alternative model to HepG2 cells, with the implicit assumption that Huh-7 cells were superior in some respects of lipoprotein metabolism, including VLDL secretion. In this study we addressed the hypothesis that the spectrum of apoB100 lipoprotein particles secreted by Huh-7 cells more closely resembles the native state in human liver. We find that Huh-7 cells resemble HepG2 cells in the effects of exogenous lipids, microsomal triglyceride transfer protein (MTP)-inhibition, and proteasome inhibitors of apoB100 secretion, recovery, and degradation. In contrast to HepG2 cells, however, MEK-ERK inhibition does not correct the defect in VLDL secretion. Huh-7 cells do not appear to offer any advantages over HepG2 cells as a general model of human apoB100-lipoprotein metabolism.