Characterization of immunoglobulin G antibodies to Plasmodium falciparum sporozoite surface antigen MB2 in malaria exposed individuals

<p>Abstract</p> <p>Background</p> <p>MB2 protein is a sporozoite surface antigen on the human malaria parasite <it>Plasmodium falciparum</it>. MB2 was identified by screening a <it>P. falciparum </it>sporozoite cDNA expression library using immune sera from a protected donor immunized via the bites of <it>P. falciparum</it>-infected irradiated mosquitoes. It is not known whether natural exposure to <it>P. falciparum </it>also induces the anti-MB2 response and if this response differs from that in protected individuals immunized via the bites of <it>P. falciparum </it>infected irradiated mosquitoes. The anti-MB2 antibody response may be part of a robust protective response against the sporozoite.</p> <p>Methods</p> <p>Fragments of polypeptide regions of MB2 were constructed as recombinant fusions sandwiched between glutathione S-transferase and a hexa histidine tag for bacterial expression. The hexa histidine tag affinity purified proteins were used to immunize rabbits and the polyclonal sera evaluated in an <it>in vitro </it>inhibition of sporozoite invasion assay. The proteins were also used in immunoblots with sera from a limited number of donors immunized via the bites of <it>P. falciparum </it>infected irradiated mosquitoes and plasma and serum obtained from naturally exposed individuals in Kenya.</p> <p>Results</p> <p>Rabbit polyclonal antibodies targeting the non-repeat region of the basic domain of MB2 inhibited sporozoites entry into HepG2-A16 cells <it>in vitro</it>. Analysis of serum from five human volunteers that were immunized via the bites of <it>P. falciparum </it>infected irradiated mosquitoes that developed immunity and were completely protected against subsequent challenge with non-irradiated parasite also had detectable levels of antibody against MB2 basic domain. In contrast, in three volunteers not protected, anti-MB2 antibodies were below the level of detection. Sera from protected volunteers preferentially recognized a non-repeat region of the basic domain of MB2, whereas plasma from naturally-infected individuals also had antibodies that recognize regions of MB2 that contain a repeat motif in immunoblots. Sequence analysis of eleven field isolates and four laboratory strains showed that these antigenic regions of the basic domain of the <it>MB2 </it>gene are highly conserved in parasites obtained from different parts of the world. Moreover, anti-MB2 antibodies also were detected in the plasma of 83% of the individuals living in a malaria endemic area of Kenya (n = 41).</p> <p>Conclusion</p> <p>A preliminary analysis of the human humoral response against MB2 indicates that it may be an additional highly conserved target for immune intervention at the pre-erythrocytic stage of <it>P. falciparum </it>life cycle.</p

A review on substances and processes relevant for optical remote sensing of extremely turbid marine areas, with a focus on the Wadden Sea

The interpretation of optical remote sensing data of estuaries and tidal flat areas is hampered by optical complexity and often extreme turbidity. Extremely high concentrations of suspended matter, chlorophyll and dissolved organic matter, local differences, seasonal and tidal variations and resuspension are important factors influencing the optical properties in such areas. This review gives an overview of the processes in estuaries and tidal flat areas and the implications of these for remote sensing in such areas, using the Wadden Sea as a case study area. Results show that remote sensing research in extremely turbid estuaries and tidal areas is possible. However, this requires sensors with a large ground resolution, algorithms tuned for high concentrations of various substances and the local specific optical properties of these substances, a simultaneous detection of water colour and land-water boundaries, a very short time lag between acquisition of remote sensing and in situ data used for validation and sufficient geophysical and ecological knowledge of the area. © 2010 The Author(s)

Upregulation of CRABP1 in human neuroblastoma cells overproducing the Alzheimer-typical Aβ42 reduces their differentiation potential

<p>Abstract</p> <p>Background</p> <p>Alzheimer's disease (AD) is characterized by neurodegeneration and changes in cellular processes, including neurogenesis. Proteolytic processing of the amyloid precursor protein (APP) plays a central role in AD. Owing to varying APP processing, several β-amyloid peptides (Aβ) are generated. In contrast to the form with 40 amino acids (Aβ₄₀), the variant with 42 amino acids (Aβ₄₂) is thought to be the pathogenic form triggering the pathological cascade in AD. While total-Aβ effects have been studied extensively, little is known about specific genome-wide effects triggered by Aβ₄₂or Aβ₄₀derived from their direct precursor C99.</p> <p>Methods</p> <p>A combined transcriptomics/proteomics analysis was performed to measure the effects of intracellularly generated Aβ peptides in human neuroblastoma cells. Data was validated by real-time polymerase chain reaction (real-time PCR) and a functional validation was carried out using RNA interference.</p> <p>Results</p> <p>Here we studied the transcriptomic and proteomic responses to increased or decreased Aβ₄₂and Aβ₄₀levels generated in human neuroblastoma cells. Genome-wide expression profiles (Affymetrix) and proteomic approaches were combined to analyze the cellular response to the changed Aβ₄₂- and Aβ₄₀-levels. The cells responded to this challenge with significant changes in their expression pattern. We identified several dysregulated genes and proteins, but only the cellular retinoic acid binding protein 1 (CRABP1) was up-regulated exclusively in cells expressing an increased Aβ₄₂/Aβ₄₀ratio. This consequently reduced all-trans retinoic acid (RA)-induced differentiation, validated by CRABP1 knock down, which led to recovery of the cellular response to RA treatment and cellular sprouting under physiological RA concentrations. Importantly, this effect was specific to the AD typical increase in the Aβ₄₂/Aβ₄₀ratio, whereas a decreased ratio did not result in up-regulation of CRABP1.</p> <p>Conclusion</p> <p>We conclude that increasing the Aβ₄₂/Aβ₄₀ratio up-regulates CRABP1, which in turn reduces the differentiation potential of the human neuroblastoma cell line SH-SY5Y, but increases cell proliferation. This work might contribute to the better understanding of AD neurogenesis, currently a controversial topic.</p

How dusty is <i>α</i> Centauri? Excess or non-excess over the infrared photospheres of main-sequence stars

Context. Debris discs around main-sequence stars indicate the presence of larger rocky bodies. The components of the nearby, solar-type binary α Centauri have metallicities that are higher than solar, which is thought to promote giant planet formation. Aims. We aim to determine the level of emission from debris around the stars in the α Cen system. This requires knowledge of their photospheres. Having already detected the temperature minimum, Tmin, of α Cen A at far-infrared wavelengths, we here attempt to do the same for the more active companion α Cen B. Using the α Cen stars as templates, we study the possible effects that Tmin may have on the detectability of unresolved dust discs around other stars. Methods. We used Herschel-PACS, Herschel-SPIRE, and APEX-LABOCA photometry to determine the stellar spectral energy distributions in the far infrared and submillimetre. In addition, we used APEX-SHeFI observations for spectral line mapping to study the complex background around α Cen seen in the photometric images. Models of stellar atmospheres and of particulate discs, based on particle simulations and in conjunction with radiative transfer calculations, were used to estimate the amount of debris around these stars. Results. For solar-type stars more distant than α Cen, a fractional dust luminosity fd ≡ Ldust/Lstar ~ 2 × 10-7 could account for SEDs that do not exhibit the Tmin effect. This is comparable to estimates of fd for the Edgeworth-Kuiper belt of the solar system. In contrast to the far infrared, slight excesses at the 2.5σ level are observed at 24 μm for both α Cen A and B, which, if interpreted as due to zodiacal-type dust emission, would correspond to fd ~ (1-3) × 10-5, i.e. some 102 times that of the local zodiacal cloud. Assuming simple power-law size distributions of the dust grains, dynamical disc modelling leads to rough mass estimates of the putative Zodi belts around the α Cen stars, viz. ≲4 × 10-6 M☾of 4 to 1000 μm size grains, distributed according to n(a) ∝ a-3.5. Similarly, for filled-in Tmin emission, corresponding Edgeworth-Kuiper belts could account for ~ 10-3 M☾ of dust. Conclusions Our far-infrared observations lead to estimates of upper limits to the amount of circumstellar dust around the stars α Cen A and B. Light scattered and/or thermally emitted by exo-Zodi discs will have profound implications for future spectroscopic missions designed to search for biomarkers in the atmospheres of Earth-like planets. The far-infrared spectral energy distribution of α Cen B is marginally consistent with the presence of a minimum temperature region in the upper atmosphere of the star. We also show that an α Cen A-like temperature minimum may result in an erroneous apprehension about the presence of dust around other, more distant stars. Based on observations with Herschel which is an ESA space observatory with science instruments provided by European-led Principal Investigator consortia and with important participation from NASA. Also based on observations with APEX, which is a 12 m diameter submillimetre telescope at 5100 m altitude on Llano Chajnantor in Chile. The telescope is operated by Onsala Space Observatory, Max-Planck-Institut für Radioastronomie (MPIfR), and European Southern Observatory (ESO)

Retinoic Acid-Dependent Signaling Pathways and Lineage Events in the Developing Mouse Spinal Cord

Studies in avian models have demonstrated an involvement of retinoid signaling in early neural tube patterning. The roles of this signaling pathway at later stages of spinal cord development are only partly characterized. Here we use Raldh2-null mouse mutants rescued from early embryonic lethality to study the consequences of lack of endogenous retinoic acid (RA) in the differentiating spinal cord. Mid-gestation RA deficiency produces prominent structural and molecular deficiencies in dorsal regions of the spinal cord. While targets of Wnt signaling in the dorsal neuronal lineage are unaltered, reductions in Fibroblast Growth Factor (FGF) and Notch signaling are clearly observed. We further provide evidence that endogenous RA is capable of driving stem cell differentiation. Raldh2 deficiency results in a decreased number of spinal cord derived neurospheres, which exhibit a reduced differentiation potential. Raldh2-null neurospheres have a decreased number of cells expressing the neuronal marker β-III-tubulin, while the nestin-positive cell population is increased. Hence, in vivo retinoid deficiency impaired neural stem cell growth. We propose that RA has separable functions in the developing spinal cord to (i) maintain high levels of FGF and Notch signaling and (ii) drive stem cell differentiation, thus restricting both the numbers and the pluripotent character of neural stem cells