Distribution and Toxic Effects of Cadmium and Lead on Maize Roots

Two-day-old seedlings of maize (Zea mays L.) were incubated on Cd and Pb nitrate solutions at the concentrations that inhibited root growth approximately by 50% after two-day-long incubation (LC₅₀; 10^–4 and 10^–3 M, respectively) or completely terminated growth of the primary root after one-day-long incubation (LC; 5 × 10^–4 and 10^–2 M, respectively). Cd and Pb contents were measured using an anodic inversion voltammetric technique in a flow injection system and a histochemical method. At LC₅₀, Cd and Pb were discerned, by histochemical techniques, in all root apical tissues, whereas in the root hair zone, the heavy metals were primarily accumulated in the apoplast of the rhizodermis and cortex and to a lesser extent, in the vascular tissues and parenchyma cells surrounding the metaxylem vessels. Insignificant accumulation of Cd and Pb in the pericycle probably explains why root branching was tolerant to these agents. At LC, Cd and Pb were found in the apoplast of all root tissues, in accordance with the practically complete inhibition of root growth and branching. Irrespectively of Cd and Pb concentrations in the external solution, the metal contents in the root apex exceeded those in the basal region. Procion dyes were used to assess cell death inflicted by Cd and Pb. At LC, the root cap and meristematic cells perished, together with the rhizodermal cells and the outer cortical cells of the root apex, whereas only the rhizodermal cells in the root apical region died at LC₅₀. The evidence that Cd and Pb cross the endodermal barrier at LC presumes that, at lower metal concentrations, the Casparian strip and plasmalemma of the endodermis regulate the transport of these metals into the central cylinder. The authors conclude that the identical barriers control Cd and Pb transport in root tissues.     

Plasmolysis as a Tool to Reveal Lead Localization in the Apoplast of Root Cells

In order to analyze the distribution of lead between cell walls and plasmalemma, two-day-old maize seedlings (Zea mays L.) were incubated for 24 h on a solution of lead nitrate at a concentration causing 50% inhibition of root growth (10^–5 M). Using the histochemical technique (precipitation of lead dithizonate), the distribution of lead in plasmolyzed and nonplasmolyzed cells of the root cortex was compared. This allowed us to separate the lead bound by cell walls from the lead located on the protoplast surface and in the periplasmic space. The plasmolysis was conducted prior to histochemical reaction by the incubation of seedling roots in 0.6 M sucrose solution for 30 min. The lead precipitates were located in cell walls and on the surface of protoplast. A small amount of lead was found in periplasmic space of some cells in root cortex. It is suggested that the lead is bound not only to the cell wall matrix but also to the plasmalemma.     

Enhancement of nickel and lead accumulation and their toxic growth-inhibitory effects on amaranth seedlings in the presence of calcium

The effects of Ni(NO₃)₂ and Pb(NO₃)₂ on Amaranthus sp. L. root growth and the effect of calcium on heavy metal (HM) accumulation in the growing root zone and root growth inhibition were studied. The seeds were germinated in the Pb(NO₃)₂ solutions at concentrations of 50, 100, 200, 500 and 700 μM or Ni(NO₃)₂ solutions at concentrations of 10, 50, 70, 100, and 500 μM in the presence of 100 μM Ca(NO₃)₂ or without it. HM toxicity was assesses in 7 days after seed sowing by the root length. Distribution of HM over the tissues of the growing root part was examined histochemically. Ni was more toxic to root growth than Pb. In the presence of Ca, Ni and Pb accumulation in the amaranth root growing part increased markedly, and this enhanced their growth-inhibitory of action. A comparison of results obtained in this work and available from the literature permitted a conclusion that the routes of HM penetration into the root differ in different plant species, and this determines ambiguity of protective Ca action.     

A New Adenovirus Based Vaccine Vector Expressing an Eimeria tenella Derived TLR Agonist Improves Cellular Immune Responses to an Antigenic Target

Background

Adenoviral based vectors remain promising vaccine platforms for use against numerous pathogens, including HIV. Recent vaccine trials utilizing Adenovirus based vaccines expressing HIV antigens confirmed induction of cellular immune responses, but these responses failed to prevent HIV infections in vaccinees. This illustrates the need to develop vaccine formulations capable of generating more potent T-cell responses to HIV antigens, such as HIV-Gag, since robust immune responses to this antigen correlate with improved outcomes in long-term non-progressor HIV infected individuals.

Methodology/Principal Findings

In this study we designed a novel vaccine strategy utilizing an Ad-based vector expressing a potent TLR agonist derived from Eimeria tenella as an adjuvant to improve immune responses from a [E1-]Ad-based HIV-Gag vaccine. Our results confirm that expression of rEA elicits significantly increased TLR mediated innate immune responses as measured by the influx of plasma cytokines and chemokines, and activation of innate immune responding cells. Furthermore, our data show that the quantity and quality of HIV-Gag specific CD8⁺ and CD8⁻ T-cell responses were significantly improved when coupled with rEA expression. These responses also correlated with a significantly increased number of HIV-Gag derived epitopes being recognized by host T cells. Finally, functional assays confirmed that rEA expression significantly improved antigen specific CTL responses, in vivo. Moreover, we show that these improved responses were dependent upon improved TLR pathway interactions.

Conclusion/Significance

The data presented in this study illustrate the potential utility of Ad-based vectors expressing TLR agonists to improve clinical outcomes dependent upon induction of robust, antigen specific immune responses.     

Regulation of Epithelial Differentiation in Rat Intestine by Intraluminal Delivery of an Adenoviral Vector or Silencing RNA Coding for Schlafen 3

Although we stimulate enterocytic proliferation to ameliorate short gut syndrome or mucosal atrophy, less effort has been directed at enterocytic differentiation. Schlafen 3 (Slfn3) is a poorly understood protein induced during IEC-6 enterocytic differentiation. We hypothesized that exogenous manipulation of Slfn3 would regulate enterocytic differentiation in vivo. Adenoviral vector coding for Slfn3 cDNA (Ad-GFP-Slfn3) or silencing RNA for Slfn3 (siSlfn3) was introduced intraluminally into rat intestine. We assessed Slfn3, villin, sucrase-isomaltase (SI), Dpp4, and Glut2 by qRT-PCR, Western blot, and immunohistochemistry. We also studied Slfn3 and these differentiation markers in atrophic defunctionalized jejunal mucosa and the crypt-villus axis of normal jejunum. Ad-GFP-Slfn3 but not Ad-GFP increased Slfn3, villin and Dpp4 expression in human Caco-2 intestinal epithelial cells. Injecting Ad-GFP-Slfn3 into rat jejunum in vivo increased mucosal Slfn3 mRNA three days later vs. intraluminal Ad-GFP. This Slfn3 overexpression was associated with increases in all four differentiation markers. Injecting siSlfn3 into rat jejunum in vivo substantially reduced Slfn3 and all four intestinal mucosal differentiation markers three days later, as well as Dpp4 specific activity. Endogenous Slfn3 was reduced in atrophic mucosa from a blind-end Roux-en-Y anastomosis in parallel with differentiation marker expression together with AKT and p38 signaling. Slfn3 was more highly expressed in the villi than the crypts, paralleling Glut2, SI and Dpp4. Slfn3 is a key intracellular regulator of rat enterocytic differentiation. Understanding how Slfn3 works may identify targets to promote enterocytic differentiation and maintain mucosal function in vivo, facilitating enteral nutrition and improving survival in patients with mucosal atrophy or short gut syndrome.     

Vaccine platforms combining circumsporozoite protein and potent immune modulators, rEA or EAT-2, paradoxically result in opposing immune responses

Background

Malaria greatly impacts the health and wellbeing of over half of the world''s population. Promising malaria vaccine candidates have attempted to induce adaptive immune responses to Circumsporozoite (CS) protein. Despite the inclusion of potent adjuvants, these vaccines have limited protective efficacy. Conventional recombinant adenovirus (rAd) based vaccines expressing CS protein can induce CS protein specific immune responses, but these are essentially equivalent to those generated after use of the CS protein subunit based vaccines. In this study we combined the use of rAds expressing CS protein along with rAds expressing novel innate immune response modulating proteins in an attempt to significantly improve the induction of CS protein specific cell mediated immune (CMI) responses.

Methods and Findings

BALB/cJ mice were co-vaccinated with a rAd vectors expressing CS protein simultaneous with a rAd expressing either TLR agonist (rEA) or SLAM receptors adaptor protein (EAT-2). Paradoxically, expression of the TLR agonist uncovered a potent immunosuppressive activity inherent to the combined expression of the CS protein and rEA. Fortunately, use of the rAd vaccine expressing EAT-2 circumvented CS protein''s suppressive activity, and generated a fivefold increase in the number of CS protein responsive, IFNγ secreting splenocytes, as well as increased the breadth of T cells responsive to peptides present in the CS protein. These improvements were positively correlated with the induction of a fourfold improvement in CS protein specific CTL functional activity in vivo.

Conclusion

Our results emphasize the need for caution when incorporating CS protein into malaria vaccine platforms expressing or containing other immunostimulatory compounds, as the immunological outcomes may be unanticipated and/or counter-productive. However, expressing the SLAM receptors derived signaling adaptor EAT-2 at the same time of vaccination with CS protein can overcome these concerns, as well as significantly improve the induction of malaria antigen specific adaptive immune responses in vivo.     

Accumulation of Nickel by Excluder Thlaspi arvense and Hyperaccumulator Noccaea caerulescens upon Short-Term and Long-Term Exposure

Russian Journal of Plant Physiology - The ability to accumulate nickel (Ni) was compared in hyperaccumulator Noccaea сaerulescens F.K. Mey and excluder Thlaspi arvense L. after a...     

Genotyping of rubella virus circulating in Western Siberia of Russia during 2004-2006 epidemic period

Twenty one strains of rubella virus were isolated in the Western Siberia during 2004-2006 epidemic period. Genotyping of isolated strains was performed by partial sequencing of glycoprotein E1 gene. Phylogenetic analysis showed that 20 out of 21 isolated in the Western Siberia strains of rubella virus belonged to genotype 1g, and 1 strain (isolated in Altai region in 2006)--to genotype 1E.