Functional analysis of the leading malaria vaccine candidate AMA-1 reveals an essential role for the cytoplasmic domain in the invasion process

A key process in the lifecycle of the malaria parasite Plasmodium falciparum is the fast invasion of human erythrocytes. Entry into the host cell requires the apical membrane antigen 1 (AMA-1), a type I transmembrane protein located in the micronemes of the merozoite. Although AMA-1 is evolving into the leading blood-stage malaria vaccine candidate, its precise role in invasion is still unclear. We investigate AMA-1 function using live video microscopy in the absence and presence of an AMA-1 inhibitory peptide. This data reveals a crucial function of AMA-1 during the primary contact period upstream of the entry process at around the time of moving junction formation. We generate a Plasmodium falciparum cell line that expresses a functional GFP-tagged AMA-1. This allows the visualization of the dynamics of AMA-1 in live parasites. We functionally validate the ectopically expressed AMA-1 by establishing a complementation assay based on strain-specific inhibition. This method provides the basis for the functional analysis of essential genes that are refractory to any genetic manipulation. Using the complementation assay, we show that the cytoplasmic domain of AMA-1 is not required for correct trafficking and surface translocation but is essential for AMA-1 function. Although this function can be mimicked by the highly conserved cytoplasmic domains of P. vivax and P. berghei, the exchange with the heterologous domain of the microneme protein EBA-175 or the rhoptry protein Rh2b leads to a loss of function. We identify several residues in the cytoplasmic tail that are essential for AMA-1 function. We validate this data using additional transgenic parasite lines expressing AMA-1 mutants with TY1 epitopes. We show that the cytoplasmic domain of AMA-1 is phosphorylated. Mutational analysis suggests an important role for the phosphorylation in the invasion process, which might translate into novel therapeutic strategies.

New Compounds Bearing [M(S2O7)3]2– Anions (M = Si, Ge, Sn): Syntheses and Characterization of A2[Si(S2O7)3] (A = Na, K, Rb), A2[Ge(S2O7)3] (A = Li, Na, K, Rb, Cs), A2[Sn(S2O7)3] (A = Na, K), and the Unique Germanate Hg2[Ge(S2O7)3]Cl2 with Cationic 1∞[

The reaction of the group 14 tetrachlorides MCl4 (M = Si, Ge, Sn) with oleum (65 % SO3) at elevated temperatures led to the unique anionic complexes [M(S2O7)3]2– that show the central M atoms in coordination of three chelating S2O72– groups. The mean distances M–O within the complexes increase from 175 pm (M = Si) via 186 pm (M = Ge) up to 200 pm (M = Sn). The charge balance for the [M(S2O7)3]2– anions is achieved by alkaline metal ions A+ (A = Li, Na, K, Rb, Cs) which were implemented in the syntheses in form of their sulfates. The size of the A+ ions, i.e. their coordination requirement causes the crystallographic differences in the crystal structures, while the structure of the complex [M(S2O7)3]2– anions remains essentially unaffected. Furthermore, we were able to characterize the unique germanate Hg2[Ge(S2O7)3]Cl2 which forms when HgCl2 is added as a source for the counter cation. The Hg2+ and the Cl– ions form infinite cationic chains according to 1∞[HgCl2/2]+ which take care for the charge compensation. For selected examples of the compounds the thermal behavior has been monitored by means of thermal analyses and X-ray powder diffraction. For A being an alkaline metal the decomposition product is a mixture of the sulfates A2SO4 and the dioxides MO2, whereas Hg2[Ge(S2O7)3]Cl2 shows a more complicated decomposition. The tris-(disulfato)-silicate Na2[Si(S2O7)3] has additionally been examined by solid state 29Si and 23Na NMR spectroscopic measurements.

Comparação entre os intumescimentos de borracha natural e polibutadieno com alto teor 1,4-cis em solventes (n-octano, ciclo-hexano e esqualeno) e em soluções poliméricas

Amostras de borracha natural foram reticuladas por meio de radiação gama (doses de 2, 4, 6 e 8 MRad) e intumescidas: a) em n-octano, ciclo-hexano e esqualeno; b) em polibutadieno líquido (Mn=1830 g/mol; alto teor 1,4- cis) a 45°C; C) em solução de polibutadieno de baixo peso molecular (M,= 1830 g/mol; alto teor 1,4-cis) em n-octano e ciclo-hexano; d) em solução de polibutadieno de alto peso molecular (Mn= 175.000 g/rnol; alto teor 1,4-cis) emn-octano e ciclo-hexano ; e) em solução de borracha natural (Mn=779.000 g/mol) em ciclo- hexano, a 25°C. Amostras de polibutadieno (alto teor 1,4-cis) foram reticuladas por meio de radiação gama (doses de 1,s; 18 e 25 MRad) e intumescidas: a) em n-octano, ciclo-hexano e esquaieno. Salvo quando mencionado em contrário, as amostras de borracha natural e de polibutadieno foram intumescidas nas temperaturas de 25, 35 e 45°C. A partir dos ensaios de intumescirnento acima mencionados, foram determinadas as solubilidades de borracha natural e de polibutadieno em n-octano, ciclo-hexano e esqualeno através dos parâmetros de Flory-Huggins, calculados com o emprego da equação de Flory-Rehner. Verificou-se a influência da temperatura e da concentração de polimero no gel sobre o valor do parâmetro de Flory-Huggins. Também através desse parâmetro procurou-se determinar a compatibilidade entre borracha natural e polibutadieno. Foi demonstrado que cadeias lineares de baixo peso molecular de polibutadieno conseguem penetrar em amostras reticuladas de borracha natural, quando o peso molecular das cadeias lineares for bem inferior ao do arco de rede Mc das amostras reticuladas. Cadeias lineares de polibutadieno e de borracha natural com peso molecular superior ao do arco de rede da borracha natural reticulada, entretanto, não conseguem penetrar no retículo. Quranto mais alta a concentração da solução externa em moléculas de alto peso molecular, tanto menor é o grau de intumescimento da amostra reticulada.

Oxidation of melatonin and its catabolites, N-1-acetyl-N (2)-formyl-5-methoxykynuramine and N-1-acetyl-5-methoxykynuramine, by activated leukocytes

N-1-acetyl-N-2-formyl-5-methoxykynuramine (AFMK) and N-1-acetyl-5-methoxykynuramine (AMK), two melatonin catabolites, have been described as potent antioxidants. We aimed to follow the kinetics of AFMK and AMK formation when melatonin is oxidized by phorbol myristate acetate (PMA) and lipopolysaccharide (LPS)-activated leukocytes. An HPLC-based method was used for AFMK and AMK determination in neutrophil and peripheral blood mononuclear cell cultures supernatants. Samples were separated isocratically on a C18 reverse-phase column using acetonitrile/H2O (25:75) as the mobile phase. AFMK was detected by fluorescence (excitation 340 nm and emission 460 nm) and AMK by UV-VIS absorbance (254 nm). Activation of neutrophils and mononuclear cells with PMA produces larger amounts of AFMK than activation with LPS, probably due to the lower levels of reactive oxygen species formation and myeloperoxidase (MPO) degranulation that occurs when cells are stimulated with LPS. The concentration of AMK found in the supernatant was about 5-10% (from 18-hr cultures) compared with AFMK. This result may reflect its reactivity. Indeed AMK, but not AFMK, is easily oxidized by activated neutrophils in a MPO and hydrogen peroxide-dependent reaction. In conclusion, we defined a simple procedure for the determination of AFMK and AMK in biological samples and demonstrated the capacity of leukocytes to oxidize melatonin and AMK.

Measurement of the t(t)over-bar production cross section in p(p)over-bar collisions at root s=1.96 TeV using kinematic characteristics of lepton plus jets events

We present a measurement of the top quark pair production cross section in p (p) over bar collisions at root s=1.96 TeV utilizing 425 pb(-1) of data collected with the D0 detector at the Fermilab Tevatron Collider. We consider the final state of the top quark pair containing one high-p(T) electron or muon and at least four jets. We exploit specific kinematic features of t (t) over bar events to extract the cross section. For a top quark mass of 175 GeV, we measure sigma(t (t) over bar)=6.4(-1.2)(+1.3)(stat)+/- 0.7(syst)+/- 0.4(lum) pb, in good agreement with the standard model prediction.

Measurement of the t(t)over-bar production cross section in p(p)over-bar collisions at root s=1.96 TeV using lepton plus jets events with lifetime b-tagging

We present a measurement of the top quark pair (t (t) over bar) production cross section (sigma(t (t) over bar)) in pp collisions at root s = 1.96 TeV using 230 pb(-1) of data collected by the DO experiment at the Fermilab Tevatron Collider. We select events with one charged lepton (electron or muon), missing transverse energy, and jets in the final state. We employ lifetime-based b-jet identification techniques to further enhance the t F purity of the selected sample. For a top quark mass of 175 GeV, we measure sigma(t (t) over bar) 8.6(-1.5)(+1.6) (stat. + syst.) +/- 0.6(lumi.) pb, in agreement with the standard model expectation. (c) 2005 Published by Elsevier B.V.

Measurement of the t(t)over-bar production cross section in p(p)over-bar collisions at root s=1.96 TeV using kinematic characteristics of lepton plus jets events

We present a measurement of the top quark pair (t (t) over bar) production cross section (sigma(t (t) over bar)) in pp collisions at a center-of-mass energy of 1.96 TeV using 230 pb(-1) of data collected by the DO detector at the Fermilab Tevatron Collider. We select events with one charged lepton (electron or muon), large missing transverse energy, and at least four jets, and extract the t (t) over bar content of the sample based on the kinematic characteristics of the events. For a top quark mass of 175 GeV, we measure sigma(t (t) over bar) 6.7(-1.3)(+1.4)(stat)(-1.1)(+1.6)(syst) +/- 0.4(lumi) pb, in good agreement with the standard model prediction. (c) 2005 Elsevier B.V. All rights reserved.

Measurement of the p(p)over-bar -> t(t)over-bar + X production cross section at root s=1.96 TeV in the fully hadronic decay channel

A measurement of the top quark pair production cross section in proton antiproton collisions at an interaction energy of root s=1.96 TeV is presented. This analysis uses 405 +/- 25 pb(-1) of data collected with the D0 detector at the Fermilab Tevatron Collider. Fully hadronic t (t) over bar decays with final states of six or more jets are separated from the multijet background using secondary vertex tagging and a neural network. The t (t) over bar cross section is measured as sigma(t (t) over bar)=4.5(-1.9)(+2.0)(stat)(-1.1)(+1.4)(syst)+/- 0.3(lumi) pb for a top quark mass of m(t)=175 GeV/c(2).

Search for associated Higgs boson production WH -> WWW*-> l(+/-)nu l('+/-)nu(')+X in p(p)over-bar collisions at root S=1.96 TeV

We present a search for associated Higgs boson production in the process p (p) over bar -> WH -> WWW*-> l(+/-)nu l('+/-)nu(')+X in final states containing two like-sign isolated electrons or muons (e(+/-)e(+/-), e(+/-)mu(+/-), or mu(+/-)mu(+/-)). The search is based on D0 run II data samples corresponding to integrated luminosities of 360-380 pb(-1). No excess is observed over the predicted standard model background. We set 95% C.L. upper limits on sigma ->(p (p) over bar WH) x Br(H -> WW*) between 3.2 and 2.8 pb for Higgs boson masses from 115 to 175 GeV.

Measurement of the tt̄ production cross section in pp̄ collisions at √s = 1.96 TeV in dilepton final states

We present a measurement of the top quark pair (tt̄) production cross section in pp̄ collisions at √s=1.96 TeV using events with two charged leptons in the final state. This analysis utilizes an integrated luminosity of 224-243 pb-1 collected with the DØ detector at the Fermilab Tevatron Collider. We observe 13 events in the e+e -, eμ and μ+μ- channels with an expected background of 3.2±0.7 events. For a top quark mass of 175 GeV, we measure a tt̄ production cross section of σtt̄=8. 6-2.7 +3.2(stat)±1.1(syst)±0.6(lumi) pb, consistent with the standard model prediction. © 2005 Elsevier B.V. All rights reserved.

Search for associated Higgs boson production WH→WWW *→l±νl′±ν′+X in pp̄ collisions at s=1.96TeV

We present a search for associated Higgs boson production in the process pp̄→WH→WWW*→l±νl′±ν′ +X in final states containing two like-sign isolated electrons or muons (e±e±, e±μ±, or μ±μ±). The search is based on D0 run II data samples corresponding to integrated luminosities of 360-380pb-1. No excess is observed over the predicted standard model background. We set 95% C.L. upper limits on σ(pp̄→WH) ×Br(H→WW*) between 3.2 and 2.8 pb for Higgs boson masses from 115 to 175 GeV. © 2006 The American Physical Society.

Sensibilidade da técnica de reação em cadeia da polimerase para HIV-1 em relação à técnica de ensaio imunoenzimático

O vírus da imunodeficiência humana (HIV) é o agente etiológico da Síndrome da Imunodeficiência Adquirida, AIDS, uma doença de grande preocupação médica. O genoma deste vírus encontra-se arranjado em nove genes individuais e por duas estruturas idênticas denominadas de repetições terminais nas extremidades 5' e 3'. Tivemos como objetivo analisar a sensibilidade do teste da reação em cadeia da polimerase (PCR) e o teste de ensaio imunoenzimático (ELISA), como teste para triagem de doadores de sangue para HIV. Foram analisadas 200 amostras de doadores e pacientes, da Fundação HEMOPA, com padrão positivo e indeterminado no teste ELISA. Na triagem sorológica pelo ELISA tiveram como resultado 35 amostras positivas, 75 amostras negativas e 90 amostras indeterminadas as quais foram submetidas ao teste pela PCR. Vinte e cinco amostras tiveram resultado positivo e 175 amostras negativas. Com estes resultados concluímos que a reação de PCR apresenta-se positiva somente nas amostras em que o teste ELISA apresenta relação DO/cutoff, acima de 3.

N=1 super sinh-Gordon model with defects revisited

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)