Boosting of Replication Competent NYVAC Primed HIV-1-Specific T-Cell Responses by HIV Synthetic Long Peptides (SLP)

Background: To enhance the induction of insert specific immune responses, a new generation of replication competent poxvirus vectors was designed and evaluated against non-replicating poxvirus vectors in a HIV vaccine study in non human primates.Methods: Rhesus macaques were immunized with either the non-replicating variant NYVAC-GagPolNef HIV-1 clade C or the replicating NYVAC-GagPolNef-C-KC, boosted with HIVGag- PolEnv-SLP and immune responses were monitored.Results: Gag-specific T-cell responses were only detected in animals immunized with the replicating NYVAC-GagPolNef-C-KC variant. Further enhancement and broadening of the immune response was studied by boosting the animals with novel T-cell immunogens HIVconsv synthetic long peptides (SLP), which direct vaccine-induced responses to the most conserved regions of HIV and contain both CD4 T-helper and CD8 CTL epitopes. The adjuvanted (Montanide ISA-720) SLP divided into subpools and delivered into anatomically separate sites enhanced the Gag-specific T-cell responses in 4 out of 6 animals, to more than 1000 SFC/106 PBMC in some animals. Furthermore, the SLP immunization broadened the immune response in 4 out of 6 animals to multiple Pol epitopes. Even Env-specific responses, to which the animals had not been primed, were induced by SLP in 2 out of 6 animals.Conclusion: This new immunization strategy of priming with replicating competent poxvirus NYVAC-HIVGagPolNef and boosting with HIVGagPolEnv-SLP, induced strong and broad Tcell responses and provides a promising new HIV vaccine approach. This study was performed within the Poxvirus T-cell Vaccine Discovery Consortium (PTVDC) which is part of the CAVD program.

Phase I safety and immunogenicity trial of Plasmodium vivax CS derived long synthetic peptides adjuvanted with montanide ISA 720 or montanide ISA 51.

We assessed the safety, tolerability, and immunogenicity of a mixture of three synthetic peptides derived from the Plasmodium vivax circumsporozoite protein formulated in Montanide ISA 720 or Montanide ISA 51. Forty healthy malaria-naive volunteers were allocated to five experimental groups (A-E): four groups (A-D) were immunized intramuscularly with 50 and 100 μg/dose injections of a mixture of N, R, and C peptides formulated in the two different adjuvants at 0, 2, and 4 months and one group was administered placebo. Vaccines were immunogenic, safe, well tolerated, and no serious adverse events related to the vaccine occurred. Seroconversion occurred in > 90% of the vaccines and antibodies recognized the sporozoite protein on immunofluorescent antibody test. Vaccines in Montanide ISA 51 showed a higher sporozoite protein recognition and interferon production. Results encourage further testing of the vaccine protective efficacy.

Estimation of surface roughness in a semiarid region from C-band ERS-1 synthetic aperture radar data

In this study, we investigated the feasibility of using the C-band European Remote Sensing Satellite (ERS-1) synthetic aperture radar (SAR) data to estimate surface soil roughness in a semiarid rangeland. Radar backscattering coefficients were extracted from a dry and a wet season SAR image and were compared with 47 in situ soil roughness measurements obtained in the rocky soils of the Walnut Gulch Experimental Watershed, southeastern Arizona, USA. Both the dry and the wet season SAR data showed exponential relationships with root mean square (RMS) height measurements. The dry C-band ERS-1 SAR data were strongly correlated (R² = 0.80), while the wet season SAR data have somewhat higher secondary variation (R² = 0.59). This lower correlation was probably provoked by the stronger influence of soil moisture, which may not be negligible in the wet season SAR data. We concluded that the single configuration C-band SAR data is useful to estimate surface roughness of rocky soils in a semiarid rangeland.

A template-assembled synthetic protein surface mimetic of the von Willebrand factor A1 domain inhibits botrocetin-induced platelet aggregation.

Platelet adhesion, the initial step of platelet activation, is mediated by the interaction of von Willebrand factor (VWF) with its platelet receptor, the GPIb-IX complex. The binding of VWF to GPIb-IX is induced either by increased shear stress or by exogenous modulators, such as botrocetin. At a molecular level, this interaction takes place between the A1 domain of VWF and the GPIb alpha chain of the GPIb-IX complex. We report here the design and functional characteristics of a VWF template-assembled synthetic protein (TASP), a chimeric four-helix-bundle TASP scaffold mimicking the surface of the A1 domain. Twelve residues located on helices alpha 3 and alpha 4 in the native A1 domain were grafted onto a surface formed by two neighboring helices of the TASP. VWF TASP was found to inhibit specifically botrocetin-induced platelet aggregation and to bind both botrocetin and GPIb alpha. However, in contrast to the native A1 domain, VWF TASP did not bind simultaneously to both ligands. Modeling studies revealed that the relative orientation of the alpha helices in VWF TASP led to a clash of bound botrocetin and GPIb alpha. These results demonstrate that a chimeric four-helix-bundle TASP as a scaffold offers a suitable surface for presenting crucial residues of the VWF A1 domain; the potential of the TASP approach for de novo protein design and mimicry is thereby illustrated.

Obtenció d'estruvita a partir de la fracció líquida digerida de purins de vaca

Actualment a Catalunya existeixen zones amb importants limitacions per l’aplicació de purins al sòl, pel que és imprescindible trobar alternatives de gestió i tractament que permetin l’aprofitament adequat dels recursos continguts a les dejeccions ramaderes sense afectar el medi. La digestió anaeròbia és una de les tècniques utilitzades en el tractament de les dejeccions ramaderes. L’efluent líquid que s’obté d’aquest tractament no modifica el contingut de nitrogen i fòsfor i per tant ha de ser gestionat correctament. L’objectiu general d’aquest projecte és avaluar la precipitació d’estruvita (sal de magnesi, amoni i fosfat) com una alternativa de gestió de l’efluent líquid d’una planta de digestió anaeròbia i compostatge que tracta dejeccions ramaderes conjuntament amb altres residus orgànics. S’han avaluat els efectes dels diferents paràmetres operacionals en la formació d’estruvita (pH, temperatura, velocitat d’agitació, alcalinitat), mitjançant assaigs en discontinu amb solució sintètica. A continuació s’ha procedit a obtenir estruvita a partir de la fracció líquida digerida de purí (FLD), en assaigs en discontinu per estudiar l’efecte del contingut de matèria orgànica i sòlids Totals (ST), així com el contingut en fosfats i el pH de reacció. Finalment, s’han optimitzat els paràmetres de procés en continu, mitjançant la posada en marxa d’un reactor a escala de laboratori i estudi de l’efecte de la velocitat d’agitació i de la introducció del stripping de CO2, tant amb solució sintètica com amb la fracció líquida digerida del purí. Dels resultats obtinguts es pot concloure que els factors que tenen una major influència en el procés d’obtenció d’estruvita són el pH (el pH òptim es situa al voltant de 9), i la presència de matèria orgànica i sòlids ens suspensió, que interfereix de forma quantitativa i qualitativa en la formació de l’estruvita. En el procés en continu s’ha aconseguit reduccions d’un 84% i 98% d’amoni i fòsfor respectivament, obtenintse estruvita que pot ser utilitzada com a fertilitzant d’alliberació lenta. Es pot concloure que la precipitació d’estruvita és una bona alternativa per millorar la gestió de les dejeccions ramaderes alhora que permet recuperar nutrients i tancar cicles. La combinació amb un tractament previ que elimini la matèria orgànica, com podria ser la digestió anaeròbia, i una separació de fases, per eliminar els sòlids en suspensió, es presenta com una configuració amb molts avantatges.

The synthetic Plasmodium falciparum circumsporozoite peptide PfCS102 as a malaria vaccine candidate: a randomized controlled phase I trial.

BACKGROUND: Fully efficient vaccines against malaria pre-erythrocytic stage are still lacking. The objective of this dose/adjuvant-finding study was to evaluate the safety, reactogenicity and immunogenicity of a vaccine candidate based on a peptide spanning the C-terminal region of Plasmodium falciparum circumsporozoite protein (PfCS102) in malaria naive adults. METHODOLOGY AND PRINCIPAL FINDINGS: Thirty-six healthy malaria-naive adults were randomly distributed into three dose blocks (10, 30 and 100 microg) and vaccinated with PfCS102 in combination with either Montanide ISA 720 or GSK proprietary Adjuvant System AS02A at days 0, 60, and 180. Primary end-point (safety and reactogenicity) was based on the frequency of adverse events (AE) and of abnormal biological safety tests; secondary-end point (immunogenicity) on P. falciparum specific cell-mediated immunity and antibody response before and after immunization. The two adjuvant formulations were well tolerated and their safety profile was good. Most AEs were local and, when systemic, involved mainly fatigue and headache. Half the volunteers in AS02A groups experienced severe AEs (mainly erythema). After the third injection, 34 of 35 volunteers developed anti-PfCS102 and anti-sporozoite antibodies, and 28 of 35 demonstrated T-cell proliferative responses and IFN-gamma production. Five of 22 HLA-A2 and HLA-A3 volunteers displayed PfCS102 specific IFN-gamma secreting CD8(+) T cell responses. Responses were only marginally boosted after the 3(rd) vaccination and remained stable for 6 months. For both adjuvants, the dose of 10 microg was less immunogenic in comparison to 30 and 100 microg that induced similar responses. AS02A formulations with 30 microg or 100 microg PfCS102 induced about 10-folds higher antibody and IFN-gamma responses than Montanide formulations. CONCLUSIONS/SIGNIFICANCE: PfCS102 peptide was safe and highly immunogenic, allowing the design of more advanced trials to test its potential for protection. Two or three immunizations with a dose of 30 microg formulated with AS02A appeared the most appropriate choice for such studies. TRIAL REGISTRATION: Swissmedic.ch 2002 DR 1227.

Effluent particle removal by microirrigation system filters

The aim of this work was to determine whether the filters used in microirrigation systems can remove potentially emitter-clogging particles. The particle size and volume distributions of different effluents and their filtrates were established, and the efficiency of the removal of these particles and total suspended solids by screen, disc and sand filters determined. In most of the effluents and filtrates, the number of particles with a diameter > 20 μm was minimal. By analysing the particle volume distribution it was found that particles larger than the disc and screen filter pores appeared in the filtrates. However, the sand filter was able to retain particles larger than the pore size. The filtration efficiency depended more on the type of effluent than on the filter. It was also found that the particle size distribution followed a potential law. Analysis of the β exponents showed that the filters did not significantly modify the particle size distribution of the effluents

Activity of phosphorus of synthetic Fe-K-P compounds in superphosphate fertilizers as affected by pH and ionic strength

The concentration of orthophosphate ions released from Fe-K-P compounds (Fe3KH8(PO4)6 .6H2O and Fe3KH14(PO4)8 .4H2O) present in superphosphates increases with pH, which initially suggests that the agronomic effectiveness of P fertilizers containing high amounts of these compounds would also increase with soil pH but studies considering activity, instead of concentration, are necessary. With this purpose, both compounds were synthesized under laboratory conditions, characterized by elemental chemical analysis, optical microscopy, X ray diffractometry, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), and used in a solubility study. Solutions of 0.01, 0.05 and 0.1 mol L-1 NaCl with pH adjusted to 3.0, 4.0, 5.0, 5.5, 6.0, 6.5, 7.0 and 7.5 were prepared for the solubility study of H8-syn, H14-syn and a phosphate rock (PR) from Brazil. The orthophosphate activity as H2PO4- and HPO4(2-) was calculated in each situation as related to pH and ionic strength using software MINTEQ. The remaining precipitates after equilibrium were chemically analyzed and subjected to X ray, SEM and EDS. Results of chemical analysis and instrumental techniques confirmed the preparation method. The activity of orthophosphate ions of both compounds tended to decrease under increasing pH and/or ionic strength of the solution, which in turn suggests that an increase in the solution pH does not necessarily promote an increase in the P bioavailability for plant uptake. This can be important when evaluating agronomic data of P fertilizers with high contents of these two Fe-K-P compounds.

Cristallochemical characterization of synthetic Zn-substituted maghemites (g-Fe2-xZn xO3)

Maghemite (g-Fe2O3) is the most usually found ferrimagnetic oxide in red basalt-derived soils. The variable degrees of ionic substitution of Fe3+ for different metals (e.g. Ti4+, Al3+, Mg2+, Zn2+, and Mn2+) and non-metals in the maghemite structure influence some cristallochemical features of this iron oxide. In this study, synthetic Zn-substituted maghemites were prepared by co-precipitation in alkaline aqueous media of FeSO4.7H2O with increasing amounts of ZnSO4.7H2O to obtain the following sequence of Fe3+ for Zn2+ substitutions: 0.0, 0.025, 0.05, 0.10, 0.15, 0.20, and 0.30 mol mol-1. The objective of this work was to evaluate the cristallochemical alterations of synthetic Zn-substituted maghemites. The dark black synthetic precipitated material was heated to 250 °C during 4 h forming a brownish maghemite that was characterized by chemical analysis as well as X ray diffraction (XRD), specific surface area and mass-specific magnetic susceptibility. The isomorphic substitution levels observed were of 0.0013, 0.0297, 0.0590, 0.1145, 0.1764, 0.2292 and 0.3404 mol mol-1, with the formation of a series of maghemites from Fe2Zn0O3 to Fe(1.49)Zn(0.770)O3 . The increase in Fe3+ for Zn2+ substitution, [Zn mol mol-1] increased the dimension a0 of the cubic unit cells of the studied maghemites according to the regression equation: a0 = 0.8343 + 0.02591Zn (R² = 0.98). On the other hand, the mean crystallite dimension and mass-specific magnetic susceptibility of the studied maghemites decreased with increasing isomorphic substitution.

Transport of a biocontrol Pseudomonas fluorescens through 2.5-m deep outdoor lysimeters and survival in the effluent water

Application of wild-type or genetically-modified bacteria to the soil environment entails the risk of dissemination of these organisms to the groundwater. To measure vertical transport of bacteria under natural climatic conditions, Pseudomonas fluorescens strain CHA0 was released together with bromide as a mobile tracer at the surface of large outdoor lysimeters. Two experiments, one starting in autumn 1993 and the other in spring 1994 were performed. Shortly after a heavy rainfall in late spring 1994, the released bacteria were detected for the first time in effluent water from the 2.5-m-deep lysimeters in both experiments, i.e. 210 d and 21 d, respectively, after inoculation. Only a 10−9 to 10−8 fraction of the inoculum was recovered as culturable cells in the effluent water, but a larger fraction of the CHA0 cells was in a non-culturable state as detected with immunofluorescence microscopy. As much as 50% of the mobile tracer percolated through the lysimeters, indicating that, compared with bromide, bacterial cells were retained in soil. In the second part of this study, persistence of CHA0 in groundwater microcosms consisting of lysimeter effluent water was studied for 380 d. Survival of the inoculant as culturable cells was better under anaerobic than under aerobic conditions. However, a large fraction of the cells became non-culturable in both cases. When the experiment was performed with filter-sterilized effluent water, the total count of introduced bacteria did not decline with time. In conclusion, the biocontrol strain was transported in low numbers to a potential groundwater level under natural climatic conditions, but could persist for an extended period in groundwater microcosms.

Intranasal administration of the synthetic polypeptide from the C-terminus of the circumsporozoite protein of Plasmodium berghei with the modified heat-labile toxin of Escherichia coli (LTK63) induces a complete protection against malaria challenge.

Needle-free procedures are very attractive ways to deliver vaccines because they diminish the risk of contamination and may reduce local reactions, pain or pain fear especially in young children with a consequence of increasing the vaccination coverage for the whole population. For this purpose, the possible development of a mucosal malaria vaccine was investigated. Intranasal immunization was performed in BALB/c mice using a well-studied Plasmodium berghei model antigen derived from the circumsporozoite protein with the modified heat-labile toxin of Escherichia coli (LTK63), which is devoid of any enzymatic activity compared to the wild type form. Here, we show that intranasal administration of the two compounds activates the T and B cell immune response locally and systemically. In addition, a total protection of mice is obtained upon a challenge with live sporozoites.

Particle dispersion in synthetic turbulent flows

We study particle dispersion advected by a synthetic turbulent flow from a Lagrangian perspective and focus on the two-particle and cluster dispersion by the flow. It has been recently reported that Richardson¿s law for the two-particle dispersion can stem from different dispersion mechanisms, and can be dominated by either diffusive or ballistic events. The nature of the Richardson dispersion depends on the parameters of our flow and is discussed in terms of the values of a persistence parameter expressing the relative importance of the two above-mentioned mechanisms. We support this analysis by studying the distribution of interparticle distances, the relative velocity correlation functions, as well as the relative trajectories.

Langevin approach to generate synthetic turbulent flows

We present an analytical scheme, easily implemented numerically, to generate synthetic Gaussian turbulent flows by using a linear Langevin equation, where the noise term acts as a stochastic stirring force. The characteristic parameters of the velocity field are well introduced, in particular the kinematic viscosity and the spectrum of energy. As an application, the diffusion of a passive scalar is studied for two different energy spectra. Numerical results are compared favorably with analytical calculations.