Molecular and epigenetic regulations and functions of the LAFL transcriptional regulators that control seed development.

The LAFL (i.e. LEC1, ABI3, FUS3, and LEC2) master transcriptional regulators interact to form different complexes that induce embryo development and maturation, and inhibit seed germination and vegetative growth in Arabidopsis. Orthologous genes involved in similar regulatory processes have been described in various angiosperms including important crop species. Consistent with a prominent role of the LAFL regulators in triggering and maintaining embryonic cell fate, their expression appears finely tuned in different tissues during seed development and tightly repressed in vegetative tissues by a surprisingly high number of genetic and epigenetic factors. Partial functional redundancies and intricate feedback regulations of the LAFL have hampered the elucidation of the underpinning molecular mechanisms. Nevertheless, genetic, genomic, cellular, molecular, and biochemical analyses implemented during the last years have greatly improved our knowledge of the LALF network. Here we summarize and discuss recent progress, together with current issues required to gain a comprehensive insight into the network, including the emerging function of LEC1 and possibly LEC2 as pioneer transcription factors.

Enzymatic gelation to prepare chitosan gels: Study of gelation kinetics and identification of limiting parameters for controlled gel morphology.

Chitosan gels with well-controlled morphology were prepared by an enzymatic chitosan gelation process using in-situ production of ammonia through the urea enzymatic hydrolysis by urease. The reaction kinetics were studied by four complementary methods (rheological, colorimetric, flowing and pH-metric measurements) to define an advanced gelation pH (and the corresponding advanced gelation time) enabling to obtain homogeneous chitosan gels with well-controlled morphology and strong enough to be suitable for a future membrane preparation. A comparative study of chitosan gelation kinetics in water and aqueous chitosan solution highlighted differences mainly related to ammonia consumption when chitosan was present. The key parameters evidenced for the control of the gelation kinetics were the temperature and the urease concentration. The operating parameters required to elaborate gels with sufficient mechanical resistance to be handled from 2.5% w/v chitosan concentration were defined: a 3.5umL-1 urease concentration, a 75mM urea concentration and a temperature of 5°C.

Removal of metal ions from aqueous effluents involving new thermosensitive polymeric sorbent.

In this work, new thermosensitive copolymers bearing phosphonated groups were synthesized and used to remove metal pollution. Sorption properties are brought by hydrolyzed (dimethoxyphosphoryl)methyl 2-methylacrylate (hMAPC1) monomer. N-n-propylacrylamide (NnPAAm) led to the thermoresponsive properties of the copolymers. Low lower critical solution temperature (LCST) values were observed, ranging between 20 and 25 °C depending on the molar ratio of each monomer in the copolymer. Sorption properties of these copolymers towards nickel ions were evaluated for increasing temperatures (10-40 °C), Ni ion concentrations of 20 mg L(-1) and pH values between 3 and 7. Best results were observed for temperatures just lower than the LCST (20 °C), when the copolymer was fully soluble in water solution. For temperature higher than the LCST, phosphonic diacid groups accessibility was considerably reduced by the precipitation of the thermosensitive part of the copolymer leading to lower sorption properties. In these conditions, the highest Ni removal by the copolymer was observed for pH = 7, when there was almost no competition between the sorption of H(+) and Ni(2+) ions on the phosphonic acid groups. These optimal conditions enabled removal of about 70% of the nickel in the synthetic effluent.

Preparation and characterization of chitin hydrogels by water vapor induced gelation route.

A novel method of chitin hydrogel preparation, called vapor induced gelation, using exposure of chitin/N-methyl-pyrrolidone/LiCl solution to water vapors is presented. Compared to gelation induced by direct immersion in water, hydrogels are characterized by smaller deformation during gelation (area shrinkage is 20% instead of 65%), larger water volume fraction (75 instead of 62%, v/v) and 10 times higher apparent compression moduli. Their nanostructure consists of thicker and larger crystalline platelets network (thickness=37 Å, apparent coherent crystalline size L020=145 Å) comparatively to direct immersion gels (25 Å and L020=95 Å). Drug delivery potential of chitin hydrogels was determined for non-interactive low molecular molecules.

Molecular identification of Rickettsia felis in ticks and fleas from an endemic area for Brazilian Spotted Fever.

Rickettsioses are arthropod-borne diseases caused by parasites from the Order Rickettsiales. The most prevalent rickettsial disease in Brazil is Brazilian Spotted Fever (BSF). This work intends the molecular detection of those agents in ectoparasites from an endemic area of BSF in the state of Espírito Santo. A total of 502 ectoparasites, among them Amblyomma cajennense, Amblyomma dubitatum (A. cooperi), Riphicephalus sanguineus, Anocentor nitens and Ctenocephalides felis, was collected from domestic animals and the environment and separated in 152 lots according to the origin. Rickettsia sp. was detected in pools of all collected species by amplification of 17 kDa protein-encoding gene fragments. The products of PCR amplification of three samples were sequenced, and Rickettsia felis was identified in R. sanguineus and C. felis. These results confirm the presence of Rickettsia felis in areas previously known as endemic for BSF, disease caused by Rickettsia rickettsii. Moreover, they show the needing of further studies for deeper knowledge of R. felis-spotted fever epidemiology and differentiation of these diseases in Brazil.

Molecular identification of Rickettsia felis in ticks and fleas from an endemic area for Brazilian Spotted Fever

Rickettsioses are arthropod-borne diseases caused by parasites from the Order Rickettsiales. The most prevalent rickettsial disease in Brazil is Brazilian Spotted Fever (BSF). This work intends the molecular detection of those agents in ectoparasites from an endemic area of BSF in the state of Espírito Santo. A total of 502 ectoparasites, among them Amblyomma cajennense, Amblyomma dubitatum (A. cooperi), Riphicephalus sanguineus, Anocentor nitens and Ctenocephalides felis, was collected from domestic animals and the environment and separated in 152 lots according to the origin. Rickettsia sp. was detected in pools of all collected species by amplification of 17kDa protein-encoding gene fragments. The products of PCR amplification of three samples were sequenced, and Rickettsia felis was identified in R. sanguineus and C. felis. These results confirm the presence of Rickettsia felis in areas previously known as endemic for BSF, disease caused by Rickettsia rickettsii. Moreover, they show the needing of further studies for deeper knowledge of R. felis-spotted fever epidemiology and differentiation of these diseases in Brazil.

Elaboration of chitosan/activated carbon composites for the removal of organic micropollutants from waters.

Composite hydrogels were prepared by a wet-casting process by blending a biopolymer, chitosan, with activated carbon (AC) for use in water treatment. Adsorption properties of the composite gels for an organic micro-pollutant (phenol) which may be encountered in wastewaters was studied with an experimental design approach as a function of: - the concentration of raw materials and thus the AC weight within the chitosan matrix. - the accessibility of AC in the polymeric matrix, which is assumed to be related to the coating and thus to the pH of the immersion bath. ESEM observations showed that at a higher pH of gelation (pH = 14), AC particles were entrapped at the surface of the polymer matrix because of a faster gelation kinetic than at a lower pH (13.3). Adsorption kinetic tests showed that phenol adsorption occurred according to two mechanisms. During the first step, phenol molecules were adsorbed by the AC particles located at the surface. The second step corresponded to a slow diffusion through chitosan chains leading to an adsorption by AC particles entrapped within the polymeric matrix coupled to an adsorption on to the chitosan. A mass transfer model was used to describe this two-step adsorption phenomenon. However, due to a heterogeneous coating of AC by chitosan, this phenomenon was not supported by experimental design results: the initial kinetic coefficients were associated with a high experimental error which didn't allow for an analysis of the influence of elaboration parameters on kinetic coefficients. Regardling equilibrium adsorption properties, it was shown that composite gels were good adsorbents for phenol with removal ranging from 94% to 98% corresponding to adsorption capacities from 30 to 41 mg g(-1). The pH of the immersion bath had no influence on equilibrium adsorption properties, contrary to the AC weight within the chitosan matrix which wasdemonstrated to influence significantly adsorption capacities. Because carbon particles may improve mechanical properties, mechanical tests were carried out on the composite gels. For a total amount of dried matter in the compositekept constant, the increase in chitosan content led to an increase in the mechanical properties, because of an increase in thenumber of interactions between chitosan chains. The influence of sonication during the stirring step, leading to a better homogenisation of AC particules within the polymer matrix, was also examined.

Identification of Ctenocephalides felis fleas as a host of Rickettsia felis, the agent of a spotted fever rickettsiosis in Yucatan, Mexico.

In search for the vector of the recently recognized spotted fever rickettsiosis of the Yucatán, ticks, fleas, and lice were collected from vegetation and dogs in localities where seropositive persons had been found. The arthropods were examined by polymerase chain reaction (PCR) using primers for the genus-specific 17-kDa protein gene followed by restriction fragment length polymorphism (RFLP) and DNA sequencing. Eleven (20%) of 54 pools of Ctenocephalides felis fleas contained DNA of Rickettsia felis. None of 219 Amblyomma cajennense, 474 Rhiphicephalus sanguineus, 258 Boophilus sp. ticks, and 33 Poliplax species lice contained DNA of Rickettsia. The identity of the rickettsial DNA was confirmed as R. felis by PCR/RFLP for the citrate synthase and outer membrane protein A genes and by DNA sequencing. The results indicate that the host of R. felis in Yucatán is C. felis and suggest that the spotted fever rickettsiosis that has infected >5% of the population of the Yucatán and can present as a dengue-like illness is likely to be caused by R. felis.

CPR5 is involved in cell proliferation and cell death control and encodes a novel transmembrane protein.

Plants often respond to pathogens by sacrificing cells at the infection site. This type of programmed cell death is mimicked by the constitutive pathogene response5 (cpr5) mutant in Arabidopsis in the absence of pathogens, suggesting a role for CPR5 in programmed cell death control. The analysis of the cellular phenotypes of two T-DNA-tagged cpr5 alleles revealed an additional role for CPR5 in the regulation of endoreduplication and cell division. In cpr5 mutant trichomes, endoreduplication cycles stop after two rounds instead of four, and trichome cells have fewer branches than normal. Eventually, cpr5 trichomes die, the nucleus disintegrates, and the cell collapses. Similarly, leaf growth stops earlier than in wild-type, and, frequently, regions displaying spontaneous cell death are observed. The cloning of the CPR5 gene revealed a novel putative transmembrane protein with a cytosolic domain containing a nuclear-targeting sequence. The dual role of CPR5 in cell proliferation and cell death control suggests a regulatory link between these two processes.

Cell polarity in Arabidopsis trichomes.

Arabidopsis leaf trichomes are unicellular hairs that display a highly characteristic cell form that has a fixed orientation with respect to the basal-distal leaf axis. The genetic, molecular and cell biological analysis of trichome morphogenesis reveal that various cellular processes need to be coordinated including regulation of the cell cycle, the cell size and the actin and tubulin cytoskeleton. Here we will focus on what is known about the establishment and maintenance of positional information during trichome formation.

Rickettsia felis: molecular characterization of a new member of the spotted fever group.

In this report, placement of Rickettsia felis in the spotted fever group (SFG) rather than the typhus group (TG) of Rickettsia is proposed. The organism, which was first observed in cat fleas (Ctenocephalides felis) by electron microscopy, has not yet been reported to have been cultivated reproducibly, thereby limiting the standard rickettsial typing by serological means. To overcome this challenge, several genes were selected as targets to be utilized for the classification of R. felis. DNA from cat fleas naturally infected with R. felis was amplified by PCR utilizing primer sets specific for the 190 kDa surface antigen (rOmpA) and 17 kDa antigen genes. The entire 5,513 bp rompA gene was sequenced, characterized and found to have several unique features when compared to the rompA genes of other Rickettsia. Phylogenetic analysis of the partial sequence of the 17 kDa antigen gene indicated that R. felis is less divergent from the SFG rickettsiae than from the TG rickettsiae. The data corroborate results from previous reports that analysed the citrate synthase, 16S rRNA, rompB (135 kDa surface antigen), metK, ftsY, polA and dnaE genes that placed R. felis as a member of the SFG. The organism is passed trans-stadially and transovarially, and infection in the cat flea has been observed in the midgut, tracheal matrix, muscle, hypodermis, ovaries and testes.

Phylogenetic analysis of the rompB genes of Rickettsia felis and Rickettsia prowazekii European-human and North American flying-squirrel strains.

The rickettsial outer membrane protein B (rompB) gene encodes the major surface antigens of Rickettsia species. We undertook sequencing and molecular analysis of the rompB gene of Rickettsia felis and a comparison with its homologs in spotted fever group (SFG) and typhus group (TG) rickettsiae, including the complete sequences of two North American flying squirrel strains and two European human strains of Rickettsia prowazekii. We sequenced 5,226 base pairs (bp) of the R. felis rompB, encoding a protein of 1,654 amino acids. We also sequenced 5,015 bp of rompB of the flying squirrel strains, encoding a protein of 1,643 amino acids. Analysis of the R. felis rompB gene sequence showed 10-13% divergence from SFG rickettsiae and 18% divergence from the TG rickettsiae. The rompB of all sequenced strains of R. prowazekii showed an overall similarity of 99.7-99.9%.