Molecular phylogeny of the fern genus Elaphoglossum (Elaphoglossaceae) based on chloroplast non-coding DNA sequences: contributions of species from the Indian Ocean area

We performed a phylogenetic analysis of the fern genus Elaphoglossum using two non-coding chloroplast spacers: trnL-trnF and rps4-trnS. The sampling includes 123 species, of which 80 have not been previously sequenced, and for the first time includes species from Africa and the Indian Ocean area. The results of this expanded study largely agree with an earlier molecular study based on a smaller group of neotropical species and with the morphology-based classification of Mickel and Atehortua. We found, however, that some infrageneric groups such as section Elaphoglossum are not monophyletic. Besides section Elaphoglossum pro parte, we recognize six sections: two new monospecific, unnamed sections, and the previously established sections Lepidoglossa, Squamipedia, Amygdalifolia, and "Subulate-scaled clade." We divide the subulate-scaled clade into subsection Setosa (hydathodes present) and Polytrichia (hydathodes absent), and section Elaphoglossum is divided into subsections Platyglossa and Pachyglossa, two groups that do not appear to be supported by any single morphological character. In general, however, the main clades are supported by morphology. Finally, we discuss the species of the Indian Ocean region and their affinities with the neotropical ones. Out of the 11 species pairs postulated by Moran and Smith on the basis of morphology, two are well supported (E. eximium-E. aubertii; E. piloselloides-E. spatulatum) and three are not supported (E. ciliatum-E. humbertii; E. muscosum-E. poolii; E. paleaceum-E. deckenii), and two remain unresolved (E. erinaceum-E. hybridum; E. glabellum-E. acrostichoides) because our molecular markers were not variable enough. Four species pairs could not be tested because specimens were lacking. Unsupported species pairs are best interpreted as morphological convergences. Two additional species pairs are proposed: E. cuspidatum-E. succisaefolium; E. doanense-E. hornei. Placement of the species from the Indian Ocean suggests that at least 13 long-distance dispersal events occurred between the Neotropics and the Indian Ocean-Africa.     

A HPTLC densitometric determination of flavonoids from Passiflora alata, P. edulis, P. incarnata and P. caerulea and comparison with HPLC method

A high-performance thin layer chromatographic (HPTLC) method was developed in order to determine quantitatively the flavonoids in leaves of Passiflora alata, P. edulis, P. caerulea and P. incarnata. The content of orientin and isoorientin was determined, and the results were compared with those obtained using a quantitative HPLC-UV method. The latter employed rutin as standard and was developed to analyse flavonoid content from Passiflora leaves for the purpose of ensuring the quality of Passiflora phytomedicines. The results obtained using the two methods indicate that there are qualitative and quantitative differences in the flavonoids of the reference Passiflora species studied. The two methods were also employed to analyse commercial samples to illustrate their application in qualitative ('fingerprint') and quantitative determination, demonstrating their feasibility in the quality control of flavonoids from crude Passiflora drugs and phytomedicines. The HPLC conditions used are also suitable for the quantitative analysis of aqueous extracts (Passiflora infusions).     

In vivo kinematic evaluation and design considerations related to high flexion in total knee arthroplasty

In designing a posterior-stabilized total knee arthroplasty (TKA) it is preferable that when the cam engages the tibial spine the contact point of the cam move down the tibial spine. This provides greater stability in flexion by creating a greater jump distance and reduces the stress on the tibial spine. In order to eliminate edge loading of the femoral component on the posterior tibial articular surface, the posterior femoral condyles need to be extended. This provides an ideal femoral contact with the tibial articular surface during high flexion angles. To reduce extensor mechanism impingement in deep flexion, the anterior margin of the tibial articular component should be recessed. This provides clearance for the patella and patella tendon. An in vivo kinematic analysis that determined three dimensional motions of the femorotibial joint was performed during a deep knee bend using fluoroscopy for 20 subjects having a TKA designed for deep flexion. The average weight-bearing range-of-motion was 125 degrees . On average, TKA subjects experienced 4.9 degrees of normal axial rotation and all subjects experienced at least -4.4 mm of posterior femoral rollback. It is assumed that femorotibial kinematics can play a major role in patellofemoral kinematics. In this study, subjects implanted with a high-flexion TKA design experienced kinematic patterns that were similar to the normal knee. It can be hypothesized that forces acting on the patella were not substantially increased for TKA subjects compared with the normal subjects.     

Failure of the interaction between presenilin 1 and the substrate of gamma-secretase to produce Abeta in insect cells

Aggregates of beta-amyloid peptide (Abeta) are the major component of the amyloid core of the senile plaques observed in Alzheimer's disease (AD). Abeta results from the amyloidogenic processing of its precursor, the amyloid precursor protein (APP), by beta- and gamma-secretase activities. If beta-secretase has recently been identified and termed BACE, the identity of gamma-secretase is still obscure. Studies with knock-out mice showed that presenilin 1 (PS1), of which mutations are known to be the first cause of inherited AD, is mandatory for the gamma-secretase activity. However, the proteolytic activity of PS1 remains a matter of debate. Here we used transfected Sf9 insect cells, a cellular model lacking endogenous beta- and/or gamma-secretase activities, to characterize the role of BACE and PS1 in the amyloidogenic processing of human APP. We show that, in Sf9 cells, BACE performs the expected beta-secretase cleavage of APP, generating C99. We also show that C99, which is a substrate of gamma-secretase, tightly binds to the human PS1. Despite this interaction, Sf9 cells still do not produce Abeta. This strongly argues against a direct proteolytic activity of PS1 in APP processing, and points toward an implication of PS1 in trafficking/presenting its substrate to the gamma-secretase.     

What is the role of PACAP in gonadotrope function?

Strong evidence in favor of a direct action of hypothalamic PACAP at the pituitary to modulate gonadotrope function has been acquired mainly by in vitro studies using cultured pituitary cells or gonadotrope cell lines. In particular, PACAP has been shown to cooperate with GnRH, the primary regulator of gonadotropes, to regulate/modulate gonadotropin subunit gene expression, gonadotropin release as well as gonadotrope responsiveness. These effects of PACAP appear to be due essentially to its high potent stimulatory action on the cAMP/protein kinase pathway. Ensuing mechanisms include signaling cross-talk and/or enhanced gene expression within gonadotropes. PACAP may also indirectly operate on these cells through paracrine mechanisms. While PACAP has long been viewed as a hypophysiotropic factor, a locally produced PACAP has also been described. Interestingly, both appear similarly up-regulated at proestrus of the reproductive cycle in female rats. Further in vivo investigation is now necessary to ascertain the physiological relevance of the observed pituitary PACAP effects and especially to evaluate the respective contribution of hypothalamic and pituitary PACAP in the dynamic control of gonadotrope function.     

Comparison between a bovine and a human enterohaemorrhagic Escherichia coli strain of serogroup O26 by suppressive subtractive hybridization reveals the presence of atypical factors in EHEC and EPEC strains

Enterohaemorrhagic Escherichia coli (EHEC) strains are responsible for food poisoning in humans in developed countries via consumption of vegetal and animal foodstuffs contaminated by ruminant feces. The clinical conditions caused by EHEC strains vary from undifferentiated diarrhea to hemorrhagic colitis with, in a few cases, the appearance of the hemolytic uremic syndrome, which can lead to death. Most EHEC strains can be found in the gut of healthy ruminants, but some of the strains, belonging to O26, O111, O118 serogroups, for example, are also responsible for digestive disorders in calves. The aim of this research was to study the genomic differences between two EHEC strains of serogroup O26 isolated from a young calf and a human with diarrhea, to identify specific sequences of the bovine strain that could be implicated in initial adherence or host specificity. No sequence implicated in host specificity was found during our study. Finally, several factors, not usually present in EHEC strains of serogroup O26, were identified in the bovine strain. One of them, the PAI I(CL3) locus initially presented as a marker for LEE-negative VTEC strains, was found in 11.3% of EPEC and EHEC strains.     

Effect of gelation temperature and gel-dissolving solution on cell viability and recovery of two Pseudomonas putida strains co-immobilized within calcium alginate or κ-carrageenan gel beads

Summary Since a lethal effect of an increased temperature (42°C) on Pseudomonas putida strains PaW8 and PaW130 was demonstrated, strictly ionotropic gels such as calcium alginate or -carrageenan type X 0909 were used for cell co-immobilization, rather than a thermoionotropic -carrageenan gel. Among the variety of gel-dissolving solutions tested, only a 0.05M Na₂CO₃/0.02M citric acid solution was able to preserve around 100 % of the cell viability. A complete cell recovery was obtained from calcium alginate gel beads, while only 6 % of viable cells was recovered from the ionotropic -carrageenan gel.     

First evidence of somatic embryogenesis from needles of 1-year-old Picea abies plants

Summary Somatic embryogenesis was obtained from hypocotyls and cotyledons of one month old plantlets of Picea abies. Embryogenic yield was higher with expiants from somatic embryo-derived plantlets (80 %) than with plantlets issued from zygotic embryos (10 %). This report also describes production of embryogenic calli from needles of 14 month old somatic embryo-derived plants cultivated in greenhouse. The influence of the physiological status and genotype of the mother plant on somatic embryogenic potential is discussed.Abbreviations ABA abscisic acid - (±) ABA racemic ABA - BAP 6-benzylaminopurine - CI callus inducing culture medium - NAA 1-naphtaleneacetic acid     

Lithium chloride increases the production of amyloid-beta peptide independently from its inhibition of glycogen synthase kinase 3

Glycogen synthase kinase 3 (GSK3) is able to phosphorylate tau at many sites that are found to be phosphorylated in paired helical filaments in Alzheimer disease. Lithium chloride (LiCl) efficiently inhibits GSK3 and was recently reported to also decrease the production of amyloid-beta peptide (Abeta) from its precursor, the amyloid precursor protein. Therefore, lithium has been proposed as a combined therapeutic agent, inhibiting both the hyperphosphorylation of tau and the production of Abeta. Here, we demonstrate that the inhibition of GSK3 by LiCl induced the nuclear translocation of beta-catenin in Chinese hamster ovary cells and rat cultured neurons, in which a decrease in tau phosphorylation was observed. In both cellular models, a nontoxic concentration of LiCl increased the production of Abeta by increasing the beta-cleavage of amyloid precursor protein, generating more substrate for an unmodified gamma-secretase activity. SB415286, another GSK3 inhibitor, induced the nuclear translocation of beta-catenin and slightly decreased Abeta production. It is concluded that the LiCl-mediated increase in Abeta production is not related to GSK3 inhibition.