Floral ontogeny of Lecointea, Zollernia, Exostyles, and Harleyodendron (Leguminosae: Papilionoideae: Swartzieae s.l)

Floral initiation and development were examined using scanning electron microscopy in Exostyles venusta, Harleyodendron unifoliolatum, Lecointea hatschbachii, and Zollernia ilicifolia. Common features include (1) unidirectional sepal initiation, (2) simultaneous petal initiation, (3) unidirectional initiation of each stamen whorl (except in the antesepalous whorl in Lecointea and Exostyles), (4) overlap in time of initiation of the two stamen whorls, and (5) initiation of the carpel concurrently with petals. Significant developmental features include (1) the first sepal median abaxial in all except Lecointea where it is non-median abaxial; (2) intraspecific variation in petal aestivation in Exostyles, Harleyodendron, and Lecointea; (3) initiation of antepetalous stamens before the antesepalous ones in Zollernia, Exostyles, and Lecointea; and (4) ovule initiation before the carpel margins are fused in Exostyles. The stamen sequence has not been found in any other legumes. The following late developmental events distinguish the four genera from each other: copious hairs hold the anthers together as a domelike structure at anthesis in Harleyodendron; zygomorphy in Zollernia results from differing petal reflexion; late hypanthium in Exostyles, Lecointea, and Holocalyx (no hypanthium in Harleyodendron or Zollernia); and reflexed sepal lobes in Exostyles, Harleyodendron, and Zollernia but not in Holocalyx and Lecointea. The genera studied here are ontogenetically more similar to taxa of Sophoreae than to other Swartzieae that have been investigated. None of the taxa studied here has a ring meristem, the structure that characterizes the remaining swartzioid taxa studied elsewhere.     

Inhibitory effect of sulfur dioxide and other stress compounds in wine on the ATPase activity of Oenococcus oeni

Malolactic fermentation (MLF) is carried out by Oenococcus oeni under very harsh conditions. This paper shows that stress compounds in wine such as SO(2), fatty acids and copper have an inhibitory effect on cell growth and MLF duration, and relates this effect to an inhibition of ATPase activity. Of the stress compounds, SO(2) and dodecanoic acid had the strongest effect, decreasing the ATPase specific activity to 37% and 58%, respectively. It can be concluded that ATPase is a good indicator of the physiological state of the cells and their ability to lead MLF.     

A stuttering model for paramyxovirus P mRNA editing.

Paramyxovirus P genes are transcribed into two mRNAs which differ from each other by either one (measles and Sendai virus) or two (SV5 and mumps virus) G insertions, and which code for either the P or V proteins. The G insertions always occur within a short run of Gs, and a stuttering mechanism for the insertions has been suggested in which the viral polymerase reiteratively copies a template C residue during mRNA synthesis. Support for this mechanism was obtained by varying the reaction conditions during Sendai virus mRNA synthesis in vitro. A stuttering model is proposed which accounts for how the ratio of inserted to uninserted mRNAs is controlled, and why some paramyxoviruses insert one G and others two Gs when insertions occur.     

Regeneration of transgenic plants by Agrobacterium-mediated transformation of somatic embryos of juvenile and mature Quercus robur

A protocol was developed for genetic transformation of somatic embryos derived from juvenile and mature Quercus robur trees. Optimal transformation conditions were evaluated on the basis of the results of transient GUS expression assays with five oak embryogenic lines and a strain of Agrobacterium tumefaciens (EHA105) harbouring a p35SGUSINT plasmid containing a nptII and a uidA (GUS) genes. For stable transformation, embryo clumps at globular/torpedo stages (4–10 mg) were inoculated with EHA105:p35SGUSINT bacterial cultures, cocultivated for 4 days and selected in proliferation medium with 75 mg/l of kanamycin. Putatively transformed masses appeared after 20–30 weeks of serial transfers to selective medium. Histochemical and molecular analysis (PCR and Southern blot) confirmed the presence of nptII and uidA genes in the plant genomes. Transformation efficiencies ranged from up to 2% in an embryogenic line derived from a 300-year-old tree, to 6% in a juvenile genotype. Twelve independent transgenic lines were obtained from these oak genotypes, and transgenic plantlets were recovered and acclimatized into the soil. This is the first demonstration of the production of transformed somatic embryos and regenerated plants from juvenile and mature trees of Q. robur and suggests the possibility of introducing other genetic constructions to develop trees that are tolerant/resistant to pathogens and/or biotic stresses.     

Screening of ESTs from Mytilus for the detection of SSR markers in Mytilus californianus

A set of expressed sequence tag‐simple sequence repeat (EST‐SSR) markers for the genus Mytilus was developed through bioinformatic mining of the GenBank public database. A total of 33 782 EST sequences from GenBank were downloaded and screened for di‐, tri‐ and tetranucleotide, with 1274 EST containing SSR markers. Nine microsatellite markers were characterized in Mytilus californianus with a number of alleles per locus ranging from two to six, and total observed and expected heterozygosities ranging from 0.490 to 0.730 and from 0.510 to 0.860 respectively. Cross‐species amplification was achieved in several other species, confirming the usefulness of these markers in Mytilus genetics.     

Calcium-independent phospholipase A2 and apoptosis

Apoptosis or programmed cell death is associated with changes in glycerophospholipid metabolism. Cells undergoing apoptosis generally release free fatty acids including arachidonic acid, which parallels the reduction in cell viability. The involvement of cytosolic group IVA phospholipase A(2)alpha (cPLA(2)alpha) in apoptosis has been the subject of numerous studies but a clear picture of the role(s) played by this enzyme is yet to emerge. More recently, the importance of lipid products generated by the action of a second phospholipase A(2), the group VIA calcium-independent phospholipase A(2) (iPLA(2)-VIA) in apoptosis has begun to be unveiled. Current evidence suggests that iPLA(2)-VIA-derived lysophosphatidylcholine may play a prominent role in mediating the chemoattractant and recognition/engulfment signals that accompany the process of apoptotic cell death, and gives possibility to the efficient clearance of dying cells by circulating phagocytes. Other lines of evidence suggest that perturbations in the control of free arachidonic acid levels within the cells, a process that may implicate iPLA(2)-VIA as well, may provide important cellular signals for the onset of apoptosis.     

Early chloroplastic alterations analysed by optical coherence tomography during a harpin-induced hypersensitive response

The hypersensitive response has been mostly studied by molecular and biochemical methods after sample destruction. The development of imaging techniques allows the monitoring of physiological changes before any signs of cell death. Here, we follow the early steps of a hypersensitive-like response induced by the bacterial elicitor harpin in Nicotiana sp. We describe cytological modifications after inoculation of the harpin protein, using confocal fluorescence microscopy (CFM) and optical coherence tomography (OCT), an interferometric-based microscopy. The changes detected by CFM occurred 5 h after harpin infiltration and corresponded to a redistribution of the chloroplasts from the upper to the inner regions of the palisade mesophyll cells which could be related to a perturbation in the microtubule network. Using OCT, we were able to detect a decrease in chloroplast backscattered signal as early as 30 min after harpin infiltration. A simple physical model, which accounted for the structure and distribution of thylakoid membranes, suggested that this loss of scattering could be associated with a modification in the refractive index of the thylakoid membranes. Our OCT observations were correlated with a decrease in photosynthesis, emphasizing changes in chloroplast structure as one of the earliest hallmarks of plant hypersensitive cell death.     

Metric and morphological study of the upper cervical spine from the Sima de los Huesos site (Sierra de Atapuerca, Burgos, Spain)

In this article, the upper cervical spine remains recovered from the Sima de los Huesos (SH) middle Pleistocene site in the Sierra de Atapuerca (Burgos, Spain) are described and analyzed. To date, this site has yielded more than 5000 human fossils belonging to a minimum of 28 individuals of the species Homo heidelbergensis. At least eleven individuals are represented by the upper cervical (C1 and C2) specimens: six adults and five subadults, one of which could represent an adolescent individual. The most complete adult vertebrae (three atlases and three axes) are described, measured, and compared with other fossil hominins and modern humans. These six specimens are associated with one another and represent three individuals. In addition, one of these sets of cervical vertebrae is associated with Cranium 5 (Individual XXI) from the site. The metric analysis demonstrates that the Sima de los Huesos atlases and axes are metrically more similar to Neandertals than to our modern human comparative sample. The SH atlases share with Neandertals a sagittally elongated canal. The most remarkable feature of the SH (and Neandertal) axes is that they are craniocaudally low and mediolaterally wide compared to our modern male sample. Morphologically, the SH sample shares with Neandertals a higher frequency of caudally projected anterior atlas arch, which could reflect greater development of the longus colli muscle. In other features, such as the frequency of weakly developed tubercles for the attachment of the transverse ligament of the atlas, the Sima de los Huesos fossils show intermediate frequencies between our modern comparative samples and the Neandertals, which could represent the primitive condition. Our results are consistent with the previous phylogenetic interpretation of H. heidelbergensis as an exclusively European species, ancestral only to H. neanderthalensis.     

Discovery of inhibitors of the channel-activating protease prostasin (CAP1/PRSS8) utilizing structure-based design

Structure-based design was utilized to guide the early stage optimization of a substrate-like inhibitor to afford potent peptidomimetic inhibitors of the channel-activating protease prostasin. The first X-ray crystal structures of prostasin with small molecule inhibitors bound to the active site are also reported.     

Serum and Lymphocytic Neurotrophins Profiles in Systemic Lupus Erythematosus: a Case-Control Study

Background

Neurotrophins play a central role in the development and maintenance of the nervous system. However, neurotrophins can also modulate B and T cell proliferation and activation, especially via autocrine loops. We hypothesized that both serum and lymphocytic neurotrophin levels may be deregulated in systemic Lupus erythematosus (SLE) and may reflect clinical symptoms of the disease.

Methods

Neurotrophins in the serum (ELISA tests) and lymphocytes (flow cytometry) were measured in 26 SLE patients and 26 control subjects. Th1 (interferon-γ) and Th2 (IL-10) profiles and serum concentration of BAFF were assessed by ELISA in the SLE and control subjects.

Findings

We have demonstrated that both NGF and BDNF serum levels are higher in SLE patients than healthy controls (p=0.003 and p<0.001), independently of Th1 or Th2 profiles. Enhanced serum NT-3 levels (p=0.003) were only found in severe lupus flares (i.e. SLEDAI ≥ 10) and significantly correlated with complement activation (decreased CH 50, Γ=-0.28, p=0.03). Furthermore, there was a negative correlation between serum NGF levels and the number of circulating T regulatory cells (Γ=0.48, p=0.01). In circulating B cells, production of both NGF and BDNF was greater in SLE patients than in healthy controls. In particular, the number of NGF-secreting B cells correlated with decreased complement levels (p=0.05). One month after SLE flare treatment, BDNF levels decreased; in contrast, NGF and NT-3 levels remained unchanged.

Conclusion

This study demonstrates that serum and B cell levels of both NGF and BDNF are increased in SLE, suggesting that the neurotrophin production pathway is deregulated in this disease. These results must be confirmed in a larger study with naive SLE patients, in order to avoid the potential confounding influence of prior immune-modulating treatments on neurotrophin levels.