Heterogeneity and cell type-specific localization of a cell wall glycoprotein from carrot suspension cells

EP1, an extracellular protein from carrot (Daucus carota) cell suspensions, has been partially characterized by means of an antiserum and a cDNA clone. In both embryo and suspension cultures different molecular mass EP1 proteins were detected, some of which (31, 32, 52, and 54 kilodaltons) were bound to the cell wall and released into the medium, whereas others (49, 60, and 62 kilodaltons) were more firmly bound to the cell wall and could be extracted with a salt solution. Immunoprecipitation of in vitro translation products revealed a single primary translation product of 45 kilodaltons, suggesting that EP1 heterogeneity is due to differential posttranslational modification. In seedlings organ-specific modification of EP1 proteins was observed, a phenomenon which did not persist in suspension cultures initiated from different seedling organs. In culture EP1 proteins were only found to be associated with vacuolated, nonembryogenic cells, and on these cells they were localized in loosely attached, pectin-containing cell wall material. Purified 52/54 kilodaltons EP1 proteins did not alleviate the inhibitory effect of the glycosylation inhibitor tunicamycin on somatic embryogenesis.     

Development of the peripheral glial cells in Drosophila

In complex organisms the nervous system comprises two cell types: neurons and glial cells. Their correct interplay is of crucial importance during both the development of the nervous system and for later function of the nervous system. In recent years tools have been developed for Drosophila that enable genetic approaches to understanding glial development and differentiation. Focusing on peripheral glial cells we first summarize wild-type development, then introduce some of the relevant genes that have been identified. Despite obvious differences between Drosophila and mammalian glial cells, the molecular machinery that controls terminal differentiation appears well conserved.     

Validation of vibration testing for the assessment of the mechanical properties of human lumbar motion segments

Experimental modal analysis is a non-destructive measurement technique, which applies low forces and small deformations to assess the integrity of a structure. It is therefore a promising method to study the mechanical properties of the spine in vivo. Previously, modal parameters successfully revealed artificially induced spinal injuries. The question remains however, whether experimental modal analysis can be applied successfully in human spinal segments with mechanical changes due to physiological processes. Since quasi-static mechanical testing is considered the "gold standard" for assessing intervertebral stiffness, the purpose of our study was to examine if the mechanical properties derived from vibration testing and quasi-static testing correlate. Six cadaver human spines (L1-L5) were loaded quasi-statically in bending and torsion, while an optical system measured the angular rotations of the individual motion segments. Subsequently, the polysegmental spines were divided into L2-L3 and L4-L5 segments and a shaker was used to vibrate the upper vertebra, while its response was obtained from accelerometers in anteroposterior and mediolateral directions. From the resulting frequency response function the eigenfrequencies (ratio between stiffness and mass) and vibration modes (pattern of motion) were determined. The vibration results showed clear eigenfrequencies for flexion-extension (mean 121.83Hz, SD 40.05Hz), lateroflexion (mean 132.17, SD 34.80Hz) and axial rotation (mean 236.17Hz, SD 81.45Hz). Furthermore, the correlation between static and dynamic tests was significant (r=0.73, p=0.01). In conclusion, the findings from this study show that experimental modal analysis is a valid method to assess the mechanical properties of human lumbar motion segments.     

The splicing factor crooked neck associates with the RNA-binding protein HOW to control glial cell maturation in Drosophila

In both vertebrates and invertebrates, glial cells wrap axonal processes to ensure electrical conductance. Here we report that Crooked neck (Crn), the Drosophila homolog of the yeast Clf1p splicing factor, is directing peripheral glial cell maturation. We show that crooked neck is expressed and required in glial cells to control migration and axonal wrapping. Within the cytoplasm, Crn interacts with the RNA-binding protein HOW and then translocates to the nucleus where the Crn/HOW complex controls glial differentiation by facilitating splicing of specific target genes. By using a GFP-exon trap approach, we identified some of the in vivo target genes that encode proteins localized in autocellular septate junctions. In conclusion, here we show that glial cell differentiation is controlled by a cytoplasmic assembly of splicing components, which upon translocation to the nucleus promote the splicing of genes involved in the assembly of cellular junctions.     

Adverse environmental conditions influence age-related innate immune responsiveness

Background-  

The innate immune system plays an important role in the recognition and induction of protective responses against infectious pathogens, whilst there is increasing evidence for a role in mediating chronic inflammatory diseases at older age. Despite indications that environmental conditions can influence the senescence process of the adaptive immune system, it is not known whether the same holds true for the innate immune system. Therefore we studied whether age-related innate immune responses are similar or differ between populations living under very diverse environmental conditions.     

Joint Evolutionary Trees: A Large-Scale Method To Predict Protein Interfaces Based on Sequence Sampling

The Joint Evolutionary Trees (JET) method detects protein interfaces, the core residues involved in the folding process, and residues susceptible to site-directed mutagenesis and relevant to molecular recognition. The approach, based on the Evolutionary Trace (ET) method, introduces a novel way to treat evolutionary information. Families of homologous sequences are analyzed through a Gibbs-like sampling of distance trees to reduce effects of erroneous multiple alignment and impacts of weakly homologous sequences on distance tree construction. The sampling method makes sequence analysis more sensitive to functional and structural importance of individual residues by avoiding effects of the overrepresentation of highly homologous sequences and improves computational efficiency. A carefully designed clustering method is parametrized on the target structure to detect and extend patches on protein surfaces into predicted interaction sites. Clustering takes into account residues' physical-chemical properties as well as conservation. Large-scale application of JET requires the system to be adjustable for different datasets and to guarantee predictions even if the signal is low. Flexibility was achieved by a careful treatment of the number of retrieved sequences, the amino acid distance between sequences, and the selective thresholds for cluster identification. An iterative version of JET (iJET) that guarantees finding the most likely interface residues is proposed as the appropriate tool for large-scale predictions. Tests are carried out on the Huang database of 62 heterodimer, homodimer, and transient complexes and on 265 interfaces belonging to signal transduction proteins, enzymes, inhibitors, antibodies, antigens, and others. A specific set of proteins chosen for their special functional and structural properties illustrate JET behavior on a large variety of interactions covering proteins, ligands, DNA, and RNA. JET is compared at a large scale to ET and to Consurf, Rate4Site, siteFiNDER|3D, and SCORECONS on specific structures. A significant improvement in performance and computational efficiency is shown.     

Ecologists can enable communities to implement malaria vector control in Africa

Background

Malaria imposes significant costs on households and the poor are disproportionately affected. However, cost data are often from quantitative surveys with a fixed recall period. They do not capture costs that unfold slowly over time, or seasonal variations. Few studies investigate the different pathways through which malaria contributes towards poverty. In this paper, a framework indicating the complex links between malaria, poverty and vulnerability at the household level is developed and applied using data from rural Kenya.

Methods

Cross-sectional surveys in a wet and dry season provide data on treatment-seeking, cost-burdens and coping strategies (n = 294 and n = 285 households respectively). 15 case study households purposively selected from the survey and followed for one year provide in-depth qualitative information on the links between malaria, vulnerability and poverty.

Results

Mean direct cost burdens were 7.1% and 5.9% of total household expenditure in the wet and dry seasons respectively. Case study data revealed no clear relationship between cost burdens and vulnerability status at the end of the year. Most important was household vulnerability status at the outset. Households reporting major malaria episodes and other shocks prior to the study descended further into poverty over the year. Wealthier households were better able to cope.

Conclusion

The impacts of malaria on household economic status unfold slowly over time. Coping strategies adopted can have negative implications, influencing household ability to withstand malaria and other contingencies in future. To protect the poor and vulnerable, malaria control policies need to be integrated into development and poverty reduction programmes.     

Free brownian motion of individual lipid molecules in biomembranes

The mobility of phospolipids in free-standing and supported membranes was investigated on the level of individual molecules. For the analysis of trajectories a new statistical treatment was developed that permitted us to clearly distinguish different types of diffusional motion. A freely diffusing subfraction of lipids within supported membranes was identified. Its mobility was characterized by a mean lateral diffusion constant of D(supp) = 4.6 &mgr;m(2)/s. In comparison, the mobility of lipids embedded in "free-standing" planar membranes yielded an increase in the mean diffusion constant by a factor of 4.5, D(free) = 20.6 &mgr;m(2)/s. This increase is attributed to the ultrathin (</=1 nm) lubricating water layer between membranes and glass support.     

Personality diagnosis in elderly psychiatric patients using a Delphi technique

A Delphi-technique was used as part of the development of a screening instrument to diagnose personality disorders in the elderly. Several statements regarding this subject were tested. Fifty-three Delphi-members, with expertise in the field of mental health services for the elderly and knowledge about the concept 'personality disorder', gave their opinion on the statements. In three successive rounds we aimed to get consensus as well as agreement on the contents of the statements. In the first round the Delphi-panel confirmed the importance of diagnosing personality disorders, with regard to individual therapy in older adults and psycho-educational activities. The DSM-IV Axis II criteria and related assessment-instruments do not take into account the emotional and social context of the elderly people. In the second round the Delphi-panel endorsed the importance of several information sources namely biographical information, informant information, behavioural observations and the reactions of the therapist himself. In the third round, 44 items considering diagnostics on personality were pro-pounded to the panel. There was agreement as well as consensus on 25 out of the 44 items. In conclusion, adjusting the DSM-IV Axis II criteria to the elderly will improve the quality of the diagnostics. Developments of a specific screening instrument for older adults probably will also increase the quality of the diagnostics.