In vitro culture of Aloe Barbadensis Mill.: Micropropagation from vegetative meristems

A method for rapid and highly effective plant micropropagation from vegetative meristems was established for Aloe barbadensis Mill. Plant micropropagation was achieved culturing apices on medium containing 1.1 M 2,4-dichlorophenoxyacetic acid and 2.3 M kinetin for 15–30 days. High morphogenetic ability was maintained by transferring explants (after 60 days) on media containing 0.11 M 2,4-dichlorophenoxyacetic acid and 2.2 M 6-benzylaminopurine.     

Lysosomal fusion and SNARE function are impaired by cholesterol accumulation in lysosomal storage disorders

The function of lysosomes relies on the ability of the lysosomal membrane to fuse with several target membranes in the cell. It is known that in lysosomal storage disorders (LSDs), lysosomal accumulation of several types of substrates is associated with lysosomal dysfunction and impairment of endocytic membrane traffic. By analysing cells from two severe neurodegenerative LSDs, we observed that cholesterol abnormally accumulates in the endolysosomal membrane of LSD cells, thereby reducing the ability of lysosomes to efficiently fuse with endocytic and autophagic vesicles. Furthermore, we discovered that soluble N‐ethylmaleimide‐sensitive factor attachment protein (SNAP) receptors (SNAREs), which are key components of the cellular membrane fusion machinery are aberrantly sequestered in cholesterol‐enriched regions of LSD endolysosomal membranes. This abnormal spatial organization locks SNAREs in complexes and impairs their sorting and recycling. Importantly, reducing membrane cholesterol levels in LSD cells restores normal SNARE function and efficient lysosomal fusion. Our results support a model by which cholesterol abnormalities determine lysosomal dysfunction and endocytic traffic jam in LSDs by impairing the membrane fusion machinery, thus suggesting new therapeutic targets for the treatment of these disorders.     

Comparison of visual and objective quantification of elbow and shoulder movement in children with obstetric brachial plexus palsy

Background

Animal studies have demonstrated complex cortical reorganization following peripheral nerve lesion. Central projection fields of intact nerves supplying skin areas which border denervated skin, extended into the deafferentiated cortical representation area. As a consequence of nerve lesions and subsequent reorganization an increase of the somatosensory evoked potentials (SEPs) was observed in cats when intact neighbouring nerves were stimulated. An increase of SEP-components of patients with nerve lesions may indicate a similar process of posttraumatic plastic cortical reorganization.

Methods

To test if a similar process of post-traumatic plastic cortical reorganization does occur in humans, the SEP of intact neighbouring hand nerves were recorded in 29 patients with hand nerve lesions. To hypothetically explain the observed changes of SEP-components, SEP recording following paired stimulation of the median nerve was performed in 12 healthy subjects.

Results

Surprisingly 16 of the 29 patients (55.2%) showed a reduction or elimination of N35, P45 and N60. Patients with lesions of two nerves showed more SEP-changes than patients with a single nerve lesion (85.7%; 6/7 nerves; vs. 34.2%; 13/38 nerves; Fisher's exact test, p < 0.05). With paired stimulation a suppression of the amplitude of N20, P25 and P45 (p < 0.05; sign test), and a marked increment of N35 (p < 0.05; sign test) and N60 (not significant; sign test) of the second response could be observed.

Conclusion

The results of the present investigation do not provide evidence of collateral innervation of peripherally denervated cortical neurons by neurons of adjacent cortical representation areas. They rather suggest that secondary components of the excitatory response to nerve stimulation are lost in cortical areas, which surround the denervated region.     

Osteopontin and systemic lupus erythematosus association: a probable gene-gender interaction

Osteopontin (SPP1) is an important bone matrix mediator found to have key roles in inflammation and immunity. SPP1 genetic polymorphisms and increased osteopontin protein levels have been reported to be associated with SLE in small patient collections. The present study evaluates association between SPP1 polymorphisms and SLE in a large cohort of 1141 unrelated SLE patients [707 European-American (EA) and 434 African-American (AA)], and 2009 unrelated controls (1309 EA and 700 AA). Population-based case-control association analyses were performed. To control for potential population stratification, admixture adjusted logistic regression, genomic control (GC), structured association (STRAT), and principal components analysis (PCA) were applied. Combined analysis of 2 ethnic groups, showed the minor allele of 2 SNPs (rs1126616T and rs9138C) significantly associated with higher risk of SLE in males (P = 0.0005, OR = 1.73, 95% CI = 1.28-2.33), but not in females. Indeed, significant gene-gender interactions in the 2 SNPs, rs1126772 and rs9138, were detected (P = 0.001 and P = 0.0006, respectively). Further, haplotype analysis identified rs1126616T-rs1126772A-rs9138C which demonstrated significant association with SLE in general (P = 0.02, OR = 1.30, 95%CI 1.08-1.57), especially in males (P = 0.0003, OR = 2.42, 95%CI 1.51-3.89). Subgroup analysis with single SNPs and haplotypes also identified a similar pattern of gender-specific association in AA and EA. GC, STRAT, and PCA results within each group showed consistent associations. Our data suggest SPP1 is associated with SLE, and this association is especially stronger in males. To our knowledge, this report serves as the first association of a specific autosomal gene with human male lupus.     

A processing product of the Plasmodium falciparum reticulocyte binding protein RH1 shows a close association with AMA1 during junction formation

Plasmodium falciparum responsible for the most virulent form of malaria invades human erythrocytes through multiple ligand‐receptor interactions. The P. falciparum reticulocyte binding protein homologues (PfRHs) are expressed at the apical end of merozoites and form interactions with distinct erythrocyte surface receptors that are important for invasion. Here using a range of monoclonal antibodies (mAbs) against different regions of PfRH1 we have investigated the role of PfRH processing during merozoite invasion. We show that PfRH1 gets differentially processed during merozoite maturation and invasion and provide evidence that the different PfRH1 processing products have distinct functions during invasion. Using in‐situ Proximity Ligation and FRET assays that allow probing of interactions at the nanometre level we show that a subset of PfRH1 products form close association with micronemal proteins Apical Membrane Antigen 1 (AMA1) in the moving junction suggesting a critical role in facilitating junction formation and active invasion. Our data provides evidence that time dependent processing of PfRH proteins is a mechanism by which the parasite is able to regulate distinct functional activities of these large processes. The identification of a specific close association with AMA1 in the junction now may also provide new avenues to target these interactions to prevent merozoite invasion.     

Population recovery of alien black rats Rattus rattus: A test of reinvasion theory

Reinvasion of pest animals after incomplete control is a major challenge for invasive species management, yet little is known about the behavioural and demographic categories of reinvaders or the mechanisms that drive population‐level responses to control. To understand the fine‐scale mechanisms of reinvasion, we examined changes in demography, movements and activity patterns of reinvading alien black rats Rattus rattus in the short (4 weeks) and longer term (3 months) following localised experimental pest removal. Using recovery and invasion theory, we tested three hypothesised mechanisms of reinvasion: the ‘in situ effect’, the ‘trickle effect’ and the ‘vacuum effect’. We created space for reinvasion by removing black rats from the core of replicate 1‐ha plots (short‐term experiment) and later by removing animals from the entire plot (longer‐term experiment). Reinvaders were characterised as dispersing juveniles, floaters or neighbours. Radio‐tracking quantified home range changes for adjacent resident animals (short‐term experiment only). In the short term, there was no net influx of rats after targeted removal. Radio‐tracked residents’ movements were highly variable and displayed no directional changes after nearby conspecifics were removed. However, in the longer term, removal led to slow population recovery through a mix of reinvading floaters, dispersing juveniles and shifting residents. These responses best support a hypothesis of reinvasion through a trickle effect, with rats being extremely mobile and having a high degree of population turnover, even in untreated sites. Our findings provide the first test of reinvasion theory at a small scale, demonstrating the importance of understanding the differing categories of reinvaders and mechanisms of reinvasion after population control. These mechanisms drive the rate of population recovery and, in turn, should help determine which strategy of pest control should be used, and the frequency with which they are implemented, in order to slow the recovery of pest populations.     

Genomic imbalances are confined to non-proliferating cells in paediatric patients with acute myeloid leukaemia and a normal or incomplete karyotype

Leukaemia is often associated with genetic alterations such as translocations, amplifications and deletions, and recurrent chromosome abnormalities are used as markers of diagnostic and prognostic relevance. However, a proportion of acute myeloid leukaemia (AML) cases have an apparently normal karyotype despite comprehensive cytogenetic analysis. Based on conventional cytogenetic analysis of banded chromosomes, we selected a series of 23 paediatric patients with acute myeloid leukaemia and performed whole genome array comparative genome hybridization (aCGH) using DNA samples derived from the same patients. Imbalances involving large chromosomal regions or entire chromosomes were detected by aCGH in seven of the patients studied. Results were validated by fluorescence in situ hybridization (FISH) to both interphase nuclei and metaphase chromosomes using appropriate bacterial artificial chromosome (BAC) probes. The majority of these copy number alterations (CNAs) were confirmed by FISH and found to localize to the interphase rather than metaphase nuclei. Furthermore, the proliferative states of the cells analyzed by FISH were tested by immunofluorescence using an antibody against the proliferation marker pKi67. Interestingly, these experiments showed that, in the vast majority of cases, the changes appeared to be confined to interphase nuclei in a non-proliferative status.     

Impact of the leaf miner <Emphasis Type="Italic">Cameraria ohridella</Emphasis> on whole-plant photosynthetic productivity of <Emphasis Type="Italic">Aesculus hippocastanum</Emphasis>: insights from a model

The leaf miner Cameraria ohridella causes premature defoliation of Aesculus hippocastanum trees. In order to assess the whole-plant loss of productivity caused by the parasite, we monitored seasonal changes of leaf gas exchange and leaf area losses in horse chestnut trees freely infested or chemically treated to prevent moth infestation (controls). Data were integrated in a model and the annual loss of net primary productivity (NPP) was calculated for infested trees with respect to controls. Measurements showed marked vertical stratification of C. ohridella attacks, with lower crown strata being more infested than higher ones. Leaf gas exchange was maximum between May and early June, but it strongly decreased starting from mid-June even in controls. Model calculations showed that NPP loss of infested trees was about 30% on an annual basis (when the first moth attack is recorded at the end of April). Model simulations showed that postponing the start day of attack would have important positive effects on plants NPP. For example, if the start day of attack was postponed to 20 May, the annual loss of NPP would be about 15%. Our study suggests that A. hippocastanum trees attacked by C. ohridella are not facing serious risks of decline, especially if methods are adopted to postpone the start day of attack (e.g. removal of fallen leaves in autumn). Our data do not support the view that plants need to be totally protected from the parasite by application of insecticides.