BMP-4 is proteolytically activated by furin and/or PC6 during vertebrate embryonic development.

Bone morphogenetic protein-4 (BMP-4) is a multifunctional developmental regulator. BMP-4 is synthesized as an inactive precursor that is proteolytically activated by cleavage following the amino acid motif -Arg-Ser-Lys-Arg-. Very little is known about processing and secretion of BMPs. The proprotein convertases (PCs) are a family of seven structurally related serine endoproteases, at least one of which, furin, cleaves after the amino acid motif -Arg-X-Arg/Lys-Arg-. To examine potential roles of PCs during embryonic development we have misexpressed a potent protein inhibitor of furin, alpha1-antitrypsin Portland (alpha1-PDX) in early Xenopus embryos. Ectopic expression of alpha1-PDX phenocopies the effect of blocking endogenous BMP activity, leading to dorsalization of mesoderm and direct neural induction. alpha1-PDX-mediated neural induction can be reversed by co-expression of downstream components of the BMP-4 signaling pathway. Thus, alpha1-PDX can block BMP activity upstream of receptor binding, suggesting that it inhibits an endogenous BMP-4 convertase(s). Consistent with this hypothesis, alpha1-PDX prevents cleavage of BMP-4 in an oocyte translation assay. Using an in vitro digestion assay, we demonstrate that four members of the PC family have the ability to cleave BMP-4, but of these, only furin and PC6B are sensitive to alpha1-PDX. These studies provide the first in vivo evidence that furin and/or PC6 proteolytically activate BMP-4 during vertebrate embryogenesis.     

Calcium and S100B Regulation of p53-Dependent Cell Growth Arrest and Apoptosis

In glial C6 cells constitutively expressing wild-type p53, synthesis of the calcium-binding protein S100B is associated with cell density-dependent inhibition of growth and apoptosis in response to UV irradiation. A functional interaction between S100B and p53 was first demonstrated in p53-negative mouse embryo fibroblasts (MEF cells) by sequential transfection with the S100B and the temperature-sensitive p53Val135 genes. We show that in MEF cells expressing a low level of p53Val135, S100B cooperates with p53Val135 in triggering calcium-dependent cell growth arrest and cell death in response to UV irradiation at the nonpermissive temperature (37.5°C). Calcium-dependent growth arrest of MEF cells expressing S100B correlates with specific nuclear accumulation of the wild-type p53Val135 conformational species. S100B modulation of wild-type p53Val135 nuclear translocation and functions was confirmed with the rat embryo fibroblast (REF) cell line clone 6, which is transformed by oncogenic Ha-ras and overexpression of p53Val135. Ectopic expression of S100B in clone 6 cells restores contact inhibition of growth at 37.5°C, which also correlates with nuclear accumulation of the wild-type p53Val135 conformational species. Moreover, a calcium ionophore mediates a reversible G₁ arrest in S100B-expressing REF (S100B-REF) cells at 37.5°C that is phenotypically indistinguishable from p53-mediated G₁ arrest at the permissive temperature (32°C). S100B-REF cells proceeding from G₁ underwent apoptosis in response to UV irradiation. Our data support a model in which calcium signaling and S100B cooperate with the p53 pathways of cell growth inhibition and apoptosis.     

Synthesis and biological activity of N-aroyl-tetrahydro-γ-carbolines

Research on dual inhibitors of both 5-LOX and COXs gained interest due to the overexpressions of these enzymes during the malignant state of the evolution of prostate cancer. In order to take part in this research, new N-aroyl-tetrahydro-γ-carbolines issued from the modification of Indomethacin have been synthesised. As for the NSAIDs, the compounds have been tested for their activity against COX₁, COX₂ plus against 5-LOX and against the proliferation of malignant prostate cancer. Interesting cytotoxic activities and selectivities of some tetrahydro-γ-carboline derivatives have been obtained.     

Material & equipment,procurement & maintenance: Impact on blood safety

Blood Transfusion Safety is dependent on effectively organised and managed blood services, which have adequate financial resources, skilled manpower, appropriate infrastructure and quality management systems in place.80% of the world's population has access to 20% of the supply blood products, of which little is consistently safe.HIV highlighted the importance of blood safety. The lack of effective blood services in low human development index (LHDI), developing countries, has lead to international funding and capacity building for more than three decades. The initial strategies focused on providing HIV testing reagents to prevention transmission, however this only addresses one part of blood safety.Blood safety is not only dependent on preventing HIV transmission. In many populations there are other infectious agents, which have a higher prevalence. Ensuring the correct blood is provided to the patient depends on: well managed services with effective leadership and adequate budgets; capacity building and retention of skilled experienced staff; availability of laboratory equipment, correctly maintained; blood cold chain systems; procedures for tendering, purchasing and ensuring an unbroken supply of reagents and consumables; and quality management systems.Barriers for simplified effective tendering, procurement and contracting require urgent attention and coordination of all funding organisations to ensure an unbroken supply of reagents.     

INTRASPECIFIC VARIATION IN THE DINOFLAGELLATE PERIDINIUM VOLZIP1

Clonal isolates of Peridinium volzii Lemmerman were analyzed morphologically and biochemically. Morphological observations at the light microscope level show the clones to be different varieties and forms of the same species. Biochemical analysis by enzyme electrophoresis and flow cytometric determination of nuclear DNA quantities indicates that these isolates are genetically heterogeneous without any clear correlation existing between morphological variation and biochemical variation. Isozyme analysis, however, indicates that strains from the same location are generally more related to each other than they are to isolates with other geographical origins. In general, our results suggest the presence of genetic redundancy and a multiclonal origin for individuals of the same species present in the same locale.     

Enantiomeric conservation of the male‐produced sex pheromone facilitates monitoring of threatened European hermit beetles (Osmoderma spp.)

Hermit beetles of the genus Osmoderma (Coleoptera: Scarabaeidae: Cetoniinae) are known for their fruity odour, which is released in large amounts by males. Two species of the genus occur in Europe, the eastern Osmoderma barnabita (Motschulsky) and the western Osmoderma eremita (Scopoli). Previous studies on Swedish populations of O. eremita showed that the compound responsible for the characteristic scent, γ‐decalactone, functions as a sex pheromone for the attraction of conspecific females. Male O. eremita only release the (R)‐enantiomer of the lactone, and both sexes are anosmic to the opposite enantiomer. As the distribution areas of the two hermit beetle species partly overlap, it may be expected that they use different enantiomeric compositions of γ‐decalactone as pheromones to promote species discrimination. This paper reports on the identification of the sex pheromone of O. barnabita. Surprisingly, males from a Polish population produce only the (R)‐enantiomer of γ‐decalactone, and conspecific females show equal attraction to the (R)‐enantiomer and a racemic mixture of the compound, indicating that O. barnabita is anosmic to the (S)‐enantiomer, similarly to what was observed for O. eremita. A mtDNA sequence analysis of the cytochrome oxidase subunit I gene of Polish and Swedish beetles confirmed their taxonomical status as O. barnabita and O. eremita, respectively, with an average sequence divergence of 10.5% between beetles from the two studied areas. Although genetic data suggest that these species diverged several million years ago, they still rely on the same enantiomer of γ‐decalactone for mate finding. Thus, the male‐produced pheromone in Osmoderma spp. may be regarded as a territorial signal being exploited by females, rather than a cue for determining species identity. Our data show that the same compound can be used to facilitate monitoring of both beetle species, which are considered indicator species of the species‐rich fauna of saproxylic insects in Europe.     

Haemolytic activity,cytotoxicity and membrane cell permeabilization of semi-synthetic and natural lupane- and oleanane-type saponins

The haemolysis of red blood cells inducing toxicity in most animals including humans is a major drawback for the clinical development of saponins as antitumour agents. In this study, the haemolytic and cytotoxic activities as well as the membrane cell permeabilization property of a library of 31 semi-synthetic and natural lupane- and oleanane-type saponins were evaluated and the structure–activity relationships were established. It was shown that lupane-type saponins do not exhibit any haemolytic activity and membrane cell permeabilization property at the maximum concentration tested (100 μM) independently of the nature of the sugar moieties. While oleanane-type saponins such as β-hederin (25) and hederacolchiside A₁ (27) cause the death of cancer cell lines by permeabilizing the cellular membranes, lupane-type saponins seem to proceed via another mechanism, which could be related to the induction of apoptosis. Altogether, the results indicate that the cytotoxic lupane-type glycosides 10 and 22 bearing an α-l-rhamnopyranose moiety at the C-3 position represent promising antitumour agents for further studies on tumour-bearing mice since they are devoid of toxicity associated with the haemolysis of red blood cells.