Immunogenicity of human embryonic stem cells

Human embryonic stem cells (HESC) are pluripotent stem cells isolated from the inner cell mass of human blastocysts. With the first successful culturing of HESC, a new era of regenerative medicine was born. HESC can differentiate into almost any cell type and, in the future, might replace solid organ transplantation and even be used to treat progressive degenerative diseases such as Parkinson’s disease. Although this sounds promising, certain obstacles remain with regard to their clinical use, such as culturing HESC under well-defined conditions without exposure to animal proteins, the risk of teratoma development and finally the avoidance of immune rejection. In this review, we discuss the immunological properties of HESC and various strategic solutions to circumvent immune rejection, such as stem cell banking, somatic cell nuclear transfer and the induction of tolerance by co-stimulation blockade and mixed chimerism.     

Demethoxylation of [O14CH3]-labelled lignin model compounds by the brown-rot fungi Gloeophyllum trabeum and Poria (Postia) placenta

Demethoxylation reactions in the cultures of the brown-rot fungi Gloeophyllum trabeum and Poria placenta were studied by determining the evolution of (14)CO(2) from a non-phenolic lignin model, beta-O-4 dimer, [O(14)CH(3)]-labelled at position 4 in the A ring (model I), and from [O(14)CH(3)]-labelled vanillic acid (model II). The fungi were grown under oxygen or air atmosphere on an agar medium with or without spruce sapwood blocks. The dimeric model (I) was impregnated onto agar or wood block in cultures to clarify the possible effect of wood as growth substrate. In the case of vanillic acid (model II), birch wood was used. The effect of supplemented nutrient nitrogen (2 mM N) and glucose (0.1 or 1.0% w/v) on demethoxylation was also studied. G. trabeum enhanced the production of (14)CO(2) from the dimer in the presence of spruce wood blocks. It released (14)CO(2) from the methoxyl groups giving 30-60% of the applied activity in 8 weeks. P. placenta produced almost 30% (14)CO(2 )from vanillic acid (model II) in 9 weeks under oxygen, but from the methoxyl group of the dimer only 3% of (14)CO(2) was evolved in 4 weeks. The biomasses determined as ergosterol assay showed variation from 14 to 226 microg g(-1) dry weight of agar, and 2 to 9 microg g(-1 )of wood, but they did not correlate with the production of (14)CO(2). The results indicate that these brown-rot fungi possess different mechanisms for demethoxylation.     

Borrelia burgdorferi inhibits human neutrophil functions

Outer surface proteins OspA and OspB are among the most prominent Borrelia burgdorferi surface molecules. We constructed OspAB and OspA complementation mutants of B. burgdorferi Osp-less strain B313 and investigated the role of these surface proteins in the interactions of B. burgdorferi, human neutrophils and the complement system. We found that (1) OspB inhibits the phagocytosis and oxidative burst of human neutrophils at low serum concentrations, whereas OspA induces the oxidative burst in neutrophils; (2) OspB may have an inhibiting role in serum sensitivity and complement activation; (3) all studied strains inhibit the chemotaxis of human neutrophils specifically towards fMLP but not towards C5a, regardless of their Osp expression. These results suggest that although OspA and OspB are co-ordinately transcribed, they differ in their effects on human neutrophil functions. Our findings suggest that B. burgdorferi exploits a wide variety of immune evasion mechanisms, besides previously documented complement resistance, to survive in the vertebrate host.     

Effect of sampling grain on patterns of species richness and turnover in Amazonian forests

Grain (size of sampling units) affects the spatial resolution at which ecological patterns can be observed and analysed, and potentially has an important effect on the results of broad‐scale studies on diversity gradients. Here we examine the effect of grain on patterns of species richness and turnover in lowland rainforests of western Amazonia (Peru and Colombia). We inventoried pteridophytes (ferns and lycophytes), melastomes (Melastomataceae) and palms (Arecaceae) in four line transects of 22–29 km length. Different grains were obtained by aggregating original 100‐m‐long sampling units into larger segments up to 19.2 km long. With any given grain and plant group, local species richness varied considerably both within and among transects, and a transect segment that was species‐rich with one grain could be relatively species‐poor with another. Which transect had the highest vs lowest mean species richness per sampling unit (α richness) differed among plant groups. It also varied to some degree with grain, as transects differed in how rapidly local species richness increased with increasing grain. Patterns of species turnover were more consistently correlated among plant groups than patterns of species richness were, and NMDS ordinations were rather similar with all grains and plant groups. Floristic heterogeneity within the Amazonian terra firme rainforest seems to contain a general compositional pattern that is sufficiently robust to be detectable with various sampling schemes, but patterns of species richness appear more case‐specific. Therefore, using one plant group as an indicator for patterns in other plant groups can be expected to work better for species turnover than for species richness.     

A rapid method for selecting suitable animal species for studying pathogen interactions with plasma protein ligands in vivo

Species tropism constitutes a serious problem for developing relevant animal models of infection. Human pathogens can express virulence factors that show specific selectivity to human proteins, while their affinity for orthologs from other species can vary significantly. Suitable animal species must be used to analyse whether virulence factors are potential targets for drug development. We developed an assay that rapidly predicts applicable animal species for studying virulence factors binding plasma proteins. We used two well‐characterized Staphylococcus aureus proteins, SSL7 and Efb, to develop an ELISA‐based inhibition assay using plasma from different animal species. The interaction between SSL7 and human C5 and the binding of Efb to human fibrinogen and human C3 was studied. Affinity experiments and Western blot analyses were used to validate the assay. Human, monkey and cat plasma interfered with binding of SSL7 to human C5. Binding of Efb to human fibrinogen was blocked in human, monkey, gerbil and pig plasma, while human, monkey, gerbil, rabbit, cat and guinea pig plasma inhibited the binding of Efb to human C3. These results emphasize the importance of choosing correct animal models, and thus, our approach is a rapid and cost‐effective method that can be used to prevent unnecessary animal experiments.     

Effects of Graded Levels of Tissue Oxygen Pressure on Dopamine Metabolism in the Striatum of Newborn Piglets

Abstract: The effect of graded levels of tissue hypoxia on the extracellular levels of dopamine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, and 5-hydroxyindoleacetic acid has been monitored in vivo by microdialysis. Reproducible levels of decreased oxygen in the brain were obtained by increasing the rate of ventilation from the control value of 25/min to as high as 95/min. With increasing ventilatory rate, the oxygen pressure in the cortex decreased from ∼40 torr to 16 torr. As the oxygen pressure decreased stepwise from 40 to 27, 22, and 16 torr, the dopamine levels in the extracellular medium rose by 70, 90, and 150%, respectively, returning to baseline within a few minutes of return to control ventilation rates. Levels of the catabolic products 3,4-dihydroxyphenylacetic acid, homovanillic acid, and 5-hydroxyindoleacetic acid decreased with decreasing tissue oxygen. Unlike the dopamine levels, these catabolite levels continued to decrease through 30 min of recovery (to 50% of control), returning to baseline only after recovery periods of 1–2 h. These data suggest that hypoxia induces long-term alterations in the neurotransmitter turnover. The marked effects of mild tissue hypoxia (decrease of oxygen from 40 torr to 26 torr) on both the extracellular dopamine concentration and dopamine metabolism indicate that the metabolic consequences of decreased tissue oxygen pressure extend to higher values than generally appreciated.     

Transformation of antisense constructs of the chalcone synthase gene superfamily into Gerbera hybrida: differential effect on the expression of family members

Suppression of gene expression using antisense technology has been successful in various applications. In this paper we report differential inhibition of gene expression of the chalcone synthase (chs) gene superfamily members in transgenic Gerbera hybrida (Asteraceae) plants. We have transformed two different cDNAs of the chs gene family, gchs 1 [4] and gchs2, in antisense orientation under control of the CaMV 35S promoter into gerbera. Gchs1 codes for an enzyme with chalcone synthase activity while gchs2 is a more diverged member of the gene family having distinct structure and expression pattern. Furthermore, gchs2 is evidently not involved in anthocyanin synthesis and encodes an enzyme with novel catalytic properties. In both cases effective blocking of the resident sense gene expression was detected. In addition, the transformation affected differentially the expression of other members of the chs gene family. The degree of inhibition appeared to depend on the sequence homology between the antisense and the target genes. In the unevenly coloured inflorescences detected among anti-gchs1 transformants during their growth, relaxation of the antisense effect was here shown to start from the most distant member of the gene family, further demonstrating the influence of sequence homology in the stability of antisense inhibition.     

Effect of dichloromethylene diphosphonate (Cl2MDP) on immune function in breast cancer patients with bone metastases

Immune function was studied in normocalcemic breast cancer patients with bone metastases treated with either dichloromethylene diphosphonate (Cl2MDP) or placebo. The results showed no significant difference between the two patient groups. This suggests that Cl2MDP does not markedly impair the host's defense mechanisms, and in this respect can be safely used in the treatment of patients with resorptive bone disease.