Mining the human autoantibody repertoire: Isolation of potent IL17A-neutralizing monoclonal antibodies from a patient with thymoma

Anti-cytokine autoantibodies have been widely reported to be present in human plasma, both in healthy subjects and in patients with underlying autoimmune conditions, such as autoimmune polyendocrinopathy candidiasis ectodermal dystrophy (APECED) or thymic epithelial neoplasms. While often asymptomatic, they can cause or facilitate a wide range of diseases including opportunistic infections. The potential therapeutic value of specific neutralizing anti-cytokine autoantibodies has not been thoroughly investigated. Here we used mammalian cell display to isolate IL17A-specific antibodies from a thymoma patient with proven high-titer autoantibodies against the same. We identified 3 distinct clonotypes that efficiently neutralized IL17A in a cell-based in vitro assay. Their potencies were comparable to those of known neutralizing antibodies, including 2, AIN457 (secukinumab) and ixekizumab that are currently in clinical development for the treatment of various inflammatory disorders. These data clearly demonstrate that the human autoantibody repertoire can be mined for antibodies with high therapeutic potential for clinical development.     

Mutational analysis of the Bradyrhizobium japonicum common nod genes and further nod box-linked genomic DNA regions

Summary By insertional and deletional marker replacement mutagenesis the common nod region of Bradyrhizobium japonicum was examined for the presence of additional, essential nodulation genes. An open reading frame located in the 800 bp large intergenic region between nodD1 and nodA did not appear to be essential for nodulation of soybean. Furthermore, a strain with a deletion of the nodI- and nodJ-like genes downstream of nodC had a Nod⁺ phenotype. A mutant with a 1.7 kb deletion immediately downstream of nodD1 considerably delayed the onset of nodulation. This region carried a second copy of nodD (nodD2). A nodD1-nodD2 double mutant had a similar phenotype to the nodD2 mutant. Using a 22-mer oligonucleotide probe partially identical to the nod box sequence, a total of six hybridizing regions were identified in B. japonicum genomic DNA and isolated from a cosmid library. Sequencing of the hybridizing regions revealed that at least three of them represented true nod box sequences whereas the others showed considerable deviations from the consensus sequence. One of the three nod box sequences was the one known to be associated with nodA, whereas the other two were located 60 to 70 kb away from nif cluster I. A deletion of one of these two sequences plus adjacent DNA material mmutant 308) led to a reduced nodulation on Vigna radiata but not on soybean. Thus, this region is probably involved in the determination of host specificity.Dedicated to Prof. Giorgio Semenza on the occasion of his 60th birthday     

ONE TWO WE—life cycle management in canteens together with suppliers,customers and guests

Purpose

Private food consumption accounts for 30 % of total environmental impacts caused by the final consumption of Swiss households. The private expenses for gastronomy and hotels account for another 6 %. Therefore, it is necessary to investigate and better understand the environmental impacts of food consumption and the possibilities for a reduction of these impacts. This was the starting point for the collaboration between the canteen operator SV Group, the life cycle assessment (LCA) consultancy ESU-services, the energy supplier ewz and the World Wide Fund for Nature (WWF) in Switzerland focusing on food consumption in canteens.

Methods

In a first step, an LCA study was used to analyse the environmental impacts of about 20 million meals served in 240 canteens in 2011. LCA data for 160 food items were linked to the food amounts of about 10,000 articles purchased in this year. This was supplemented by data on canteen operation and resulted in a full organisational LCA.

Results and discussion

The impacts of food purchases are about four times higher than the direct impacts due to the operation of the canteens. The most important product groups are meat and dairy products. Improvement potentials have been identified within 14 different themes by the project group. They include measures in the canteen operation (e.g. reduction of food waste or energy-efficient appliances); measures in the supply chain, e.g. a reduction of vegetables grown in heated greenhouses; or the abandonment of air-transported products. But also dietary choices such as a reduction of the average amount of meat per meal are considered as an option. The results and recommendations of the detailed LCA as well as information by other partners have been used by the SV Group to develop the programme ONE TWO WE. It assists the customers (companies who commission the operation of canteens in their premises) to reach improved levels of environmental performance. The programme aims for a 20 % cut on GHG emissions after full implementation in the participating canteens.

Conclusions

The programme started successfully with many customers positively convinced by the proposed changes in the provision of canteen meals. An initial reduction of greenhouse gas emissions compared to the baseline was achieved. This LCA study is a good example for the value of calculating a full organisational environmental footprint for a company in the gastronomy sector and for using the results of such a study to bring down the overall environmental impacts.

     

The permeability of thin lipid membranes to bromide and bromine

Thin lipid (optically black) membranes were made from sheep red cell lipids dissolved in n-decane. The flux of Br across these membranes was measured by the use of tracer ⁸²Br. The unidirectional flux of Br (in 50–100 mM NaBr) was 1–3 x 10^-12 mole/cm²sec. This flux is more than 1000 times the flux predicted from the membrane electrical resistance (>10⁸ ohm-cm²) and the transference number for Br^- (0.2–0.3), which was estimated from measurements of the zero current potential difference. The Br flux was not affected by changes in the potential difference imposed across the membrane (±60 mv) or by the ionic strength of the bathing solutions. However, the addition of a reducing agent, sodium thiosulfate (10^-3 M), to the NaBr solution bathing the membrane caused a 90% reduction in the Br flux. The inhibiting effect of S₂O₃ ⁼ suggests that the Br flux is due chiefly to traces of Br₂ in NaBr solutions. As expected, the addition of Br₂ to the NaBr solutions greatly stimulated the Br flux. However, at constant Br₂ concentration, the Br flux was also stimulated by increasing the Br^- concentration, in spite of the fact that the membrane was virtually impermeable to Br^-. Finally, the Br flux appeared to saturate at high Br₂ concentrations, and the saturation value was roughly proportional to the Br^- concentration. These results can be explained by a model which assumes that Br crosses the membrane only as Br₂ but that rapid equilibration of Br between Br₂ and Br^- occurs in the unstirred layers of aqueous solution bathing the two sides of the membrane. A consequence of the model is that Br^- "facilitates" the diffusion of Br across the unstirred layers.     

Proton/hydroxide conductance through phospholipid bilayer membranes: effects of phytanic acid

Mechanisms of proton/hydroxide conductance (GH/OH) were investigated in planar (Mueller-Rudin) bilayer membranes made from decane solutions of phospholipids or phospholipids plus phytanic acid (a 20-carbon, branched chain fatty acid). At neutral pH, membranes made from diphytanoylphosphatidylcholine or bacterial phosphatidylethanolamine had GH/OH values in the range of (2-5) X 10(-9) S X cm-2, corresponding to H+/OH- 'net' permeabilities of about (0.4-1.0) X 10(-5) cm X s-1. GH/OH was inhibited by serum albumin, phloretin, glycerol and low pH, but was increased by chlorodecane and voltage greater than 80 mV. Water permeability and GH/OH were not correlated, suggesting that water and H+/OH- cross the membrane by separate pathways. Addition of phytanic acid to the phospholipids caused an increase in GH/OH which was proportional to the first power of the phytanic acid concentration. In membranes containing phytanic acid, GH/OH was inhibited by albumin, phloretin, glycerol and low pH, but was increased by chlorodecane and voltages greater than 80 mV. The results suggest that phytanic acid acts as a simple (A- type) proton carrier. The qualitative similarities between the behavior of GH/OH in unmodified and phytanic-acid containing membranes suggest that phospholipids may contain weakly acidic contaminants which cause most of GH/OH at pH greater than 4. However, there is also a significant background (pH independent) GH/OH which may be due to hydrogen-bonded water chains. The ability of phytanic acid to act as a proton carrier may help to explain the toxicity of phytanic acid in Refsum's disease, a metabolic disorder in which phytanic acid accumulates to high levels in plasma, cells and tissues.     

Changes to airborne pollen counts across Europe

A progressive global increase in the burden of allergic diseases has affected the industrialized world over the last half century and has been reported in the literature. The clinical evidence reveals a general increase in both incidence and prevalence of respiratory diseases, such as allergic rhinitis (common hay fever) and asthma. Such phenomena may be related not only to air pollution and changes in lifestyle, but also to an actual increase in airborne quantities of allergenic pollen. Experimental enhancements of carbon dioxide (CO[Formula: see text]) have demonstrated changes in pollen amount and allergenicity, but this has rarely been shown in the wider environment. The present analysis of a continental-scale pollen data set reveals an increasing trend in the yearly amount of airborne pollen for many taxa in Europe, which is more pronounced in urban than semi-rural/rural areas. Climate change may contribute to these changes, however increased temperatures do not appear to be a major influencing factor. Instead, we suggest the anthropogenic rise of atmospheric CO[Formula: see text] levels may be influential.     

Relief of microRNA-mediated translational repression in human cells subjected to stress

In metazoans, most microRNAs imperfectly base-pair with the 3' untranslated region (3'UTR) of target mRNAs and prevent protein accumulation by either repressing translation or inducing mRNA degradation. Examples of specific mRNAs undergoing microRNA-mediated repression are numerous, but whether the repression is a reversible process remains largely unknown. Here we show that cationic amino acid transporter 1 (CAT-1) mRNA and reporters bearing its 3'UTR can be relieved from the microRNA miR-122-induced inhibition in human hepatocarcinoma cells subjected to different stress conditions. The derepression of CAT-1 mRNA is accompanied by its release from cytoplasmic processing bodies and its recruitment to polysomes. The derepression requires binding of HuR, an AU-rich-element binding protein, to the 3'UTR of CAT-1 mRNA. We propose that proteins interacting with the 3'UTR will generally act as modifiers altering the potential of miRNAs to repress gene expression.     

Ion Fluxes and Short-Circuit Current in Internally Perfused Cells of Valonia ventricosa

Ion transport in the giant celled marine alga, Valonia ventricosa, was studied during internal perfusion and short-circuiting of the vacuole potential. The perfusing and bathing solutions were similar to natural Valonia sap and contained the following concentrations of major ions: Na 51, K 618, and Cl 652 mM. The average short-circuit current (I _sc) was 97 pEq/cm² sec (inward positive current), and the average open-circuit potential difference (PD) was 74 mv (vacuole positive to external solution). Perfused and short-circuited cells showed a small net influx of Na (2.0 pEq/cm² sec) and large net influxes of K (80 pEq/cm² sec) and Cl (50 pEq/cm² sec). Unidirectional K influx was proportional to I _sc, but more than one-half of the I _sc remained unaccounted for. Both the I _sc and PD were partly light-dependent, declining rapidly during the first 1–2 min of darkness. Ouabain (5 x 10^-4 M) had little effect on the influx of Na or K and had no effect on I _inf or PD. Fluid was absorbed at a rate of about 93 pliter/cm² sec. Reversing the direction of fluid movement by adding mannitol to the outside solution had little effect on ion movements. The ionic and electrical properties of normal and perfused cells of Valonia are compared.