In Vitro Evolution of Allergy Vaccine Candidates, with Maintained Structure, but Reduced B Cell and T Cell Activation Capacity

Allergy and asthma to cat (Felis domesticus) affects about 10% of the population in affluent countries. Immediate allergic symptoms are primarily mediated via IgE antibodies binding to B cell epitopes, whereas late phase inflammatory reactions are mediated via activated T cell recognition of allergen-specific T cell epitopes. Allergen-specific immunotherapy relieves symptoms and is the only treatment inducing a long-lasting protection by induction of protective immune responses. The aim of this study was to produce an allergy vaccine designed with the combined features of attenuated T cell activation, reduced anaphylactic properties, retained molecular integrity and induction of efficient IgE blocking IgG antibodies for safer and efficacious treatment of patients with allergy and asthma to cat. The template gene coding for rFel d 1 was used to introduce random mutations, which was subsequently expressed in large phage libraries. Despite accumulated mutations by up to 7 rounds of iterative error-prone PCR and biopanning, surface topology and structure was essentially maintained using IgE-antibodies from cat allergic patients for phage enrichment. Four candidates were isolated, displaying similar or lower IgE binding, reduced anaphylactic activity as measured by their capacity to induce basophil degranulation and, importantly, a significantly lower T cell reactivity in lymphoproliferative assays compared to the original rFel d 1. In addition, all mutants showed ability to induce blocking antibodies in immunized mice.The approach presented here provides a straightforward procedure to generate a novel type of allergy vaccines for safer and efficacious treatment of allergic patients

Family Business Restructuring:A Review and Research Agenda

Although business restructuring occurs frequently and it is important for the prosperity of family firms across generations, research on family firms has largely evolved separately from research on business restructuring. This is a missed opportunity, since the two domains are complementary, and understanding the context, process, content, and outcome dimensions is relevant to both research streams. We address this by examining the intersection between research on business restructuring and family firms to improve our knowledge of each area and inform future research. To achieve this goal, we review and organize research across different dimensions to create an integrative framework. Building on current research, we focus on 88 studies at the intersection of family firm and business restructuring research to develop a model that identifies research needs and suggests directions for future research

The Laukunkangas nickel-copper occurrence in southeastern Finland

The Laukunkangas Ni-Cu deposit in a Svecokarelidic synorogenic plutonite is located at the southeastern end of the Kotalahti nickel belt. The mineralization is associated with the norites, olivine gabbros and peridotites of an olivine tholeiitic suite. It contains about 4.5 million tons of ore in situ averaging 0.33 % Ni and 0.10 % Cu. The intrusive is a rather small pipelike body with an elliptical surface plan. It extends to a depth of at least 300 m; its largest horizontal dimension is about 1 km and its width in the middle of the intrusive is some 200 m. The variations in the Ni and Fe contents in silicates, the MgO content in rock types and the Ni content in the sulphide phase clearly indicate the internal layering in the intrusive. Emplacement probably took place in two or three stages. The boundaries between the layers show contamination that is reflected in the distribution of MgO and trace elements. »The model crystallization temperatures» suggest that the intrusion was emplaced in three separate phases. The mineralization occurs near the contact zone at the eastern end of the intrusive. The few nickel-rich portions encountered inside the intrusive are small and discontinous lenses or slabs. The mineralization is characterized by the following parameters: Ni/Co = 17.31, Ni/Cu = 3.15, Cu/Co = 5.49, and the Ni content in sulphide phase 4.81 %

Paleomagnetism of the dike systems in Finland IV: Remanent magnetization of the dolerite and related dikes in the Åland archipelago

The dikes of the area can be divided into three groups having different directions of remanent magnetization after demagnetization in the AC field. One of these groups has a direction which coincides with that measured for the Jotnian dolerites in the Satakunta and Vaasa archipelago. The direction of the second group differs but slightly from this and the third one has a reversed orientation. The age relations and the origin of the magnetization are discussed

Directed evolution of a type I antifreeze protein expressed in Escherichia coli with sodium chloride as selective pressure and its effect on antifreeze tolerance

Both freezing tolerance and NaCl tolerance are improved when antifreeze proteins are expressed as fusion proteins with two domains of staphylococcal protein A (SPA) in Escherichia coli. To characterize these properties further we created a randomly mutated expression library in E. coli, based on the winter flounder antifreeze protein HPLC-8 component gene. Low-fidelity PCR products of this gene were fused to the spa gene encoding two domains of the SPA. The library was screened for enhanced NaCl tolerance and four clones were selected. The freezing tolerance of each of the selected clones was enhanced to varying extents. DNA sequencing of the isolated mutants revealed that the amphiphilic properties of the native antifreeze protein were essentially conserved. Furthermore, by studying the primary sequence of the randomly mutated clones, in comparison with the degree of freezing tolerance, we have identified clues which help in understanding the relationship between salt and freezing tolerance

Suppression by neuropeptide Y of capsaicin-sensitive sensory nerve-mediated contraction in guinea-pig airways.

1. In the present study we have examined whether neuropeptide Y (NPY) interferes with non-adrenergic, non-cholinergic nerve-mediated contractions and relaxations in the guinea-pig airways. In these experiments we have used ring preparations of bronchi and trachea, incubated in the presence of atropine, propranolol and indomethacin (each 1 microM). 2. The contractile response to electrical stimulation of non-adrenergic, non-cholinergic nerve fibres was suppressed by NPY and NPY 13-36 in a concentration-dependent manner, these agents having similar inhibitory potencies. NPY caused a more complete inhibition than the C terminal fragment. 3. NPY affected neither the basal tension nor the substance P-evoked contraction in the bronchi and trachea and did not interfere with nerve-mediated, non-adrenergic relaxation in the trachea. 4. On the basis of these results, it is suggested that NPY may act on the terminals of sensory neurones in the airways to prevent antidromic, excitatory neurotransmission by inhibiting transmitter release