Donor APCs are required for maximal GVHD but not for GVL

Graft-versus-host disease (GVHD) is a major source of morbidity in allogenic stem cell transplantation. We previously showed that recipient antigen-presenting cells (APCs) are required for CD8-dependent GVHD in a mouse model across only minor histocompatibility antigens (minor H antigens). However, these studies did not address the function of donor-derived APCs after GVHD is initiated. Here we show that GVHD develops in recipients of donor major histocompatibility complex class I-deficient (MHC I(-)) bone marrow. Thus, after initial priming, CD8 cells caused GVHD without a further requirement for hematopoietic APCs, indicating that host APCs are necessary and sufficient for GHVD. Nonetheless, GVHD was less severe in recipients of MHC I(-) bone marrow. Therefore, once initiated, GVHD is intensified by donor-derived cells, most probably donor APCs cross-priming alloreactive CD8 cells. Nevertheless, donor APCs were not required for CD8-mediated graft-versus-leukemia (GVL) against a mouse model of chronic-phase chronic myelogenous leukemia. These studies identify donor APCs as a new target for treating GVHD, which may preserve GVL.     

The Structure of the Cyprinid herpesvirus 3 ORF112-Zα·Z-DNA Complex Reveals a Mechanism of Nucleic Acids Recognition Conserved with E3L,a Poxvirus Inhibitor of Interferon Response

In vertebrate species, the innate immune system down-regulates protein translation in response to viral infection through the action of the double-stranded RNA (dsRNA)-activated protein kinase (PKR). In some teleost species another protein kinase, Z-DNA-dependent protein kinase (PKZ), plays a similar role but instead of dsRNA binding domains, PKZ has Zα domains. These domains recognize the left-handed conformer of dsDNA and dsRNA known as Z-DNA/Z-RNA. Cyprinid herpesvirus 3 infects common and koi carp, which have PKZ, and encodes the ORF112 protein that itself bears a Zα domain, a putative competitive inhibitor of PKZ. Here we present the crystal structure of ORF112-Zα in complex with an 18-bp CpG DNA repeat, at 1.5 Å. We demonstrate that the bound DNA is in the left-handed conformation and identify key interactions for the specificity of ORF112. Localization of ORF112 protein in stress granules induced in Cyprinid herpesvirus 3-infected fish cells suggests a functional behavior similar to that of Zα domains of the interferon-regulated, nucleic acid surveillance proteins ADAR1 and DAI.     

A genome-wide association study of the Protein C anticoagulant pathway

The Protein C anticoagulant pathway regulates blood coagulation by preventing the inadequate formation of thrombi. It has two main plasma components: protein C and protein S. Individuals with protein C or protein S deficiency present a dramatically increased incidence of thromboembolic disorders. Here, we present the results of a genome-wide association study (GWAS) for protein C and protein S plasma levels in a set of extended pedigrees from the Genetic Analysis of Idiopathic Thrombophilia (GAIT) Project. A total number of 397 individuals from 21 families were typed for 307,984 SNPs using the Infinium® 317 k Beadchip (Illumina). Protein C and protein S (free, functional and total) plasma levels were determined with biochemical assays for all participants. Association with phenotypes was investigated through variance component analysis. After correcting for multiple testing, two SNPs for protein C plasma levels (rs867186 and rs8119351) and another two for free protein S plasma levels (rs1413885 and rs1570868) remained significant on a genome-wide level, located in and around the PROCR and the DNAJC6 genomic regions respectively. No SNPs were significantly associated with functional or total protein S plasma levels, although rs1413885 from DNAJC6 showed suggestive association with the functional protein S phenotype, possibly indicating that this locus plays an important role in protein S metabolism. Our results provide evidence that PROCR and DNAJC6 might play a role in protein C and free protein S plasma levels in the population studied, warranting further investigation on the role of these loci in the etiology of venous thromboembolism and other thrombotic diseases.     

Kefir yeast technology: scale-up in SCP production using milk whey

A three-step process to scale-up kefir biomass production at a semi-industrial scale employing whey is reported. Aerobic fermentations were initially performed at laboratory scales, in 1.5- and 4-L bioreactors, yielding 79 g/L final kefir biomass (0.89 g/g of lactose utilized), in 7 h of fermentation time. The use of whey as carbon source even in solid cultures led to the formation of a granular biomass. These results encouraged scale-up at a semi-industrial-scale pilot plant employing 100- and 3,000-L bioreactors, leading to the development of a process for granular kefir biomass production. The results validated the laboratory-scale experiments and the avoidance of centrifugal separators due to granular biomass formation. Pilot-plant operations showed kefir to be highly resistant to contamination under actual industrial conditions and no serious problems in handling of raw materials and equipment were observed. Economic analysis showed a 20% higher cost of the market price of products, with added value of up to 15.9 x 10(9) within the European Union.     

Crystal structure of the soluble form of equinatoxin II, a pore-forming toxin from the sea anemone Actinia equina

BACKGROUND: Membrane pore-forming toxins have a remarkable property: they adopt a stable soluble form structure, which, when in contact with a membrane, undergoes a series of transformations, leading to an active, membrane-bound form. In contrast to bacterial toxins, no structure of a pore-forming toxin from an eukaryotic organism has been determined so far, an indication that structural studies of equinatoxin II (EqtII) may unravel a novel mechanism. RESULTS: The crystal structure of the soluble form of EqtII from the sea anemone Actinia equina has been determined at 1.9 A resolution. EqtII is shown to be a single-domain protein based on a 12 strand beta sandwich fold with a hydrophobic core and a pair of alpha helices, each of which is associated with the face of a beta sheet. CONCLUSIONS: The structure of the 30 N-terminal residues is the largest segment that can adopt a different structure without disrupting the fold of the beta sandwich core. This segment includes a three-turn alpha helix that lies on the surface of a beta sheet and ends in a stretch of three positively charged residues, Lys-30, Arg-31, and Lys-32. On the basis of gathered data, it is suggested that this segment forms the membrane pore, whereas the beta sandwich structure remains unaltered and attaches to a membrane as do other structurally related extrinsic membrane proteins or their domains. The use of a structural data site-directed mutagenesis study should reveal the residues involved in membrane pore formation.     

Morphology, anatomy and reproduction of the eastern Mediterranean coralline Tenarea tortuosa and its relationship to members of the Lithophylloideae and Mastophoroideae (Rhodophyta, Corallinales)

One of the most characteristic organisms of the Mediterranean flora, the endemic coralline Tenarea tortuosa , has been collected along the Cretan coast and its vegetative and reproductive structures are reassessed. Gametophytes are described for the first time and are rare, observed only in one locality on the south coast. Reproductive structures suggest that the sexual life history is triphasic, but the present collections indicate unequal representation of males, carposporophytes and tetrasporophytes, and a dominance of bisporophytes suggesting that the alga is reproducing mainly asexually. The frond of Tenarea is formed by the coordinated growth of two elements (branches), growing back-to-back, each showing a simple Titanodema-type of organization; i.e. a monostromatic hypothallium of sinuous cells and a single layer of obliquely borne epithallial cells. Contiguous cells of the two opposite branches are not connected by secondary pit-plugs, and easily glide apart if decalcified. Natural splitting of the lamellae into its two branches may also occur, and single branches may fold regenerating the back-to-back organization. Analogous back-to-back growths occur in species of the distantly related genera Goniolithon (Lithophylloideae) and Leptophytum (Melobe-sioideae) and apparently represent cases of convergence. The affinities of Tenarea to related genera are discussed.     

The genus Leptophytum (Melobesioideae, Corallinales, Rhodophyta) in NW Spitsbergen

Three species of the genus Leptophytum , viz. the generitype L. laeve, L. foecundum and the new Arctic endemic L. jenneborgii nov. sp. are described from localities in NW Spitsbergen. A fourth species, related to L. laeve , may also be present and its status requires further comparative studies with Lithothamnion tenue described from western Greenland. Leptophytum jenneborgii is restricted to sublittoral sites around Vasahalvøya, between 8 and 30 m depth growing on hard substrates (usually on Lithothamnion glaciate ) and forming encrusting to foliose thalli up to 10 cm in diameter. New lamellae develop in an unattached-superimposed pattern, overgrowing the parent thallus, so that individuals can reach at least 1.5 cm in thickness. Thallus organization is dorsiventral, with a noncoaxial hypothallium (producing rare patches of coaxial cells) and an ascending perithallium with short subepithallial and flattened epithallial cells. Multiporate conceptacles have convex or flattened roof and develop specialized (thinner-wider) pore cells lining the canals. This character combination supports a position in the genus Leptophytum , as recently emended by Athanasiadis & Adey (2006), and simultaneously distinguishes L. jenneborgii from its Arctic and North Atlantic congeners. Like Leptophytum arcticum nov. comb., that is only known from Uddebay on the east coast of Novaja Zemlya, L. jenneborgii shows a restricted distribution on the presumed north periphery of maximum ice cover during the latest glaciation. It is suggested that these two species represent the first evidence of algal glacial survivals in this part of the Arctic.     

The taxonomic status of Lithophyllum stìctaeforme (Rhodophyta, Corallinales) and its generic position in light of phylogenetic considerations

The type of Lithophyllum stìctaeforme (Areschoug) Hauck (‵ Melobesia stictaeformis Areschoug in J. Agardh) is re-examined and shown to be conspecific with a common Mediterranean foliose coralline recently recognized as Lithophyllum frondosum (Dufour) Furnari, Cormaci et Alongi (‵ Melobesia frondosa Dufour) and previously known by the misapplied name Lithophyllum (Pseudolithophyllum) expansum Philippi. The generic position of the species in Lithophyllum or Pseudolithophyllum is phylogenetically examined, in the Hennigian sense. It is shown that secondary perithallial outgrowths ('faux hypothalle') may represent a synapomorphy for these genera that traditionally have been segregated from each other by the presence, respectively absence, of a coaxial 'faux hypothalle'. Recognition of both Lithophyllum and Pseudolithophyllum by virtue of a single character in its two states (absent vs. present) is however not tenable, if these genera are considered to be sister-taxa.     

Population relationships in the Mediterranean revealed by autosomal genetic data (Alu and Alu/STR compound systems)

The variation of 18 Alu polymorphisms and 3 linked STRs was determined in 1,831 individuals from 15 Mediterranean populations to analyze the relationships between human groups in this geographical region and provide a complementary perspective to information from studies based on uniparental markers. Patterns of population diversity revealed by the two kinds of markers examined were different from one another, likely in relation to their different mutation rates. Therefore, while the Alu biallelic variation underlies general heterogeneity throughout the whole Mediterranean region, the combined use of Alu and STR points to a considerable genetic differentiation between the two Mediterranean shores, presumably strengthened by a considerable sub‐Saharan African genetic contribution in North Africa (around 13% calculated from Alu markers). Gene flow analysis confirms the permeability of the Sahara to human passage along with the existence of trans‐Mediterranean interchanges. Two specific Alu/STR combinations—CD4 110(?) and DM 107(?)—detected in all North African samples, the Iberian Peninsula, Greece, Turkey, and some Mediterranean islands suggest an ancient genetic background of current Mediterranean peoples. Am J Phys Anthropol 2010. © 2009 Wiley‐Liss, Inc.