The TIM translocase in T. brucei contains Rhomboid-like proteins that are essential for mitochondrial protein import

The parasitic protozoon Trypanosoma brucei is often considered as one of the earliest branching eukaryotes that have mitochondria capable of oxidative phosphorylation. Its protein import systems are therefore of great interest. Recently, it was shown that the outer mitochondrial membrane protein translocase is of similar complexity yet different composition than in other eukaryotes (1). In the inner membrane however, only a single orthologue of the pore forming Tim17/22/23 protein family was identified and termed TbTim17. Based on this finding it has been suggested that, instead of separate TIM22 and TIM23 complexes as in other eukaryotes, trypanosomes may have a single multifunctional translocase of the inner mitochondrial membrane (TIM) of reduced complexity. To elucidate the composition of the trypanosomal TIM complex we performed co-immunoprecipitations (CoIP) of epitope-tagged TbTim17 in combination with SILAC-based quantitative mass spectrometry. This led to the identification of 22 highly enriched TbTim17-interacting proteins. We tagged two of the top-scoring proteins for reciprocal CoIP analyses and recovered a set of ten proteins that are highly enriched in all three CoIPs. These proteins are excellent candidates for core subunits of the trypanosomal TIM complex. Eight of them were present in the previously determined inner membrane proteome and four show homology to small Tim chaperones. Three candidates, a novel trypanosomatid-specific 42 kDa protein, termed Tim42, and two putative orthologues of probably inactive rhomboid proteases were chosen for further analysis. All three proteins are essential in both life cycle stages and in a cell line that can grow in the absence of mitochondrial DNA. Additionally, their ablation by RNAi results in a strong protein import defect both in vivo and in vitro. Blue native PAGE reveals that Tim42, like TbTim17 is present in a high molecular weight complex. Moreover, ablation of either Tim42 or TbTim17 leads to a destabilization of the complex containing the other protein, suggesting a tight interaction of the two proteins. In summary our study shows that unlike anticipated trypanosomes have a highly complex TIM translocase that has extensively been redesigned. We have characterized three novel TIM subunits that have never been associated with mitochondrial protein import before. Two of them belong to the rhomboid protease family, a member of which recently has been implicated in the ERAD translocation system. Our study provides insight into mitochondrial evolution over large phylogenetic distances and suggests an exciting analogy between protein translocation systems of mitochondria and the ER.

Characterization of fetal antigen 1/delta-like 1 homologue expressing cells in the rat nigrostriatal system: effects of a unilateral 6-hydroxydopamine lesion.

Fetal antigen 1/delta-like 1 homologue (FA1/dlk1) belongs to the epidermal growth factor superfamily and is considered to be a non-canonical ligand for the Notch receptor. Interactions between Notch and its ligands are crucial for the development of various tissues. Moreover, FA1/dlk1 has been suggested as a potential supplementary marker of dopaminergic neurons. The present study aimed at investigating the distribution of FA1/dlk1-immunoreactive (-ir) cells in the early postnatal and adult midbrain as well as in the nigrostriatal system of 6-hydroxydopamine (6-OHDA)-lesioned hemiparkinsonian adult rats. FA1/dlk1-ir cells were predominantly distributed in the substantia nigra (SN) pars compacta (SNc) and in the ventral tegmental area. Interestingly, the expression of FA1/dlk1 significantly increased in tyrosine hydroxylase (TH)-ir cells during early postnatal development. Co-localization and tracing studies demonstrated that FA1/dlk1-ir cells in the SNc were nigrostriatal dopaminergic neurons, and unilateral 6-OHDA lesions resulted in loss of both FA1/dlk1-ir and TH-ir cells in the SNc. Surprisingly, increased numbers of FA1/dlk1-ir cells (by 70%) were detected in dopamine-depleted striata as compared to unlesioned controls. The higher number of FA1/dlk1-ir cells was likely not due to neurogenesis as colocalization studies for proliferation markers were negative. This suggests that FA1/dlk1 was up-regulated in intrinsic cells in response to the 6-OHDA-mediated loss of FA1/dlk1-expressing SNc dopaminergic neurons and/or due to the stab wound. Our findings hint to a significant role of FA1/dlk1 in the SNc during early postnatal development. The differential expression of FA1/dlk1 in the SNc and the striatum of dopamine-depleted rats could indicate a potential involvement of FA1/dlk1 in the cellular response to the degenerative processes.

Insulin-like growth factor binding protein 2 (IGFBP2) promotes glioma development and progression

Overexpression of insulin-like growth factor binding protein 2 (IGFBP2) is associated with progression and poor survival in many types of human cancer (such as prostate, ovarian, adrenocortical, breast, colorectal carcinomas, leukemia, and high-grade gliomas). We therefore hypothesize that IGFBP2 is a key regulator of tumor progression. We tested our hypothesis in gliomas using the somatic gene transfer RCAS-tva mouse model system, which permits the introduction of specific genes into specific, cell lineages, in this case glial cells (RCAS: Replication competent avian sarcomavirus, tv-a: avian RCAS virus receptor). Mice are transgenic and harbor the tv-a receptor under the control of a glial-specific promoter and study genes are cloned into the RCAS vector for post-natal intracranial delivery. For these experiments, the study genes were IGFBP2, platelet-derived growth factor B (PDGFB), K-Ras, Akt, and IIp45 (invasion inhibitory protein 45 kDa; known to bind and block IGFBP2 activity), which were delivered separately and in combination. Our results show that PDGFB signaling leads exclusively to the formation of low-grade (WHO grade II) oligodendrogliomas. PDGFB delivered in combination with IGFBP2 results in the formation of anaplastic oligodendrogliomas (WHO grade III), which are characterized by increased cellularity, vascular proliferation, small regions of necrosis, increased mitotic activity, and increased activation of the Akt pathway. IIp45 injected in combination with PDGFB and IGFBP2 ablates IGFBP2-induced tumor progression, which results in formation of low-grade oligodendrogliomas, and an overall reduction in tumor incidence. K-Ras expression was required to form astrocytomas with either IGFBP2 or Akt, indicating the activation of two separate pathways is necessary for gliomagenesis. In ex vivo experiments, blockade of Akt by an inhibitor led to decreased viability of cells co-expressing IGFBP2 versus PDGFB expression alone. This study provides definitive evidence, for the first time, that: (1) IGFBP2 plays a role in activation of the Akt pathway, (2) IGFBP2 collaborates with K-Ras or PDGFB in the development and progression of two major types of glioma, and (3) IGFBP2-induced tumor progression can be ablated by IIp45 or by specific inhibition of the Akt pathway. ^

PSV records from Tangra Yumco and Taro Co (Tibetan Plateau)

Magnetostratigraphy has been serving as a valuable tool for dating and confirming chronologies of lacustrine sediments in many parts of the world. Suitable paleomagnetic records on the Tibetan Plateau (TP) and adjacent areas are, however, extremely scarce. Here, we derive paleomagnetic records from independently radiocarbon-dated sediments from two lakes separated by 250 km on the southern central TP, Tangra Yumco and Taro Co. Studied through alternating field demagnetization of u-channel samples, characteristic remanent magnetization (ChRM) directions document similar inclination patterns in multiple sediment cores for the past 4000 years. Comparisons to an existing record from Nam Co, a lake 350 km east of Tangra Yumco, a varve-dated record from the Makran Accretionary Wedge, records from Lakes Issyk-Kul and Baikal, and a stack record from East Asia reveal many similarities in inclination. This regional similarity demonstrates the high potential of inclination to compare records over the Tibetan Plateau and eventually date other Tibetan records stratigraphically. PSV similarities over such a large area (>3000 km) suggest a large-scale core dynamic origin rather than small scale processes like drift of the non-dipole field often associated with PSV records.

Competition may explain the fine-scale spatial patterns and genetic structure of two co-occurring plant congeners

1. The spatial distribution of individual plants within a population and the population’s genetic structure are determined by several factors, like dispersal, reproduction mode or biotic interactions. The role of interspecific interactions in shaping the spatial genetic structure of plant populations remains largely unknown. 2. Species with a common evolutionary history are known to interact more closely with each other than unrelated species due to the greater number of traits they share. We hypothesize that plant interactions may shape the fine genetic structure of closely related congeners. 3. We used spatial statistics (georeferenced design) and molecular techniques (ISSR markers) to understand how two closely related congeners, Thymus vulgaris (widespread species) and T. loscosii (narrow endemic) interact at the local scale. Specific cover, number of individuals of both study species and several community attributes were measured in a 10 × 10 m plot. 4. Both species showed similar levels of genetic variation, but differed in their spatial genetic structure. Thymus vulgaris showed spatial aggregation but no spatial genetic structure, while T. loscosii showed spatial genetic structure (positive genetic autocorrelation) at short distances. The spatial pattern of T. vulgaris’ cover showed significant dissociation with that of T. loscosii. The same was true between the spatial patterns of the cover of T. vulgaris and the abundance of T. loscosii and between the abundance of each species. Most importantly, we found a correlation between the genetic structure of T. loscosii and the abundance of T. vulgaris: T. loscosii plants were genetically more similar when they were surrounded by a similar number of T. vulgaris plants. 5. Synthesis. Our results reveal spatially complex genetic structures of both congeners at small spatial scales. The negative association among the spatial patterns of the two species and the genetic structure found for T. loscosii in relation to the abundance of T. vulgaris indicate that competition between the two species may account for the presence of adapted ecotypes of T. loscosii to the abundance of a competing congeneric species. This suggests that the presence and abundance of close congeners can influence the genetic spatial structure of plant species at fine scales.

Influence of the molecular architecture on the secondary relaxations of Poly(styrene-co-methylmethacrylate) copolymers

The processes of adsorption of grafted copolymers onto negatively charged surfaces were studied using a dissipative quartz crystal microbalance (D-QCM) and ellipsometry. The control parameters in the study of the adsorption are the existence or absence on the molecular architecture of grafted polyethyleneglycol (PEG) chains with different lengths and the chemical nature of the main chain, poly(allylamine) (PAH) or poly(L-lysine) (PLL). It was found out that the adsorption kinetics of the polymers showed a complex behavior. The total adsorbed amount depends on the architecture of the polymer chains (length of the PEG chains), on the polymer concentration and on the chemical nature of the main chain. The comparison of the thicknesses of the adsorbed layers obtained from D-QCM and from ellipsometry allowed calculation of the water content of the layers that is intimately related to the grafting length. The analysis of D-QCM results also provides information about the shear modulus of the layers, whose values have been found to be typical of a rubber-like polymer system. It is shown that the adsorption of polymers with a charged backbone is not driven exclusively by the electrostatic interactions, but the entropic contributions as a result of the trapping of water in the layer structure are of fundamental importance.

The involvement of thaumatin-like proteins in plant food cross-reactivity: a multicenter study using a specific protein microarray.

Cross-reactivity of plant foods is an important phenomenon in allergy, with geographical variations with respect to the number and prevalence of the allergens involved in this process, whose complexity requires detailed studies. We have addressed the role of thaumatin-like proteins (TLPs) in cross-reactivity between fruit and pollen allergies. A representative panel of 16 purified TLPs was printed onto an allergen microarray. The proteins selected belonged to the sources most frequently associated with peach allergy in representative regions of Spain. Sera from two groups of well characterized patients, one with allergy to Rosaceae fruit (FAG) and another against pollens but tolerant to food-plant allergens (PAG), were obtained from seven geographical areas with different environmental pollen profiles. Cross-reactivity between members of this family was demonstrated by inhibition assays. Only 6 out of 16 purified TLPs showed noticeable allergenic activity in the studied populations. Pru p 2.0201, the peach TLP (41%), chestnut TLP (24%) and plane pollen TLP (22%) proved to be allergens of probable relevance to fruit allergy, being mainly associated with pollen sensitization, and strongly linked to specific geographical areas such as Barcelona, Bilbao, the Canary Islands and Madrid. The patients exhibited mayor que50% positive response to Pru p 2.0201 and to chestnut TLP in these specific areas. Therefore, their recognition patterns were associated with the geographical area, suggesting a role for pollen in the sensitization of these allergens. Finally, the co-sensitizations of patients considering pairs of TLP allergens were analyzed by using the co-sensitization graph associated with an allergen microarray immunoassay. Our data indicate that TLPs are significant allergens in plant food allergy and should be considered when diagnosing and treating pollen-food allergy.

Einstein-like geometric structures on surfaces

An AH (affine hypersurface) structure is a pair comprising a projective equivalence class of torsion-free connections and a conformal structure satisfying a compatibility condition which is automatic in two dimensions. They generalize Weyl structures, and a pair of AH structures is induced on a co-oriented non-degenerate immersed hypersurface in flat affine space. The author has defined for AH structures Einstein equations, which specialize on the one hand to the usual Einstein Weyl equations and, on the other hand, to the equations for affine hyperspheres. Here these equations are solved for Riemannian signature AH structures on compact orientable surfaces, the deformation spaces of solutions are described, and some aspects of the geometry of these structures are related. Every such structure is either Einstein Weyl (in the sense defined for surfaces by Calderbank) or is determined by a pair comprising a conformal structure and a cubic holomorphic differential, and so by a convex flat real projective structure. In the latter case it can be identified with a solution of the Abelian vortex equations on an appropriate power of the canonical bundle. On the cone over a surface of genus at least two carrying an Einstein AH structure there are Monge-Amp`ere metrics of Lorentzian and Riemannian signature and a Riemannian Einstein K"ahler affine metric. A mean curvature zero spacelike immersed Lagrangian submanifold of a para-K"ahler four-manifold with constant para-holomorphic sectional curvature inherits an Einstein AH structure, and this is used to deduce some restrictions on such immersions.

Co-creación de semiconductores

El mercado de los semiconductores está saturado de productos similares y de distribuidores con una propuesta de servicios similar. Los procesos de Co-Creación en los que el cliente colabora en la definición y desarrollo del producto y proporciona información sobre su utilidad, prestaciones y valor percibido, con el resultado de un producto que soluciona sus necesidades reales, se están convirtiendo en un paso adelante en la diferenciación y expansión de la cadena de valor. El proceso de diseño y fabricación de semiconductores es bastante complejo, requiere inversiones cada vez mayores y demanda soluciones completas. Se requiere un ecosistema que soporte el desarrollo de los equipos electrónicos basados en dichos semiconductores. La facilidad para el diálogo y compartir información que proporciona internet, las herramientas basadas en web 2.0 y los servicios y aplicaciones en la nube; favorecen la generación de ideas, el desarrollo y evaluación de productos y posibilita la interacción entre diversos co-creadores. Para iniciar un proceso de co-creación se requiere métodos y herramientas adecuados para interactuar con los participantes e intercambiar experiencias, procesos para integrar la co-creación dentro de la operativa de la empresa, y desarrollar una organización y cultura que soporten y fomenten dicho proceso. Entre los métodos más efectivos están la Netnografía que estudia las conversaciones de las comunidades en internet; colaboración con usuarios pioneros que van por delante del Mercado y esperan un gran beneficio de la satisfacción de sus necesidades o deseos; los estudios de innovación que permiten al usuario definir y a menudo crear su propia solución y la externalización a la multitud, que mediante una convocatoria abierta plantea a la comunidad retos a resolver a cambio de algún tipo de recompensa. La especialización de empresas subcontratistas en el desarrollo y fabricación de semiconductores; facilita la innovación abierta colaborando con diversas entidades en las diversas fases del desarrollo del semiconductor y su ecosistema. La co-creación se emplea actualmente en el sector de los semiconductores para detectar ideas de diseños y aplicaciones, a menudo mediante concursos de innovación. El servicio de soporte técnico y la evaluación de los semiconductores con frecuencia es fruto de la colaboración entre los miembros de la comunidad fomentada y soportada por los fabricantes del producto. Con el programa EBVchips se posibilita el acceso a empresas pequeñas y medianas a la co-creación de semiconductores con los fabricantes en un proceso coordinado y patrocinado por el distribuidor EBV. Los semiconductores configurables como las FPGAs constituyen otro ejemplo de co-creación mediante el cual el fabricante proporciona el circuito integrado y el entorno de desarrollo y los clientes crean el producto final definiendo sus características y funcionalidades. Este proceso se enriquece con bloques funcionales de diseño, IP-cores, que a menudo son creados por la comunidad de usuarios. ABSTRACT. The semiconductor market is saturated of similar products and distributors with a similar proposal for services. The processes of co-creation in which the customer collaborates in the definition and development of the product and provides information about its utility, performance and perceived value, resulting in a product that solves their real needs, are becoming a step forward in the differentiation and expansion of the value chain. The design and semiconductor manufacturing process is quite complex, requires increasingly higher investments and demands complete solutions. It requires an ecosystem that supports the development of electronic equipments based on such semiconductors. The ease of dialogue and sharing information that provides internet, web 2.0-based tools and services and applications in the cloud; favor the generation of ideas, the development and evaluation of products and allows the interaction between various co-creators. To start a process of co-creation adequate methods and tools are required to interact with the participants and exchange experiences, processes to integrate the co-creation within the operations of the company, and developing an organization and culture that support and promote such process. Among the most effective methods are the Netnography that studies the conversations of the communities on the internet; collaboration with Lead Users who are ahead of the market and expect a great benefit from the satisfaction of their needs or desires; Innovation studies that allow the user to define and often create their own solution and Crowdsourcing, an open call to the community to solve challenges in exchange for some kind of reward. The specialization of subcontractors in the development and manufacture of semiconductors; facilitates open innovation in the context of collaboration with different entities working in the different phases of the development of the semiconductor and its ecosystem. Co-creation is used currently in the semiconductor sector to detect ideas of designs and applications, often through innovation’s contests. Technical support and evaluation of semiconductors frequently is the result of collaboration between members of the community fostered and supported by the manufacturers of the product. The EBVchips program provides access to small and medium-sized companies to the co-creation of semiconductors with manufacturers in a process coordinated and sponsored by the Distributor EBV. Configurable semiconductors like FPGAs are another example of co-creation whereby the manufacturer provides the integrated circuit and the development environment and customers create the final product by defining their features and functionality. This process is enriched with IP-cores, designs blocks that are often created by the user community.

Alt a 1 from Alternaria interacts with PR5 thaumatin-like proteins

Alt a 1 is a protein found in Alternaria alternata spores related to virulence and pathogenicity and considered to be responsible for chronic asthma in children. We found that spores of Alternaria inoculated on the outer surface of kiwifruits did not develop hyphae. Nevertheless, the expression of Alt a 1 gene was upregulated, and the protein was detected in the pulp where it co-localized with kiwi PR5. Pull-down assays demonstrated experimentally that the two proteins interact in such a way that Alt a 1 inhibits the enzymatic activity of PR5. These results are relevant not only for plant defense, but also for human health as patients with chronic asthma could suffer from an allergic reaction when they eat fruit contaminated with Alternaria.

Subunit cleavage of mosquito pro-vitellogenin by a subtilisin-like convertase.

The eukaryotic convertase family plays an important role in posttranslational proteolytic processing and activation of many pro- and polypeptides that have at their cleavage sites the paired basic motif, RX(K/R)R. Recent studies have revealed that the cleavage site of insect pro-vitellogenins (pro-Vg) also contains this motif. To identify and characterize the insect pro-Vg processing enzyme, Vg convertase (VC), its cDNA was cloned from a vitellogenic female fat body cDNA library of the mosquito, Aedes aegypti. The 3735-bp-long VC cDNA has an open reading frame encoding a 115-kDa protein. In vitro transcription/translation of VC cDNA revealed that this 115-kDa protein becomes 140 kDa after co- and posttranslational modifications. The VC deduced amino acid sequence has high similarity to and a domain structure characteristic of furin-like convertases. Northern blot analysis showed that a single 4.2-kb transcript was expressed in the fat body during the first 18 hr of the Vg synthetic period. Coexpression of VC cDNA with mosquito Vg cDNA resulted in correct cleavage of pro-Vg. Thus, this newly identified convertase is, indeed, a functional fat body-specific enzyme for pro-Vg cleavage.

Louise, or, The beauty of integrity : a story for the young ; to which are added The mother's grave and No place like home.

Mode of access: Internet.

Our modern navy; illustrations of battleships, cruisers, monitors, gunboats and torpedo boats with full detail description of the material, displacement, dimensions, horse-power, speed, armor, batteries, caliber and firing capacity of the guns, and the cost of the principal vessels in the United States navy and like details of the great navies of the world.

Mode of access: Internet.

What boys and girls like to read.

Mode of access: Internet.

Poly(beta-hydroxybutyrate-co-beta-hydroxyvalerate) supports in vitro osteogenesis

Studies have demonstrated that polymeric biomaterials have the potential to support osteoblast growth and development for bone tissue repair. Poly( beta- hydroxybutyrate- co- beta- hydroxyvalerate) ( PHBV), a bioabsorbable, biocompatible polyhydroxy acid polymer, is an excellent candidate that, as yet, has not been extensively investigated for this purpose. As such, we examined the attachment characteristics, self- renewal capacity, and osteogenic potential of osteoblast- like cells ( MC3T3- E1 S14) when cultured on PHBV films compared with tissue culture polystyrene ( TCP). Cells were assayed over 2 weeks and examined for changes in morphology, attachment, number and proliferation status, alkaline phosphatase ( ALP) activity, calcium accumulation, nodule formation, and the expression of osteogenic genes. We found that these spindle- shaped MC3T3- E1 S14 cells made cell - cell and cell - substrate contact. Time- dependent cell attachment was shown to be accelerated on PHBV compared with collagen and laminin, but delayed compared with TCP and fibronectin. Cell number and the expression of ALP, osteopontin, and pro- collagen alpha 1( I) mRNA were comparable for cells grown on PHBV and TCP, with all these markers increasing over time. This demonstrates the ability of PHBV to support osteoblast cell function. However, a lag was observed for cells on PHBV in comparison with those on TCP for proliferation, ALP activity, and cbfa- 1 mRNA expression. In addition, we observed a reduction in total calcium accumulation, nodule formation, and osteocalcin mRNA expression. It is possible that this cellular response is a consequence of the contrasting surface properties of PHBV and TCP. The PHBV substrate used was rougher and more hydrophobic than TCP. Although further substrate analysis is required, we conclude that this polymer is a suitable candidate for the continued development as a biomaterial for bone tissue engineering.