Alboserpin, a Factor Xa Inhibitor from the Mosquito Vector of Yellow Fever, Binds Heparin and Membrane Phospholipids and Exhibits Antithrombotic Activity

The molecular mechanism of factor Xa (FXa) inhibition by Alboserpin, the major salivary gland anticoagulant from the mosquito and yellow fever vector Aedes albopictus, has been characterized. cDNA of Alboserpin predicts a 45-kDa protein that belongs to the serpin family of protease inhibitors. Recombinant Alboserpin displays stoichiometric, competitive, reversible and tight binding to FXa (picomolar range). Binding is highly specific and is not detectable for FX, catalytic site-blocked FXa, thrombin, and 12 other enzymes. Alboserpin displays high affinity binding to heparin (K(D) similar to 20 nM), but no change in FXa inhibition was observed in the presence of the cofactor, implying that bridging mechanisms did not take place. Notably, Alboserpin was also found to interact with phosphatidylcholine and phosphatidylethanolamine but not with phosphatidylserine. Further, annexin V (in the absence of Ca(2+)) or heparin outcompetes Alboserpin for binding to phospholipid vesicles, suggesting a common binding site. Consistent with its activity, Alboserpin blocks prothrombinase activity and increases both prothrombin time and activated partial thromboplastin time in vitro or ex vivo. Furthermore, Alboserpin prevents thrombus formation provoked by ferric chloride injury of the carotid artery and increases bleeding in a dose-dependent manner. Alboserpin emerges as an atypical serpin that targets FXa and displays unique phospholipid specificity. It conceivably uses heparin and phosphatidylcholine/phosphatidylethanolamine as anchors to increase protein localization and effective concentration at sites of injury, cell activation, or inflammation.

Laterally transferred genomic islands in Xanthomonadales related to pathogenicity and primary metabolism

Lateral gene transfer (LGT) is considered as one of the drivers in bacterial genome evolution, usually associated with increased fitness and/or changes in behavior, especially if one considers pathogenic vs. non-pathogenic bacterial groups. The genomes of two phytopathogens, Xanthomonas campestris pv. campestris and Xanthomonas axonopodis pv. citri, were previously inspected for genome islands originating from LGT events, and, in this work, potentially early and late LGT events were identified according to their altered nucleotide composition. The biological role of the islands was also assessed, and pathogenicity, virulence and secondary metabolism pathways were functions highly represented, especially in islands that were found to be recently transferred. However, old islands are composed of a high proportion of genes related to cell primary metabolic functions. These old islands, normally undetected by traditional atypical composition analysis, but confirmed as product of LGT by atypical phylogenetic reconstruction, reveal the role of LGT events by replacing core metabolic genes normally inherited by vertical processes.

ADCK3, an ancestral kinase, is mutated in a form of recessive ataxia associated with coenzyme Q(10) deficiency

Muscle coenzyme Q(10) (CoQ(10) or ubiquinone) deficiency has been identified in more than 20 patients with presumed autosomal-recessive ataxia. However, mutations in genes required for CoQ(10) biosynthetic pathway have been identified only in patients with infantile-onset multisystemic diseases or isolated nephropathy. Our SNP-based genome-wide scan in a large consanguineous family revealed a locus for autosomal-recessive ataxia at chromosome 1q41. The causative mutation is a homozygous splice-site mutation in the aarF-domain-containing kinase 3 gene (ADCK3). Five additional mutations in ADCK3 were found in three patients with sporadic ataxia, including one known to have CoQ(10) deficiency in muscle. All of the patients have childhood-onset cerebellar ataxia with slow progression, and three of six have mildly elevated lactate levels. ADCK3 is a mitochondrial protein homologous to the yeast COQ8 and the bacterial UbiB proteins, which are required for CoQ biosynthesis. Three out of four patients tested showed a low endogenous pool of CoQ(10) in their fibroblasts or lymphoblasts, and two out of three patients showed impaired ubiquinone synthesis, strongly suggesting that ADCK3 is also involved in CoQ(10) biosynthesis. The deleterious nature of the three identified missense changes was confirmed by the introduction of them at the corresponding positions of the yeast COQ8 gene. Finally, a phylogenetic analysis shows that ADCK3 belongs to the family of atypical kinases, which includes phosphomositide and choline kinases, suggesting that ADCK3 plays an indirect regulatory role in ubiquinone biosynthesis possibly as part of a feedback loop that regulates ATP production.

Replacement of the arginine biosynthesis operon in Xanthomonadales by ateral gene transfer

The role of lateral gene transfer (LGT) in prokaryotes has been shown to rapidly change the genome content, providing new gene tools for environmental adaptation. Features related to pathogenesis and resistance to strong selective conditions have been widely shown to be products of gene transfer between bacteria. The genomes of the gamma-proteobacteria from the genus Xanthomonas, composed mainly of phytopathogens, have potential genomic islands that may represent imprints of such evolutionary processes. In this work, the evolution of genes involved in the pathway responsible for arginine biosynthesis in Xanthomonadales was investigated, and several lines of evidence point to the foreign origin of the arg genes clustered within a potential operon. Their presence inside a potential genomic island, bordered by a tRNA gene, the unusual ranking of sequence similarity, and the atypical phylogenies indicate that the metabolic pathway for arginine biosynthesis was acquired through LGT in the Xanthomonadales group. Moreover, although homologues were also found in Bacteroidetes (Flavobacteria group), for many of the genes analyzed close homologues are detected in different life domains (Eukarya and Archaea), indicating that the source of these arg genes may have been outside the Bacteria clade. The possibility of replacement of a complete primary metabolic pathway by LGT events supports the selfish operon hypothesis and may occur only under very special environmental conditions. Such rare events reveal part of the history of these interesting mosaic Xanthomonadales genomes, disclosing the importance of gene transfer modifying primary metabolism pathways and extending the scenario for bacterial genome evolution.

Mycoflora and fumonisin contamination in Brazilian sorghum from sowing to harvest

BACKGROUND: The aim of this study was to characterise the mycoflora and the presence of fumonisin in sorghum grains, correlating the results with the environment and abiotic factors. RESULTS: Fifty samples (five collections of ten samples each) of sorghum were analysed. All samples were found to be contaminated with fungi, with higher frequencies of Cladosporium spp. (61.8%) and Helminthosporium spp. (33.4%). Fusarium verticillioides was isolated from 15.1% of the samples, with 38% of them being contaminated with fumonisin B(1) (FB(1)) at levels ranging from 50 to 368.78 ng g(-1). Regarding abiotic factors, temperature, water activity and rainfall showed a positive correlation with the frequency of F. verticillioides and FB(1) production. There was a significant positive correlation between relative air humidity and FB(1) production. The results obtained from sexual crosses between standard F mating tester strains and the isolated strains confirmed that the strains isolated were F. verticillioides. CONCLUSION: It can be concluded that the decrease in F. verticillioides and fumonisin contamination occurred owing to atypical climatic factors during the period of sorghum cultivation, when there was any occurrence of rain and the level of water activity of grains did not reach 0.58. (C) 2010 Society of Chemical Industry

Domestic Cats Constitute a Natural Reservoir of Human Enteropathogenic Escherichia coli Types

Feces of 70 diarrhoeic and 230 non-diarrhoeic domestic cats from Sao Paulo, Brazil were investigated for enteropathogenic (EPEC), enterohaemorrhagic (EHEC) and enterotoxigenic (ETEC) Escherichia coli types. While ETEC and EHEC strains were not found, 15 EPEC strains were isolated from 14 cats, of which 13 were non-diarrhoeic, and one diarrhoeic. None of 15 EPEC strains carried the bfpA gene or the EPEC adherence factor plasmid, indicating atypical EPEC types. The EPEC strains were heterogeneous with regard to intimin types, such as eae-theta (three strains), eae-kappa (n = 3), eae-alpha 1 (n = 2), eae-iota (n = 2), one eae-alpha 2, eae-beta 1 and eae-eta each, and two were not typeable. The majority of the EPEC isolates adhered to HEp-2 cells in a localized adherence-like pattern and were positive for fluorescence actin staining. The EPEC strains belonged to 12 different serotypes, including O111:H25 and O125:H6, which are known to be pathogens in humans. Multi locus sequence typing revealed a close genetic similarity between the O111:H25 and O125:H6 strains from cats, dogs and humans. Our results show that domestic cats are colonized by EPEC, including serotypes previously described as human pathogens. As these EPEC strains are also isolated from humans, a cycle of mutual infection by EPEC between cats and its households cannot be ruled out, though the transmission dynamics among the reservoirs are not yet understood clearly.

Biochemical characterization of serine acetyltransferase and cysteine desulfhydrase from Leishmania major

Cysteine metabolism exhibits atypical features in Leishmania parasites. The nucleotide sequence annotated as LmjF32.2640 encodes a cysteine desulfhydrase, which specifically catalyzes the breakdown of cysteine into pyruvate, NH(3) and H(2)S. Like in other pathogens, this capacity might be associated with regulatory mechanisms to control the intracellular level of cysteine, a highly toxic albeit essential amino acid, in addition to generate pyruvate for energy production. Besides, our results provide the first insight into the biochemical properties of Leishmania major serine acetyltransferase (SAT), which is likely involved in the two routes for de novo synthesis of cysteine in this pathogen. When compared with other members of SAT family, the N-terminal region of L. major homologue is uniquely extended, and seems to be essential for proper protein folding. Furthermore, unlike plant and bacterial enzymes, the carboxy-terminal-C(10) sequence stretch of L major SAT appears not to be implicated in forming a tight bi-enzyme complex with cysteine synthase. (C) 2010 Elsevier B.V. All rights reserved.

DIAGNOSTIC TECHNIQUES OF LOCAL INFLUENCE IN SPATIAL ANALYSIS OF SOYBEAN YIELD

Modeling of spatial dependence structure, concerning geoestatistics approach, is an indispensable tool for fixing parameters that define this structure, applied on interpolation of values in places that are not sampled, by kriging techniques. However, the estimation of parameters can be greatly affected by the presence of atypical observations on sampled data. Thus, this trial aimed at using diagnostics techniques of local influence in spatial linear Gaussians models, applied at geoestatistics in order to evaluate sensitivity of maximum likelihood estimators and restrict maximum likelihood to small perturbations in these data. So, studies with simulated and experimental data were performed. Those results, obtained from the study of real data, allowed us to conclude that the presence of atypical values among the sampled data can have a strong influence on thematic maps, changing, therefore, the spatial dependence. The application of diagnostics techniques of local influence should be part of any geoestatistic analysis, ensuring that the information from thematic maps has better quality and can be used with greater security by farmers.

A GUI for online presentation of steel and steelmaking ladle temperature data and simulation.

Continuous casting is a casting process that produces steel slabs in a continuous manner with steel being poured at the top of the caster and a steel strand emerging from the mould below. Molten steel is transferred from the AOD converter to the caster using a ladle. The ladle is designed to be strong and insulated. Complete insulation is never achieved. Some of the heat is lost to the refractories by convection and conduction. Heat losses by radiation also occur. It is important to know the temperature of the melt during the process. For this reason, an online model was previously developed to simulate the steel and ladle wall temperatures during the ladle cycle. The model was developed as an ODE based model using grey box modeling technique. The model’s performance was acceptable and needed to be presented in a user friendly way. The aim of this thesis work was basically to design a GUI that presents steel and ladle wall temperatures calculated by the model and also allow the user to make adjustments to the model. This thesis work also discusses the sensitivity analysis of different parameters involved and their effects on different temperature estimations.

Sports and mediatization : a case of women´s ski-jumping

Ski jumping was the only sport included in the Winter Olympics, where women were not allowed to take part until the first week of April 2011, when IOC decided to let them in to the Olympic family, a major victory for women´s ski-jumping. Since 2004 the Norwegian media had women ski jumping as one important topic in their media coverage. The third of March 2010 the new Holmenkollen ski-jumping hill was inaugurated. It had been rebuilt for the World Championships 2011. The first jump on the hill was surrounded by an intense debate. Annette Sagen was decided to be the first jumper on the hill after a poll on the social media channel Facebook that gave Annette Sagen over 40 000 votes.  However Tuesday the second of March Björn Einar Romören, a world cup jumper, did the first jump during a training session. This act started a significant chain of events. The biggest Norwegian web – magazines Verldens Gang and Aftenposten made this the head story and they invited the readers to interaction. Within 24 hours 11 000 comments were written, most of them in favor of Sagen and against Romören´s behavior. The Norwegian Ski Association excluded Romören from two World Cup competitions. What did happen during these days in the media?  I want to present the activities, which went on in two web-magazines, and analyze the articles written on the topic. Main issues are to show what an impact a social media network had on the sport in this specific case and how different actors appeared in the media and discuss if Romörens exclusion was a result of the mediatization.

Effect of exercise on protein kinase C activity and localization in human skeletal muscle

To investigate the effect of exercise on protein kinase C (PKC) activity and localization in human skeletal muscle, eight healthy men performed cycle  ergometer exercise for 40 min at 76±1% the peak pulmonary O₂ uptake (V_O2peak), with muscle samples obtained at rest and after 5 and 40 min of exercise. PKC expression, phosphorylation and activities were examined by immunoblotting and in vitro kinase assays of fractionated and whole tissue preparations. In response to exercise, total PKC activity was slightly higher at 40 min in an enriched membrane fraction, and using a pSer-PKC-substrate motif antibody it was revealed that exercise increased the serine phosphorylation of a ∼50 kDa protein. There were no changes in conventional PKC (cPKC) or PKCθ activities; however, atypical PKC (aPKC) activity was ∼70% higher at 5 and 40 min, and aPKC expression and Thr^410/403 phosphorylation were unaltered by exercise. There were no effects of exercise on the abundance of PKCα, PKCδ, PKCθ and aPKC within cytosolic or enriched membrane fractions of skeletal muscle. These data indicate that aPKC, but not cPKC or PKCθ, are activated by exercise in contracting muscle suggesting a potential role for aPKC in the regulation of skeletal muscle function and metabolism during exercise in humans.

More? You want more?..of course I do! Transsexualism and birth certificates-changing records or attitudes?

In 2004, the Victorian Government enacted legislation allowing people treated for transsexualism to correct the record of their sex on the Register of Births, Deaths and Marriages and obtain a new certificate reflecting their contemporaneous circumstances. It was the last of all the States and Territories to do so.

The legislation gave effect to some important changes to the law and was generally couched in terms more sensitive than those already in place in the other jurisdictions. In the view of the author, however, its proponents failed to both understand the import of the expert medical evidence adduced in, and to implement the common law position enunciated by, the Family Court in Re Kevin (validity of marriage of transsexual) [2001] FamCA 1074 and subsequently confirmed on appeal two years later by the Full Court.

The author argues that, while a welcome improvement to the human rights record of successive Victorian Governments, the result is still a largely disappointing piece of legislation. Rather than being truly 'beneficial' to all who need security of their personal identities, it perpetuates some of the very worst discrimination directed at people with transsexualism and their families by continuing to portray them as psychologically deluded rather than physiologically atypical and denying a small number of them their rights on the basis of legal reasoning which is no longer regarded as tenable. She asserts the legislation serves as a clear demonstration that prejudices and misconceptions about transsexualism stilI abound and explains much more is needed if real human rights, acceptance and freedom from discrimination are to be eventually obtained by those affected by the phenomenon.

The challenge of reconciling working time demands and life transitions : a policy proposal

Labour markets, like all market institutions, exhibit structural and dynamic characteristics. Both the structural and dynamic characteristics of labour markets inevitably change and evolve over time in response to a host of exogenous and endogenous factors. In the case of the Australian labour market, structural changes are reflected in significant shifts in the industry and occupational composition of employment, the decline of full-time work and the concomitant rise in part-time and atypical forms of employment, demographic changes in the labour force, as well as changes in social and individual preferences. Dynamic shifts can be found in cyclical pattern of employment and wages growth, the growth in labour mobility, and transitions between various labour market states.
The starting point for this paper is that these structural and dynamic changes have given rise to an increase in the likelihood that individuals will experience a transition between various labour market states, and a greater diversity in the range of transitions they may experience over their working life. This acceleration in the rate of transition generates ‘transition costs’ for both employers and employees, as well the likelihood of mismatch between employer and employee working time preferences. As a consequence, existing labour market policy regimes, based on the traditional model of labour market participation over the life course may not provide adequate protection for most workers today.
Gunther Schmid (1998) and others have proposed institutional reforms which promote ‘transitional labour markets’. Transitional labour market institutions are those that allow individuals (and firms) to successfully adjust to critical events. While transitional labour market institutions may consist of traditional ‘active labour market policy’ mechanisms, Schmid and others have proposed a range of innovative policy responses which allow individuals (and firms) to adjust the intensity of their abour market participation over the life cycle. In this paper we use the general approach of advocated by the transitional labour market concept to do three things. First, we investigate the processes by which the nature of labour market transitions has changed over time. Second, we review the range of policy options available to government to smooth labour market dysfunctions associated with labour market transitions, with the objective of ensuring labour markets operate more efficiently and more equitably. Third, we focus on one possible way in which an existing labour market institution, Long Service Leave (LSL), could be reformed to make way for a more comprehensive transitional labour market institution in the form of a ‘working time bank’.

An exploration of the components of relational bonds to an arts festival brand

This research investigates the nature of the bonds that consumers form with a brand that provides highly uncertain outcomes, and is only available intermittently. The research model draws upon elements of Keller’s (2001) conceptualisation of brand resonance, and extends McAlexander, Kim, and Roberts’ (2003), and Muniz and O’Guinn’s (2001) brand community construct, testing these in an atypical service environment. Qualitative research suggested the need for a broader view of the bond formed in these circumstances, specifically one comprising measures of anticipation of usage, social attraction, commitment, loyalty, and trust. This paper reports on analysis undertaken to develop such a construct, which has been labelled “brand affinity”. Tests for discriminant validity suggest that the brand affinity construct is a distinct construct that can be used to measure consumer attitudes toward a highly uncertain, intermittently available product.

The case of "difficult" brands

Purpose – To examine an atypical member of the brand family, “difficult” brands, defined as brands that have intermittent availability, and have inherent performance risks, that, contrary to traditional marketing teachings, may have a positive influence on consumer evaluation and brand loyalty. Findings – Through an examination of the underlying social and psychological reasons that consumers might be attracted to difficult brands, an alternative approach to the marketing of these types of brands might be warranted. Practical implications – The conceptualisation of difficult brands provides marketers with a means to identify potential antecedents to brand bonding under atypical marketing conditions. Originality/value – Suggests how researchers, and marketing practitioners might go about understanding the nature of the bond that consumers form with these types of brands, and therefore, how they can take advantage of consumers' desire to develop a relationship with a difficult brand.