Reference gene selection and RNA preservation protocol in the cat flea, Ctenocephalides felis, for gene expression studies

This work was supported by a Knowledge Transfer Network BBSRC Industrial Case (#414 BB/L502467/1) studentship in association Zoetis Inc.

Phenological sensitivity to climate across taxa and trophic levels

Peer reviewed

Substitutions in the glycogenin-1 gene are associated with the evolution of endothermy in sharks and tunas

We thank Sean Tracey and Jaime McAllister for supplying albacore and southern bluefin tuna samples, Eva Giacomello for collecting the skipjack tuna sample, Elena Sarropoulou for providing the Atlantic bonito assembly, Helen Hipperson for assistance in the lab, Barbara Block and Ziheng Yang for advice, the editors and reviewers for comments, and the Leverhulme Trust and BBSRC for funding

Sortase as a Tool in Biotechnology and Medicine

We have harnessed two reactions catalyzed by the enzyme sortase A and applied them to generate new methods for the purification and site-selective modification of recombinant protein therapeutics.

We utilized native peptide ligation —a well-known function of sortase A— to attach a small molecule drug specifically to the carboxy-terminus of a recombinant protein. By combining this reaction with the unique phase behavior of elastin-like polypeptides, we developed a protocol that produces homogenously-labeled protein-small molecule conjugates using only centrifugation. The same reaction can be used to produce unmodified therapeutic proteins simply by substituting a single reactant. The isolated proteins or protein-small molecule conjugates do not have any exogenous purification tags, eliminating the potential influence of these tags on bioactivity. Because both unmodified and modified proteins are produced by a general process that is the same for any protein of interest and does not require any chromatography, the time, effort, and cost associated with protein purification and modification is greatly reduced.

We also developed an innovative and unique method that attaches a tunable number of drug molecules to any recombinant protein of interest in a site-specific manner. Although the ability of sortase A to carry out native peptide ligation is widely used, we demonstrated that Sortase A is also capable of attaching small molecules to proteins through an isopeptide bond at lysine side chains within a unique amino acid sequence. This reaction —isopeptide ligation— is a new site-specific conjugation method that is orthogonal to all available protein-small conjugation technologies and is the first site-specific conjugation method that attaches the payload to lysine residues. We show that isopeptide ligation can be applied broadly to peptides, proteins, and antibodies using a variety of small molecule cargoes to efficiently generate stable conjugates. We thoroughly assessed the site-selectivity of this reaction using a variety of analytical methods and showed that in many cases the reaction is site-specific for lysines in flexible, disordered regions of the substrate proteins. Finally, we showed that isopeptide ligation can be used to create clinically-relevant antibody-drug conjugates that have potent cytotoxicity towards cancerous cells

Characterization of in vitro effects of patulin on intestinal epithelial and immune cells

The intestinal mucosa is the first biological barrier encountered by natural toxins, and could possibly be exposed to high amounts of dietary mycotoxins. Patulin (PAT), a mycotoxin produced by Penicillium spp. during fruit spoilage, is one of the best known enteropathogenic mycotoxins able to alter functions of the intestine (Maresca et al., 2008). This study evaluated the effects of PAT on barrier function of the gut mucosa utilizing the intestinal epithelial cell model Caco-2, and scrutinized immunomodulatory effects using human peripheral blood mononuclear cells (PBMC) and human blood monocyte-derived dendritic cells (moDCs) as test systems. PAT exposure reduced Caco-2 cell viability at concentrations above 12 mM. As expected, the integrity of a polarized Caco-2 monolayer was affected by PAT exposure, as demonstrated by a decrease in TER values, becoming more pronounced at 50 mM. No effects were detected on the expression levels of the tight junction proteins occludin, claudin-1 and claudin-3 at 50 mM. However, the expression of zonula occludens-1 (ZO-1) and myosin light chain 2 (MLC2) declined. Also, levels of phospho-MLC2 (p-MLC2) increased after 24 h of exposure to 50 mM of PAT. T cell proliferation was highly sensitive to PAT with major effects for concentrations above 10 nM of PAT. The same conditions did not affect the maturation of moDC. PAT causes a reduction in Caco-2 barrier function mainly by perturbation of ZO-1 levels and the phosphorylation of MLC. Low doses of PAT strongly inhibited T cell proliferation induced by a polyclonal activator, but had no effect on the maturation of moDC. These results provide new information that strengthens the concept that the epithelium and immune cells of the intestinal mucosa are important targets for the toxic effects of food contaminants like mycotoxins

Challenges in imaging and predictive modeling of rhizosphere processes

Background Plant-soil interaction is central to human food production and ecosystem function. Thus, it is essential to not only understand, but also to develop predictive mathematical models which can be used to assess how climate and soil management practices will affect these interactions. Scope In this paper we review the current developments in structural and chemical imaging of rhizosphere processes within the context of multiscale mathematical image based modeling. We outline areas that need more research and areas which would benefit from more detailed understanding. Conclusions We conclude that the combination of structural and chemical imaging with modeling is an incredibly powerful tool which is fundamental for understanding how plant roots interact with soil. We emphasize the need for more researchers to be attracted to this area that is so fertile for future discoveries. Finally, model building must go hand in hand with experiments. In particular, there is a real need to integrate rhizosphere structural and chemical imaging with modeling for better understanding of the rhizosphere processes leading to models which explicitly account for pore scale processes.

Biotechnology and crop protection in Brazil.

The process of modernization of Brazilian agriculture aimed at increasing the productivity in response to the high demand for agricultural products in the world market and it was based on the intensive use of inputs such as agrochemicals, intense mechanisation and breeding of new varieties. Among these, pesticides were incorporated in almost all production systems. Over reliance on pesticide use has produced many negative effects on both biotic and abiotic components of the environment, generating chemical contamination of soil and water, decrease in biological diversity of agroecosystems, disruption of natural cycles, pest resistance, intoxication of growers, among others. The consumption of pesticides in Brazil was 151.8 thousand tonnes in 1989, and today the country is the fifth world market of these products. The use of pesticides increased from 16 thousand tonnes (a.i.) in 1964 to 60.2 thousand tonnes in 1991, while the area planted to crops grew from 28.4 to 50.0 million ha in the same period. This means an increase of 276.2% in consumption of pesticides compared to an increase of 76% in planted area. Even with this large increase in the use of pesticides, the losses caused by pests have not been significantly reduced, and the net gain in crop productivity has been low. On the other hand, problems with food contamination, environmental degradation of growers have considerably mounted. It is possible to define two classes of crops regarding intense use of pesticides. One is represented by those crops that occupy large areas, and therefore contribute to a large amont of pesticides used for pest control in a country basis. The other class comprises crops that require large amounts of pesticides per unit of area, but not necessarily represent large amounts of pesticides used coutry-wide. Based on the classes proposed, citrus, soybean and sugarcane stand as crops with a nationally great consumption of pesticides, while tomato, potato and citrus are important as intensive users of pesticides. In this paper the biotechnologies in use, the biotechnologies in advanced stages of development, the main constraints to the development and use of biotechnlology and the impact of pesticed on the environment are discussed.

A multilevel study of area socio-economic status and food purchasing behaviour

Objective: This study examined the association between area socioeconomic status (SES) and food purchasing behaviour.----- Setting: Melbourne city, Australia, 2003.----- Participants: Residents of 2,564 households located in 50 small areas.----- Design: Data were collected by mail survey (64.2% response rate). Area SES was indicated by the proportion of households in each area earning less than Aus$400 per week, and individual-level socioeconomic position was measured using education, occupation, and household income. Food purchasing was measured on the basis of compliance with dietary guideline recommendations (for grocery foods) and variety of fruit and vegetable purchase. Multilevel regression examined the association between area SES and food purchase after adjustment for individual-level demographic (age, sex, household composition) and socioeconomic factors.----- Results: Residents of low SES areas were significantly less likely than their counterparts in advantaged areas to purchase grocery foods that were high in fibre and low in fat, salt, and sugar; and they purchased a smaller variety of fruits. There was no evidence of an association between area SES and vegetable variety.----- Conclusions In Melbourne, area SES was associated with some food purchasing behaviours independent of individual-level factors, suggesting that areas in this city may be differentiated on the basis of food availability, accessibility, and affordability, making the purchase of some types of foods more difficult in disadvantaged areas.

Best practices in prediction for decision-making : lessons from the atmospheric and earth sciences

Predictions that result from scientific research hold great appeal for decision-makers who are grappling with complex and controversial environmental issues, by promising to enhance their ability to determine a need for and outcomes of alternative decisions. A problem exists in that decision-makers and scientists in the public and private sectors solicit, produce, and use such predictions with little understanding of their accuracy or utility, and often without systematic evaluation or mechanisms of accountability. In order to contribute to a more effective role for ecological science in support of decision-making, this paper discusses three ``best practices'' for quantitative ecosystem modeling and prediction gleaned from research on modeling, prediction, and decision-making in the atmospheric and earth sciences. The lessons are distilled from a series of case studies and placed into the specific context of examples from ecological science.

Enhancing the teaching of family and consumer sciences : the role of graphic organisers

In a world of constant and rapid change there are greater demands placed on learners to not only gain content knowledge, but also to develop learning skills and to adopt new strategies that will enable them to produce better and faster learning outcomes. Especially in internationally advancing nations like Kuwait this will be a major challenge of the future. This literature review examines theoretical frameworks that enhance Kuwaiti teachers’ knowledge and skill to adopt culturally relevant reform practices across a number of disciplines and provide guidance in an exploration and use of newer pedagogical tools like graphic organisers. It analyses the effects of graphic organisers on higher order learning and evaluates how they can effect professional development and pedagogical change in Kuwait.

Democracy, Crime and Justice. Annals of the American Academy of Political and Social Sciences

As a growing number of nations embark on a path to democracy, criminologists have become increasingly interested and engaged in the challenges, concerns, and questions connecting democracy with both crime and criminal justice. Rising levels of violence and street crime, white collar crime and corruption both in countries where democracy is securely in place and where it is struggling, have fuelled a deepening skepticism as to the capacity of democracy to deliver on its promise of security and justice for all citizens. What role does crime and criminal justice play in the future of democracy and for democratic political development on a global level? The editors of this special volume of The Annals realized the importance of collecting research from a broad spectrum of countries and covering a range of problems that affect citizens, politicians, and criminal justice officials. The articles here represent a solid balance between mature democracies like the U.S. and U.K. as well as emerging democracies around the globe – specifically in Latin America, Africa and Eastern Europe. They are based on large and small cross-national samples, regional comparisons, and case studies. Each contribution addresses a seminal question for the future of democratic political development across the globe. What is the role of criminal justice in the process of building democracy and instilling confidence in its institutions? Is there a role for unions in democratizing police forces? What is the impact of widespread disenfranchisement of felons on democratic citizenship and the life of democratic institutions? Under what circumstances do mature democracies adopt punitive sentencing regimes? Addressing sensitive topics such as relations between police and the Muslim communities of Western Europe in the wake of terrorist attacks, this volume also sheds light on the effects of terrorism on mature democracies under increasing pressure to provide security for their citizens. By taking a broad vantage point, this collection of research delves into complex topics such as the relationship between the process of democratization and violent crime waves; the impact of rising crime rates on newly established as well as secure democracies; how crime may endanger the transition to democracy; and how existing practices of criminal justice in mature democracies affect their core values and institutions. The collection of these insightful articles not only begins to fill a gap in criminological research but also addresses issues of critical interest to political scientists as well as other social and behavioral scientists and scholars. Taking a fresh approach to the intersection of crime, criminal justice, and democracy, this volume of The Annals is a must-read for criminologists and political scientists and provides a solid foundation for further interdisciplinary research.