Antibody-induced internalization of viral glycoproteins in pseudorabies virus-infected monocytes and role of the cytoskeleton: a confocal study

Addition of pseudorabies virus (PrV)-specific polyclonal immunoglobulins to PrV-infected monocytes induces internalization of plasma membrane anchored viral glycoproteins. This process may interfere with antibody-dependent cell lysis and resembles the well-studied physiological endocytosis process. A confocal study was designed to investigate whether the major cellular components, involved in physiological endocytosis (clathrin, actin, dynein and microtubules), play a role in this virological internalization process. In order to visualize the interaction of endosomes, which contain the internalized viral glycoproteins, with clathrin, actin, dynein and microtubules, a double labeling of viral glycoproteins and different cellular proteins was performed. Porcine monocytes were inoculated with the PrV-strain 89V87 at a multiplicity of infection of 50 for 13h. After the addition of FITC-labeled porcine polyclonal PrV-specific antibodies, cells were fixed with para-formaldehyde at different time points and afterwards permeabilized. The different cellular components were visualized with monoclonal antibodies and a Texas Red-conjugate, with the exception of actin, which was stained with phalloidin-Texas Red. The cells were analyzed by confocal microscopy. A clear co-localization was observed between the viral glycoproteins and clathrin and dynein during the internalization process. The microtubules were in close contact with the internalized vesicles. For actin no co-localization could be observed. It can be stated that clathrin, dynein and microtubules, important components during physiological endocytosis, are also of importance during the antibody-induced internalization of viral glycoproteins.     

The UK Women's Cohort Study: comparison of vegetarians, fish-eaters and meat-eaters

BACKGROUND: This paper describes the development of the UK Women's Cohort Study and presents cohort baseline characteristics. METHODS: In total, 35,372 women, aged 35-69 years at recruitment, were selected to ensure a wide range of dietary intakes. Diet was assessed by a 217-item food-frequency questionnaire (FFQ). Detailed lifestyle information was collected by postal questionnaire. Vegetarians, fish-eaters and meat-eaters were compared. RESULTS: The cohort women are mainly white, well-educated, middle-class and married with children. They are health-conscious with only 11% current smokers and 58% taking dietary supplements. Twenty-eight per cent of subjects self-report as being vegetarian and 1% as vegan. However, only 18% are defined as 'vegetarian' from the FFQ. Fat provides 32% of energy; vitamin and mineral intakes are high, with a broad range of intakes. Meat-eaters are older, with a higher body mass index (BMI) and the lowest intakes of carbohydrate, fibre, vitamin C, folate, iron and calcium. Other fish-eaters are similar to vegetarians. Vegetarians have the lowest intakes of protein, fat and saturated fat. Oily fish-eaters have the lowest BMI; are the least likely to smoke or use full-fat milk; and are the most likely to use dietary supplements and consume the most fruit and vegetables. Oily fish-eaters have the highest total energy intake and vegetarians the lowest. Semi-skimmed milk, bread, potatoes, wine, bananas and muesli are important contributors to energy for all groups. CONCLUSION: A large cohort of middle-aged women has been created encompassing a wide range of different eating patterns, including diets currently of interest to research into protection against cancer and coronary heart disease. Participants will be followed up to study the effects of different food and nutrient intakes on long-term health outcomes.     

Oral infection with a Shiga toxin-negative Escherichia coli O157:H7 strain elicits humoral and cellular responses but does not protect sheep from colonisation with the homologous strain

We have previously shown that rectally inoculated sheep excrete Escherichia coli O157:H7 during weeks to months without developing a clear antibody response. However, antibodies against this bacterium were observed in naturally infected sheep, which most likely became orally infected. To understand this difference, sheep were orally inoculated with the same Shiga toxin-negative E. coli O157:H7 strain that was used for the rectal inoculation. A primary oral inoculation resulted in shedding of E. coli O157:H7 in the faeces and detection of antibody responses against intimin, EspA and EspB. The antibody titres waned as shedding decreased. A secondary inoculation resulted in longer shedding, even though a booster antibody response occurred. Cellular responses followed a similar pattern as the antibody levels, albeit with a lower secondary response. The presence of antigen-specific antibody-secreting cells indicates involvement of both a systemic response in the spleen and a local immune response in the terminal rectum. These results suggest that E. coli O157:H7 has to pass the small intestine to evoke antibody responses.     

Investigation of the prevalence of neurologic equine herpes virus type 1 (EHV-1) in a 23-year retrospective analysis (1984–2007)

A single nucleotide polymorphism in the equine herpesvirus 1 (EHV-1) DNA polymerase gene (ORF30 A₂₂₅₄ to G) has been associated with clinical signs of equine herpes myeloencephalopathy (EHM). The purpose of our study was to determine the odds ratio for this genetic marker and EHM using a panel of field isolates from North America collected over the past twenty-three years. EHV-1 isolates cultured at the Cornell University Animal Health Diagnostic Laboratory from 1984 to 2007 were retrieved along with their clinical histories. DNA was extracted from these EHV-1 cultures and allelic discrimination was performed using real-time PCR. The results were confirmed by sequencing of the target region in ORF30. PCR and sequencing were in 100% agreement and showed that 19 out of the 176 isolates had the ORF30 G₂₂₅₄ allele (11%), of which16 were EHM cases and 3 respiratory or abortion cases. The odds of having neurologic disease with the ORF30 G₂₂₅₄ genotype were computed as 162 times greater than those with the opposite allele ORF30 A₂₂₅₄ (95% confidence interval: 35–742). Despite this strong statistical significance, 24% (5/21) of horses with neurologic disease in our study population harbored the “non-neurologic” form of the allele (ORF30 A₂₂₅₄), suggesting that other factors may also contribute to the onset of EHM.     

Nutritional value of bread: Influence of processing,food interaction and consumer perception

The nearly ubiquitous consumption of cereals all over the world gives cereals an important position in international nutrition. Besides the high starch content as energy source, cereals provide dietary fibre, nutritious protein and lipids rich in essential fatty acids. Important micronutrients present in cereals are vitamins, especially many B vitamins, minerals, antioxidants and phytochemicals. In general, cereals provide important amounts of most nutrients. However, processing may decrease or increase the levels of the bioactive components in grains and also modify the bioavailability of these components. In addition, interactions between bread and companion foods have effects on the nutritional quality. The aim of this paper is to review the existing literature on the effects of processing techniques and interactions with other food components in a mixed meal on the nutritional quality of bread. Furthermore, research findings on the consumer perception of bread in Belgium are included. This information can help health professionals and policy-makers to give clear and targeted advice about the positive effects of bread in the human nutrition during consultations and information campaigns.     

The Evaluation of Aerial Application Systems1

There is no single measurement technique suitable for the evaluation of all aerial application systems, since the type of measurement required, and the technique used to obtain it, are dependent on the application method. In this paper, techniques used by the authors to assess atomiser performance, droplet dispersion, deposition, and aircraft swath width are described. The applicability of each technique to various application methods is described.