Mathilda J. M. Toussaint

The acute phase response of haptoglobin and serum amyloid A (SAA) in cattle undergoing experimental infection with bovine respiratory syncytial virus.

Abstract The ability of a pure virus infection to induce an acute phase protein response is of interest as viral infections are normally considered to be less efficient in inducing an acute phase protein response than bacterial infections. This was studied in a bovine model for infection with bovine respiratory syncytial virus (BRSV), analysing the induction of the two most dominant bovine acute phase proteins haptoglobin and serum amyloid A (SAA). Strong and reproducible acute phase responses were detected for both proteins, peaking at around 7–8 days after inoculation of BRSV, while no response was seen in mock-inoculated control animals. The serum concentrations reached for SAA and haptoglobin during the BRSV-induced acute phase response were generally the same or higher than previously reported for bacterial infections in calves. The magnitude and the duration of the haptoglobin response was found to correlate well with the severity of clinical signs (fever) and with the extent of lung consolidation while SAA responded most rapidly to infection.

Cognitive disturbances in old dogs suffering from the canine counterpart of Alzheimer's disease

In geriatric dogs, Alzheimer-like behavior is frequently observed. This behavior has been classified by several authors using questionnaires and a correlation has been described between cognitive dysfunctions and Alzheimer-like pathology. In the present study, cognitive performance was correlated with brain pathology for 30 dogs of varying ages. Within these animals, two age-matched groups of old dogs with and without behavioral changes were compared. The behavioral changes were analyzed and scored with questionnaires and necropsy was performed to rule out any other cause for changed behavior. Measurements, (immuno)-histochemical staining and fluorescence microscopy were used to detect cortex atrophy, amyloid, rest-products of oxidative damage, demyelination and accumulations of macrophages in the brains of these dogs. Spearman rank correlation coefficients (r) were calculated and adjusted according to Bonferonni. In the whole group (young to very old dogs), the age of the animal showed a significant correlation with various behavioral changes (r = 0.7 to 0.9, P < 0.01). The dementia score correlated significantly (r = 0.6 to 0.8, P < 0.01) with all the brain lesions studied, except one, i.e. demyelination (r = -0.4, P > 0.05). These results suggest that a questionnaire can be used to diagnose Alzheimer-like changes in canine practice. Oxidative damage on a cellular and a nuclear level plays an important role in behavior changes.

E2F8 is essential for polyploidization in mammalian cells

Polyploidization is observed in all mammalian species and is a characteristic feature of hepatocytes, but its molecular mechanism and biological significance are unknown. Hepatocyte polyploidization in rodents occurs through incomplete cytokinesis, starts after weaning and increases with age. Here, we show in mice that atypical E2F8 is induced after weaning and required for hepatocyte binucleation and polyploidization. A deficiency in E2f8 led to an increase in the expression level of E2F target genes promoting cytokinesis and thereby preventing polyploidization. In contrast, loss of E2f1 enhanced polyploidization and suppressed the polyploidization defect of hepatocytes deficient for atypical E2Fs. In addition, E2F8 and E2F1 were found on the same subset of target promoters. Contrary to the long-standing hypothesis that polyploidization indicates terminal differentiation and senescence, we show that prevention of polyploidization through inactivation of atypical E2Fs has, surprisingly, no impact on liver differentiation, zonation, metabolism and regeneration. Together, these results identify E2F8 as a repressor and E2F1 as an activator of a transcriptional network controlling polyploidization in mammalian cells.

An overview of swine influenza.

Summary Swine influenza is a highly infectious viral disease of pigs, causing considerable economic impact. The causative agent is known as a type A orthomyxovirus with a segmented RNA genome. Influenza type A virus is a highly contagious pathogen among a limited number of birds and mammals. The objective of this review is to summarize the current knowledge in swine influenza infection in pigs with emphasizing on epidemiology, pathogenesis, diagnostic techniques and control measures.

Optimal combinations of acute phase proteins for detecting infectious disease in pigs

The acute phase protein (APP) response is an early systemic sign of disease, detected as substantial changes in APP serum concentrations and most disease states involving inflammatory reactions give rise to APP responses. To obtain a detailed picture of the general utility of porcine APPs to detect any disease with an inflammatory component seven porcine APPs were analysed in serum sampled at regular intervals in six different experimental challenge groups of pigs, including three bacterial (Actinobacillus pleuropneumoniae, Streptococcus suis, Mycoplasma hyosynoviae), one parasitic (Toxoplasma gondii) and one viral (porcine respiratory and reproductive syndrome virus) infection and one aseptic inflammation. Immunochemical analyses of seven APPs, four positive (C-reactive protein (CRP), haptoglobin (Hp), pig major acute phase protein (pigMAP) and serum amyloid A (SAA)) and three negative (albumin, transthyretin, and apolipoprotein A1 (apoA1)) were performed in the more than 400 serum samples constituting the serum panel. This was followed by advanced statistical treatment of the data using a multi-step procedure which included defining cut-off values and calculating detection probabilities for single APPs and for APP combinations. Combinations of APPs allowed the detection of disease more sensitively than any individual APP and the best three-protein combinations were CRP, apoA1, pigMAP and CRP, apoA1, Hp, respectively, closely followed by the two-protein combinations CRP, pigMAP and apoA1, pigMAP, respectively. For the practical use of such combinations, methodology is described for establishing individual APP threshold values, above which, for any APP in the combination, ongoing infection/inflammation is indicated.

Serum amyloid A and transferrin in chicken. A preliminary investigation of using acute-phase variables to assess diseases in chickens.

The concentrations of serum amyloid A protein (SAA) and transferrin in blood samples from broilers in various stages of natural Staphylococcus aureus infection, from healthy counterparts, and from turpentine- or saline-injected pullets were measured using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting, and densitometry. SAA was not detected in healthy chickens but was detected in turpentine-injected pullets and in S. aureus-infected broilers. Relative percentages of transferrin in turpentine- and saline-injected pullets were not different. Broilers with a S. aureus infection had a two-fold higher transferrin level than did their unaffected counterparts. There was also a difference between the transferrin levels of healthy broilers and healthy pullets, which indicates that other variables, such as breed or breeding conditions, may influence the transferrin level. This preliminary study showed that SAA is an acute-phase protein and a more reliable variable for diagnosing lesions in chickens than transferrin.

Implication of clinical pathology in assessment of animal health and in animal production and meat inspection

Clinical, zootechnical and industrial developments of the last decades have led to new ideas on monitoring systems for animal production and meat inspection. Quality assessment systems, integrated monitoring, risk assessment concerning consumers health and monitoring for infectious animal diseases, are more relevant today than pathomorphological control of individual subjects. Published papers on investigations to assess slaughtered animals by blood variables of classical clinical pathology and by measuring acute phase reactants, are mentioned. Most papers deal with ruminants giving good perspectives for the acute phase proteins in that species. Only limited literature is available regarding acute phase proteins in swine; zinc and iron have been shown to be of little value for general health monitoring in swine.Preliminary studies on limited numbers of finishing pigs showed good prospects for the development of species specific assays for acute phase proteins for future practical use in the slaughterhouse. Isolation procedures for porcine haptoglobin (HP), serum amyloid A (SAA) and C reactive protein (CRP) are described. It is suggested that for monitoring individual animals, the signal of acute phase variable can be amplified considerably by applying an acute phase index (API) combining positive × negative reactants−1. Calculations on the original data from a former investigation on 233 nonhealthy and 21 control cattle gave excellent results for an API. Similar studies are in progress for swine.

Standardization of diagnostic assays for animal acute phase proteins.

This chapter discusses the standardization of diagnostic assays for animal acute phase proteins. Monitoring the plasma concentration of acute phase proteins in animals is now established as providing valuable diagnostic information in conditions involving inflammation, infection, or trauma. The investigation of these proteins is becoming even more widespread as commercial diagnostic kit producers provide veterinary laboratories with assay systems that have been validated for particular species. In addition, it has been suggested that the determination of the plasma concentration of these proteins is of particular value in farm animal production as an indicator of clinical and subclinical diseases and as an aid to meat inspection when carcasses of potential risk to public health can be identified. Development of assay methods to quantify acute phase proteins in animals has been pioneered by a number of laboratories on at least three continents. It is important for veterinary medicine that assays for each species should be as accurate as possible. International harmonization of calibration by use of a common reference preparation would be an important step toward this goal.

Deletion of the serotonin transporter in rats disturbs serotonin homeostasis without impairing liver regeneration.

The serotonin transporter is implicated in the uptake of the vasoconstrictor serotonin from the circulation into the platelets, where 95% of all blood serotonin is stored and released in response to vascular injury. In vivo studies indicated that platelet-derived serotonin mediates liver regeneration after partial hepatectomy. We have recently generated serotonin transporter knockout rats and demonstrated that their platelets were almost completely depleted of serotonin. Here we show that these rats exhibit impaired hemostasis and contain about 1-6% of wild-type serotonin levels in the blood. Despite the marked reduction of serotonin levels in blood and platelets, efficient liver regeneration and collagen-induced platelet aggregation occur in rats lacking the serotonin transporter. These results provide evidence that liver regeneration is not dependent on the release of serotonin from platelets. Our findings indicate that very low levels of serotonin in blood are sufficient for liver regeneration.

Pig-MAP, porcine acute phase proteins and standardisation of assays in Europe

The pattern of plasma proteins changes greatly following infection, inflammation or tissue injury. The concentration of some proteins referred to as acute phase proteins (APPs) significantly increases within hours or days after the onset of these processes. In contrast, the concentration of other proteins, such as albumin (Alb), called negative acute phase proteins, decreases. APPs have been extensively studied in man and rat, but less so in other species. In recent work, the APPs have been characterised in pigs in response to inflammation following turpentine injection. The concentrations of C reactive protein (CRP) and haptoglobin (Hp) increase 5–7 times 48 h after the injection. Porcine Alb, α-lipoprotein, fetuin and transferrin were negative APP. Finally, the concentration of α1-acid glycoprotein and α1-protease inhibitor (α1-antitrypsin) did not change significantly during the inflammation. In addition to CRP and Hp, a serum α2-globulin was observed to be the major acute phase (MAP) protein in pigs. Pig-MAP is a new mammalian plasma protein, which is the pig counterpart of a recently cloned human serum protein denominated PK-120 or IHRP. Pig-MAP shows promise as a prominent positive APP and has been shown to be a good marker of different pig pathologies.Recent collaboration in an EU-Concerted Action Project (AIR3-CT94-2255) has joined several laboratories involved in pig APP studies in Europe. These groups have been working on the approach that APP are markers for the presence of infectious, inflammatory and pathological lesions in animals. The ability to monitor the APP concentration in serum of pigs will improve the quality and safety of the meat produced as well as provide important diagnostic information for animal health and welfare. The serum concentration of APP are altered in several diseases supporting that APP can be used as markers of pig pathologies. Antibody-based techniques (for example, ELISA and immunonephelometric assays) have been and will be further developed for the selected porcine APP to accomplish these objectives. However, an important effort should be made in the future between laboratories and between countries in Europe to standardise assays for porcine proteins and to harmonise the different methods currently used for measuring the porcine APPs.