Comparison of the IFAT and Iscom-ELISA response in bovine foetuses with Neospora caninum infection

The study was carried out to evaluate the efficacy of foetal serology in the diagnosis of Neospora-associated bovine abortions. Fluids from 14 foetuses of cows with confirmed neosporosis (Group A), seven foetuses with confirmed bovine viral diarrhoea virus (BVD infection) (Group B) and 11 aborted foetuses without demonstrable infection (Group C) were examined. The age of the foetuses ranged from 4.5 months to 9 months. Albumin concentration (measured by Rocket Immunoelectrophoresis) was not significantly different in Group A compared with that in both Groups B and C, while that in Group B was significantly lower than in Group C. Levels of total IgG ranged from 0.01 to 1.78 (mg IgG) ml(-1) measured by single radial immunodiffusion technique. A measurable level of total IgG was found in all foetuses from Groups A and B, with no significant difference between levels in the two groups. Only one foetus in Group C had a detectable level of IgG. All foetuses in Group A had a specific IgG response (titre> or = 20) against Neospora caninum using the IFAT, while no positive responses in IFAT were found in Groups B and C. Measurement of specific IgG1 and IgG2 by Iscom-ELISA showed one and three false-negative results, respectively, in Group A. The IgG1 and IgG2 response in Group A was correlated according to the Spearman test (r = 0.66). Increasing age of the foetuses correlated significantly with the foetal IgG concentration, the specific IgG and IgG1 + IgG2. On the basis of the results obtained, it was concluded that the IFAT with a cut-off titre of 1:20, was a specific method for diagnosis of neosporosis in foetuses older than 4.5 months. The Iscom-ELISA also showed promising results as a method for screening specific antibodies against N. caninum in foetal fluid.     

Melanocortin 1-receptor (MC1R) mutations are associated with plumage colour in chicken

The co-segregation of plumage colour and sequence polymorphism in the melanocortin 1-receptor gene (MC1R) was investigated using an intercross between the red junglefowl and White Leghorn chickens. The results provided compelling evidence that the Extended black (E) locus controlling plumage colour is equivalent to MC1R. E/MC1R was assigned to chromosome 11 with overwhelming statistical support. Sequence analysis indicated that the E92K substitution, causing a constitutively active receptor in the sombre mouse, is the most likely causative mutation for the Extended black allele carried by the White Leghorn founders in this intercross. The MC1R sequence associated with the recessive buttercup (ebc) allele indicated that this allele evolved from a dominant Extended black allele as it shared the E92K and M71T substitutions with some E alleles. It also carried a third missense mutation H215P which thus may interfere with the constitutive activation of the receptor caused by E92K (and possibly M71T).     

Engineering of bone tissue with porcine bone marrow stem cells in three-dimensional trabecular metal: in vitro and in vivo studies

The aim of this study was to investigate capability of cell attachment and ectopic bone formation in pigs after either ex vivo transplantation and expansion of bone marrow stem cells (BMSc) into three-dimensional porous tantalum, or porous tantalum supplemented with BMSc. After 24 hours incubation, cells adhering to the porous tantalum discs were quantified by means of scintillation counting of 3H-thymidine-labeled cells. After 7 days of incubation, the cell-loaded porous tantalum discs were harvested for histological analysis or implanted in the infrasternal muscle; an empty disc and disc implanted immediately after cell loading served as controls. All implants were taken out after 8 weeks of implantation and histological examination was performed. The results of in vitro cell attachment to the porous tantalum discs were not improved significantly with gelatin, collagen or fibronectin coatings. Histological analysis of cell loaded discs in vitro demonstrated viable BMSc within the 3-D tantalum structure. In vivo bone induction was demonstrated when the porous tantalum discs were cultured with BMSc. Our findings indicated that porous tantalum was suitable for cell attachment, and ectopic bone formation in pigs was achieved by means of BMSc cultured with porous tantalum. The present study suggests that cell-mediated hard bone tissue repair technology makes it possible to prefabricate autologous BMSc into three-dimensional trabecular metal in order to engineer bone tissue.     

Effects of physical properties of feed on microbial ecology and survival of Salmonella enterica serovar Typhimurium in the pig gastrointestinal tract

A two-by-two factorial experiment with pigs was conducted to study the effect of feed grinding (fine and coarse) and feed processing (pelleted and nonpelleted) on physicochemical properties, microbial populations, and survival of Salmonella enterica serovar Typhimurium DT12 in the gastrointestinal tracts of pigs. Results demonstrated a strong effect of diet on parameters measured in the stomachs of the pigs, whereas the effect was less in the other parts of the gastrointestinal tract. Pigs fed the coarse nonpelleted (C-NP) diet showed more solid gastric content with higher dry matter content than pigs fed the fine nonpelleted (F-NP), coarse pelleted (C-P), or fine pelleted (F-P) diet. Pigs fed the C-NP diet also showed significantly increased number of anaerobic bacteria (P < 0.05), increased concentrations of organic acids, and reduced pH in the stomach. In addition, pigs fed the C-NP diet showed increased in vitro death rate of S. enterica serovar Typhimurium DT12 in content from the stomach (P < 0.001). Pigs fed the C-NP diet had a significantly higher concentration of undissociated lactic acid in gastric content than pigs fed the other diets (P < 0.001). A strong correlation between the concentration of undissociated lactic acid and the death rate of S. enterica serovar Typhimurium DT12 was found. In the distal small intestine, cecum, and midcolon, significantly lower numbers of coliform bacteria were observed in pigs fed the coarse diets than in pigs fed the fine diets (P < 0.01). Pigs fed the C-NP diet showed the lowest number of coliform bacteria in these segments of the gastrointestinal tract. Pigs fed the coarse diets showed increased concentration of butyric acid in the cecum (P < 0.05) and colon (P < 0.10) compared with pigs fed the fine diets. It was concluded that feeding a coarsely ground meal feed to pigs changes the physicochemical and microbial properties of content in the stomach, which decreases the survival of Salmonella during passage through the stomach. In this way the stomach acts as a barrier preventing harmful bacteria from entering and proliferating in the lower part of the gastrointestinal tract.     

Interaction between growth hormone and insulin in the regulation of lipoprotein metabolism in the rat

The importance of insulin for the in vivo effects of growth hormone (GH) on lipid and lipoprotein metabolism was investigated by examining the effects of GH treatment of hypophysectomized (Hx) female rats with and without concomitant insulin treatment. Hypophysectomy-induced changes of HDL, apolipoprotein (apo)E, LDL, and apoB levels were normalized by GH treatment but not affected by insulin treatment. The hepatic triglyceride secretion rate was lower in Hx rats than in normal rats and increased by GH treatment. This effect of GH was blunted by insulin treatment. The triglyceride content in the liver changed in parallel with the changes in triglyceride secretion rate, indicating that the effect of the hormones on triglyceride secretion was dependent on changed availability of triglycerides for VLDL assembly. GH and insulin independently increased editing of apoB mRNA, but the effects were not additive. The expression of fatty-acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD-1), and sterol regulatory element-binding protein-1c (SREBP-1c) was increased by GH treatment. Insulin and GH had no additive effects on these genes; instead, insulin blunted the effect of GH on SREBP-1c mRNA. In contrast to the liver, adipose tissue expression of SREBP-1c, FAS, or SCD-1 mRNA was not influenced by GH. In conclusion, the increased hepatic expression of lipogenic enzymes after GH treatment may be explained by increased expression of SREBP-1c. Insulin does not mediate the effects of GH but inhibits the stimulatory effect of GH on hepatic SREBP-1c expression and triglyceride secretion rate.     

Increasing winter temperatures explain body size decrease in wintering bird populations of Northern Europe—But response patterns vary along the spatioclimatic gradient

Aim

Recent evidence has shown changes in body size and shape of individuals, which are suggested to be a result of global warming caused by climate change. Here, we explored the spatiotemporal changes in wing length and body mass of 24 wintering bird species in Northern Europe and how these relate to temperature anomaly.

Location

Finland and Sweden, Europe.

Time Period

50 years, 1970 to 2020.

Major Taxa Studied

Birds, 24 species.

Methods

We used site-specific, long-term winter ringing data containing wing length and body mass measurements from across Sweden and Finland for 24 bird species. We modelled wing length and body mass change over time, in relation to the spatioclimatic gradient and as response to temperature anomalies (of [i] the same winter as the ringing took place, [ii] the previous winter and [iii] the previous spring) by accounting for phylogenetic relatedness between species and their species-specific responses to each predictor of interest.

Results

We show that across all species, body size has decreased since the 1970s, with a negative relationship between wing length and temperature anomalies of previous winters, suggesting carry-over effects likely linked to body size-related survival or dispersal. Body mass was negatively related to the temperature anomaly of the same winter, indicating more immediate effects related to reduced fat reserves during mild winters.

Main Conclusions

Our results highlight a climate-driven decrease in body size across several species and its association with positive anomalies in winter temperature in the high latitudes. However, the responses are not spatially uniform and there is considerable species-specific variation, emphasizing the importance of conducting multispecies studies when investigating responses to climate change. The mechanisms of decreasing wing length and body mass seem to differ and underline the immediate and carry-over effects of temperature warming during the nonbreeding season.     

Naloxone inhibits diazepam-induced feeding in rats.

Diazepam will induce feeding in rats. This feeding has a rather stable latency and does not wane with repeated doses. Naloxone hydrochloride, in doses as small as 0.25 mg/kg, subcutaneously, inhibited the feeding induced by diazepam, 25 mg/kg, intragastrically.     

The functional unit of calcium-plus-magnesium-ion-dependent adenosine triphosphatase from sarcoplasmic reticulum. The aggregational state of the deoxycholate-solubilized protein in an enzymically active form

Vesicles consisting of (Ca²⁺+Mg²⁺)-dependent ATPase (adenosine triphosphatase), and lipid were prepared from sarcoplasmic reticulum of rabbit skeletal muscle. As with non-ionic detergents [le Maire, Møller & Tanford (1976) Biochemistry 15, 2336–2342] the (Ca²⁺+Mg²⁺)-dependent ATPase after solubilization by deoxycholate showed a pronounced tendency to form oligomers in gel-chromatographic experiments, when eluted in the presence of deoxycholate and phosphatidylcholine. To evaluate the functional significance of oligomer formation the properties of enzymically active preparations of ATPase, solubilized by deoxycholate, were studied. Such preparations were obtained at a protein concentration of 2.5mg/ml in the presence of a high salt concentration (0.4m-KCl) and sucrose (0.3m) in the solubilization medium. Analytical ultracentrifugation of solubilized ATPase showed one protein boundary moving at the same rate as gel-chromatographically prepared monomeric ATPase (s_20,w=6.0S). From simultaneous measurements of the diffusion coefficient an apparent molecular weight of 133000 was calculated, consistent with solubilization of ATPase in predominantly monomeric form. The enzymic activity of deoxycholate-solubilized ATPase when measured directly in the solubilization medium at optimal Ca²⁺ and MgATP concentrations was about 35–50% of that of vesicular ATPase. The dependence of enzymic activity on MgATP concentration indicated that the solubilized ATPase retained high-affinity binding of MgATP, but the presence of high concentrations of the nucleotide did not stimulate activity further, in contrast with that of vesicular ATPase. The dependence of enzymic activity on the free Ca²⁺ concentration was essentially the same for both solubilized and vesicular forms, indicating that interaction of ATPase with more than one molecule of Ca²⁺ is required for enzyme activity. Solubilized enzyme at 20°C was phosphorylated to about the same degree as vesicular ATPase. It is concluded that the catalytic activity of monomeric ATPase retains most of the features of vesicular ATPase and that extensive oligomer formation in gel-chromatographic experiments in the presence of deoxycholate probably reflects processes taking place during inactivation and delipidation of the protein.