FBXL2 is a ubiquitin E3 ligase subunit that triggers mitotic arrest

Mitotic progression is regulated by ubiquitin E3 ligase complexes to carefully orchestrate eukaryotic cell division. Here, we show that a relatively new E3 ligase component belonging to the SCF (Skip-Cullin1-F-box protein) E3 ligase family, SCF^FBXL2, impairs cell proliferation by mediating cyclin D3 polyubiquitination and degradation. Both cyclin D3 and FBXL2 colocalize within the centrosome. FBXL2 overexpression led to G₂/M-phase arrest in transformed epithelia, resulting in the appearance of supernumerary centrosomes, tetraploidy and nuclei where condensed chromosomes are arranged on circular monopolar spindles typical of mitotic arrest. RNAi-mediated knockdown of cyclin D3 recapitulated effects of SCF^FBXL2 expression. SCF^FBXL2 impaired the ability of cyclin D3 to associate with centrosomal assembly proteins [Aurora A, polo-like kinase 4 (Plk4), CDK11]. Thus, these results suggest a role for SCF^FBXL2 in regulating the fidelity of cellular division.Key words: F-box protein, centrosome, mitosis, cyclin D3, Aurora A     

Response of milk fat concentration and yield to nutrient supply in dairy cows

Dietary changes alter dairy cow milk fat concentration (MFC) and yield (MFY) through modifications in the supply of nutrients, which act as precursors or inhibitors of mammary fat synthesis. The current models used to formulate dairy cow diets cannot predict changes in milk fat. The knowledge of the effects of the nutrients on milk fat would help to progress toward this prediction. To this end, we quantified and compared the milk fat responses to variations in the supply of seven nutrients derived from digestion: volatile fatty acids, glucose, proteins, long-chain fatty acids (LCFA) and t10,c12-conjugated linoleic acid (CLA). A database was compiled from studies involving digestive infusions of these nutrients in dairy cows. It included 147 comparisons between a nutrient infusion and a control treatment. The nutrient infusions were limited to the range of physiological variations to mimic nutrient changes after dietary modifications. We established models for the response of MFC, MFY and milk fatty acid (FA) composition to the supply of each nutrient. MFC and MFY responses to the nutrients were significant and linear, except for the MFC response to glucose that was curvilinear. The nutrients differed in their effects on MFC and MFY: acetate, butyrate and LCFA increased MFC and MFY, whereas propionate, glucose and t10,c12-CLA decreased them. Protein infusions increased MFY and decreased MFC because of an increase in milk yield. The effects of numerous interfering factors related to animals, diets or experimental conditions were tested on the residuals of the response models. The responses of milk FA percentages are also provided. When adjusted to the in vivo variations in the nutrients observed after dietary changes, the effects of the different nutrients were moderate. Finally, this study showed that several of these nutrients could contribute to the changes in milk fat production and composition observed after dietary changes. This is a first step toward predicting milk fat response to changes in nutrient supply.     

Digestion of fatty acids in ruminants: a meta-analysis of flows and variation factors: 2. C18 fatty acids

In ruminants, dietary lipids are extensively hydrogenated by rumen micro-organisms, and the extent of this biohydrogenation is a major determinant of long-chain fatty acid profiles of animal products (milk, meat). This paper reports on the duodenal flows of C18 fatty acids and their absorption in the small intestine, using a meta-analysis of a database of 77 experiments (294 treatments). We established equations for the prediction of duodenal flows of various 18-carbon (C18) fatty acids as a function of the intakes of their precursors and other dietary factors (source and/or technological treatment of dietary lipids). We also quantified the influence of several factors modifying rumen metabolism (pH, forage : concentrate ratio, level of intake, fish oil supplementation). We established equations for the apparent absorption of these fatty acids in the small intestine as a function of their duodenal flows. For all C18 unsaturated fatty acids, apparent absorption was a linear function of duodenal flow. For 18:0, apparent absorption levelled off for high duodenal flows. From this database, with fatty acid flows expressed in g/kg dry matter intake, we could not find any significant differences between animal categories (lactating cows, other cattle or sheep) in terms of rumen metabolism or intestinal absorption of C18 fatty acids.     

Resistance of Aedes aegypti from the state of São Paulo,Brazil, to organophosphates insecticides

Since the reintroduction of Aedes aegypti in the state of S o Paulo, in the middle of the 1980-decade, organophosphate insecticides are being used to control the dengue vector. In 1996, an annual program for monitoring the susceptibility of Ae. aegypti to the insecticides was implemented. Some of the results of this monitoring program are presented. Ae. aegypti populations from ten localities have been submitted to bioassays with the diagnostic dose of temephos and fenitrothion. Only two (Mar lia and Presidente Prudente) remain susceptible to both insecticides and one (Santos) exhibits true resistance. Ae. aegypti from the remaining localities showed an incipient altered susceptibility. Resistance ratios varied from 1.2 to 2.9 for temephos and from 1.5 to 3.2 to fenitrothion, indicating moderate levels of resistance. Biochemical assays did not detect alterations in the enzyme acetilcholinesterase, but indicated that resistance is associated with esterases.     

Separation, characterization and hydrogel-formation of hemicellulose from aspen wood

Hemicellulose from aspen (Populus tremula) was isolated by an alkali extraction method, which was followed by hydrogen peroxide treatment, ultrafiltration and recovery by spray drying. The sugar composition and lignin content were monitored with HPLC at each step of the separation procedure. Size-exclusion chromatography showed a polymeric hemicellulose of relatively high molar mass. The product was characterized by ¹H and ¹³C NMR spectroscopy and was found to be composed of a linear (1→4)-β-linked -xylose main chain with a 4-O-methyl-- -glucuronic acid substituting the 2-position of approximately every eighth xylose unit. Lignin and O-acetyl groups had largely been removed in the separation process. The xylan was soluble in hot water, and the film forming properties were examined at various mixtures of the hemicellulose and chitosan. These films formed hydrogels with a high swelling capacity at certain compositions. The morphologies of the films were examined with wide angle X-ray spectroscopy, and a pure xylan film was found to be crystalline, which was suggested to be a consequence of the lack of O-acetyl groups. The crystallinity of the films was found to decrease with an increasing amount of chitosan, and the film of chitosan alone showed no crystallinity. The cohesive forces of the hydrogels are suggested to be the result of the crystalline arrangement of the polymers and of electrostatic interactions between acidic groups in the hemicellulose and amine groups in the chitosan.     

Crohn's disease-associated adherent-invasive E. coli are selectively favoured by impaired autophagy to replicate intracellularly

Ileal lesions in Crohn's disease (CD) patients are colonized by pathogenic adherent-invasive Escherichia coli (AIEC) able to invade and to replicate within intestinal epithelial cells. Recent genome-wide association studies have highlighted the autophagy pathway as being associated with CD risk. In the present study we investigated whether defects in autophagy enhance replication of commensal and pathogenic Escherichia coli and CD-associated AIEC. We show that functional autophagy limits intracellular AIEC replication and that a subpopulation of the intracellular bacteria is located within LC3-positive autophagosomes. In IRGM and ATG16L1 deficient cells intracellular AIEC LF82 bacteria have enhanced replication. Surprisingly autophagy deficiency did not interfere with the ability of intracellular bacteria to survive and/or replicate for any other E. coli strains tested, including non-pathogenic, environmental, commensal, or pathogenic strains involved in gastro enteritis. Together these findings demonstrate a central role for autophagy restraining Adherent-Invasive E. coli strains associated with ileal CD. AIEC infection in patients with polymorphisms in autophagy genes may have a significant impact on the outcome of intestinal inflammation.     

Antibiotic therapy for ocular infection.

Infections of the eye can rapidly damage important functional structures and lead to permanent vision loss or blindness. Broad-spectrum antibiotics should be administered to the appropriate site of infection as soon as a diagnosis is made. Topical drops are preferred for corneal and conjunctival infections. Intravitreal antibiotics, and possibly subconjunctival and parenteral antibiotics, are preferred for endophthalmitis. Parenteral antibiotics are recommended for infection in deep adnexal structures. We review specific aspects of antibiotic therapy for ocular and periocular infection.