In vivo production and molar percentages of volatile fatty acids in the rumen: a quantitative review by an empirical approach

Despite their major contribution to the energy supply of ruminants, the production of volatile fatty acids (VFA) in the rumen is still poorly predicted by rumen models. We have developed an empirical approach, based on the interpretation of large bibliographic databases gathering published in vivo measurements of ruminal VFA production rate (PR), rates of duodenal and faecal digestion and molar percentages of VFA in the rumen. These databases, covering a wide range of intake levels and dietary composition, were studied by meta-analysis using within-experiment models. We established models to quantify response laws of total VFA-PR and individual VFA molar percentages in the rumen to variations in intake level and dietary composition. The rumen fermentable organic matter (RfOM) intake, estimated from detailed knowledge of the chemical composition of diets according to INRA Feed Tables, appears as an accurate explanatory variable of measured total VFA-PR, with an average increment of 8.03 ± 0.64 mol total VFA/kg RfOM intake. Similar results were obtained when total VFA-PR was estimated from measured apparent RfOM (total VFA-PR/RfOM averaging 8.3 ± 1.2 mol/kg). The VFA molar percentages were related to dry matter intake and measured digestible organic matter (OM), digestible NDF and rumen starch digestibility, with root mean square error of 1.23, 1.45, 0.88 and 0.41 mol/100 mol total VFA for acetate, propionate, butyrate and minor VFA, respectively, with no effect of pH on the residuals. Stoichiometry coefficients were calculated from the slopes of the relationships between individual VFA production (estimated from measured apparent RfOM and individual VFA molar percentages) and measured fermented fractions. Coefficients averaged, respectively, 66, 17, 14 and 3 mol/100 mol for NDF; 41, 44, 12 and 4 mol/100 mol for starch; and 46, 35, 13 and 6 mol/100 mol for crude protein. Their use to predict VFA molar percentages appear relevant for most dietary conditions, that is, when the digested NDF/digested OM ratio exceeded 0.12. This study provides a quantitative review on VFA yield in the rumen. It contributes to the development of feed evaluation systems based on nutrient fluxes.     

NKG2D mediates NK cell hyperresponsiveness and influenza-induced pathologies in a mouse model of chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is characterized by peribronchial and perivascular inflammation and largely irreversible airflow obstruction. Acute disease exacerbations, due frequently to viral infections, lead to enhanced disease symptoms and contribute to long-term progression of COPD pathology. Previously, we demonstrated that NK cells from cigarette smoke (CS)-exposed mice exhibit enhanced effector functions in response to stimulating cytokines or TLR ligands. In this article, we show that the activating receptor NKG2D is a key mediator for CS-stimulated NK cell hyperresponsiveness, because CS-exposed NKG2D-deficient mice (Klrk1(-/-)) did not exhibit enhanced effector functions as assessed by cytokine responsiveness. NK cell cytotoxicity against MHC class I-deficient targets was not affected in a COPD model. However, NK cells from CS-exposed mice exhibit greater cytotoxic activity toward cells that express the NKG2D ligand RAET1ε. We also demonstrate that NKG2D-deficient mice exhibit diminished airway damage and reduced inflammation in a model of viral COPD exacerbation, which do not affect viral clearance. Furthermore, adoptive transfer of NKG2D(+) NK cells into CS-exposed, influenza-infected NKG2D-deficient mice recapitulated the phenotypes observed in CS-exposed, influenza-infected wild-type mice. Our findings indicate that NKG2D stimulation during long-term CS exposure is a central pathway in the development of NK cell hyperresponsiveness and influenza-mediated exacerbations of COPD.     

The Crohn's disease-associated adherent-invasive Escherichia coli strain LF82 replicates in mature phagolysosomes within J774 macrophages

Adherent-invasive Escherichia coli (AIEC) bacteria isolated from Crohn's disease patients are able to extensively replicate within macrophages in large vacuoles. The mechanism by which AIEC bacteria survive within phagocytic cells is unknown. This report describes the maturation of AIEC LF82-containing phagosomes within J774 macrophages. LF82-containing phagosomes traffic through the endocytic pathway as shown by the sequential acquisition and loss of EEA1 and Rab7 and by accumulation of Lamp-1, Lamp-2 and cathepsin D. We demonstrated that AIEC LF82-containing phagosomes mature into active phagolysosomes where bacteria are exposed to low pH and to the degradative activity of cathepsin D. Finally, we showed that an acidic environment is necessary for replication of AIEC LF82 bacteria within J774 macrophages. Thus, evidence is provided that AIEC LF82 bacteria do not escape from the endocytic pathway but undergo normal interaction with host endomembrane organelles and replicate within acidic and cathepsin D-positive vacuolar phagolysosomes.     

Tissue expression patterns identify mouse cilia genes.

In mammals, cilia are critical for development, sensation, cell signaling, sperm motility, and fluid movement. Defects in cilia are causes of several congenital syndromes, providing additional reasons to identify cilia-related genes. We hypothesized that mRNAs selectively abundant in tissues rich in highly ciliated cells encode cilia proteins. Selective abundance in olfactory epithelium, testes, vomeronasal organ, trachea, and lung proved to be an expression pattern uniquely effective in identifying documented cilia-related genes. Known and suspected cilia-related genes were statistically overrepresented among the 99 genes identified, but the majority encoded proteins of unknown function, thereby predicting new cilia-related proteins. Evidence of expression in a highly ciliated cell, the olfactory sensory neuron, exists for 73 of the genes. In situ hybridization for 17 mRNAs confirmed expression of all 17 in olfactory sensory neurons. Most were also detected in vomeronasal sensory neurons and in neighboring tissues rich in ciliated cells such as respiratory epithelium. Immunoreactivity for one of the proteins identified, Spa17, colocalized with acetylated tubulin in the cilia layer of the olfactory epithelium. In contrast, the ciliary rootlet protein, Crocc, was located in discrete structures whose position was consistent with the dendritic knobs of the olfactory sensory neurons. A compilation of >2,000 mouse genes predicted to encode cilia-related proteins revealed a strong correlation (R = 0.99) between the number of studies predicting a gene's involvement in cilia and documented evidence of such involvement, a fact that simplifies the selection of genes for further study of the physiology of cilia.     

Heme oxygenase-1 is a critical regulator of nitric oxide production in enterohemorrhagic Escherichia coli-infected human enterocytes

Enterohemorrhagic Escherichia coli (EHEC) are the causative agent of hemolytic-uremic syndrome. In the first stage of the infection, EHEC interact with human enterocytes to modulate the innate immune response. Inducible NO synthase (iNOS)-derived NO is a critical mediator of the inflammatory response of the infected intestinal mucosa. We therefore aimed to analyze the role of EHEC on iNOS induction in human epithelial cell lines. In this regard, we show that EHEC down-regulate IFN-gamma-induced iNOS mRNA expression and NO production in Hct-8, Caco-2, and T84 cells. This inhibitory effect occurs through the decrease of STAT-1 activation. In parallel, we demonstrate that EHEC stimulate the rapid inducible expression of the gene hmox-1 that encodes for the enzyme heme oxygenase-1 (HO-1). Knock-down of hmox-1 gene expression by small interfering RNA or the blockade of HO-1 activity by zinc protoporphyrin IX abrogated the EHEC-dependent inhibition of STAT-1 activation and iNOS mRNA expression in activated human enterocytes. These results highlight a new strategy elaborated by EHEC to control the host innate immune response.     

Placental lactogen production and functional differentiation of rat trophoblast cells in vitro

Cells from the labyrinth region of the developing rat chorioallantoic placenta were able to differentiate in vitro into cells capable of producing placental lactogen. Progesterone selectively inhibited placental lactogen production by labyrinth cell cultures undergoing differentiation but had no apparent effect on lactogen production by mature trophoblast giant cells. The measurement of placental lactogen production is a useful method for monitoring the functional differentiation of rat trophoblast cells in vitro.