Détermination de l’expression des gènes codant pour le TNF-α et la leptine par RT-PCR dans le sang de vaches présentant un déplacement de la caillette à gauche

The aims of this study are to evaluate the TNF-α and leptin gene expression in blood from Holstein cows with left abomasal displacement and to correlate it with induced liver injury. The TNF-α and leptin expression in blood samples was determined by RT-PCR after normalisation using the constant expression of the housekeeping GAPDH gene in cows with left abomasal displacement (LAD) (n = 20) before surgery and 7 days after as well as in healthy controls (n = 10). Plasma hepatic enzyme (AST: aspartate aminotransferase, ALT: alanine aminotransferase and ALP: alkaline phosphatase) activities were measured in parallel. Plasma AST and ALP activities dramatically increased in diseased cows during the preoperative period and then declined. Although not significantly, the leptin expression tended to decrease in LAD affected cows while the TNF-α expression tended to increase during the postoperative period. These results suggest that TNF-α may be associated with liver damage during abomasal displacement and that leptin was inversely correlated.Les objectifs de cette étude ont été d’évaluer l’expression des gènes codant pour le TNF-α et la leptine dans le sang de vaches Holstein présentant un déplacement à gauche de la caillette et de la corréler avec les lésions hépatiques induites. L’expression du TNF-α et de la leptine a été déterminée par RT-PCR après normalisation en considérant l’expression du gène de ménage GAPDH comme constante dans les échantillons sanguins provenant de vaches atteintes d’un déplacement à gauche de la caillette (n = 20) avant et 7 jours après traitement chirurgical ou provenant de vaches saines (témoins, n = 10). Les activités plasmatiques des enzymes hépatiques (AST : aspartate aminotransférase, ALT ; alanine aminotransférase et PAL : phosphatase alcaline) ont été mesurées en parallèle. Les activités plasmatiques de l’AST et de la PAL étaient considérablement augmentées chez les vaches malades avant la chirurgie puis elles ont diminué durant la période postopératoire. Bien que les variations n’aient pas été significatives, l’expression de la leptine chez les animaux malades a tendu à diminuer alors que celle du TNF-α a augmenté durant la période postopératoire. Ces résultats suggèrent que le TNF-α pourrait être associé aux lésions hépatiques associées à un déplacement de la caillette alors que la leptine serait inversement corrélée.Scientific Research Projects Commission of Mehmet Akif Ersoy Universit

Surfactant protein D inhibits HIV-1 infection of target cells via interference with gp120-CD4 interaction and modulates pro-inflammatory cytokine production

© 2014 Pandit et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Surfactant Protein SP-D, a member of the collectin family, is a pattern recognition protein, secreted by mucosal epithelial cells and has an important role in innate immunity against various pathogens. In this study, we confirm that native human SP-D and a recombinant fragment of human SP-D (rhSP-D) bind to gp120 of HIV-1 and significantly inhibit viral replication in vitro in a calcium and dose-dependent manner. We show, for the first time, that SP-D and rhSP-D act as potent inhibitors of HIV-1 entry in to target cells and block the interaction between CD4 and gp120 in a dose-dependent manner. The rhSP-D-mediated inhibition of viral replication was examined using three clinical isolates of HIV-1 and three target cells: Jurkat T cells, U937 monocytic cells and PBMCs. HIV-1 induced cytokine storm in the three target cells was significantly suppressed by rhSP-D. Phosphorylation of key kinases p38, Erk1/2 and AKT, which contribute to HIV-1 induced immune activation, was significantly reduced in vitro in the presence of rhSP-D. Notably, anti-HIV-1 activity of rhSP-D was retained in the presence of biological fluids such as cervico-vaginal lavage and seminal plasma. Our study illustrates the multi-faceted role of human SPD against HIV-1 and potential of rhSP-D for immunotherapy to inhibit viral entry and immune activation in acute HIV infection. © 2014 Pandit et al.The work (Project no. 2011-16850) was supported by Medical Innovation Fund of Indian Council of Medical Research, New Delhi, India (www.icmr.nic.in/)

Latherin: A Surfactant Protein of Horse Sweat and Saliva

Horses are unusual in producing protein-rich sweat for thermoregulation, a major component of which is latherin, a highly surface-active, non-glycosylated protein. The amino acid sequence of latherin, determined from cDNA analysis, is highly conserved across four geographically dispersed equid species (horse, zebra, onager, ass), and is similar to a family of proteins only found previously in the oral cavity and associated tissues of mammals. Latherin produces a significant reduction in water surface tension at low concentrations (≤1 mg ml−1), and therefore probably acts as a wetting agent to facilitate evaporative cooling through a waterproofed pelt. Neutron reflection experiments indicate that this detergent-like activity is associated with the formation of a dense protein layer, about 10 Å thick, at the air-water interface. However, biophysical characterization (circular dichroism, differential scanning calorimetry) in solution shows that latherin behaves like a typical globular protein, although with unusual intrinsic fluorescence characteristics, suggesting that significant conformational change or unfolding of the protein is required for assembly of the air-water interfacial layer. RT-PCR screening revealed latherin transcripts in horse skin and salivary gland but in no other tissues. Recombinant latherin produced in bacteria was also found to be the target of IgE antibody from horse-allergic subjects. Equids therefore may have adapted an oral/salivary mucosal protein for two purposes peculiar to their lifestyle, namely their need for rapid and efficient heat dissipation and their specialisation for masticating and processing large quantities of dry food material