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

Membrane Interactome of a Recombinant Fragment of Human Surfactant Protein D Reveals GRP78 as a Novel Binding Partner in PC3, a Metastatic Prostate Cancer Cell Line

ICMR- National Institute for Research in Reproductive Health ICMR-NIRRH (Accession no. 921); ICMR- National Institute for Research in Reproductive Health ICMR-NIRRH-JRF; ICMR- National Institute for Research in Reproductive Health ICMR-SRF; Researchers Supporting Project (RSP-2020/26) King Saud University, Riyadh

Human SP-D acts as an innate immune surveillance molecule against androgen-responsive and androgen-resistant prostate cancer cells

Surfactant Protein D (SP-D), a pattern recognition innate immune molecule, has been implicated in the immune surveillance against cancer. A recent report showed an association of decreased SP-D expression in human prostate adenocarcinoma with an increased Gleason score and severity. In the present study, the SP-D expression was evaluated in primary prostate epithelial cells (PrEC) and prostate cancer cell lines. LNCaP, an androgen dependent prostate cancer cell line, exhibited significantly lower mRNA and protein levels of SP-D than PrEC and the androgen independent cell lines (PC3 and DU145). A recombinant fragment of human SP-D, rfhSP-D, showed a dose and time dependent binding to prostate cancer cells via its carbohydrate recognition domain. This study, for the first time, provides evidence of significant and specific cell death of tumor cells in rfhSP-D treated explants as well as primary tumor cells isolated from tissue biopsies of metatstatic prostate cancer patients. Viability of PrEC was not altered by rfhSP-D. Treated LNCaP (p53+/+) and PC3 (p53 −/−) cells exhibited reduced cell viability in a dose and time dependent manner and were arrested in G2/M and G1/G0 phase of the cell cycle, respectively. rfhSP-D treated LNCaP cells showed a significant upregulation of p53 whereas a significant downregulation of pAkt was observed in both PC3 and LNCaP cell lines. The rfhSP-D-induced apoptosis signaling cascade involved upregulation of Bax:Bcl2 ratio, cytochrome c and cleaved products of caspase 7. The study concludes that rfhSP-D induces apoptosis in prostate tumor explants as well as in androgen dependent and independent prostate cancer cells via p53 and pAkt pathways.ICMR-NIRR

Immunodetection of surfactant proteins in human organ of Corti, Eustachian tube and kidney.

The presence of surfactant proteins was investigated in the human organ of Corti, Eustachian tube and kidney tissues. It has previously been shown that lamellar bodies are present in hairy cells of organ of Corti, in the cytoplasm of secretory and lumen of tubal glands of Eustachian tube and kidney renal basement membrane. No evidence for the presence of surfactant proteins in the organ of Corti and kidney has been presented until now. The aim of this study was to find out if surfactant proteins were expressed in other epithelia such as organ of Corti, Eustachian tube and kidney. Surfactant proteins were identified using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. On one-dimensional Western blots, bands for surfactant protein A in human Eustachian tube (SP-A, 34 kDa) and in kidney extracts, and for surfactant protein D (SP-D, 43 kDa) in Eustachian tube and in kidney extracts (SP-D, 86 kDa), and for surfactant protein B (SP-B, 8 kDa) in human Eustachian tubeand organ of Corti extracts were detected. Bands corresponded to monomeric forms of lung surfactant proteins. These results indicate the presence of SP-A and SP-D in kidney epithelium, SP-A, SP-B and SP-D in Eustachian tube and SP-B in the organ of Corti

Increased Expression of Surfactant Protein A and D in Rheumatoid Arthritic Synovial Fluid

Aim: To determine the concentrations of surfactant protein (SP)-A and SP-D in synovial fluid samples from patients with rheumatoid arthritis and healthy controls and correlate them with rheumatoid factor, C-reactive protein (CRP), immunoglobulin (Ig) A, IgM, and IgG, total protein, and total lipid content. Methods: The concentrations of SP-A, SP-D, CRP, IgA, IgM, IgG, and rheumatoid factor were measured in the synovial fluid of 7 patients with rheumatoid arthritis and 3 samples of healthy synovial fluid obtained at autopsy. The bands obtained by specific antibodies in Western blotting to determine the surfactant protein concentration were analyzed densitometrically and synovial fluid total phospholipids and protein content were analyzed spectrophotometrically. Results: Patients with rheumatoid arthritis had increased concentrations of proteins and lipids in the synovial fluid, which correlated with 3.5-fold increase in SP-A and 6.1-fold increase in SP-D concentrations. Total protein content in synovial fluid samples from patients with rheumatoid arthritis showed a 2.1-fold increase and phospholipid content showed 7.0-fold increase in comparison with samples of healthy synovial fluid. Rheumatoid factor, CRP, IgA, IgM, and IgG concentrations in synovial fluid samples from patients with rheumatoid arthritis were 40-2660 KIU/L, 4-35 mg/L, 0.10-2.70 g/L, 0.50-1.90 g/L, and 5.3-15.4 g/L respectively. Conclusion: SP-A and SP-D were present and expressed in various degrees in the synovial fluid of patients with rheumatoid arthritis. SP-A and SP-D may play role in the initiation of immune system and joint inflammation, and may be an integral component of synovial fluid and provide insights for in innate and adaptive immunity within the joint

Detection of surfactant protein A (SP-A) and surfactant protein D (SP-D) in equine synovial fluid with immunoblotting

Once considered unique to the lung, surfactant proteins have been clearly identified in the intestine and peritoneum and are suggested to exist in several other organs. In the lung, surfactant proteins assist in the formation of a monolayer of surface-active phospholipid at the liquid–air interface of the alveolar lining, reducing the surface tension at this surface. In contrast, surface-active phospholipid adsorbed to articular surfaces has been identified as the load-bearing boundary lubricant of the joint. This raises the question of whether surfactant proteins in synovial fluid (SF) are required for the formation of the adsorbed layer in normal joints. Proteins from small volumes of equine SF were resolved by 1- and 2-dimensional polyacrylamide gel electrophoresis and detected by Western blotting to investigate the presence of surfactant proteins. The study showed that surfactant proteins A and D (SP-A and SP-D) are present in the SF of normal horses. We suggest that, like surface-active phospholipid, SP-A and SP-D play a significant role in the functioning of joints. Next will be clarification of the roles of surfactant proteins as disease markers in a variety of joint diseases, such as degenerative joint disease and inflammatory problems