Altered hepatic inflammatory response in the offspring following prenatal LPS exposure

Postnatal development of lipopolysaccharide-induced inflammatory response in the brain

“…RT-PCR is a technique that has been successfully used by many studies examining the expression levels of many inflammatory mediators at the mRNA level [30,[42][43][44][45]; however, subtle differences in the expression of these neuroinflammatory mediators may fail to be detected by RT-PCR due to its lower sensitivity compared to other techniques such as real-time PCR.…”

“…After cDNA synthesis, PCR amplification was carried out using appropriate sense and antisense primers specific for rat b-actin (a housekeeping gene), IL-1b, IL-6, TNF-a, KC, MIP-1b, MCP-1, TLR-4, CD14, and Myd88 in a final volume of 20 ll containing 1 ll of cDNA, 19 PCR buffer, 0.2 lM of each sense and antisense primer, 0.2 mM of dNTPs, and 1 U of Taq DNA polymerase (Promega). The information on specific primers and PCR conditions for b-actin, IL-1b, IL-6, TNF-a, KC, MIP-1b, TLR-4, and CD14 is described in Lasala et al [19] and Surriga et al [30]. The sense and antisense primers for rat Myd88 were 5 0 -TGT-CCC-AAA-GGA-AAC-ACA-CA-3 0 and 5 0 -CCC-CTG-ACA-TGC-CTA-TCA-CT-3 0 , respectively, and the sense and antisense primers for rat MCP-1 were 5 0 -ATG-ATC-CCA-ATG-AGT-CGG-3 0 and 5 0 -ACA-GAA-GTG-CTT-GAG-GTG-3 0 , respectively.…”

“…Total protein was extracted with lysis buffer containing 50 mM HEPES (pH 7.5), 5 mM EDTA, 50 mM NaCl, 10 mM NaPPi, 50 mM NaF, 1% IGEPAL, 2 mM Na 3 VO 4 , and 19 protease inhibitor cocktail (Roche, Indianapolis, IN, USA) as described in Surriga et al [30]. Protein concentration was then determined using the bicinchoninic acid (BCA) Protein Assay Reagent from Pierce Biotechnology Inc. (Rockford, IL, USA) following the manufacturer's instructions.…”

“…After scanning, membranes were stripped and reprobed with an antibody immunoreactive to actin (Sigma, St. Louis, MO, USA). The scanned images were then digitized using ImageQuant software (Molecular Dynamics) [30].…”

Alleviation of zearalenone toxicity by modified halloysite nanotubes in the immune response of swine

“…The results reported by Urakubo et al (2001) show that maternal exposure to infection alters the inflammatory cytokine levels in the foetal brain. Lipopolysaccharide (LPS) stimulation during pregnancy alters the immune response in developing offspring (Surriga et al 2009). Reduced litter size, abnormal foetal development and decreased fertility have been major problems associated with livestock production in China.…”