Can a combination of vaccination, probiotic and organic acid treatment in layer hens protect against early life exposure to Salmonella Typhimurium and challenge at sexual maturity?

Day old layer chicks were challenged with Salmonella Typhimurium using a seeder bird technique. Treatment groups were untreated control, administration of a probiotic in drinking water weekly, vaccination by intramuscular injection of a live aro-A deletion mutant vaccine at 10 weeks of age (woa) followed by an oral dose at 16 woa, probiotic administration plus vaccination, vaccination plus the administration of an organic acid preparation in feed from 16 woa and a combination of probiotic, vaccine and organic acid. Faecal shedding was monitored by culture at 1, 2, 3, 4, 8, 12, 15, 17, 20, 21, 23 and 25 woa and in dust from settle plates by PCR at intervals from 8 woa. Birds from each group were separated at 17 and 18 woa and challenged orally with 106 CFU of S. Typhimurium. Both untreated and probiotic groups shed Salmonella until 56 days. Salmonella was also detected in dust from 8 until 12 woa but little after this. After vaccination, from sexual maturity (18 woa) all groups except those that were vaccinated with and without probiotic re-excreted Salmonella. The probiotic alone was ineffective against this re-excretion and all groups receiving organic acids shed Salmonella. At 17 woa, unchallenged controls were fully susceptible to caecal colonization, however all other groups showed reduced susceptibility, including the untreated challenged group. However, at 18 woa (sexual maturity) only the groups that were vaccinated with or without probiotic showed reduced susceptibility to colonization. The organic acid treated groups (including the vaccinated group) did not show a difference to the untreated controls. S. Typhimurium demonstrated an ability to re-emerge at sexual maturity, similar to other serovars. The vaccine assisted in limiting the re-excretion at sexual maturity and decreased susceptibility to subsequent challenge. Use of a probiotic augmented the vaccine's protective capacity.

Statins impair survival of primary human mesenchymal progenitor cells via mevalonate depletion, NF-κB signaling, and Bnip3.

Circulating progenitor cells of bone marrow origin have been implicated in transplant cardiac allograft vasculopathy (CAV) and cardiac fibrosis. HMG-CoA reductase inhibitors, called "statins," have been shown to impair the progression of CAV and improve patient survival. We examined the in vitro effects of three HMG-CoA reductase inhibitors atorvastatin, simvastatin, and pravastatin on the viability of MSCs and expression of nuclear factor kappa B (NF-κB). Mesenchymal stem cells (MSCs) isolated from human patients were treated with atorvastatin, simvastatin, and pravastatin at 0.1, 1.0, or 10 µM ± mevalonate. Human MSC treatment with 1 and 10 µM simvastatin or atorvastatin resulted in progressively reduced cell viability, which was associated with a decline in NF-κB p65. Viability was rescued by co-incubation with mevalonate or by pretreatment with Inhibitor of nuclear factor kappa-B kinase subunit beta (Iκκ-ß). Pravastatin did not affect MSC viability or NF-κB expression. Mevalonate depletion through HMG-CoA reductase inhibition impairs the viability of primary human MSC through down-regulating NF-κB.

Human mesenchymal stem cells express a myofibroblastic phenotype in vitro: comparison to human cardiac myofibroblasts.

Cardiac fibrosis accompanies a variety of myocardial disorders, and is induced by myofibroblasts. These cells may be composed of a heterogeneous population of parent cells, including interstitial fibroblasts and circulating progenitor cells. Direct comparison of human bone marrow-derived mesenchymal stem cells (BM-MSCs) and cardiac myofibroblasts (CMyfbs) has not been previously reported. We hypothesized that BM-MSCs readily adopt a myofibroblastic phenotype in culture. Human primary BM-MSCs and human CMyfbs were isolated from patients undergoing open heart surgery and expanded under standard culture conditions. We assessed and compared their phenotypic and functional characteristics by examining their gene expression profile, their ability to contract collagen gels and synthesize collagen type I. In addition, we examined the role of non-muscle myosin II (NMMII) in modulating MSC myogenic function using NMMII siRNA knockdown and blebbistatin, a specific small molecule inhibitor of NMMII. We report that, while human BM-MSCs retain pluripotency, they adopt a myofibroblastic phenotype in culture and stain positive for the myofibroblast markers α-SMA, vimentin, NMMIIB, ED-A fibronectin, and collagen type 1 at each passage. In addition, they contract collagen gels in response to TGF-ß1 and synthesize collagen similar to human CMyfbs. Moreover, inhibition of NMMII activity with blebbistatin completely attenuates gel contractility without affecting cell viability. Thus, human BM-MSCs share and exhibit similar physiological and functional characteristics as human CMyfbs in vitro, and their propensity to adopt a myofibroblast phenotype in culture may contribute to cardiac fibrosis.

Models to study airway smooth muscle contraction in vivo, ex vivo and in vitro: implications in understanding asthma.

Asthma is a chronic obstructive airway disease characterised by airway hyperresponsiveness (AHR) and airway wall remodelling. The effector of airway narrowing is the contraction of airway smooth muscle (ASM), yet the question of whether an inherent or acquired dysfunction in ASM contractile function plays a significant role in the disease pathophysiology remains contentious. The difficulty in determining the role of ASM lies in limitations with the models used to assess contraction. In vivo models provide a fully integrated physiological response but ASM contraction cannot be directly measured. Ex vivo and in vitro models can provide more direct assessment of ASM contraction but the loss of factors that may modulate ASM responsiveness and AHR, including interaction between multiple cell types and disruption of the mechanical environment, precludes a complete understanding of the disease process. In this review we detail key advantages of common in vivo, ex vivo and in vitro models of ASM contraction, as well as emerging tissue engineered models of ASM and whole airways. We also highlight important findings from each model with respect to the pathophysiology of asthma.