Epithelial Cells Induce a Cyclo-Oxygenase-1-Dependent Endogenous Reduction in Airway Smooth Muscle Contractile Phenotype.

Airway smooth muscle cells (ASMCs) are phenotypically regulated to exist in either a proliferative or a contractile state. However, the influence of other airway structural cell types on ASMC phenotype is largely unknown. Although epithelial cells are known to drive ASM proliferation, their effects on the contractile phenotype are uncertain. In the current study, we tested the hypothesis that epithelial cells reduce the contractile phenotype of ASMCs. To do so, we measured force production by traction microscopy, gene and protein expression, as well as calcium release by Fura-2 ratiometric imaging. ASMCs incubated with epithelial-derived medium produced less force after histamine stimulation. We observed reduced expression of myocardin, α-smooth muscle actin, and calponin within ASMCs after coculture with epithelial cells. Peak calcium release in response to histamine was diminished, and depended on the synthesis of cyclo-oxygenase-1 products by ASM and on prostaglandin E receptors 2 and 4. Together, these in vitro results demonstrate that epithelial cells have the capacity to coordinately reduce ASM contraction by functional antagonism and by reduction of the expression of certain contractile proteins.

ROCK insufficiency attenuates ozone-induced airway hyperresponsiveness in mice.

Ozone causes airway hyperresponsiveness (AHR) and pulmonary inflammation. Rho kinase (ROCK) is a key regulator of smooth muscle cell contraction and inflammatory cell migration. To determine the contribution of the two ROCK isoforms ROCK1 and ROCK2 to ozone-induced AHR, we exposed wild-type, ROCK1(+/-), and ROCK2(+/-) mice to air or ozone (2 ppm for 3 h) and evaluated mice 24 h later. ROCK1 or ROCK2 haploinsufficiency did not affect airway responsiveness in air-exposed mice but significantly reduced ozone-induced AHR, with a greater reduction in ROCK2(+/-) mice despite increased bronchoalveolar lavage (BAL) inflammatory cells in ROCK2(+/-) mice. Compared with wild-type mice, ozone-induced increases in BAL hyaluronan, a matrix protein implicated in ozone-induced AHR, were lower in ROCK1(+/-) but not ROCK2(+/-) mice. Ozone-induced increases in other inflammatory moieties reported to contribute to ozone-induced AHR (IL-17A, osteopontin, TNFα) were not different in wild-type vs. ROCK1(+/-) or ROCK2(+/-) mice. We also observed a dose-dependent reduction in ozone-induced AHR after treatment with the ROCK1/ROCK2 inhibitor fasudil, even though fasudil was administered after induction of inflammation. Ozone increased pulmonary expression of ROCK2 but not ROCK1 or RhoA. A ROCK2 inhibitor, SR3677, reduced contractile forces in primary human airway smooth muscle cells, confirming a role for ROCK2 in airway smooth muscle contraction. Our results demonstrate that ozone-induced AHR requires ROCK. Whereas ROCK1-dependent changes in hyaluronan may contribute to ROCK1's role in O3-induced AHR, the role of ROCK2 is downstream of inflammation, likely at the level of airway smooth muscle contraction.

Processing speed and neurodevelopment in adolescent-onset psychosis: cognitive slowing predicts social function.

Onset of psychosis may be associated with abnormal adolescent neurodevelopment. Here we examined the neurocognitive profile of first-episode, adolescent onset psychosis (AOP) as compared to typically developing adolescents, and asked whether neurocognitive performance varied differentially as a function of age in the cases compared with controls. A comprehensive neuropsychological battery was administered to 35 patients experiencing a first-episode of a DSM-IV psychotic disorder and to 31 matched controls. Clinicians also rated subjects' social and role functioning, both at the time of neuropsychological assessment and 1 year later. Although patients displayed a wide range of impairments relative to controls, their most pronounced deficits included verbal memory, sensorimotor dexterity and cognitive processing speed. Among these, only processing speed showed a significant group-by-age interaction, consistent with an aberrant developmental course among AOP patients. Processing speed also accounted for substantial variance in other areas of deficit, and predicted social functioning 1 year later. AOP patients fail to show normal age-related increases in processing speed, which in turn predicts poorer functional outcomes. This pattern is consistent with the view that adolescent brain developmental processes, such as myelination, may be disrupted in these patients.

Substrate stiffening promotes endothelial monolayer disruption through enhanced physical forces.

A hallmark of many, sometimes life-threatening, inflammatory diseases and disorders is vascular leakage. The extent and severity of vascular leakage is broadly mediated by the integrity of the endothelial cell (EC) monolayer, which is in turn governed by three major interactions: cell-cell and cell-substrate contacts, soluble mediators, and biomechanical forces. A potentially critical but essentially uninvestigated component mediating these interactions is the stiffness of the substrate to which the endothelial monolayer is adherent. Accordingly, we investigated the extent to which substrate stiffening influences endothelial monolayer disruption and the role of cell-cell and cell-substrate contacts, soluble mediators, and physical forces in that process. Traction force microscopy showed that forces between cell and cell and between cell and substrate were greater on stiffer substrates. On stiffer substrates, these forces were substantially enhanced by a hyperpermeability stimulus (thrombin, 1 U/ml), and gaps formed between cells. On softer substrates, by contrast, these forces were increased far less by thrombin, and gaps did not form between cells. This stiffness-dependent force enhancement was associated with increased Rho kinase activity, whereas inhibition of Rho kinase attenuated baseline forces and lessened thrombin-induced inter-EC gap formation. Our findings demonstrate a central role of physical forces in EC gap formation and highlight a novel physiological mechanism. Integrity of the endothelial monolayer is governed by its physical microenvironment, which in normal circumstances is compliant but during pathology becomes stiffer.

Decreased bone mineral density of femur in patients with cervical cancer.

AIMS: We compared the bone mineral density (BMD) of spinal bone and the femur between patients with cervical cancer and a control group. METHODS: We retrospectively analyzed the BMD of spinal bone and the femur in 40 patients with cervical cancer and 72 control women. The control women were patients treated with surgery for benign disease. RESULTS: Age, height, bodyweight and body mass index were not significantly different between the two groups. Total femoral BMD in patients with cervical cancer was significantly lower (P = 0.006) than that of control women. In multiple regression analysis, the bodyweight of patients with cervical cancer was a statistically significant predictor positively correlated with the BMD of the total femur. CONCLUSION: The results suggest that patients with cervical cancer have low bone mass, especially in the femur, and an increased risk of developing osteoporosis.