Bacterial clearance of Pseudomonas aeruginosa is enhanced by the inhibition of COX-2

Prostanoids generated by COX-2 are involved in the regulation of inflammation but their exact role in the innate immune response has not been defined. We investigated whether COX-2 is involved in host defense against Pseudomonas aeruginosa pneumonia. In vitro studies, in a macrophage cell line, showed that cytotoxic strain of P aeruginosa (PA103) induced significant COX-2 protein expression and enzymatic function. In vivo data showed that infection with PA103 increased COX-2 protein production in whole lung tissue compared to mice that were infected with mutant bacteria that lack ExoU (DeltaU) or ExoU and ExoT (DeltaUT). COX-2(-/-) mice had accentuated clearance of cytotoxic P. aeruginosa from the lungs. We further tested the effects of COX-2 products such as prostaglandin E(2) on the function of phagocytic cells. Our studies indicate that prostaglandin E(2) may be involved through interacting with the EP2 receptors in modulating the host response because treatment of macrophages with prostaglandin E(2) suppressed production of reactive oxygen species. Furthermore there was enhanced bacterial clearance in EP2 receptor(-/-) mice compared to the wild-type controls. Thus it is possible that inhibition of COX-2 or EP2 receptors could be an effective adjunctive treatment for severe or resistant P. aeruginosa pneumonia.     

Aquaporin 1 regulates GTP-induced rapid gating of water in secretory vesicles

The swelling of secretory vesicles has been implicated in exocytosis, but the underlying mechanism of vesicle swelling remains largely unknown. Zymogen granules (ZGs), the membrane-bound secretory vesicles in exocrine pancreas, swell in response to GTP mediated by a G(alpha)i3 protein. Evidence is presented here that the water channel aquaporin-1 (AQP1) is present in the ZG membrane and participates in rapid GTP-induced vesicular water gating and swelling. Isolated ZGs exhibit low basal water permeability. However, exposure of granules to GTP results in a marked potentiation of water entry. Treatment of ZGs with the known water channel inhibitor Hg2+ is accompanied by a reversible loss in both the basal and GTP-stimulatable water entry and vesicle swelling. Introduction of AQP1-specific antibody raised against the carboxyl-terminal domain of AQP1 blocks GTP-stimulable swelling of vesicles. Our results demonstrate that AQP1 associated at the ZG membrane is involved in basal as well as GTP-induced rapid gating of water in ZGs of the exocrine pancreas.     

Differential G protein-coupled cannabinoid receptor signaling by anandamide directs blastocyst activation for implantation

Mammalian fertility absolutely depends on synchronized development of the blastocyst to the stage when it is competent to implant, and the uterus to the stage when it is receptive to implantation. However, the molecular basis for the reciprocal interaction between the embryo and the uterus remains largely unexplored. One potentially important mechanism involves signaling between an evolutionarily conserved G protein-coupled protein cannabinoid receptor, CB1, that is expressed at high levels on the surface of the trophectoderm and anandamide (N-arachi-donoylethanolamine), an endocannabinoid ligand found to be produced at higher levels by the uterus before implantation and then down-regulated at the time of implantation. Using genetic, pharmacological, and physiological approaches, we show here that anandamide within a very narrow range regulates blastocyst function and implantation by differentially modulating mitogen-activated protein kinase signaling and Ca2+ channel activity via CB1 receptors. Anandamide at a low concentration (7 nM) induces extracellular regulated kinase phosphorylation and nuclear translocation in trophectoderm cells without influencing Ca2+ channels, and renders the blastocyst competent for implantation in the receptive uterus. In contrast, anandamide at a higher concentration (28 nM) inhibits Ca2+ channel activity and blastocyst competency for implantation without influencing mitogen-activated protein kinase signaling. Besides uncovering a potentially important regulatory mechanism for synchronizing blastocyst and uterine competency to implantation, this observation has high clinical relevance, because elevated levels of anandamide induce spontaneous pregnancy loss in women.     

Machinability Investigation of Nitronic 60 Steel Turning Using SiAlON Ceramic Tools under Different Cooling/Lubrication Conditions

The machining of nickel-based super alloys is challenging, owing to the generation of high cutting temperatures, as well as difficulty in maintaining dimensional accuracy and minimizing surface roughness, which compels the use of cutting fluids for reducing these issues due to efficient cooling/lubrication strategies. The present work investigates the comparative performance of four cooling/lubrication techniques: dry cutting, wet, minimum quantity lubricant (MQL) and compressed-air modes in turning Nitronic 60 steel using a new-generation SiAlON ceramic inserts. Several machinability parameters were analyzed for performance evaluation. For this purpose, 16 cycles of turning trials were performed based on Taguchi’s L₁₆ orthogonal array experimental design by varying cutting conditions and lubrication modes. MQL exhibits beneficial effects as compared to the other lubrication conditions concerning low cutting force, improved surface finish, decreased cutting temperature, longer tool life, and lower white layer thickness on machined surface. Burr formation on the saw-tooth chip surface, as well as friction, greatly influenced the tool flank wear due to improper cooling and poor lubrication approach in dry, wet, and compressed-air-cooled machining environments in comparison to MQL-machining. From an economical perspective, the tool life in MQL machining improved by 11%, 72%, and 138% in the comparison with flooded, compressed-air, and dry conditions, respectively. The results of the study demonstrate that using the MQL system can help with heat extraction capability, and provide some promising outcomes.     

FKBP52 deficiency-conferred uterine progesterone resistance is genetic background and pregnancy stage specific

Immunophilin FKBP52 serves as a cochaperone to govern normal progesterone (P(4)) receptor (PR) function. Using Fkbp52(-/-) mice, we show intriguing aspects of uterine P(4)/PR signaling during pregnancy. Implantation failure is the major phenotype found in these null females, which is conserved on both C57BL6/129 and CD1 backgrounds. However, P(4) supplementation rescued implantation and subsequent decidualization in CD1, but not C57BL6/129, null females. Surprisingly, experimentally induced decidualization in the absence of blastocysts failed in Fkbp52(-/-) mice on either background even with P(4) supplementation, suggesting that embryonic signals complement uterine signaling for this event. Another interesting finding was that while P(4) at higher than normal pregnancy levels conferred PR signaling sufficient for implantation in CD1 null females, these levels were inefficient in maintaining pregnancy to full term. However, elevating P(4) levels further restored PR signaling to a level optimal for successful term pregnancy with normal litter size. Collectively, the results show that the indispensability of FKBP52 in uterine P(4)/PR signaling is a function of genetic disparity and is pregnancy stage specific. Since there is evidence for a correlation between P(4) supplementation and reduced risks of P(4)-resistant recurrent miscarriages and remission of endometriosis, these findings have clinical implications for genetically diverse populations of women.     

Long‐term management issues in restless legs syndrome

Restless legs syndrome is a neurologic movement and sleep disorder with lifelong symptoms causing considerable morbidity. Several short‐term and some long‐term open‐label and double‐blind clinical trials have demonstrated the efficacy and safety of dopaminergic treatment in restless legs syndrome. Long‐term treatment, however, is associated with the emergence of vexing long‐term side effects that pose a challenge for physicians. These long‐term complications can be broadly categorized as disease‐related (impact on sleep and acute exacerbation of restless legs syndrome symptoms), and medication issues (augmentation, sleep attacks, impulse control disorders, addiction and dependence, site reaction, occasionally sleep apnea, fibrotic complications, and weight gain). © 2011 Movement Disorder Society