ABSTRACT
The roles of cell division control protein 42 homologue (CDC42) and actin polymerisation in regulating the phenotype of superficial-zone chondrocytes (SZCs) have been demonstrated in vitro; however, the signalling pathway(s) downstream have yet to be fully elucidated. The study hypothesis was that Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) act downstream to regulate proteoglycan 4 (PRG4) and tenascin C (TNC). Bovine SZCs grown in monolayer were treated with ML141 (CDC42 inhibitor) or the actin depolymerising agents, latrunculin B and cytochalasin D, to determine the effect on YAP/TAZ. Verteporfin (YAP/TAZ inhibitor) and YAP/TAZ siRNA-mediated knockdown were used to determine their role in regulating PRG4 and TNC. ML141 treatment reduced total YAP/TAZ protein, nuclear TAZ levels and the YAP/TAZ target gene, connective tissue growth factor (CTGF) mRNA levels. Latrunculin B decreased nuclear TAZ, while cytochalasin D treatment trended towards increased nuclear TAZ (p = 0.06), correlating with decreased and increased CTGF mRNA levels, respectively. Verteporfin treatment decreased PRG4 and TNC expression, with no effect on actin polymerisation. siRNA-mediated knockdown of YAP/TAZ revealed that PRG4 was regulated by YAP/TAZ while TNC was regulated by TAZ only. As cytochalasin D can activate myocardin-related transcription factor-A (MRTF-A), siRNA-mediated knockdown was performed to determine the role of MRTF-A in regulating YAP/TAZ. Although nuclear TAZ decreased, no significant changes in total protein levels were observed. Findings suggested that CDC42 and actin polymerisation regulated SZCs through multiple actin-regulated pathways. Understanding the regulation of these chondroprotective molecules may have important implications for prevention/treatment of osteoarthritis.
Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Antigens/metabolism , Chondrocytes/metabolism , Intracellular Signaling Peptides and Proteins/metabolism , Proteoglycans/metabolism , Tenascin/metabolism , Actin Cytoskeleton/drug effects , Actin Cytoskeleton/metabolism , Animals , Cattle , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Chondrocytes/drug effects , Phenotype , Protein Transport/drug effects , Trans-Activators/metabolism , Verteporfin/pharmacology , cdc42 GTP-Binding Protein/antagonists & inhibitors , cdc42 GTP-Binding Protein/metabolismABSTRACT
Proteoglycan 4 (PRG4/lubricin) is secreted by cells that reside in articular cartilage and line the synovial joint. Lubricin may play a role in modulating inflammatory responses through interaction with CD44. This led us to examine if lubricin could be playing a larger role in the modulation of inflammation/immunity through interaction with Toll-like receptors (TLRs). Human Embryonic Kidney (HEK) cells overexpressing TLRs 2, 4 or 5 and surface plasmon resonance were employed to determine if full length recombinant human lubricin was able to bind to and activate TLRs. Primary human synovial fibroblasts were also examined using flow cytometry and Luminex multiplex ELISA. A rat destabilization model of osteoarthritis (OA) was used to determine if lubricin injections were able to regulate pain and/or inflammation in vivo. Lubricin can bind to and regulate the activity of TLRs, leading to downstream changes in inflammatory signalling independent of HA. We confirmed these findings in vivo through intra-articular injections of lubricin in a rat OA model where the inhibition of systemic inflammatory signaling and reduction in pain were observed. Lubricin plays an important role in regulating the inflammatory environment under both homeostatic and tissue injury states.
Subject(s)
Glycoproteins/metabolism , Toll-Like Receptors/metabolism , Adult , Animals , CHO Cells , Cricetinae , Cricetulus , Cytokines/metabolism , Disease Models, Animal , Fibroblasts/drug effects , Fibroblasts/metabolism , Fibroblasts/pathology , Humans , Hyaluronic Acid/metabolism , Inflammation/pathology , Lipopolysaccharides/pharmacology , NF-kappa B/metabolism , Osteoarthritis/pathology , Protein Binding/drug effects , Protein Transport/drug effects , RatsABSTRACT
BACKGROUND: Cancer during pregnancy is rare, occurring one in every 1,000 pregnancies. Cancer itself rarely harms the baby and some cancer treatments are safe during pregnancy. However, treatment dilemmas often occur. METHODS: Descriptive study was conducted at B. P. Koirala memorial cancer hospital. Case records of women with cancer and pregnancy from January 2001 to February 2012 were analyzed regarding their clinical details, treatment, follow-up and feto-maternal outcome. RESULTS: Nineteen women, of 17 to 40 years had cancer with pregnancy. Observed cancers with pregnancy were: leukemia (4), head and neck (3), ovary (3), cervix (2), rectum (2), breast (1), Non-Hodgkin's lymphoma (1), osteosarcoma (1), spinal cord (1) and vulva (1). Seven women (36%) presented in the second trimester and six women (32%) presented in the first and third trimester each. Seven (36%) women opted for termination of pregnancy for definitive treatment, five (26%) deferred treatment until delivery. Among the seven (36%) that accepted definitive treatment along with pregnancy, fetal demise occurred in three and delivery of healthy baby occurred in four. Nine babies born to mothers with cancer during pregnancy till date have normal growth and development. Total 10 (52%) of the mothers are in remission, six (32%) have died from disease. CONCLUSIONS: Cancers during pregnancy, more common in younger women, posed treatment challenges. Definitive cancer treatment could have greater fetal risk during the first trimester but could be offered with more acceptable risk in the second and third trimesters.
Subject(s)
Pregnancy Complications, Neoplastic/epidemiology , Adolescent , Adult , Female , Fetal Mortality , Gestational Age , Humans , Male , Maternal Mortality , Medical Audit , Nepal/epidemiology , Outcome Assessment, Health Care , Pregnancy , Pregnancy Complications, Neoplastic/classification , Pregnancy Complications, Neoplastic/therapy , Pregnancy Trimesters , Remission Induction , Young AdultABSTRACT
PIP: It is argued that projects and programs designed to meet the practical needs of men, women, and children in communities should also focus on meeting the strategic gender needs of women. This paper shows what project planners can do to ensure women¿s participation in the design and maintenance of development projects without increasing their workloads, and with the goal of raising their status in the family and society, as well as challenging men's prejudice. It utilizes the framework of strategic and practical gender needs in the context of the drinking-water sector, to argue that understanding how these needs are linked is important to the sustainability of drinking-water projects. Overall, this paper pointed out the great need to involve women in the management of water projects in order for it to be effective in reducing the burdens of the people. Moreover, all development initiatives, including improvements in water supply, should have explicit focus on improving women status and increasing their confidence. Meeting such gender needs requires real commitment from concerned individuals on all levels as well as budgetary provision to enhance the capacity of the involved sector, and heighten their awareness. Fulfillment of their strategic gender needs, will in turn, contribute to the sustainability of water projects.^ieng
