Effect of a small molecule inhibitor of nuclear factor-κB nuclear translocation in a murine model of arthritis and cultured human synovial cells

A small cell-permeable compound, dehydroxymethylepoxyquinomicin (DHMEQ), does not inhibit phosphorylation and degradation of IκB (inhibitor of nuclear factor-κB [NF-κB]) but selectively inhibits nuclear translocation of activated NF-κB. This study aimed to demonstrate the antiarthritic effect of this novel inhibitor of the NF-κB pathway in vivo in a murine arthritis model and in vitro in human synovial cells. Collagen-induced arthritis was induced in mice, and after onset of arthritis the mice were treated with DHMEQ (5 mg/kg body weight per day). Using fibroblast-like synoviocyte (FLS) cell lines established from patients with rheumatoid arthritis (RA), NF-κB activity was examined by electrophoretic mobility shift assays. The expression of molecules involved in RA pathogenesis was determined by RT-PCR, ELISA, and flow cytometry. The proliferative activity of the cells was estimated with tritiated thymidine incorporation. After 14 days of treatment with DHMEQ, mice with collagen-induced arthritis exhibited decreased severity of arthritis, based on the degree of paw swelling, the number of swollen joints, and radiographic and histopathologic scores, compared with the control mice treated with vehicle alone. In RA FLS stimulated with tumor necrosis factor-α, activities of NF-κB components p65 and p50 were inhibited by DHMEQ, leading to suppressed expression of the key inflammatory cytokine IL-6, CC chemokine ligand-2 and -5, matrix metalloproteinase-3, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1. The proliferative activity of the cells was also suppressed. This is the first demonstration of an inhibitor of NF-κB nuclear translocation exhibiting a therapeutic effect on established murine arthritis, and suppression of inflammatory mediators in FLS was thought to be among the mechanisms underlying such an effect

Building resilience to climate change: MGNREGS, drought and flooding in Odisha

"The Mahatma Gandhi National Rural Employment Guarantee Scheme (MGNREGS) is one of India’s flagship social protection programmes. This paper is part of a series of briefings that analyse how MGNREGS builds the resilience of rural households to different climate shocks. The goal of the series is to identify options for Indian policymakers to integrate climate risk management into MGNREGS. These findings can also provide global policymakers with evidence on how to mainstream climate risk management into social protection programmes, or combine and layer social protection instruments with climate risk management instruments to address poverty in the context of climate change.

The development and characterization of neural-specific continuous cell lines in Drosophila melanogaster

Neurodegenerative diseases affect millions of Americans and are caused by the deterioration of neurons in the central nervous system (CNS). The mutations that cause these neurodegenerative diseases have been modeled in Drosophila melanogaster (fruit fly) transgenic lines that produce flies with neurological defects mirroring the human pathology. Meanwhile, continuous cell lines have transformed biomedical research, making it possible to study biological systems on a scale requiring large numbers of similar cells. However, there is a lack of continuous cell lines of neuronal lineage in Drosophila melanogaster available in the scientific community that would allow for the modeling of neurodegenerative diseases and the detailed study of specific mechanisms within the nervous system. Beyond modeling a neurodegenerative disease phenotype in a fly, continuous cell lines of neuronal lineage would offer a large, homogenous population of cells to potentially study disease pathways and gene function, to test pharmacological targets, and to examine processes occurring before differentiation. The primary objective of this study was to utilize the expression of oncogenic RasV12 and inhibition of the Notch signaling pathway to produce neuronal cell lines in Drosophila melanogaster. In the first approach, the overexpression of RasV12 was limited to cells of neuronal lineage in order to promote their survival over other cell types in the culture. In the second approach, the Notch signaling necessary for segregation of progenitor cells into neuroblasts or epithelial cells was inhibited throughout the Drosophila embryo; without Notch signaling, these progenitor cells all become neuroblasts. For the second objective of the study, cell lines previously established in the lab from constitutive RasV12 expression and that appeared to exhibit neuronal properties were characterized with neuronal lineage markers. This study tests the efficacy of utilizing RasV12 or the inhibition of the Notch signaling pathway for the production of continuous cell lines of neuronal lineage to model and develop therapies for neurodegenerative diseases.2015 and 2016 Undergraduate Research Scholarships to NH, 2014 NSF grant to ASNo embargoAcademic Major: Biolog

Building resilience to climate change: MGNREGS, drought and flooding in Odisha

"The Mahatma Gandhi National Rural Employment Guarantee Scheme (MGNREGS) is one of India’s flagship social protection programmes. This paper is part of a series of briefings that analyse how MGNREGS builds the resilience of rural households to different climate shocks. The goal of the series is to identify options for Indian policymakers to integrate climate risk management into MGNREGS. These findings can also provide global policymakers with evidence on how to mainstream climate risk management into social protection programmes, or combine and layer social protection instruments with climate risk management instruments to address poverty in the context of climate change.

Building resilience to climate change MGNREGS and climate-induced droughts in Sikkim

"The Mahatma Gandhi National Rural Employment Guarantee Scheme (MGNREGS) is one of India’s flagship social protection programmes. This paper is part of a series of briefings that analyse how the scheme builds the resilience of rural households to different climate shocks. The goal of the series is to identify options for Indian policymakers to integrate climate risk management into MGNREGS. It will also provide evidence for global policymakers on how to mainstream climate risk management into social protection programmes, or converge and layer social protection and climate risk management instruments to address poverty in the context of climate change.

NF-κB inhibitor dehydroxymethylepoxyquinomicin suppresses osteoclastogenesis and expression of NFATc1 in mouse arthritis without affecting expression of RANKL, osteoprotegerin or macrophage colony-stimulating factor

Inhibition of NF-κB is known to be effective in reducing both inflammation and bone destruction in animal models of arthritis. Our previous study demonstrated that a small cell-permeable NF-κB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), suppresses expression of proinflammatory cytokines and ameliorates mouse arthritis. It remained unclear, however, whether DHMEQ directly affects osteoclast precursor cells to suppress their differentiation to mature osteoclasts in vivo. The effect of DHMEQ on human osteoclastogenesis also remained elusive. In the present study, we therefore examined the effect of DHMEQ on osteoclastogenesis using a mouse collagen-induced arthritis model, and using culture systems of fibroblast-like synovial cells obtained from patients with rheumatoid arthritis, and of osteoclast precursor cells from peripheral blood of healthy volunteers. DHMEQ significantly suppressed formation of osteoclasts in arthritic joints, and also suppressed expression of NFATc1 along the inner surfaces of bone lacunae and the eroded bone surface, while serum levels of soluble receptor activator of NF-κB ligand (RANKL), osteoprotegerin and macrophage colony-stimulating factor were not affected by the treatment. DHMEQ also did not suppress spontaneous expression of RANKL nor of macrophage colony-stimulating factor in culture of fibroblast-like synovial cells obtained from patients with rheumatoid arthritis. These results suggest that DHMEQ suppresses osteoclastogenesis in vivo, through downregulation of NFATc1 expression, without significantly affecting expression of upstream molecules of the RANKL/receptor activator of NF-κB/osteoprotegerin cascade, at least in our experimental condition. Furthermore, in the presence of RANKL and macrophage colony-stimulating factor, differentiation and activation of human osteoclasts were also suppressed by DHMEQ, suggesting the possibility of future application of NF-κB inhibitors to rheumatoid arthritis therapy

Activation of fibroblast-like synoviocytes derived from rheumatoid arthritis via lysophosphatidic acid-lysophosphatidic acid receptor 1 cascade

INTRODUCTION: Lysophosphatidic acid (LPA) is a bioactive lipid that binds to G protein–coupled receptors (LPA(1–6)). Recently, we reported that abrogation of LPA receptor 1 (LPA(1)) ameliorated murine collagen-induced arthritis, probably via inhibition of inflammatory cell migration, Th17 differentiation and osteoclastogenesis. In this study, we examined the importance of the LPA–LPA(1) axis in cell proliferation, cytokine/chemokine production and lymphocyte transmigration in fibroblast-like synoviocytes (FLSs) obtained from the synovial tissues of rheumatoid arthritis (RA) patients. METHODS: FLSs were prepared from synovial tissues of RA patients. Expression of LPA(1–6) was examined by quantitative real-time RT-PCR. Cell surface LPA(1) expression was analyzed by flow cytometry. Cell proliferation was analyzed using a cell-counting kit. Production of interleukin 6 (IL-6), vascular endothelial growth factor (VEGF), chemokine (C-C motif) ligand 2 (CCL2), metalloproteinase 3 (MMP-3) and chemokine (C-X-C motif) ligand 12 (CXCL12) was measured by enzyme-linked immunosorbent assay. Pseudoemperipolesis was evaluated using a coculture of RA FLSs and T or B cells. Cell motility was examined by scrape motility assay. Expression of adhesion molecules was determined by flow cytometry. RESULTS: The expression of LPA(1) mRNA and cell surface LPA(1) was higher in RA FLSs than in FLSs from osteoarthritis tissue. Stimulation with LPA enhanced the proliferation of RA FLSs and the production of IL-6, VEGF, CCL2 and MMP-3 by FLSs, which were suppressed by an LPA(1) inhibitor (LA-01). Ki16425, another LPA(1) antagonist, also suppressed IL-6 production by LPA-stimulated RA FLSs. However, the production of CXCL12 was not altered by stimulation with LPA. LPA induced the pseudoemperipolesis of T and B cells cocultured with RA FLSs, which was suppressed by LPA(1) inhibition. In addition, LPA enhanced the migration of RA FLSs and expression of vascular cell adhesion molecule and intercellular adhesion molecule on RA FLSs, which were also inhibited by an LPA(1) antagonist. CONCLUSIONS: Collectively, these results indicate that LPA–LPA(1) signaling contributes to the activation of RA FLSs

Familial pancreatic cancer with PALB2 and NBN pathogenic variants: a case report

Background Family history is one of the risk factors for pancreatic cancer. It is suggested that patients with pancreatic cancer who have a familial history harbor germline pathogenic variants of BRCA1 and/or BRCA2 (BRCA1/2), PALB2, or ATM. Recently, some germline variants of familial pancreatic cancers (FPCs), including PALB2, have been detected. Several countries, including Japan, perform screening workups and genetic analysis for pancreatic cancers. We have been carrying out active surveillance for FPC through epidemiological surveys, imaging analyses, and genetic analysis. Case presentation Here, we present the case of a female patient harboring pathogenic variants of PALB2 and NBN, with a family history of multiple pancreatic cancer in her younger brother, her aunt, and her father. Moreover, her father harbored a PALB2 pathogenic variant and her daughter harbored the same NBN pathogenic variant. Given the PALB2 and NBN variants, we designed surveillance strategies for the pancreas, breast, and ovary. Conclusions Further studies are required to develop strategies for managing FPCs to facilitate prompt diagnosis before their progression