The role of adipose tissue-associated macrophages and T lymphocytes in the pathogenesis of inflammatory bowel disease

Inflammatory bowel diseases (IBDs) are chronic inflammatory disorders of the gastrointestinal tract that affect more than 3 million people worldwide, but the pathological etiology is still unknown. The overall purpose of our investigations was to elucidate the possibility of pathological causes of IBD, and therefore, we determined the difference of inflammatory cytokine profiles in adipose tissue macrophages (ATMs) and T lymphocytes (ATTs) obtained near active lesions of IBD; investigated whether the alteration in ATM activation induces genes involved in collagen formation; and evaluated the effects of fatty acid oxidation inhibitors on factors involved in inflammation and collagen production by ATMs in IBD. Adipose tissues (ATs) were collected near active lesions and also at the margin of resected segments of the bowel from IBD patients with ulcerative colitis (UC) and CD (n = 14/group). Normal appearing ATs from control subjects (n = 14) who had colon resection for adenocarcinoma were collected as far away from the cancer lesion as possible to rule out possible changes. Compared with inactive disease lesions, ATMs and ATTs from active lesions released more IL-6, IL-4 and IL-13. Treatments of cytokine IL-4 and/or IL-13 to ATMs reduced iNOS expression but increased Arg-I expression which were exacerbated when treated with T cell- and adipocyte-conditioned medium. However, fatty acid oxidation inhibitors prevented the effects of cytokines IL-4 and/or IL-13 on iNOS and Arg-I expressions. This study was the first to show the effect of IL-4 and IL-13 on collagen formation, through iNOS and Arg-I expressions, that was exacerbated in a condition that mimics in vivo condition of active lesions. Moreover, our study was the first to provide potential benefits of fatty acid oxidation inhibitors to ATMs on preventing collagen formation; thus, providing therapeutic implications for individuals with intestinal fibrosis and stricture lesions, although future study should be guaranteed to elucidate the underlying mechanisms.     

Influence of landscape and vegetation characteristics on herpetofaunal assemblages in Gulf Coastal Plain pine forests

Longleaf pine (Pinus palustris) savanna characterized by open-canopy, diverse herbaceous vegetation, and high amounts of bare soil once covered much of the southeastern United States Coastal Plain. The unique structural and vegetative conditions of this ecosystem support endemic reptiles and amphibians that have declined as longleaf pine forests have been lost or degraded. Private working pine (Pinus spp.) forests managed for timber production now occur throughout the southeastern United States and have replaced much of the historical longleaf pine savanna. The examination of herpetofaunal (reptile, amphibian) communities in private working loblolly pine (P. taeda) landscapes, particularly in the western Gulf Coastal Plain is lacking. Using repeated field surveys and hierarchical community occupancy models, we examined occupancy and species richness of herpetofauna across 81 sites spanning gradients of management practices, vegetative conditions, and soil composition in northwestern Louisiana, USA, 2017–2019. Young pine stands (<6 yr) exhibited structural characteristics most similar to mature longleaf pine reference sites (>30 yr), while mid-aged stands (13–26 yr) often featured closed canopy and dense midstory. Vegetation conditions varied widely depending on landscape characteristics and site-specific disturbance regimes. We documented 43 species of herpetofauna, including 9 open-pine-associated species. Occupancy of open-pine-associated herpetofauna was positively associated with open-canopy and understory conditions, and sandy soil area. Sites providing open-canopy conditions were often occupied by open-pine-associated species regardless of overstory type and disturbance method. Overall richness of herpetofauna was greatest at sites with moderate canopy cover outside of sandy soil regions. Working pine landscapes in the western Gulf Coastal Plain can support diverse herpetofaunal assemblages, including open-pine-associated species, when management practices maintain open-canopy conditions on sandy, upland soils. More broadly, our results provide insight into how forest management practices affect herpetofauna and may guide practices that can contribute to conservation value of working pine forests.     

Lipid Droplet-Binding Protein TIP47 Regulates Hepatitis C Virus RNA Replication through Interaction with the Viral NS5A Protein

The nonstructural protein NS5A has emerged as a new drug target in antiviral therapies for Hepatitis C Virus (HCV) infection. NS5A is critically involved in viral RNA replication that takes place at newly formed membranes within the endoplasmic reticulum (membranous web) and assists viral assembly in the close vicinity of lipid droplets (LDs). To identify host proteins that interact with NS5A, we performed a yeast two-hybrid screen with the N-terminus of NS5A (amino acids 1–31), a well-studied α-helical domain important for the membrane tethering of NS5A. Our studies identified the LD-associated host protein, Tail-Interacting Protein 47 (TIP47) as a novel NS5A interaction partner. Coimmunoprecipitation experiments in Huh7 hepatoma cells confirmed the interaction of TIP47 with full-length NS5A. shRNA-mediated knockdown of TIP47 caused a more than 10-fold decrease in the propagation of full-length infectious HCV in Huh7.5 hepatoma cells. A similar reduction was observed when TIP47 was knocked down in cells harboring an autonomously replicating HCV RNA (subgenomic replicon), indicating that TIP47 is required for efficient HCV RNA replication. A single point mutation (W9A) in NS5A that disrupts the interaction with TIP47 but preserves proper subcellular localization severely decreased HCV RNA replication. In biochemical membrane flotation assays, TIP47 cofractionated with HCV NS3, NS5A, NS5B proteins, and viral RNA, and together with nonstructural viral proteins was uniquely distributed to lower-density LD-rich membrane fractions in cells actively replicating HCV RNA. Collectively, our data support a model where TIP47—via its interaction with NS5A—serves as a novel cofactor for HCV infection possibly by integrating LD membranes into the membranous web.