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Myelodysplastic syndromes (MDS) and myelodysplastic/myeloproliferative neoplasms (MDS/MPN) are bone marrow disorders characterized by cytopenias and progression to acute myeloid leukemia. Hypomethylating agents (HMAs) are Food and Drug Administration-approved therapies for MDS and MDS/MPN patients. HMAs have improved patients' survival and quality of life when compared with other therapies. Although HMAs are effective in MDS and MDS/MPN patients, they are associated with significant toxicities that place a large burden on patients. Our goal is to develop a safer and more effective HMA from natural products. We previously reported that black raspberries (BRBs) have hypomethylating effects in the colon, blood, spleen, and bone marrow of mice. In addition, BRBs exert hypomethylating effects in patients with colorectal cancer and familial adenomatous polyposis. In the current study, we conducted a pilot clinical trial to evaluate the hypomethylating effects of BRBs in patients with low-risk MDS or MDS/MPN. Peripheral blood mononuclear cells (PBMCs) were isolated before and after three months of BRB intervention. CD45

Little is known about the distribution and ecology of juvenile green sturgeon throughout their range and specifically within California’s Central Valley and the San Francisco Bay Estuary. The objective of our study was to identify rearing movement patterns of juvenile green sturgeon and their interaction with Delta and Bay habitats. Acoustic biotelemetry was used to record juvenile movements and residence within different regions of this highly modified watershed. We document a diversity of movements exhibited by 31 acoustically tagged juvenile green sturgeon monitored over a 9-month period. The juvenile sturgeon either (1) remained in the Delta or moved (2) into the Carquinez Straits, (3) into San Pablo Bay, (4) into San Pablo Bay but returned to Delta, (5) through the estuary and likely left through the mouth of the bay, (6) or left the estuary only to return later. Spatiotemporal habitat quality varies within and between Delta and Bay habitats. Variation in water year and hydrologic connectivity are likely to influence habitat quality and quantity, and behaviors may be differentially optimized for fitness in response to environmental variation. Our results provide initial evidence supporting multiple behavioral movement patterns and a broad use of regional habitats for green sturgeon in the California Central Valley. This apparent behavioral diversity observed through movement patterns may ensure long-term success of all population segments over time in an extremely variable environment, ultimately supporting overall population stability (i.e., the portfolio effect). Enhanced conservation efforts may be needed to protect juvenile green sturgeon and the habitats they rely upon.

Period life expectancy is defined as the average number of additional years someone of a given age would live if current age-specific mortality rates were to stay the same for the remainder of their life. It is one of the world's most widely used population health metrics to summarize, compare and rank the mortality situation of countries, forming the basis for various public health, life insurance and retirement policies (Luy et al., 2020). This is because a country's life expectancy reflects, among other things, its existing socio-economic conditions and the quality of its public health and healthcare infrastructure (Ho & Hendi, 2018). Although immigration and emigration events are routinely factored into estimates of life expectancy, the potential impact of the unique – and typically low mortality – of international migrants (Aldridge et al., 2018; Shor & Roelfs, 2021) on national life expectancy has received little attention. A handful of studies have so far been limited to Australia (Page et al., 2007) and the United States (US) (Hendi & Ho, 2021; Mehta et al., 2016; Preston & Elo, 2014). In the context of rising shares of migrants in many countries (United Nations, 2019), alongside their “ageing in place” (Ciobanu et al., 2017), the extent to which the mortality of international migrants affects national population health demands attention.Here, we aim to understand whether and how the mortality of international migrants affect the estimation and comparison of national life expectancy in four Nordic countries. Denmark, Finland, Norway and Sweden are all high-income countries in the latest stage of health transition. They have a long tradition of collaboration with shared features of social policy and universal welfare (Knudsen et al., 2019). Despite this, health inequality gaps remain large in the region relative to countries with less developed welfare systems (Mackenbach et al., 2016). International migration has been the major driving force behind population growth in the Nordic region in the past few decades. All four countries have experienced large increases in their absolute and relative numbers of migrants, alongside a transformation in migrant inflows from principally intra-Nordic flows to flows from all over the world (Karlsdottir et al., 2018). In recent years, life expectancy gains have slowed within the region; national life expectancies have also converged, although men and women in Denmark and men in Finland continue to lag some way behind Norway and Sweden (Knudsen et al., 2019).

BackgroundUnderstanding movement patterns of anadromous fishes is critical to conservation management of declining wild populations and preservation of habitats. Yet, infrequent observations of individual animals fundamentally constrain accurate descriptions of movement dynamics.MethodsIn this study, we synthesized over a decade (2006–2018) of acoustic telemetry tracking observations of green sturgeon (Acipenser medirostris) in the Sacramento River system to describe major anadromous movement patterns.ResultsWe observed that green sturgeon exhibited a unimodal in-migration during the spring months but had a bimodal distribution of out-migration timing, split between an ‘early’ out-migration (32%) group during May - June, or alternatively, holding in the river until a ‘late’ out-migration (68%), November - January. Focusing on these out-migration groups, we found that river discharge, but not water temperature, may cue the timing of migration, and that fish showed a tendency to maintain out-migration timing between subsequent spawning migration events.ConclusionsWe recommend that life history descriptions of green sturgeon in this region reflect the distinct out-migration periods described here. Furthermore, we encourage the continued use of biotelemetry to describe migration timing and life history variation, not only this population but other green sturgeon populations and other species.

Obesity is a universal epidemic and underlies comorbidities including type 2 diabetes and cardiovascular disease. Adipocytes store dietary metabolites within lipid droplets to avoid lipotoxicity and to provide energy to the body through metabolic processes such as lipid and glucose metabolism. Collaborative studies demonstrated that the extracellular matrix protein procollagen C-endopeptidase enhancer 2 (PCPE2) mediates the ability of scavenger receptor BI (SR-BI) to transport cholesterol into hepatocytes and adipocytes.It remains unknown if SR-BI and PCPE2 also play roles in other adipocyte metabolic processes and if their cooperative partnership is conserved. This knowledge gap prompted us to test the hypothesis that PCPE2 facilitates SR-BI’s ability to regulate metabolic processes in adipocytes. We validated a differentiated adipocyte model system using mesenchymal stem cells isolated from ears of wild-type (WT), SR-BI -/- , and PCPE2 - /- mice by demonstrating increased lipid droplet formation, mRNA expression of adipogenesis markers, and secretion of adiponectin over a 9-day post-differentiation period across genotypes. We used this model system to test the roles SR-BI and PCPE2in glucose and lipid metabolism. Compared to WT adipocytes, glucose uptake in SR-BI -/- and PCPE2 -/- adipocytes was decreased by 5.4% and 12.6%, respectively, despite increases in surface GLUT4 levels. We also observed that adipocytes lacking PCPE2 had an increased response to insulin through AKT signaling, which may be responsible for the increased surface expression of GLUT4. Loss of SR-BI or PCPE2 in adipocytes did not affect palmitic acid uptake, despite elevated mRNA expression of fatty acid transporters ( Cd36, Fabp4 ) compared to WT adipocytes. SR-BI -/- and PCPE2 -/- adipocytes showed an inverse response to insulin when measuring phosphorylation of fatty acid regulatory proteins (AMPK, ACC) compared to WT adipocytes, suggesting altered lipid metabolism in adipocytes lacking SR-BI or PCPE2. Together, our data suggest that, individually, SR-BI or PCPE2 is important for glucose uptake and fatty acid regulation, but not fatty acid uptake. Further investigations will determine whether a partnership is required for optimal metabolic function in adipocytes.

Cellular membranes are not homogenous mixtures of proteins; rather, they are segregated into microdomains on the basis of preferential association between specific lipids and proteins. These microdomains, called lipid rafts, are well known for their role in receptor signaling on the plasma membrane (PM) and are essential to such cellular functions as signal transduction and spatial organization of the PM. A number of disease states, including atherosclerosis and other cardiovascular disorders, may be caused by dysfunctional maintenance of lipid rafts. Lipid rafts do not occur only in the PM but also have been found in intracellular membranes and extracellular vesicles (EVs). Here, we focus on discussing newly discovered functions of lipid rafts and microdomains in intracellular membranes, including lipid and protein trafficking from the ER, Golgi bodies, and endosomes to the PM, and we examine lipid raft involvement in the production and composition of EVs. Because lipid rafts are small and transient, visualization remains challenging. Future work with advanced techniques will continue to expand our knowledge about the roles of lipid rafts in cellular functioning.

A series of habitat suitability models were created based upon 2-dimensional tracking of Green Sturgeon and hydraulic simulations. This is an effort to better understand the relationship between the population decline, habitat suitability, and knowledge of the remaining post-dam era habitat available to Green Sturgeon. Records of the movements of Green Sturgeon were collected using a refined acoustic telemetry system (Vemco Ltd., Vemco Positioning System [VPS]) in three pools, the first at rkm 377.0 at the confluence with the Sacramento River of Antelope Creek, the second at rkm 407.5 at the confluence with Inks Creek, and the third at rkm 426.0 at the confluence with Paynes Creek near Red Bluff over a period of two years. The Flow and Sediment Transport with Morphologic Evolution of Channels (FaSTMECH) model was used to simulate depth and velocity. Previously developed habitat suitability curves for spawning Green Sturgeon within the study area were coupled to two-dimensional hydraulic simulations to estimate Weighted Usable Area (WUA), a metric of suitable habitat area within each of the studied reaches. The effect of changing river discharges on suitable spawning habitat for Green Sturgeon was examined over a six-year period, the first of which had normal rain conditions and the following years drought conditions. The peak amounts of spawning habitat in the pool on the Sacramento River at the rkm 377 was the same for all six years, roughly 8000 square meters. The constancy in the amount of WUA, in the face of decreasing rates of discharge may explain why these sites are occupied from year to year. The amount of spawning habitat in the pool at rkm 426, decreased during the four-year drought period, from 2012 to 2015. This may make it less favorable for occupation by Green Sturgeon.

Swimming behavior of Chinook Salmon (Oncorhynchus tshawytscha) smolts affects transit time, route selection and survival in complex aquatic ecosystems. Behavior quantified at the river reach and junction scale is of particular importance for route selection and predator avoidance, though few studies have developed field-based approaches for quantifying swimming behavior of juvenile migratory fishes at this fine spatial scale. Two-dimensional acoustic fish telemetry at a river junction was combined with a three-dimensional hydrodynamic model to estimate in situ emigration swimming behavior of federally-threatened juvenile Chinook salmon smolts. Fish velocity over ground was estimated from telemetry, while the hydrodynamic model supplied simultaneous, colocated water velocities, with swimming velocity defined by the vector difference of the two velocities. Resulting swimming speeds were centered around 2 body lengths/second, and included distinct behaviors of positive rheotaxis, negative rheotaxis, lateral swimming, and passive transport. Lateral movement increased during the day, and positive rheotaxis increased in response to local hydrodynamic velocities. Swim velocity estimates were sensitive to the combination of vertical shear in water velocities and vertical distribution of fish.

Description, Altered ubiquitin conjugation is a mechanism by which mammalian cells can respond to fluctuating pCO2., The identification of CO2-binding proteins is crucial to understanding CO2-regulated molecular processes. CO2 can form a reversible posttranslational modification through carbamylation of neutral N-terminal α-amino or lysine ε-amino groups. We have previously developed triethyloxonium (TEO) ion as a chemical proteomics tool for covalent trapping of carbamates, and here, we deploy TEO to identify ubiquitin as a mammalian CO2-binding protein. We use 13C-NMR spectroscopy to demonstrate that CO2 forms carbamates on the ubiquitin N terminus and ε-amino groups of lysines 6, 33, 48, and 63. We demonstrate that biologically relevant pCO2 levels reduce ubiquitin conjugation at lysine-48 and down-regulate ubiquitin-dependent NF-κB pathway activation. Our results show that ubiquitin is a CO2-binding protein and demonstrates carbamylation as a viable mechanism by which mammalian cells can respond to fluctuating pCO2.

Obesity continues to be an epidemic in the United States, placing individuals at a higher risk of developing other comorbidities such as type 2 diabetes and cardiovascular disease. Adipocytes accumulate excess triglycerides and undergo hypertrophic expansion as a protective mechanism to avoid lipotoxicity. Adipocytes are also highly dynamic cells that perform many key metabolic processes including lipid and glucose metabolism. Recent collaborative studies demonstrated that SR-BI function may require the extracellular matrix protein procollagen C-endopeptidase enhancer 2 (PCPE2) in to regulate cholesterol transport in hepatocytes and adipocytes. However, the roles of SR-BI and PCPE2 in other adipocyte metabolic processes remain unclear. This knowledge gap prompted our novel hypothesis that PCPE2 facilitates SR-BI’s ability to regulate metabolic processes in adipocytes. To test our hypothesis, we have optimized an adipocyte model system where mesenchymal stem cells isolated from the outer ears of wild-type (WT), SR-BI knockout (SR-BI -/- ), and PCPE2 -/- mice were differentiated into adipocyte-like cells. We validated our adipocyte model system by demonstrating increased Oil Red O staining, mRNA expression of adipogenesis markers, and adiponectin secretion over a 9-day post-differentiation period for all three genotypes. This model system was then used to test the roles of SR-BI and PCPE2 on various metabolic functions. Preliminary data suggests that absence of SR-BI or PCPE2 slightly impairs fatty acid uptake into adipocytes despite elevated gene expression of fatty acid transporters (FABP4, CD36) compared to WT. These data further suggest that SR-BI may rely on PCPE2 to aid in transport of fatty acids. Adipocytes lacking SR-BI or PCPE2 also displayed higher levels of mitochondrial reactive oxygen species, indicating that SR-BI and PCPE2 may be important for maintaining healthy mitochondria in adipocytes. These changes in function do not appear to depend on SR-BI oligomerization, as BS 3 crosslinking studies demonstrated the presence of SR-BI oligomers in adipocytes despite the loss of PCPE2. Altogether, these data suggest that SR-BI and PCPE2 may be reliant on each other and are important for mediating metabolic processes in adipocytes.

RATIONALE: Single nucleotide polymorphisms (SNPs) near the ILRUN gene are genome-wide significantly associated with plasma lipid traits and coronary artery disease (CAD) but the biological basis of this association is unknown. OBJECTIVE: To investigate the role of ILRUN in plasma lipid and lipoprotein metabolism. METHODS AND RESULTS: ILRUN encodes a protein that contains a ubiquitin-associated (UBA)-like domain, suggesting that it may interact with ubiquitinylated proteins. We generated mice globally deficient for Ilrun (IlrunKO) and found they had significantly lower plasma cholesterol levels resulting from reduced liver lipoprotein production. Liver transcriptome analysis uncovered altered transcription of genes downstream of lipid-related transcription factors, particularly PPARα, and livers from IlrunKO mice had increased PPARα protein. Human ILRUN was shown to bind to ubiquitinylated proteins including PPARα, and the UBA-like domain of ILRUN was found to be required for its interaction with PPARα. CONCLUSIONS: These findings establish ILRUN as a novel regulator of lipid metabolism that promotes hepatic lipoprotein production. Our results also provide functional evidence that ILRUN may be the casual gene underlying the observed genetic associations with plasma lipids at 6p21 in human.

We used acoustic telemetry to determine the spatial and temporal overlap between adult Green Sturgeon movements and areas affected by dredging within the San Francisco Estuary. Autonomous receivers were deployed for 3 years within the lower Estuary at priority locations to assess the potential for adverse effects on Green Sturgeon. Green Sturgeon were present at the designated placement sites during all months of the year but more were detected during two time periods (February–March and June–September). Of the 134 tagged fish detected in the estuary, 109 (81%) were detected at one or more dredged or dredged material placement sites. The median duration of residence at dredged material placement sites was 72.5 min near the Carquinez Strait, 141.1 min in San Pablo Bay, and 37.1 min near Alcatraz Island in San Francisco Bay. The median duration of residence at the dredged San Pablo Channel was 77.5 min. Nine fish were detected with depth sensing transmitters. The majority of detections (95.2%) from these fish were at depths greater than five meters. Combined with information regarding the specific impacts of dredging on Green Sturgeon (e.g., suspended sediments, toxicity, entrainment, and behavior changes), these spatiotemporal data could be used to make recommendations for reassessing best management practices.

Objective The aim of this study was to calculate the societal economic burden of shoulder pain in patients on the orthopaedic waiting list at an Australian public hospital and calculate the cost (from the government’s perspective) of care delivered by the hospital for those patients. Methods A cost-of-illness analysis was undertaken in a cohort of 277 orthopaedic patients on the Gold Coast in Australia. Outcomes included a health care costs and impacts questionnaire, work absenteeism, presenteeism questionnaires (Work Limitations Questionnaire (WLQ) and Work Productivity and Activity Impairment Questionnaire (WPAI)) and hospital care provision over a 2-year period. Results The mean societal cost of healthcare and domestic support was AU$20.72 per day (AU$7563 annually) per patient on the orthopaedic waiting list. When absenteeism and presenteeism were included, the cost per patient who was employed was AU$38.04 per day (AU$13 885 annually) calculated with the WLQ and AU$61.31 per day (AU$22 378 annually) calculated with the WPAI. The mean per-patient cost to government of public hospital care was AU$2622 in Year 1 and AU$3835.78 (s.d. 4961.28) over 2 years. The surgical conversion rate was 22%, and 51% of hospital care cost was attributable to outpatient services. Conclusions Public orthopaedic shoulder waiting lists create a large economic burden for society; few referrals require surgery and just over half the hospital care costs are for out-patient services. New models of care that better manage shoulder pain and identify surgical candidates before orthopaedic referral could reduce this burden. What is known about the topic? Little is known about the cost of shoulder pain in Australia, or the cost of patients referred for public orthopaedic care. What does this paper add? This article quantifies the costs of shoulder pain and the value of lost production from shoulder pain. The time spent waiting for public hospital orthopaedic appointments and the costs associated with waiting demonstrate that the time spent on a waiting list is a key driver of the economic burden. What are the implications for practitioners? Greater resourcing to reduce public orthopaedic shoulder waiting lists may be helpful, but system change is also required. Earlier and more accurate identification of surgical cases could reduce inefficient referrals and improve hospital productivity. Collaboration between clinicians and policy makers is needed to design more economically efficient shoulder care.

Vast sections of the Sacramento River have been listed as critical habitat by the National Marine Fisheries Service for green sturgeon spawning (Acipenser medirostris), yet spawning is known to occur at only a few specific locations. This study reveals the range of physical habitat variables selected by adult green sturgeon during their spawning period. We integrated fine-scale fish positions, physical habitat characteristics, discharge, bathymetry, and simulated velocity and depth using a two-dimensional hydraulic model (FaSTMECH). The objective was to create habitat suitability curves for depth, velocity, and substrate type within three known spawning locations over two years. An overall cumulative habitat suitability score was calculated that averaged the depth, velocity, and substrate scores over all fish, sites, and years. A weighted usable area index was calculated throughout the sampling periods for each of the three sites. Cumulative results indicate that the microhabitat characteristics most preferred by green sturgeon in these three spawning locations were velocities between 1.0 and 1.1 m/s, depths of 8–9 m, and gravel and sand substrate. This study provides guidance for those who may in the future want to increase spawning habitat for green sturgeon within the Sacramento River.

Polarization of macrophages to proinflammatory M1 and to antiinflammatory alternatively activated M2 states has physiological implications in the development of experimental hypertension and other pathological conditions. 12/15-Lipoxygenase (12/15-LO) and its enzymatic products 12(S)- and 15(S)-hydroxyeicosatetraenoic acid (HETE) are essential in the process since disruption of the gene encoding 12/15-LO renders the mice unsusceptible to hypertension. The objective was to test the hypothesis that M2 macrophages catabolize 12(S)-HETE into products that are incapable of promoting vasoconstriction. Cultured M2 macrophages metabolized externally added [(14)C]12(S)-HETE into more polar metabolites, while M1 macrophages had little effect on the catabolism. The major metabolites were identified by mass spectrometry as (ω-1)-hydroxylation and β-oxidation products. The conversion was inhibited by both peroxisomal β-oxidation inhibitor, thioridazine, and cytochrome P450 inhibitors. Quantitative PCR analysis confirmed that several cytochrome P450 enzymes (CYP2E1 and CYP1B1) and peroxisomal β-oxidation markers were upregulated upon M2 polarization. The identified 12,19-dihydroxy-5,8,10,14-eicosatetraenoic acid and 8-hydroxy-6,10-hexadecadienoic acid metabolites were tested on abdominal aortic rings for biological activity. While 12(S)-HETE enhanced vasoconstrictions to angiotensin II from 15% to 25%, the metabolites did not. These results indicate that M2, but not M1, macrophages degrade 12(S)-HETE into products that no longer enhance the angiotensin II-induced vascular constriction, supporting a possible antihypertensive role of M2 macrophages.

Apolipoprotein (apo)A-I is an organizing scaffold protein that is critical to high-density lipoprotein (HDL) structure and metabolism, probably mediating many of its cardioprotective properties. However, HDL biogenesis is poorly understood, as lipid-free apoA-I has been notoriously resistant to high-resolution structural study. Published models from low-resolution techniques share certain features but vary considerably in shape and secondary structure. To tackle this central issue in lipoprotein biology, we assembled a team of structural biologists specializing in apolipoproteins and set out to build a consensus model of monomeric lipid-free human apoA-I. Combining novel and published cross-link constraints, small-angle X-ray scattering (SAXS), hydrogen-deuterium exchange (HDX) and crystallography data, we propose a time-averaged model consistent with much of the experimental data published over the last 40 years. The model provides a long-sought platform for understanding and testing details of HDL biogenesis, structure and function.

HDL normally transports about 50–70% of plasma sphingosine 1-phosphate (S1P), and the S1P in HDL reportedly mediates several HDL-associated biological effects and signaling pathways. The HDL receptor, SR-BI, as well as the cell surface receptors for S1P (S1PRs) may be involved partially and/or completely in these HDL-induced processes. Here we investigate the nature of the HDL-stimulated interaction between the HDL receptor, SR-BI, and S1PR1 using a protein-fragment complementation assay and confocal microscopy. In both primary rat aortic vascular smooth muscle cells and HEK293 cells, the S1P content in HDL particles increased intracellular calcium concentration, which was mediated by S1PR1. Mechanistic studies performed in HEK293 cells showed that incubation of cells with HDL led to an increase in the physical interaction between the SR-BI and S1PR1 receptors that mainly occurred on the plasma membrane. Model recombinant HDL (rHDL) particles formed in vitro with S1P incorporated into the particle initiated the internalization of S1PR1, whereas rHDL without supplemented S1P did not, suggesting that S1P transported in HDL can selectively activate S1PR1. In conclusion, these data suggest that S1P in HDL stimulates the transient interaction between SR-BI and S1PRs that can activate S1PRs and induce an elevation in intracellular calcium concentration.

Green Sturgeon (Acipenser medirostris) and White Sturgeon (A. transmontanus) are closely related species inhabiting the San Francisco Estuary (SFE) and Sacramento River. Both spawn in the Sacramento River. White Sturgeon typically remain within SFE throughout their lives while subadult and adult Green Sturgeon make marine migrations. These broad-scale patterns are known. However, the reach-specific timing of spawning migrations is not, and little is known about seasonal movements of juveniles and subadults. We tagged 41 Green and 160 White Sturgeon with acoustic transmitters (2010–2012). Analyses also included 167 previously tagged sturgeon. Over 300 receivers deployed throughout the Sacramento River, SFE, and nearshore Pacific Coast detected movements. Adult Green Sturgeon moved quickly though SFE to their spawning reach, using the mainstem through the Delta. 5% of spawning Green Sturgeon remained on the spawning reach for over 300 days. White Sturgeon spent significantly less time in their spawning reach before exiting (32 ± 48.2 days vs. 197 ± 97.3 days) and instead selected northern Delta sloughs over the mainstem for pre-spawning migrations. Even at earlier life stages, Green Sturgeon had more marine distributions than White Sturgeon. Juveniles and subadults of both species moved between the San Francisco Bay and Delta throughout the year, but only White Sturgeon overwintered in the Delta. Additionally, we refine several life history parameters for sturgeon in California. We provide a comprehensive characterization of Green and White Sturgeon movements within the SFE and Sacramento River across life stages. These analyses will inform reach-specific management of each species.

Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface.

Cellular membranes are not homogenous mixtures of proteins; rather, they are segregated into microdomains on the basis of preferential association between specific lipids and proteins. These microdomains, called lipid rafts, are well known for their role in receptor signaling on the plasma membrane (PM) and are essential to such cellular functions as signal transduction and spatial organization of the PM. A number of disease states, including atherosclerosis and other cardiovascular disorders, may be caused by dysfunctional maintenance of lipid rafts. Lipid rafts do not occur only in the PM but also have been found in intracellular membranes and extracellular vesicles (EVs). Here, we focus on discussing newly discovered functions of lipid rafts and microdomains in intracellular membranes, including lipid and protein trafficking from the ER, Golgi bodies, and endosomes to the PM, and we examine lipid raft involvement in the production and composition of EVs. Because lipid rafts are small and transient, visualization remains challenging. Future work with advanced techniques will continue to expand our knowledge about the roles of lipid rafts in cellular functioning.

Southern distinct population segment (sDPS) juvenile Green Sturgeon are thought to rear in the Sacramento-San Joaquin Delta for two to four years. The southern population segment has been listed as “threatened” under the Endangered Species Act. Habitat loss is cited as a significant driver in declines of the population. Specific to juvenile rearing, much of the historic floodplain habitat has been lost and replaced by leveed channelized waterways. In addition to geomorphic alterations, other direct and indirect activities such as water export, dredging, and the introduction of invasive species may have significant impacts to the viability of the species. The objective of this tracking study was to provide a first of its kind look at the movements and behaviors of wild caught juvenile Green Sturgeon within the putative nursery grounds of the Sacramento-San Joaquin Delta. We acoustically tagged and continuously tracked six yearling juvenile Green Sturgeon with mean total length (TL) of 50.6cm (SD = 5.3) and mean mass of 563.3g (SD = 151.7). We performed continuous mobile tracking for a period of up to five days per individual while recording GPS coordinates, depths, and water temperature at the sturgeon’s location. We used first-passage times, defined as the amount of time spent within a given area, for which a radius was determined post hoc. First passage times were used as a metric for parsing bouts of localized movements from directed movements. Our results indicate that juvenile Green Sturgeon largely oriented at or near the bottom. Depths utilized by all sturgeon ranged from >3m to 20m across all tracks. Individuals exhibited fidelity to the San Joaquin River Channel with only one individual leaving the main channel. Juvenile green sturgeon were shown to exhibit both positive and negative rheotaxis during sustained directed movements under independent ebb tidal cycles. This study provides the first detailed examination into the early life movements and behaviors of a rare species within their putative nursery grounds.

Despite the popular use of hummingbird feeders, there are limited studies evaluating the effects of congregation, sharing food resources and increased contact when hummingbirds visit feeders in urban landscapes. To evaluate behavioral interactions occurring at feeders, we tagged 230 individuals of two species, Anna's and Allen's Hummingbirds, with passive integrated transponder tags and recorded their visits with RFID transceivers at feeders. For detecting the presence of tagged birds, we developed an RFID equipped feeding station using a commercially available antenna and RFID transceiver. Data recorded included the number of feeder visits, time spent at the feeder, simultaneous feeder visitation by different individuals, and identifying which feeders were most commonly visited by tagged birds. For the study period (September 2016 to March 2018), 118,017 detections were recorded at seven feeding stations located at three California sites. The rate of tagged birds returning to RFID equipped feeders at least once was 61.3% (141/230 birds). Females stayed at feeders longer than males per visit. We identified primary, secondary and tertiary feeders at Sites 2 and 3, according to the frequency of visitation to them, with a mean percentage of 86.9% (SD±19.13) visits to a primary feeder for each tagged hummingbird. During spring and summer, hummingbirds visited feeders most often in morning and evening hours. Feeder visits by males overlapped in time with other males more frequently than other females. The analysis of the contact network at the feeders did not distinguish any significant differences between age or sex. Although most hummingbirds visited the feeders during the daytime, our system recorded night feeder visitations (n = 7 hummingbirds) at one site. This efficient use of RFID technology to characterize feeder visitations and contact networks of hummingbirds in urban habitats could be used in the future to elucidate behaviors, population dynamics and community structure of hummingbirds visiting feeders.

Objective— Aim of this study was to evaluate changes in LCAT (lecithin:cholesterol acyltransferase) concentration and activity in patients with an acute coronary syndrome, to investigate if these changes are related to the compromised capacity of HDL (high-density lipoprotein) to promote endothelial nitric oxide (NO) production, and to assess if rhLCAT (recombinant human LCAT) can rescue the defective vasoprotective HDL function. Approach and Results— Thirty ST-segment–elevation myocardial infarction (STEMI) patients were enrolled, and plasma was collected at hospital admission, 48 and 72 hours thereafter, at hospital discharge, and at 30-day follow-up. Plasma LCAT concentration and activity were measured and related to the capacity of HDL to promote NO production in cultured endothelial cells. In vitro studies were performed in which STEMI patients’ plasma was added with rhLCAT and HDL vasoprotective activity assessed by measuring NO production in endothelial cells. The plasma concentration of the LCAT enzyme significantly decreases during STEMI with a parallel significant reduction in LCAT activity. HDL isolated from STEMI patients progressively lose the capacity to promote NO production by endothelial cells, and the reduction is related to decreased LCAT concentration. In vitro incubation of STEMI patients’ plasma with rhLCAT restores HDL ability to promote endothelial NO production, possibly related to significant modification in HDL phospholipid classes. Conclusions— Impairment of cholesterol esterification may be a major factor in the HDL dysfunction observed during acute coronary syndrome. rhLCAT is able to restore HDL-mediated NO production in vitro, suggesting LCAT as potential therapeutic target for restoring HDL functionality in acute coronary syndrome.

Nogo-B receptor (NgBR) was identified as a specific receptor for binding Nogo-B and is essential for the stability of Niemann-Pick type C2 protein (NPC2) and NPC2-dependent cholesterol trafficking. Here, we report that NgBR expression levels decrease in the fatty liver and that NgBR plays previously unrecognized roles in regulating hepatic lipogenesis through NPC2-independent pathways. To further elucidate the pathophysiological role of NgBR in mammals, we generated NgBR liver-specific knockout mice and investigated the roles of NgBR in hepatic lipid homeostasis. The results showed that NgBR knockout in mouse liver did not decrease NPC2 levels or increase NPC2-dependent intracellular cholesterol levels. However, NgBR deficiency still resulted in remarkable cellular lipid accumulation that was associated with increased free fatty acids and triglycerides in hepatocytes in vitro and in mouse livers in vivo. Mechanistically, NgBR deficiency specifically promotes the nuclear translocation of the liver X receptor alpha (LXRα) and increases the expression of LXRα-targeted lipogenic genes. LXRα knockout attenuates the accumulation of free fatty acids and triglycerides caused by NgBR deficiency. In addition, we elucidated the mechanisms by which NgBR bridges the adenosine monophosphate–activated protein kinase alpha signaling pathway with LXRα nuclear translocation and LXRα-mediated lipogenesis. Conclusion: NgBR is a specific negative regulator for LXRα-dependent hepatic lipogenesis. Loss of NgBR may be a potential trigger for inducing hepatic steatosis. (Hepatology 2016;64:1559-1576)

Animal fitness is influenced by diverse assemblages of internal and external symbionts. These assemblages often change throughout host ontogeny, but the mechanisms that underlie these changes and their consequences for host fitness are seldom revealed. Here we examine a cleaning symbiosis between crayfish and an assemblage of ectosymbiotic branchiobdellidan worms to uncover what mechanisms drive changes in symbiont composition during host ontogeny and the consequences of these changes for both the host and symbionts. In surveys of a North Carolina river, the dominant worm species shifted from Cambarincola philadelphicus to Cambarincola ingens as crayfish (Cambarus bartonii) increased in size. We demonstrate that this shift is a function of host regulation by small crayfish and exclusion by a dominant symbiont on large crayfish. In a controlled lab experiment, small crayfish often removed their symbionts but C. ingens was removed at a higher rate than C. philadelphicus. In contrast, C. ingens had higher survivorship and reproduction than C. philadelphicus on large crayfish. We also measured the effect of each worm species on crayfish growth through ontogeny; neither worm species had an effect on small crayfish but both species had similar positive effects on the growth of large crayfish relative to controls. Evidence from another experiment suggested that intraguild predation by C. ingens caused a decline in C. philadelphicus on large crayfish. We have shown that shifts in partner fitness are a function of host size and that these shifts can involve the succession of symbionts. Further, our results suggest that changes in the outcome of symbioses can remain robust throughout host ontogeny despite interactive mechanisms that lead to shifts in symbiont community structure.

Outmigration survival of acoustic-tagged, hatchery-origin, late-fall-run Chinook salmon (Oncorhynchus tshawytscha) smolts from the Sacramento River was estimated for 5 years (2007–2011) using a receiver array spanning the entire outmigration corridor, from the upper river, through the estuary, and into the coastal ocean. The first 4 years of releases occurred during below-average river flows, while the fifth year (2011) occurred during above-average flows. In 2011, overall outmigration survival was two to five times higher than survival in the other 4 years. Regional survival estimates indicate that most of the improved survival seen in 2011 occurred in the riverine reaches of the outmigration corridor, while survival in the brackish portions of the estuary did not significantly differ among the 5 years. For the 4 low-flow years combined, survival rate in the river was lower in the less anthropogenically modified upper reaches; however, across all regions, survival rate was lowest in the brackish portion of the estuary. Even in the high-flow year, outmigration survival was substantially lower than yearling Chinook salmon populations in other large rivers. Potential drivers of these patterns are discussed, including channelization, water flow, and predation. Finally, management strategies are suggested to best exploit survival advantages described in this study.

Recent evidence suggests that the vasoprotective effects of HDL are impaired in patients during an acute coronary syndrome (ACS). In particular, HDL from ACS patients are defective in promoting nitric oxide (NO) release from cultured endothelial cells (ECs). Lecithin:cholesterol acyltransferase (LCAT) plays a key role in HDL maturation and remodeling, and it is the enzyme responsible for cholesterol esterification in plasma. Very few studies have suggested that LCAT activity is decreased during an acute myocardial infarction, but the possible link between LCAT mass/activity and the HDL dysfunction observed in ACS has never been tested. To test this hypothesis, plasma from 30 STEMI patients was collected at admission, 48 and 72 hours and 30 days after event. LCAT concentration and activity significantly decreased 48 hours after event (-0.49±0.11 μg/ml, PIn vitro studies were performed in which STEMI patients’ plasma was added with rhLCAT and HDL vasoprotective activity assessed by measuring NO production in ECs. In vitro incubation of STEMI patients’ plasma with rhLCAT remodels phospholipids and protein content in HDL and restores HDL ability to promote endothelial NO production (+25%, P=0.03). Impairment of cholesterol esterification may be a major factor in the HDL dysfunction observed during ACS. rhLCAT is able to restore HDL-mediated NO production in vitro, suggesting LCAT as potential therapeutic target for restoring HDL functionality in ACS.

Adipose tissue undergoes distinct structural remodeling in response to increases in fat mass, as in obesity. During this process, the extracellular matrix (ECM) plays a pivotal role with respect to both adipocyte receptor signaling and structural remodeling. One ECM protein, procollagen endopeptidase enhancer 2 (PCPE2), has been shown by our lab to work in partnership with the scavenger receptor class B type I (SR-BI) to regulate adipocyte cholesterol homeostasis. To investigate the functional impact of the partnership between SR-BI and PCPE2, we studied mature adipocytes differentiated from murine embryonic fibroblasts (MEF) and compared to our adipose specific PCPE2 knockout mice. Our studies showed the presence of an SR-BI-PCPE2 protein complex based on the co-immunoprecipitation of PCPE2 with SR-BI in both differentiated MEFs and mouse adipose tissue. Furthermore, stimulated emission depletion (STED) microscopy showed SR-BI and PCPE2 localized on the lipid droplet surface separated by distances ranging from 24 – 90 nm, suggesting that the SR-BI-PCPE2 complex may contain other proteins. Most significantly, a loss of SR-BI function was shown in PCPE2-deficient adipocytes by the SR-BI mediated HDL binding and cholesteryl ester (CE) uptake assays. Compared to differentiated MEFs from Ldlr -/- mice, binding of HDL to differentiated MEFs from Ldlr -/- Pcpe2 -/- mice was reduced by 75%, and HDL cholesteryl oleyl ether uptake was reduced by 50%. Disruption of adipocyte SR-BI function in the absence of PCPE2 also reduced free cholesterol efflux indicating substantial disruption to intracellular cholesterol homeostasis. Moreover, analysis of RNAseq from several human clinical studies show that PCPE2 mRNA expression is positively correlated with fat mass in specific adipose depots, while negatively correlated with plasma insulin, glucose and triglyceride levels, with the latter two more robust in type 2 diabetic populations (FDR < 0.004). Overall, these results suggest that the partnership between SR-BI and PCPE2 in adipocytes plays an important role in cholesterol trafficking and homeostasis which influences fat mass expansion, glucose tolerance and insulin resistance.

Regulatory T (Treg) cells contribute to the anti-inflammatory response during atherogenesis. Here we show that during atherogenesis Treg cells lose Foxp3 expression and their immunosuppressive function, leading to the conversion of a fraction of these cells into T follicular helper (Tfh) cells. We show that Tfh cells are pro-atherogenic and that their depletion reduces atherosclerosis. Mechanistically, the conversion of Treg cells to Tfh cells correlates with reduced expression of IL-2Rα and pSTAT5 levels and increased expression of IL-6Rα. In vitro, incubation of naive T cells with oxLDL prevents their differentiation into Treg cells. Furthermore, injection of lipid-free Apolipoprotein AI (ApoAI) into ApoE−/− mice reduces intracellular cholesterol levels in Treg cells and prevents their conversion into Tfh cells. Together our results suggest that ApoAI, the main protein in high-density lipoprotein particles, modulates the cellular fate of Treg cells and thus influences the immune response during atherosclerosis., Regulatory T (Treg) cells contribute to the anti-inflammatory response during atherogenesis. Here Gaddis et al. show that Apolipoprotein AI prevents the conversion of Treg cells into pro-atherogenic T follicular helper cells, and thus regulates the immune response during atherogenesis.

The formation of the atherosclerotic plaque that is characterized by the accumulation of abnormal amounts of cholesterol-loaded macrophages in the artery wall is mediated by both inflammatory events and alterations of lipid/lipoprotein metabolism. Reverse transport of cholesterol opposes the formation and development of atherosclerotic plaque by promoting high density lipoprotein (HDL)-mediated removal of cholesterol from peripheral macrophages and its delivery back to the liver for excretion into the bile. Although an inverse association between HDL plasma levels and the risk of cardiovascular disease (CVD) has been demonstrated over the years, several studies have recently shown that the antiatherogenic functions of HDL seem to be mediated by their functionality, not always associated with their plasma concentrations. Therefore, assessment of HDL function, evaluated as the capacity to promote cell cholesterol efflux, may offer a better prediction of CVD than HDL levels alone. In agreement with this idea, it has recently been shown that the assessment of serum cholesterol efflux capacity (CEC), as a metric of HDL functionality, may represent a predictor of atherosclerosis extent in humans. The purpose of this narrative review is to summarize the current evidence concerning the role of cholesterol efflux capacity that is important for evaluating CVD risk, focusing on pharmacological evidences and its relationship with inflammation. We conclude that HDL therapeutics are a promising area of investigation but strategies for identifying efficacy must move beyond the idea of simply raising static HDL-cholesterol levels and toward methods of measuring the dynamics of HDL particle remodeling and the generation of lipid-free apolipoprotein A-I (apoA-I). In this way, apoA-I, unlike mature HDL, can promote the greatest extent of cholesterol efflux relieving cellular cholesterol toxicity and the inflammation it causes.

Despite the popular use of hummingbird feeders, there are limited studies evaluating the effects of congregation, sharing food resources and increased contact when hummingbirds visit feeders in urban landscapes. To evaluate behavioral interactions occurring at feeders, we tagged 230 individuals of two species, Anna's and Allen's Hummingbirds, with passive integrated transponder tags and recorded their visits with RFID transceivers at feeders. For detecting the presence of tagged birds, we developed an RFID equipped feeding station using a commercially available antenna and RFID transceiver. Data recorded included the number of feeder visits, time spent at the feeder, simultaneous feeder visitation by different individuals, and identifying which feeders were most commonly visited by tagged birds. For the study period (September 2016 to March 2018), 118,017 detections were recorded at seven feeding stations located at three California sites. The rate of tagged birds returning to RFID equipped feeders at least once was 61.3% (141/230 birds). Females stayed at feeders longer than males per visit. We identified primary, secondary and tertiary feeders at Sites 2 and 3, according to the frequency of visitation to them, with a mean percentage of 86.9% (SD±19.13) visits to a primary feeder for each tagged hummingbird. During spring and summer, hummingbirds visited feeders most often in morning and evening hours. Feeder visits by males overlapped in time with other males more frequently than other females. The analysis of the contact network at the feeders did not distinguish any significant differences between age or sex. Although most hummingbirds visited the feeders during the daytime, our system recorded night feeder visitations (n = 7 hummingbirds) at one site. This efficient use of RFID technology to characterize feeder visitations and contact networks of hummingbirds in urban habitats could be used in the future to elucidate behaviors, population dynamics and community structure of hummingbirds visiting feeders.

Atherosclerosis is a chronic inflammatory disease of large and medium-sized arteries and the underlying cause of cardiovascular disease, a major cause of mortality worldwide. The over-accumulation of modified cholesterol-containing low-density lipoproteins (e.g. oxLDL) in the artery wall and the subsequent recruitment and activation of macrophages contributes to the development of atherosclerosis. The excessive uptake of modified-LDL by macrophages leads to a lipid-laden "foamy" phenotype and pro-inflammatory cytokine production. Modified-LDLs promote foam cell formation in part by stimulating de novo lipid biosynthesis. However, it is unknown if lipid biosynthesis directly regulates foam cell pro-inflammatory mediator production. Lipin-1, a phosphatidate phosphohydrolase required for the generation of diacylglycerol during glycerolipid synthesis has recently been demonstrated to contribute to bacterial-induced pro-inflammatory responses by macrophages. In this study we present evidence demonstrating the presence of lipin-1 within macrophages in human atherosclerotic plaques. Additionally, reducing lipin-1 levels in macrophages significantly inhibits both modified-LDL-induced foam cell formation in vitro, as observed by smaller/fewer intracellular lipid inclusions, and ablates modified-LDL-elicited production of the pro-atherogenic mediators tumor necrosis factor-α, interleukin-6, and prostaglandin E2. These findings demonstrate a critical role for lipin-1 in the regulation of macrophage inflammatory responses to modified-LDL. These data begin to link the processes of foam cell formation and pro-inflammatory cytokine production within macrophages.

Purpose of review The purpose of this study is to provide an update on the role HDL apolipoprotein A-I plays in reducing the risk of cardiovascular disease (CVD) and how it relates to reverse cholesterol transport (RCT). Recent findings Despite numerous studies showing that plasma HDL cholesterol concentrations are correlated with a reduced risk of CVD, pharmacologic elevation of HDL has not shown any beneficial effects to date. In contrast, studies correlating the measure of an individual's plasma cholesterol efflux capacity show greater promise as a tool for assessing CVD risk. Although ATP-binding cassette transporter 1-mediated lipidation of apoA-I is considered the principal source of plasma HDL, it represents only one side of the RCT pathway. Equally important is the second half of the RCT pathway in which the liver scavenger receptor class B1 selectively removes HDL cholesteryl esters for excretion. The combined action of the two enzyme systems is reflected in the overall steady-state concentration of plasma HDL cholesterol. For example, reduced ATP-binding cassette transporter 1-mediated production of nascent HDL lowers plasma HDL concentration, just as an increase in cholesteryl ester uptake by scavenger receptor class B1 reduces HDL levels. Thus, the complexity of intravascular HDL metabolism suggests that steady-state plasma HDL concentrations do not provide adequate information regarding an individual's HDL quality or function. Herein, we describe a new player, procollagen C-endopeptidase enhancer 2, which shows atheroprotective function and influences both sides of RCT by enhancing production and catabolism of HDL cholesteryl esters. Summary The discovery of a new molecule, procollagen C-endopeptidase enhancer 2, implicated in the regulation of HDL cholesteryl ester concentrations suggests that the extracellular matrix and the proteins that regulate its function represent a new and as yet unexplored realm of HDL cholesterol metabolism.

The ectosymbiotic branchiobdellidan Cambarincola ingens Hoffman, 1963 can increase the growth and reduce mortality of the crayfish Cambarus chasmodactylus James, 1966. These results suggest that these two species are engaged in a cleaning symbiosis mutualism. However, the potential benefits C. ingens derives from the association, specifically its reproductive dependence on the crayfish, have not been examined experimentally. We conducted two experiments to determine if C. ingens was reproductively dependent on C. chasmodactylus. In the experiments, worms only laid cocoons on live crayfish. Also, significantly more cocoons were laid on larger crayfish which was consistent with a survey of worm and cocoon abundance on crayfish collected from a local river. These results suggest that C. ingens is dependent on its crayfish host for reproduction. The association between C. chasmodactylus and C. ingens is a mutualism, at least under environmental conditions in which crayfish survival and growth are enhanced by the worms.