Functional identification and characterizationof the diuretic hormone 31 (DH31) signaling system in the Green shore crab, Carcinus maenas

<p>The functional characterization of crustacean neuropeptides and their cognate receptors has not kept pace with the recent advances in sequence determination and, therefore, our understanding of the physiological roles of neuropeptides in this important arthropod sub-phylum is rather limited. We identified a candidate receptor-ligand pairing for diuretic hormone 31 (DH31) in a neural transcriptome of the crab, Carcinus maenas. In insects, DH31 plays species -specific but central roles in many facets of physiology, including fluid secretion, myoactivity, and gut peristalsis but little is known concerning its functions in crustaceans. The C. maenas DH31 transcript codes for a 147 amino acid prepropeptide, and a single receptor transcript translates to a secretin-like (Class B1) G protein-coupled receptor (GPCR). We used an in vitro aequorin luminescence Ca²⁺ mobilization assay to demonstrate that this candidate DH31R is activated byCarcinus and insect DH31s in a dose-dependent manner (EC₅₀ 15–30 nM). Whole mount immunohistochemical and in situ hybridization localization revealed extensive DH31 expressing neurons throughout the central nervous system, most notably in the abdominal ganglion where large, unpaired cells give rise to medial nerves, which terminate in extensive DH31 immunopositive dendritic fields intimately associated with oesophageal musculature. This system constitutes a large and hitherto undescribed neurohemal area adjacent to key muscle groups associated with the gastric system. DH31 expressing neurons were also seen in the cardiac, commissural, oesophageal, and stomatogastric ganglia and intense labeling was seen in dendrites innervating fore- and hindgut musculature but not with limb muscles. These labeling patterns, together with measurement of DH31R mRNA in the heart and hindgut, prompted us test the effects of DH31 on semi-isolated heart preparations. Cardiac superfusion with peptide evoked increased heart rates (10–100 nM). The neuroanatomical distribution of DH31 and its receptor transcripts, particularly that associated with gastric and cardiac musculature, coupled with the cardio- acceleratory effects of the peptide implicate this peptide in key myoactive roles, likely related to rhythmic coordination.</p

The effect of high hydrostatic pressure acclimation on acute temperature tolerance and phospholipid fatty acid composition in the shallow-water shrimp Palaemon varians

Extant deep-sea fauna, including hydrothermal vent endemics such as bresiliid shrimp, are descended from shallow-water ancestors. Previous studies have demonstrated the capacity of shallow-water shrimp to acclimate to hydrostatic pressure representative of the vent environment. It has been proposed that this hyperbaric acclimation depends in part on shifts in phospholipid fatty acid composition to maintain biomembrane function. These shifts are also predicted to reduce critical temperature tolerance, potentially limiting the possibility of direct colonisation of the hydrothermal vent environment. Here, we present evidence that acclimation to high hydrostatic pressure (10MPa≈1000m water depth) decreases acute temperature tolerance from 30.2°C to 27.1°C in the shallow-water shrimp Palaemon varians acclimated to 10°C. Statistically significant shifts in phospholipid fatty acid composition occurred during exposure to high hydrostatic pressure, suggesting that homeoviscous modifications support shifts in environmental tolerances during hyperbaric acclimation. Despite the reduction in temperature tolerance, P.varians retains sufficient thermal scope to tolerate the thermal regime in the hydrothermal vent environment, allowing for the possibility of direct deep-sea hydrothermal vent colonisation by shallow-water shrimp

Functional characterization and signalling systems of corazonin and red pigment concentrating hormone in the green shore crab, Carcinus maenas

Neuropeptides play a central role as neurotransmitters, neuromodulators and hormones in orchestrating arthropod physiology. The post-genomic surge in identified neuropeptides and their putative receptors has not been matched by functional characterization of ligand-receptor pairs. Indeed, until very recently no G protein-coupled receptors (GPCRs) had been functionally defined in any crustacean. Here we explore the structurally-related, functionally-diverse gonadotropin-releasing hormone paralogs, corazonin (CRZ) and red-pigment concentrating hormone (RPCH) and their G-protein coupled receptors (GPCRs) in the crab, Carcinus maenas. Using aequorin luminescence to measure in vitro Ca2+ mobilization we demonstrated receptor-ligand pairings of CRZ and RPCH. CRZR-activated cell signaling in a dose-dependent manner (EC50 0.75 nM) and comparative studies with insect CRZ peptides suggest that the C-terminus of this peptide is important in receptor-ligand interaction. RPCH interacted with RPCHR with extremely high sensitivity (EC50 20 pM). Neither receptor bound GnRH, nor the AKH/CRZ-related peptide. Transcript distributions of both receptors indicate that CRZR expression was, unexpectedly, restricted to the Y-organs (YO). Application of CRZ peptide to YO had no effect on ecdysteroid biosynthesis, excepting a modest stimulation in early post-molt. CRZ had no effect on heart activity, blood glucose levels, lipid mobilization or pigment distribution in chromatophores, a scenario that reflected the distribution of its mRNA. Apart from the well-known activity of RPCH as a chromatophorotropin, it also indirectly elicited hyperglycemia (which was eyestalk-dependent). RPCHR mRNA was also expressed in the ovary, indicating possible roles in reproduction. The anatomy of CRZ and RPCH neurons in the nervous system is described in detail by immunohistochemistry and in situ hybridization. Each peptide has extensive but non-overlapping distribution in the CNS, and neuroanatomy suggests that both are possibly released from the post-commissural organs. This study is one of the first to deorphanize a GPCR in a crustacean and to provide evidence for hitherto unknown and diverse functions of these evolutionarily-related neuropeptides

Optimising scale and deployment of community health workers in Sierra Leone: a geospatial analysis

Little is known about strategies for optimising the scale and deployment of community health workers (CHWs) to maximise geographic accessibility of primary healthcare services.We used data from a national georeferenced census of CHWs and other spatial datasets in Sierra Leone to undertake a geospatial analysis exploring optimisation of the scale and deployment of CHWs, with the aim of informing implementation of current CHW policy and future plans of the Ministry of Health and Sanitation

Pigment Dispersing Factors and Their Cognate Receptors in a Crustacean Model, With New Insights Into Distinct Neurons and Their Functions

Pigment dispersing factors (PDFs, or PDHs in crustaceans) form a structurally related group of neuropeptides found throughout the Ecdysozoa and were first discovered as pigmentary effector hormones in crustaceans. In insects PDFs fulfill crucial neuromodulatory roles, most notably as output regulators of the circadian system, underscoring their central position in physiological and behavioral organization of arthropods. Intriguingly, decapod crustaceans express multiple isoforms of PDH originating from separate genes, yet their differential functions are still to be determined. Here, we functionally define two PDH receptors in the crab Carcinus maenas and show them to be selectively activated by four PDH isoforms: PDHR 43673 was activated by PDH-1 and PDH-2 at low nanomolar doses whilst PDHR 41189 was activated by PDH-3 and an extended 20 residue e-PDH. Detailed examination of the anatomical distribution of all four peptides and their cognate receptors indicate that they likely perform different functions as secreted hormones and/or neuromodulators, with PDH-1 and its receptor 43,673 implicated in an authentic hormonal axis. PDH-2, PDH-3, and e-PDH were limited to non-neurohemal interneuronal sites in the CNS; PDHR 41189 was largely restricted to the nervous system suggesting a neuromodulatory function. Notably PDH-3 and e-PDH were without chromatophore dispersing activity. This is the first report which functionally defines a PDHR in an endocrine system in a crustacean and to indicate this and other putative roles of this physiologically pivotal peptide group in these organisms. Thus, our findings present opportunities to further examine the endocrine and circadian machinery in this important arthropod phylum

Beyond Red vs. Blue: The Political Typology - 2021

Partisan polarization remains the dominant, seemingly unalterable condition of American politics. Republicans and Democrats agree on very little – and when they do, it often is in the shared belief that they have little in common.Yet the gulf that separates Republicans and Democrats sometimes obscures the divisions and diversity of views that exist within both partisan coalitions – and the fact that many Americans do not fit easily into either one.Republicans are divided on some principles long associated with the GOP: an affinity for businesses and corporations, support for low taxes and opposition to abortion. Democrats face substantial internal differences as well – some that are long-standing, such as on the importance of religion in society, others more recent. For example, while Democrats widely share the goal of combating racial inequality in the United States, they differ on whether systemic change is required to achieve that goal.These intraparty disagreements present multiple challenges for both parties: They complicate the already difficult task of governing in a divided nation. In addition, to succeed politically, the parties must maintain the loyalty of highly politically engaged, more ideological voters, while also attracting support among less engaged voters – many of them younger – with weaker partisan ties.Pew Research Center's new political typology provides a road map to today's fractured political landscape. It segments the public into nine distinct groups, based on an analysis of their attitudes and values. The study is primarily based on a survey of 10,221 adults conducted July 8-18, 2021; it also draws from several additional interviews with these respondents conducted since January 2020

ParkIndex: Validation and Application of a Pragmatic Measure of Park Access and Use

Composite metrics integrating park availability, features, and quality for a given address or neighborhood are lacking. The purposes of this study were to describe the validation, application, and demonstration of ParkIndex in four diverse communities. This study occurred in Fall 2018 in 128 census block groups within Seattle(WA), Brooklyn(NY), Raleigh(NC), and Greenville County(SC). All parks within a half-mile buffer were audited to calculate a composite park quality score, and select households provided data about use of proximal parks via an online, map-based survey. For each household, the number of parks, total park acreage, and average park quality score within one half-mile were calculated using GIS. Logistic regression was used to identify a parsimonious model predicting park use. ParkIndex values (representing the probability of park use) were mapped for all study areas and after scenarios involving the addition and renovation/improvement of parks. Out of 360 participants, 23.3% reported visiting a park within the past 30 days. The number of parks (OR = 1.36, 95% CI = 1.15–1.62), total park acreage (OR = 1.13, 95% CI = 1.07–1.19), and average park quality score (OR = 1.04, 95% CI = 1.01–1.06) within one half-mile were all associated with park use. Composite ParkIndex values across the study areas ranged from 0 to 100. Hypothetical additions of or renovations to study area parks resulted in ParkIndex increases of 22.7% and 19.2%, respectively. ParkIndex has substantial value for park and urban planners, citizens, and researchers as a common metric to facilitate awareness, decision-making, and intervention planning related to park access, environmental justice, and community health

Total- and Monomethyl-Mercury and Major Ions in Coastal California Fog Water: Results from Two Years of Sampling on Land and at Sea

Marine fog water samples were collected over two summers (2014–2015) with active strand collectors (CASCC) at eight coastal sites from Humboldt to Monterey counties in California, USA, and on four ocean cruises along the California coastline in order to investigate mercury (Hg) cycling at the ocean-atmosphere-land interface. The mean concentration of monomethylmercury (MMHg) in fog water across terrestrial sites for both years was 1.6 ± 1.9 ng L-1 (\u3c0.01–10.4 ng L-1, N = 149), which corresponds to 5.7% (2.0–10.8%) of total Hg (HgT) in fog. Rain water samples from three sites had mean MMHg concentrations of 0.20 ± 0.12 ng L-1 (N = 5) corresponding to 1.4% of HgT. Fog water samples collected at sea had MMHg concentrations of 0.08 ± 0.15 ng L-1 (N = 14) corresponding to 0.4% of HgT. Significantly higher MMHg concentrations in fog were observed at terrestrial sites next to the ocean relative to a site 40 kilometers inland, and the mean difference was 1.6 ng L-1. Using a rate constant for photo-demethylation of MMHg of -0.022 h-1 based on previous demethylation experiments and a coastal-inland fog transport time of 12 hours, a mean difference of only 0.5 ng L-1 of MMHg was predicted between coastal and inland sites, indicating other unknown source and/or sink pathways are important for MMHg in fog. Fog water deposition to a standard passive 1.00 m2 fog collector at six terrestrial sites averaged 0.10 ± 0.07 L m-2 d-1, which was ∼2% of typical rainwater deposition in this area. Mean air-surface fog water fluxes of MMHg and HgT were then calculated to be 34 ± 40 ng m-2 y-1 and 546 ± 581 ng m-2 y-1, respectively. These correspond to 33% and 13% of the rain fluxes, respectively

Longitudinal population analysis of dual infection with recombination in two strains of HIV type 1 subtype B in an individual from a phase 3 HIV vaccine efficacy trial

This study documents a case of coinfection (simultaneous infection of an individual with two or more strains) of two HIV-1 subtype B strains in an individual from a Phase 3 HIV-1 vaccine efficacy trial, conducted in North American and the Netherlands. We examined 86 full-length gp120 (env) gene sequences from this individual collected from nine different time points over a 20-month period. We estimated evolutionary relationships using maximum likelihood and Bayesian methods and inferred recombination breakpoints and recombinant sequences using phylogenetic and substitutional methods. These analyses identified two strongly supported monophyletic clades (clades A and B) of 14 and 69 sequences each and a small paraphyletic recombinant clade of three sequences. We then studied the genetic characteristics of these lineages by comparing estimates of genetic diversity generated by mutation and recombination and adaptive selection within a coalescent and maximum likelihood framework. Our results suggest significant differences on the evolutionary dynamics of these strains. We then discuss the implications of these results for vaccine development