IL-4Rα Signaling in Keratinocytes and Early IL-4 Production Are Dispensable for Generating a Curative T Helper 1 Response in Leishmania major-Infected C57BL/6 Mice.

Experimental infection with the protozoan parasite Leishmania major has been extensively used to understand the mechanisms involved in T helper cell differentiation. Following infection, C57BL/6 mice develop a small self-healing cutaneous lesion and they are able to control parasite burden, a process linked to the development of T helper (Th) 1 cells. The local presence of IL-12 has been reported to be critical in driving Th1 cell differentiation. In addition, the early secretion of IL-4 was reported to potentially contribute to Th1 cell differentiation. Following infection with L. major, early keratinocyte-derived IL-4 was suggested to contribute to Th1 cell differentiation. To investigate a putative autocrine role of IL-4 signaling on keratinocytes at the site of infection, we generated C57BL/6 mice deficient for IL-4Rα expression selectively in keratinocytes. Upon infection with L. major, these mice could control their inflammatory lesion and parasite load correlating with the development of Th1 effector cells. These data demonstrate that IL-4 signaling on keratinocytes does not contribute to Th1 cell differentiation. To further investigate the source of IL-4 in the skin during the first days after L. major infection, we used C57BL/6 IL-4 reporter mice allowing the visualization of IL-4 mRNA expression and protein production. These mice were infected with L. major. During the first 3 days after infection, skin IL-4 mRNA expression was observed selectively in mast cells. However, no IL-4 protein production was detectable locally. In addition, early IL-4 blockade locally had no impact on subsequent Th1 cell differentiation and control of the disease. Taken together, the present data rule out a major role for skin IL-4 and keratinocyte IL-4Rα signaling in the development of a Th1 protective immune response following experimental infection with L. major

The quorum-sensing molecules farnesol/homoserine lactone and dodecanol operate via distinct modes of action in Candida albicans

Living as a commensal, Candida albicans must adapt and respond to environmental cues generated by the mammalian host and by microbes comprising the natural flora. These signals have opposing effects on C. albicans, with host cues promoting the yeast-to-hyphal transition and bacteria-derived quorum-sensing molecules inhibiting hyphal development. Hyphal development is regulated through modulation of the cyclic AMP (cAMP)/protein kinase A (PKA) signaling pathway, and it has been postulated that quorum-sensing molecules can affect filamentation by inhibiting the cAMP pathway. Here, we show that both farnesol and 3-oxo-C12-homoserine lactone, a quorum-sensing molecule secreted by Pseudomonas aeruginosa, block hyphal development by affecting cAMP signaling; they both directly inhibited the activity of the Candida adenylyl cyclase, Cyr1p. In contrast, the 12-carbon alcohol dodecanol appeared to modulate hyphal development and the cAMP signaling pathway without directly affecting the activity of Cyr1p. Instead, we show that dodecanol exerted its effects through a mechanism involving the C. albicans hyphal repressor, Sfl1p. Deletion of SFL1 did not affect the response to farnesol but did interfere with the response to dodecanol. Therefore, quorum sensing in C. albicans is mediated via multiple mechanisms of action. Interestingly, our experiments raise the possibility that the Burkholderia cenocepacia diffusible signal factor, BDSF, also mediates its effects via Sfl1p, suggesting that dodecanol's mode of action, but not farnesol or 3-oxo-C12-homoserine lactone, may be used by other quorum-sensing molecules

Pan-Arctic diel vertical migration during the polar night

Diel vertical migration (DVM) has generally been assumed to cease during the polarnight in the high Arctic, although recent studies have shown the occurrence of lunar vertical migrations (LVMs) and shallow DVMs. Here, we quantified when and where full-depth (>20 m), solar-mediated DVM exists on a pan-Arctic scale. We observed the scattering population, most likely to be comprised of zooplankton, using 300 kHz acoustic Doppler current profilers (ADCPs). We quantified the presence/absence of DVM, and found that DVM continues throughout the year to at least 20 m at all locations south of 74° N. North of 77° N, DVM ceases for a period of time duringthe polar night. The dates of this cessation accurately align with the date of the winter solstice (±2 d). Between 74 and 77° N, DVM presence/absence is variable. Acoustic data sampled at 89° N, however, showed no evidence of DVM at any time during the year — a new observation. Using indicators of presence/absence of sea ice from ADCPs and satellite-derived sea ice concentration data, we revealed that local variations in sea ice cover directly determine the continuation or cessationof DVM during the polar night. Earlier-forming and higher-concentration sea ice causes a cessation in DVM, whereas low-concentration or late-forming sea ice results in continuous DVM when comparing migrations at similar latitudes

Arctic Oceanography - Oceanography: Atmosphere-Ocean Exchange, Biogeochemistry & Physics

The Arctic Ocean is, on average, the shallowest of Earth’s oceans. Its vast continental shelf areas, which account for approximately half of the Arctic Ocean’s total area, are heavily influenced by the surrounding land masses through river run-off and coastal erosion. As a main area of deep water formation, the Arctic is one of the main «engines» of global ocean circulation, due to large freshwater inputs, it is also strongly stratified. The Arctic Ocean’s complex oceanographic configuration is tightly linked to the atmosphere, the land, and the cryosphere. The physical dynamics not only drive important climate and global circulation patterns, but also control biogeochemical cycles and ecosystem dynamics. Current changes in Arctic sea-ice thickness and distribution, air and water temperatures, and water column stability are resulting in measurable shifts in the properties and functioning of the ocean and its ecosystems. The Arctic Ocean is forecast to shift to a seasonally ice-free ocean resulting in changes to physical, chemical, and biological processes. These include the exchange of gases across the atmosphere-ocean interface, the wind-driven ciruclation and mixing regimes, light and nutrient availability for primary production, food web dynamics, and export of material to the deep ocean. In anticipation of these changes, extending our knowledge of the present Arctic oceanography and these complex changes has never been more urgent

Deletion of Interleukin-4 Receptor Alpha-Responsive Keratinocytes in BALB/c Mice Does Not Alter Susceptibility to Cutaneous Leishmaniasis.

The skin microenvironment at the site of infection plays a role in the early events that determine protective T helper 1/type 1 immune responses during cutaneous leishmaniasis (CL) infection. During CL in nonhealing BALB/c mice, early interleukin-4 (IL-4) can instruct dendritic cells for protective Th1 immunity. Additionally, keratinocytes, which are the principal cell type in the skin epidermis, have been shown to secrete IL-4 early after Leishmania major infection. Here, we investigated whether IL-4/IL-13 signaling via the common IL-4 receptor alpha chain (IL-4Rα) on keratinocytes contributes to susceptibility during experimental CL. To address this, keratinocyte-specific IL-4Rα-deficient (KRT14 <sup>cre</sup> IL-4Rα <sup>-/lox</sup> ) mice on a BALB/c genetic background were generated by gene targeting and site-specific recombination (Cre/loxP) under the control of the keratinocyte-specific krt14 locus. Following high-dose infection with L. major IL-81 and LV39 promastigotes subcutaneously in the footpad, footpad swelling, parasite burden, IFN-γ/IL-4/IL-13 cytokine production, and type 1 and type 2 antibody responses were similar between KRT14 <sup>cre</sup> IL-4Rα <sup>-/lox</sup> and littermate control IL-4Rα <sup>-/lox</sup> BALB/c mice. An intradermal infection with low-dose L. major IL-81 and LV39 promastigotes in the ear showed results in infected KRT14 <sup>cre</sup> IL-4Rα <sup>-/lox</sup> BALB/c mice similar to those of littermate control IL-4Rα <sup>-/lox</sup> BALB/c mice, with the exception of a significant decrease observed in parasite burden only at the site of LV39 infection in the ear. Collectively, our results show that autocrine and paracrine signaling of IL-4/IL-13 through the IL-4Rα chain on keratinocytes does not influence the establishment of a nonhealing Th2 immune response in BALB/c mice during L. major infection

Redundant Notch1 and Notch2 Signaling Is Necessary for IFNγ Secretion by T Helper 1 Cells During Infection with Leishmania major

The protective immune response to intracellular parasites involves in most cases the differentiation of IFNγ-secreting CD4+ T helper (Th) 1 cells. Notch receptors regulate cell differentiation during development but their implication in the polarization of peripheral CD4+ T helper 1 cells is not well understood. Of the four Notch receptors, only Notch1 (N1) and Notch2 (N2) are expressed on activated CD4+ T cells. To investigate the role of Notch in Th1 cell differentiation following parasite infection, mice with T cell-specific gene ablation of N1, N2 or both (N1N2ΔCD4Cre) were infected with the protozoan parasite Leishmania major. N1N2ΔCD4Cre mice, on the C57BL/6 L. major-resistant genetic background, developed unhealing lesions and uncontrolled parasitemia. Susceptibility correlated with impaired secretion of IFNγ by draining lymph node CD4+ T cells and increased secretion of the IL-5 and IL-13 Th2 cytokines. Mice with single inactivation of N1 or N2 in their T cells were resistant to infection and developed a protective Th1 immune response, showing that CD4+ T cell expression of N1 or N2 is redundant in driving Th1 differentiation. Furthermore, we show that Notch signaling is required for the secretion of IFNγ by Th1 cells. This effect is independent of CSL/RBP-Jκ, the major effector of Notch receptors, since L. major-infected mice with a RBP-Jκ deletion in their T cells were able to develop IFNγ-secreting Th1 cells, kill parasites and heal their lesions. Collectively, we demonstrate here a crucial role for RBP-Jκ-independent Notch signaling in the differentiation of a functional Th1 immune response following L. major infection

Postglacial expansion of the arctic keystone copepod calanus glacialis

Calanus glacialis, a major contributor to zooplankton biomass in the Arctic shelf seas, is a key link between primary production and higher trophic levels that may be sensitive to climate warming. The aim of this study was to explore genetic variation in contemporary populations of this species to infer possible changes during the Quaternary period, and to assess its population structure in both space and time. Calanus glacialis was sampled in the fjords of Spitsbergen (Hornsund and Kongsfjorden) in 2003, 2004, 2006, 2009 and 2012. The sequence of a mitochondrial marker, belonging to the ND5 gene, selected for the study was 1249 base pairs long and distinguished 75 unique haplotypes among 140 individuals that formed three main clades. There was no detectable pattern in the distribution of haplotypes by geographic distance or over time. Interestingly, a Bayesian skyline plot suggested that a 1000-fold increase in population size occurred approximately 10,000 years before present, suggesting a species expansion after the Last Glacial Maximum.GAME from the National Science Centre, the Polish Ministry of Science and Higher Education Iuventus Plus [IP2014 050573]; FCT-PT [CCMAR/Multi/04326/2013]; [2011/03/B/NZ8/02876

Glacier Calving Rates Due to Subglacial Discharge, Fjord Circulation, and Free Convection

Tidewater glacier calving provides the most direct mechanism of ice transfer from land to the ocean. However, the physical melt processes influencing calving remain challenging to constrain. In this study we focus on calving rates at Kongsbreen, a tidewater glacier in Svalbard, due to three mechanisms of submarine melt: (1) free convection, (2) horizontal fjord circulation, and (3) meltwater discharge. To calculate an overall calving rate, we measure glacier velocity and terminus change using Sentinel imagery. We calculate free convection, fjord circulation, and meltwater discharge calving using mooring data for mid-fjord ocean temperature (30–80 m depth), reanalysis results for meltwater runoff, and georectified time-lapse imagery to track icebergs and infer surface circulation. Results show that the total glacier calving rate is highly correlated with ocean temperature during the 2016 melt season. When runoff was present, we found that subglacial discharge accounted for calving rates an order of magnitude greater than the maximum calving rates assigned to the other two melting mechanisms combined. Further, subglacial discharge at Kongsbreen was more efficient in inducing calving later in the season than earlier in the season, implying that the increase in ocean temperatures, the timing of meltwater discharge within a melt season, and/or the development of discrete meltwater exit channels are critical components to calving rates. As the recent atmospheric warming trend and subsequent increase in meltwater discharge is expected to continue, it is essential to understand the processes contributing to an increase in glacier calving and incorporate these processes into predictive models

Shine a light: Under-ice light and its ecological implications in a changing Arctic Ocean

The Arctic marine ecosystem is shaped by the seasonality of the solar cycle, spanning from 24-h light at the sea surface in summer to 24-h darkness in winter. The amount of light available for under-ice ecosystems is the result of different physical and biological processes that affect its path through atmosphere, snow, sea ice and water. In this article, we review the present state of knowledge of the abiotic (clouds, sea ice, snow, suspended matter) and biotic (sea ice algae and phytoplankton) controls on the underwater light field. We focus on how the available light affects the seasonal cycle of primary production (sympagic and pelagic) and discuss the sensitivity of ecosystems to changes in the light field based on model simulations. Lastly, we discuss predicted future changes in under-ice light as a consequence of climate change and their potential ecological implications, with the aim of providing a guide for future research

Neutrophils Reduce the Parasite Burden in Leishmania (Leishmania) amazonensis-Infected Macrophages

Background: Studies on the role of neutrophils in Leishmania infection were mainly performed with L. (L) major, whereas less information is available for L. (L) amazonensis. Previous results from our laboratory showed a large infiltrate of neutrophils in the site of infection in a mouse strain resistant to L. (L.) amazonensis (C3H/HePas). in contrast, the susceptible strain (BALB/c) displayed a predominance of macrophages harboring a high number of amastigotes and very few neutrophils. These findings led us to investigate the interaction of inflammatory neutrophils with L. (L.) amazonensis-infected macrophages in vitro.Methodology/Principal Findings: Mouse peritoneal macrophages infected with L. (L.) amazonensis were co-cultured with inflammatory neutrophils, and after four days, the infection was quantified microscopically. Data are representative of three experiments with similar results. the main findings were 1) intracellular parasites were efficiently destroyed in the co-cultures; 2) the leishmanicidal effect was similar when cells were obtained from mouse strains resistant (C3H/HePas) or susceptible (BALB/c) to L. (L.) amazonensis; 3) parasite destruction did not require contact between infected macrophages and neutrophils; 4) tumor necrosis factor alpha (TNF-alpha), neutrophil elastase and platelet activating factor (PAF) were involved with the leishmanicidal activity, and 5) destruction of the parasites did not depend on generation of oxygen or nitrogen radicals, indicating that parasite clearance did not involve the classical pathway of macrophage activation by TNF-alpha, as reported for other Leishmania species.Conclusions/Significance: the present results provide evidence that neutrophils in concert with macrophages play a previously unrecognized leishmanicidal effect on L. (L.) amazonensis. We believe these findings may help to understand the mechanisms involved in innate immunity in cutaneous infection by this Leishmania species.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, São Paulo, BrazilWeb of Scienc