Carbon dynamics in a Boreal land-stream-lake continuum during the spring freshet of two hydrologically contrasting years

We studied in 2013 and 2014 the spring carbon dynamics in a Boreal landscape consisting of a lake and 15 inflowing streams and an outlet. The first year had weather and a hydrological regime typical of past years with a distinct spring freshet connected with the thaw of the average snowpack. The latter year had higher air temperatures which did not permit snow accumulation, despite similar winter precipitation. As such, there was hardly any spring freshet in 2014, and stream discharge peaked in January, i.e., the conditions resembled those predicted in the future climate. Despite the hydrological differences between the years, there were only small interannual differences in the stream CO2 and DOC concentrations. The relationship between the concentrations and discharge was stronger in the typical year. CO2 concentrations in medium-sized streams correlated negatively with the discharge, indicating dilution effect of melting snowpacks, while in large-sized streams the correlation was positive, suggesting stronger groundwater influence. The DOC pathway to these streams was through the subsurface soil layers, not the groundwater. The total amount of carbon transported into the lake was ca. 1.5-fold higher in the typical year than in the year with warm winter. In 2013, most of the lateral inputs took place during spring freshet. In 2014, the majority of inputs occurred earlier, during the winter months. The lateral CO2 signal was visible in the lake at 1.5 m depth. DOC dominated the carbon transport, and in both years, 12% of the input C was in inorganic form.Peer reviewe

Buprenorphine-naloxone, buprenorphine, and methadone throughout pregnancy in maternal opioid use disorder

IntroductionCurrent WHO guidelines recommend using methadone or buprenorphine as maintenance treatments for maternal opioid use disorder. However, buprenorphine-naloxone, with a lower abuse risk than buprenorphine monotherapy or methadone, offers a potentially beneficial alternative, but scientific evidence on its effects on pregnancies, fetuses, and newborns is scarce. This paper compares the outcomes of the pregnancies, deliveries, and newborns of women on buprenorphine-naloxone, buprenorphine, or methadone maintenance treatments. According to the hypothesis, as a maintenance treatment, buprenorphine-naloxone does not have more adverse effects than buprenorphine, whereas methadone is more complicated. Material and methodsIn this population-based study, 172 pregnant women on medical-assisted treatments were followed-up at Helsinki University Women's Hospital (Finland). Women receiving the same opioid maintenance treatment from conception to delivery and their newborns were included. Consequently, 67 mother-child dyads met the final inclusion criteria. They were divided into three groups based on their opioid pharmacotherapy. The outcomes were compared among the groups and, where applicable, with the Finnish population. ResultsThe buprenorphine-naloxone and buprenorphine groups showed similar outcomes and did not significantly differ from each other in terms of maternal health during pregnancies, deliveries, or newborns. Illicit drug use during the pregnancy was common in all groups, but in the methadone group it was most common (p = 0.001). Most neonates (96%) were born full-term with good Apgar scores. They were of relatively small birth size, with those in the methadone group tending to be the smallest. Of the neonates 63% needed pharmacological treatment for neonatal opioid withdrawal syndrome. The need was lower in the buprenorphine-based groups than in the methadone group (p = 0.029). ConclusionsBuprenorphine-naloxone seems to be as safe for pharmacotherapy for maternal opioid use disorder as buprenorphine monotherapy for both mother and newborn. Hence it could be a choice for oral opioid maintenance treatment during pregnancy, but larger studies are needed before changing the official recommendations. Women on methadone treatment carry multifactorial risks and require particularly cautious follow up. Furthermore, illicit drug use is common in all treatment groups and needs to be considered for all patients with opioid use disorder.Peer reviewe

Enhanced Antibody Production in Clever-1/Stabilin-1-Deficient Mice

Clever-1, encoded by the Stab1 gene, is a scavenger and leukocyte trafficking receptor expressed by subsets of vascular and lymphatic endothelial cells and immunosuppressive macrophages. Monocyte Clever-1 also modulates T cell activation. However, nothing is known about the possible links between B cell function and Clever-1. Here, we found that Stab1 knockout mice (Stab1(-/-)) lacking the Clever-1 protein from all cells present with abnormally high antibody levels under resting conditions and show enhanced humoral immune responses after immunization with protein and carbohydrate antigens. Removal of the spleen does not abolish the augmented basal and post-immunization antibody levels in Clever-1-deficient mice. The increased IgG production is also present in mice in which Clever-1 is selectively ablated from macrophages. When compared to wildtype macrophages, Clever-1-deficient macrophages show increased TNF-alpha synthesis. In co-culture experiments, monocytes/macrophages deficient of Clever-1 support higher IgM production by B cells, which is blocked by TNF-alpha depletion. Collectively, our data show that the excessive inflammatory activity of monocytes/macrophages in the absence of Clever-1 results in augmented humoral immune responses in vivo

Methane dynamics in different boreal lake types

This study explores the variability in concentrations of dissolved CH<sub>4</sub> and annual flux estimates in the pelagic zone in a statistically defined sample of 207 lakes in Finland. The lakes were situated in the boreal zone, in an area where the mean annual air temperature ranges from −2.8 to 5.9°C. We examined how lake CH<sub>4</sub> dynamics related to regional lake types assessed according to the EU water framework directive. Ten lake types were defined on the basis of water chemistry, color, and size. Lakes were sampled for dissolved CH<sub>4</sub> concentrations four times per year, at four different depths at the deepest point of each lake. We found that CH<sub>4</sub> concentrations and fluxes to the atmosphere tended to be high in nutrient rich calcareous lakes, and that the shallow lakes had the greatest surface water concentrations. Methane concentration in the hypolimnion was related to oxygen and nutrient concentrations, and to lake depth or lake area. The surface water CH<sub>4</sub> concentration was related to the depth or area of lake. Methane concentration close to the bottom can be viewed as proxy of lake status in terms of frequency of anoxia and nutrient levels. The mean pelagic CH<sub>4</sub> release from randomly selected lakes was 49 mmol m<sup>−2</sup> a<sup>−1</sup>. The sum CH<sub>4</sub> flux (storage and diffusion) correlated with lake depth, area and nutrient content, and CH<sub>4</sub> release was greatest from the shallow nutrient rich and humic lakes. Our results support earlier lake studies regarding the regulating factors and also the magnitude of global emission estimate. These results propose that in boreal region small lakes have higher CH<sub>4</sub> fluxes per unit area than larger lakes, and that the small lakes have a disproportionate significance regarding to the CH<sub>4</sub> release

Thymocytes in Lyve1-CRE/S1pr1(f/f) Mice accumulate in the Thymus due to cell-intrinsic loss of sphingosine-1-Phosphate receptor expression

T cell emigration from the thymus is essential for immunological homeostasis. While stromal cell-produced sphingosine-1-phosphate (S1P) has been shown to promote thymocyte egress via the S1P receptor, S1PR1, the significance of S1P/S1PR1 signaling in the thymic stromal cells that surround T cells remains unclear. To address this issue, we developed conditional knockout mice (Lyve1-CRE/S1pr1f/f mice) in which S1pr1 was selectively targeted in cells expressing the lymphatic endothelial cell marker, Lyve1. In these mice, T cells were significantly reduced in secondary lymphoid tissues, and CD62L(+) mature CD4 and CD8 single-positive (SP) T cells accumulated in the medulla failed to undergo thymus egress. Using a Lyve1 reporter strain in which Lyve1 lineage cells expressed tdTomato fluorescent protein, we unexpectedly found that a considerable proportion of the thymocytes were fluorescently labeled, indicating that they belonged to the Lyve1 lineage. The CD4 and CD8 SP thymocytes in Lyve1-CRE/S1pr1f/f mice exhibited an egress-competent phenotype (HSA(low), CD62L(high), and Qa-2(high)), but were CD69(high) and lacked S1PR1 expression. In addition, CD4 SP thymocytes from these mice were unable to migrate to the periphery after their intrathymic injection into wild-type (WT) mice. In contrast, WT T cells could migrate to the periphery in both WT and Lyve1-CRE/S1pr1f/f thymuses. These results demonstrated that thymocyte egress is mediated by T cell-expressed, but not stromal cell-expressed, S1PR1 and caution against using the Lyve1-CRE system for selectively gene deletion in lymphatic endothelial cells

Transcytosis route mediates rapid delivery of intact antibodies to draining lymph nodes

Lymph nodes (LNs) filter lymph to mount effective immune responses. Small soluble lymph-borne molecules from the periphery enter the draining LNs via a reticular conduit system. Intact antibodies and other larger molecules, in contrast, are physically unable to enter the conduits, and they are thought to be transported to the LNs only within migratory DCs after proteolytic degradation. Here, we discovered that lymph-borne antibodies and other large biomolecules enter within seconds into the parenchyma of the draining LN in an intact form. Mechanistically, we found that the uptake of large molecules is a receptor-independent, fluid-phase process that takes place by dynamin-dependent vesicular transcytosis through the lymphatic endothelial cells in the subcapsular sinus of the LN. Physiologically, this pathway mediates a very fast transfer of large protein antigens from the periphery to LN-resident DCs and macrophages. We show that exploitation of the transcytosis system allows enhanced whole-organ imaging and spatially controlled lymphocyte activation by s.c. administered antibodies in vivo. Transcytosis through the floor of the subcapsular sinus thus represents what we believe to be a new physiological and targetable mode of lymph filtering.</p

Gene-expression profiling of different arms of lymphatic vasculature identifies candidates for manipulation of cell traffic

Afferent lymphatic vessels bring antigens and diverse populations of leukocytes to draining lymph nodes, whereas efferent lymphatics allow only lymphocytes and antigens to leave the nodes. Despite the fundamental importance of afferent vs. efferent lymphatics in immune response and cancer spread, the molecular characteristics of these different arms of the lymphatic vasculature are largely unknown. The objective of this work was to explore molecular differences behind the distinct functions of afferent and efferent lymphatic vessels, and find possible molecules mediating lymphocyte traffic. We used laser-capture microdissection and cell sorting to isolate lymphatic endothelial cells (LECs) from the subcapsular sinus (SS, afferent) and lymphatic sinus (LS, efferent) for transcriptional analyses. The results reveal marked differences between afferent and efferent LECs and identify molecules on lymphatic vessels. Further characterizations of Siglec-1 (CD169) and macrophage scavenger receptor 1 (MSR1/CD204), show that they are discriminatively expressed on lymphatic endothelium of the SS but not on lymphatic vasculature of the LS. In contrast, endomucin (EMCN) is present on the LS endothelium and not on lymphatic endothelium of the SS. Moreover, both murine and human MSR1 on lymphatic endothelium of the SS bind lymphocytes and in in vivo studies MSR1 regulates entrance of lymphocytes from the SS to the lymph node parenchyma. In conclusion, this paper reports surprisingly distinct molecular profiles for afferent and efferent lymphatics and a function for MSR1. These results may open avenues to explore some of the now-identified molecules as targets to manipulate the function of lymphatic vessels

Fetal-derived macrophages dominate in adult mammary glands

Macrophages serve multiple functions including immune regulation, morphogenesis, tissue homeostasis and healing reactions. The current paradigm holds that mammary gland macrophages first arise postnatally during the prepubertal period from the bone marrow-derived monocytes. Here we delineate the origins of tissue-resident mammary gland macrophages using high-dimension phenotypic analyses, cell-fate mapping experiments, gene-deficient mice lacking selective macrophage subtypes, and antibody-based depletion strategies. We show that tissue-resident macrophages are found in mammary glands already before birth, and that the yolk sac-derived and fetal liver-derived macrophages outnumber the adult-derived macrophages in the mammary gland also in the adulthood. In addition, fetal-derived mammary gland macrophages have a characteristic phenotype, display preferential periductal and perivascular localization, and are highly active in scavenging. These findings identify fetal-derived macrophages as the predominant leukocyte type in the adult mammary gland stroma, and reveal previously unknown complexity of macrophage biology in the breast

KP-LAB Knowledge Practices Laboratory -- Specification of end-user applications

deliverablesThe present deliverable provides a high-level view on the new specifications of end user applications defined in the WPII during the M37-M46 period of the KP-Lab project. This is the last in the series of four deliverables that cover all the tools developed in the project, the previous ones being D6.1, D6.4 and D6.6. This deliverable presents specifications for the new functionalities for supporting the dedicated research studies defined in the latest revision of the KP-Lab research strategy. The tools addressed are: the analytic tools (Data export, Time-line-based analyser, Visual analyser), Clipboard, Search, Versioning of uploadable content items, Visual Model Editor (VME) and Visual Modeling Language Editor (VMLE). The main part of the deliverable provides the summary of tool specifications and the description of the Knowledge Practices Environment architecture, as well as an overview of the revised technical design process, of the tools’ relationship with the research studies, and of the driving objectives and the high-level requirements relevant for the present specifications. The full specifications of tools are provided in the annexes 1-9