A MyD88-dependent IFNγR-CCR2 signaling circuit is required for mobilization of monocytes and host defense against systemic bacterial challenge

Monocytes are mobilized to sites of infection via interaction between the chemokine MCP-1 and its receptor, CCR2, at which point they differentiate into macrophages that mediate potent antimicrobial effects. In this study, we investigated the mechanisms by which monocytes are mobilized in response to systemic challenge with the intracellular bacterium Francisella tularensis. We found that mice deficient in MyD88, interferon-γ (IFNγ)R or CCR2 all had defects in the expansion of splenic monocyte populations upon F. tularensis challenge, and in control of F. tularensis infection. Interestingly, MyD88-deficient mice were defective in production of IFNγ, and IFNγR-deficient mice exhibited defective production of MCP-1, the ligand for CCR2. Transplantation of IFNγR-deficient bone marrow (BM) into wild-type mice further suggested that mobilization of monocytes in response to F. tularensis challenge required IFNγR expression on BM-derived cells. These studies define a critical host defense circuit wherein MyD88-dependent IFNγ production signals via IFNγR expressed on BM-derived cells, resulting in MCP-1 production and activation of CCR2-dependent mobilization of monocytes in the innate immune response to systemic F. tularensis challenge.     

Responses of Ecosystem Carbon Cycling to Climate Change Treatments Along an Elevation Gradient

Global temperature increases and precipitation changes are both expected to alter ecosystem carbon (C) cycling. We tested responses of ecosystem C cycling to simulated climate change using field manipulations of temperature and precipitation across a range of grass-dominated ecosystems along an elevation gradient in northern Arizona. In 2002, we transplanted intact plant?Csoil mesocosms to simulate warming and used passive interceptors and collectors to manipulate precipitation. We measured daytime ecosystem respiration (ER) and net ecosystem C exchange throughout the growing season in 2008 and 2009. Warming generally stimulated ER and photosynthesis, but had variable effects on daytime net C exchange. Increased precipitation stimulated ecosystem C cycling only in the driest ecosystem at the lowest elevation, whereas decreased precipitation showed no effects on ecosystem C cycling across all ecosystems. No significant interaction between temperature and precipitation treatments was observed. Structural equation modeling revealed that in the wetter-than-average year of 2008, changes in ecosystem C cycling were more strongly affected by warming-induced reduction in soil moisture than by altered precipitation. In contrast, during the drier year of 2009, warming induced increase in soil temperature rather than changes in soil moisture determined ecosystem C cycling. Our findings suggest that warming exerted the strongest influence on ecosystem C cycling in both years, by modulating soil moisture in the wet year and soil temperature in the dry year.     

The association of the sulphogalactosylglycerolipid of rat brain with myelination

The sulphogalactosylglycerolipid of rat brain is closely associated with the process of myelination, as demonstrated by the following observations. 1. The lipid is barely detectable in rat brain before 10 days of age, accumulates rapidly between age 10 and 25 days, and remains relatively constant in amount (between 0.3 and 0.4mumol per brain) thereafter into adult life. 2. The activity of adenosine 3'-phosphate 5'-sulphatophosphate-galactosyldiacylglycerol sulphotransferase is almost absent before 10 days of age, attains a maximum at age 20 days, and slowly decreases thereafter with increasing age. This developmental pattern correlates well with that of other myelin-specific metabolites. 3. Both the concentration of the sulphogalactosylglycerolipid and the activity of sulphotransferase are greatly decreased in the non-myelinating jimpy mouse. 4. The myelin fraction of rat brain contains most of the sulphogalactosylglycerolipid. The lipid occurs in a diacyl and an alkylacyl form. Determinations of the relative amount of each type in brain showed about a 1:1 mixture in both 21-day-old and adult rats. Rats injected with H(2) (35)SO(4) at 20 days of age lost (35)S from the diacyl form at a higher rate than from the alkylacyl compound over a 21-day period. These data suggest that the diacyl form has a higher turnover than the alkylacyl derivative. The percentage of the total sulpholipid content of brain contributed by the sulphogalactosylglycerolipid is 16% in 21-day-old rats and 8.4% in adult rats.     

Plasma proteome analysis in HTLV-1-associated myelopathy/tropical spastic paraparesis

Background

A new subgroup of HIV-1, designated Group P, was recently detected in two unrelated patients of Cameroonian origin. HIV-1 Group P phylogenetically clusters with SIVgor suggesting that it is the result of a cross-species transmission from gorillas. Until today, HIV-1 Group P has only been detected in two patients, and its degree of adaptation to the human host is largely unknown. Previous data have shown that pandemic HIV-1 Group M, but not non-pandemic Group O or rare Group N viruses, efficiently antagonize the human orthologue of the restriction factor tetherin (BST-2, HM1.24, CD317) suggesting that primate lentiviruses may have to gain anti-tetherin activity for efficient spread in the human population. Thus far, three SIV/HIV gene products (vpu, nef and env) are known to have the potential to counteract primate tetherin proteins, often in a species-specific manner. Here, we examined how long Group P may have been circulating in humans and determined its capability to antagonize human tetherin as an indicator of adaptation to humans.

Results

Our data suggest that HIV-1 Group P entered the human population between 1845 and 1989. Vpu, Env and Nef proteins from both Group P viruses failed to counteract human or gorilla tetherin to promote efficient release of HIV-1 virions, although both Group P Nef proteins moderately downmodulated gorilla tetherin from the cell surface. Notably, Vpu, Env and Nef alleles from the two HIV-1 P strains were all able to reduce CD4 cell surface expression.

Conclusions

Our analyses of the two reported HIV-1 Group P viruses suggest that zoonosis occurred in the last 170 years and further support that pandemic HIV-1 Group M strains are better adapted to humans than non-pandemic or rare Group O, N and P viruses. The inability to antagonize human tetherin may potentially explain the limited spread of HIV-1 Group P in the human population.     

Redefining Phrymaceae: the placement of Mimulus, tribe Mimuleae, and Phryma

Chloroplast trnL/F and nuclear ribosomal ITS and ETS sequence data were used to analyze phylogenetic relationships among members of tribe Mimuleae (Scrophulariaceae) and other closely related families in Lamiales. The results of these analyses led to the following conclusions. (1) The Australian genera Glossostigma and Peplidium and the taxonomically isolated Phryma join four genera of tribe Mimuleae to form a well-supported clade that is distinct from other families in the Lamiales. We refer to that clade as the subfamily Phrymoideae. (2) The genera Mazus and Lancea (tribe Mimuleae) together form a well-supported clade that we recognize as the subfamily Mazoideae. Mazoideae is weakly supported as sister to Phrymoideae. We assign Mazoideae and Phrymoideae to a redefined family Phrymaceae. (3) Mimulus is not monophyletic, because members of at least six other genera have been derived from within it. In light of the molecular evidence, it is clear that species of Phrymaceae (about 190 species) have undergone two geographically distinct radiations; one in western North America (about 130 species) and another in Australia (about 30 species). Phylogenetic interpretations of morphological evolution and biogeographical patterns are discussed.     

Do Leptin Levels Predict Weight Gain?—A 5-Year Follow-Up Study in Mauritius

Objective : To investigate whether relative baseline leptin levels predict long-term changes in adiposity and/or its distribution. Research Methods and Procedures : In a longitudinal study of 2888 nondiabetic Mauritians aged 25 years to 74 years who participated in population-based surveys in 1987 and 1992, changes in body mass index (BMI), waist/hip ratio (WHR), and waist circumference were compared between “hyperleptinemic,” “normoleptinemic,” and “hypoleptinemic” groups. “Relative leptin levels” were calculated as standardized residuals from the regression of log₁₀ leptin on baseline BMI to provide a leptin measure independent of BMI. Analyses were performed within each sex. A linear regression model was used to assess the effect of standardized residuals on changes in BMI, WHR, and waist circumference, independent of baseline BMI, age, fasting insulin, and ethnicity. Results : After adjusting for age and baseline BMI by analysis of covariance, there was no difference in changes in BMI, WHR, or waist circumference between men with low, normal, or high relative leptin levels. Among women, there was a significant difference in ΔWHR across leptin groups, such that the largest increase occurred in the “normal” leptin group. For both men and women, the linear regression models explained ?10% of variation in dependent variables, and the only significant independent variables were age, BMI, and being of Chinese origin, compared with Indian origin. Discussion : These findings do not support a role for leptin concentration in predicting weight gain or changes in fat distribution in adults over a 5-year period.     

Culture-Independent Investigation of the Microbiome Associated with the Nematode Acrobeloides maximus

Background

Symbioses between metazoans and microbes are widespread and vital to many ecosystems. Recent work with several nematode species has suggested that strong associations with microbial symbionts may also be common among members of this phylu. In this work we explore possible symbiosis between bacteria and the free living soil bacteriovorous nematode Acrobeloides maximus.

Methodology

We used a soil microcosm approach to expose A. maximus populations grown monoxenically on RFP labeled Escherichia coli in a soil slurry. Worms were recovered by density gradient separation and examined using both culture-independent and isolation methods. A 16S rRNA gene survey of the worm-associated bacteria was compared to the soil and to a similar analysis using Caenorhabditis elegans N2. Recovered A. maximus populations were maintained on cholesterol agar and sampled to examine the population dynamics of the microbiome.

Results

A consistent core microbiome was extracted from A. maximus that differed from those in the bulk soil or the C. elegans associated set. Three genera, Ochrobactrum, Pedobacter, and Chitinophaga, were identified at high levels only in the A. maximus populations, which were less diverse than the assemblage associated with C. elegans. Putative symbiont populations were maintained for at least 4 months post inoculation, although the levels decreased as the culture aged. Fluorescence in situ hybridization (FISH) using probes specific for Ochrobactrum and Pedobacter stained bacterial cells in formaldehyde fixed nematode guts.

Conclusions

Three microorganisms were repeatedly observed in association with Acrobeloides maximus when recovered from soil microcosms. We isolated several Ochrobactrum sp. and Pedobacter sp., and demonstrated that they inhabit the nematode gut by FISH. Although their role in A. maximus is not resolved, we propose possible mutualistic roles for these bacteria in protection of the host against pathogens and facilitating enzymatic digestion of other ingested bacteria.     

Transient Combination Therapy Targeting the Immune Synapse Abrogates T Cell Responses and Prolongs Allograft Survival in Mice

T cells play a major role in allograft rejection, which occurs after T cell activation by the engagement of several functional molecules to form an immune synapse with alloantigen presenting cells. In this study, the immune synapse was targeted using mAbs directed to the TCR beta-chain (TCRβ) and lymphocyte function-associated antigen−1 (LFA1) to induce long-term allograft survival. Evaluation of antigen-specific T cell responses was performed by adoptively transferring CFSE labeled transgenic OT-II cells into wild-type mice and providing OVA peptide by intravenous injection. Graft survival studies were performed in mice by transplanting BALB/c ear skins onto the flanks of C57BL/6 recipients. The anti-TCRβ plus anti-LFA1 mAb combination (but not either mAb alone) abrogated antigen-specific T cell responses invitro and invivo. Transient combination therapy with these agents resulted in significantly prolonged skin allograft survival in mice (51±10 days; p<0.01) when compared to treatment with either anti-TCRβ mAb (24±5 days) or anti-LFA1 mAb (19±3 days) alone or no treatment (10±1 days). When lymphoid tissues from these mice were analyzed at different times post-transplant, only those receiving the combination of anti-TCRβ and anti-LFA1 mAbs demonstrated long-lasting reductions in total T cell numbers, cellular and humoral anti-donor responses, and expression of CD3 on the surface of T cells. These results demonstrate that transient anti-TCRβ and anti-LFA1 mAb combination therapy abrogates antigen-reactive T cell responses with long-lasting effects that significantly prolong allograft survival.     

Spatial and Temporal Variation of Roots, Arbuscular Mycorrhizal Fungi, and Plant and Soil Nutrients in a Mature Pinot Noir (Vitis vinifera L.) Vineyard in Oregon, USA

Soil and crop management practices may influence biomass growth and yields of cotton (Gossypium hirsutum L.) and sorghum (Sorghum bicolorL.) and sequester significant amount of atmospheric CO₂in plant biomass and underlying soil, thereby helping to mitigate the undesirable effects of global warming. This study examined the effects of three tillage practices [no-till (NT), strip till (ST), and chisel till (CT)], four cover crops [legume (hairy vetch) (Vicia villosa roth), nonlegume (rye) (Secale cerealeL), hairy vetch/rye mixture, and winter weeds orno covercrop], and three N fertilization rates (0, 60–65, and 120–130 kg N ha ^–1) on the amount of C sequestered in cotton lint (lint + seed), sorghum grain, their stalks (stems + leaves) and roots, and underlying soil from 2000 to 2002 in central Georgia, USA. A field experiment was conducted on a Dothan sandy loam (fine-loamy, kaolinitic, thermic, Plinthic Kandiudults). In 2000, C accumulation in cotton lint was greater in NT with rye or vetch/rye mixture but in stalks, it was greater in ST with vetch or vetch/rye mixture than in CT with or without cover crops. Similarly, C accumulation in lint was greater in NT with 60 kg N ha ^–1 but in stalks, it was greater in ST with 60 and 120 kg N ha ^–1 than in CT with 0 kg N ha ^–1. In 2001, C accumulation in sorghum grains and stalks was greater in vetch and vetch/rye mixture with or without N rate than in rye without N rate. In 2002, C accumulation in cotton lint was greater in CT with or without N rate but in stalks, it was greater in ST with 60 and 120 kg N ha ^–1 than in NT with or without N rate. Total C accumulation in the above- and belowground biomass in cotton ranged from 1.7 to 5.6 Mg ha ^–1 and in sorghum ranged from 3.4 to 7.2 Mg ha ^–1. Carbon accumulation in cotton and sorghum roots ranged from 1 to 14% of the total C accumulation in above- and belowground biomass. In NT, soil organic C at 0–10 cm depth was greater in vetch with 0 kg N ha ^–1 or in vetch/rye with 120–130 kg N ha ^–1 than in weeds with 0 and 60 kg N ha ^–1 but at 10–30 cm, it was greater in rye with 120–130 kg N ha ^–1 than in weeds with or without rate. In ST, soil organic C at 0–10 cm was greater in rye with 120–130 kg N ha ^–1 than in rye, vetch, vetch/rye and weeds with 0 and 60 kg N ha ^–1. Soil organic C at 0–10 and 10–30 cm was also greater in NT and ST than in CT. Since 5 to 24% of C accumulation in lint and grain were harvested, C sequestered in cotton and sorghum stalks and roots can be significant in the terrestrial ecosystem and can significantly increase C storage in the soil if these residues are left after lint or grain harvest, thereby helping to mitigate the effects of global warming. Conservation tillage, such as ST, with hairy vetch/rye mixture cover crops and 60–65 kg N ha ^–1 can sustain C accumulation in cotton lint and sorghum grain and increase C storage in the surface soil due to increased C input from crop residues and their reduced incorporation into the soil compared with conventional tillage, such as CT, with no cover crop and N fertilization, thereby maintaining crop yields, improving soil quality, and reducing erosion.