RESUMO
Trophic ecology of detrital-based food webs is still poorly understood. Abyssal plains depend entirely on detritus and are among the most understudied ecosystems, with deposit feeders dominating megafaunal communities. We used compound-specific stable isotope ratios of amino acids (CSIA-AA) to estimate the trophic position of three abundant species of deposit feeders collected from the abyssal plain of the Northeast Pacific (Station M; ~ 4000 m depth), and compared it to the trophic position of their gut contents and the surrounding sediments. Our results suggest that detritus forms the base of the food web and gut contents of deposit feeders have a trophic position consistent with primary consumers and are largely composed of a living biomass of heterotrophic prokaryotes. Subsequently, deposit feeders are a trophic level above their gut contents making them secondary consumers of detritus on the abyssal plain. Based on δ13C values of essential amino acids, we found that gut contents of deposit feeders are distinct from the surrounding surface detritus and form a unique food source, which was assimilated by the deposit feeders primarily in periods of low food supply. Overall, our results show that the guts of deposit feeders constitute hotspots of organic matter on the abyssal plain that occupy one trophic level above detritus, increasing the food-chain length in this detritus-based ecosystem.
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Much of what we know about speciation comes from detailed studies of well-known model systems. Although there have been several important syntheses on speciation, few (if any) have explicitly compared speciation among major groups across the Tree of Life. Here, we synthesize and compare what is known about key aspects of speciation across taxa, including bacteria, protists, fungi, plants, and major animal groups. We focus on three main questions. Is allopatric speciation predominant across groups? How common is ecological divergence of sister species (a requirement for ecological speciation), and on what niche axes do species diverge in each group? What are the reproductive isolating barriers in each group? Our review suggests the following patterns. (i) Based on our survey and projected species numbers, the most frequent speciation process across the Tree of Life may be co-speciation between endosymbiotic bacteria and their insect hosts. (ii) Allopatric speciation appears to be present in all major groups, and may be the most common mode in both animals and plants, based on non-overlapping ranges of sister species. (iii) Full sympatry of sister species is also widespread, and may be more common in fungi than allopatry. (iv) Full sympatry of sister species is more common in some marine animals than in terrestrial and freshwater ones. (v) Ecological divergence of sister species is widespread in all groups, including ~70% of surveyed species pairs of plants and insects. (vi) Major axes of ecological divergence involve species interactions (e.g. host-switching) and habitat divergence. (vii) Prezygotic isolation appears to be generally more widespread and important than postzygotic isolation. (viii) Rates of diversification (and presumably speciation) are strikingly different across groups, with the fastest rates in plants, and successively slower rates in animals, fungi, and protists, with the slowest rates in prokaryotes. Overall, our study represents an initial step towards understanding general patterns in speciation across all organisms.
Assuntos
Ecossistema , Especiação Genética , Animais , Insetos , Filogenia , PlantasRESUMO
Plant-soil feedbacks (PSFs) drive plant community diversity via interactions between plants and soil microbes. However, we know little about how frequently PSFs affect plants at the seed stage, and the compositional shifts in fungi that accompany PSFs on germination.We conducted a pairwise PSF experiment to test whether seed germination was differentially impacted by conspecific versus heterospecific soils for seven grassland species. We used metagenomics to characterize shifts in fungal community composition in soils conditioned by each plant species. To investigate whether changes in the abundance of certain fungal taxa were associated with multiple PSFs, we assigned taxonomy to soil fungi and identified putative pathogens that were significantly more abundant in soils conditioned by plant species that experienced negative or positive PSFs.We observed negative, positive, and neutral PSFs on seed germination. Although conspecific and heterospecific soils for pairs with significant PSFs contained host-specialized soil fungal communities, soils with specialized microbial communities did not always lead to PSFs. The identity of host-specialized pathogens, that is, taxa uniquely present or significantly more abundant in soils conditioned by plant species experiencing negative PSFs, overlapped among plant species, while putative pathogens within a single host plant species differed depending on the identity of the heterospecific plant partner. Finally, the magnitude of feedback on germination was not related to the degree of fungal community differentiation between species pairs involved in negative PSFs. Synthesis. Our findings reveal the potential importance of PSFs at the seed stage. Although plant species developed specialized fungal communities in rhizosphere soil, pathogens were not strictly host-specific and varied not just between plant species, but according to the identity of plant partner. These results illustrate the complexity of microbe-mediated interactions between plants at different life stages that next-generation sequencing can begin to unravel.
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Around the world, many species are confined to "Sky Islands," with different populations in isolated patches of montane habitat. How does this pattern arise? One scenario is that montane species were widespread in lowlands when climates were cooler, and were isolated by local extinction caused by warming conditions. This scenario implies that many montane species may be highly susceptible to anthropogenic warming. Here, we test this scenario in a montane lizard (Sceloporus jarrovii) from the Madrean Sky Islands of southeastern Arizona. We combined data from field surveys, climate, population genomics, and physiology. Overall, our results support the hypothesis that this species' current distribution is explained by local extinction caused by past climate change. However, our results for this species differ from simple expectations in several ways: (a) their absence at lower elevations is related to warm winter temperatures, not hot summer temperatures; (b) they appear to exclude a low-elevation congener from higher elevations, not the converse; (c) they are apparently absent from many climatically suitable but low mountain ranges, seemingly "pushed off the top" by climates even warmer than those today; (d) despite the potential for dispersal among ranges during recent glacial periods (~18,000 years ago), populations in different ranges diverged ~4.5-0.5 million years ago and remained largely distinct; and (e) body temperatures are inversely related to climatic temperatures among sites. These results may have implications for many other Sky Island systems. More broadly, we suggest that Sky Island species may be relevant for predicting responses to future warming.
Assuntos
Mudança Climática , DNA Mitocondrial/genética , Lagartos/genética , Filogeografia , Animais , Arizona , Ecossistema , Variação Genética/genética , Ilhas , FilogeniaRESUMO
We respond to seven criticisms made by Vermeij et al. () regarding Miller & Wiens (). Their criticisms generally reflect misunderstandings, unsupported speculations, and topics that were explicitly addressed in our paper.
Assuntos
BiodiversidadeRESUMO
Phylogenomic approaches offer a wealth of data, but a bewildering diversity of methodological choices. These choices can strongly affect the resulting topologies. Here, we explore two controversial approaches (binning genes into "supergenes" and inclusion of only rapidly evolving sites), using new data from hyloid frogs. Hyloid frogs encompass â¼53% of frog species, including true toads (Bufonidae), glassfrogs (Centrolenidae), poison frogs (Dendrobatidae), and treefrogs (Hylidae). Many hyloid families are well-established, but relationships among these families have remained difficult to resolve. We generated a dataset of ultraconserved elements (UCEs) for 50 ingroup species, including 18 of 19 hyloid families and up to 2214 loci spanning >800,000 aligned base pairs. We evaluated these two general approaches (binning, rapid sites only) based primarily on their ability to recover and strongly support well-established clades. Data were analyzed using concatenated likelihood and coalescent species-tree methods (NJst, ASTRAL). Binning strongly affected inferred relationships, whereas use of only rapidly evolving sites did not (indicating â¼87% of the data contributed little information). The optimal approaches for maximizing recovery and support of well-established clades were concatenated likelihood analysis and the use of a limited number of naive bins (statistical binning gave more problematic results). These two optimal approaches converged on similar relationships among hyloid families, and resolved them with generally strong support. The relationships found were very different from most previous estimates of hyloid phylogeny, and a new classification is proposed. The new phylogeny also suggests an intriguing biogeographical scenario, in which hyloids originated in southern South America before radiating throughout the world.
Assuntos
Anuros/classificação , Loci Gênicos , Genômica/métodos , Filogenia , Animais , Sequência Conservada/genética , Funções Verossimilhança , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
The marine-terrestrial richness gradient is among Earth's most dramatic biodiversity patterns, but its causes remain poorly understood. Here, we analyse detailed phylogenies of amniote clades, paleontological data and simulations to reveal the mechanisms underlying low marine richness, emphasising speciation, extinction and colonisation. We show that differences in diversification rates (speciation minus extinction) between habitats are often weak and inconsistent with observed richness patterns. Instead, the richness gradient is explained by limited time for speciation in marine habitats, since all extant marine clades are relatively young. Paleontological data show that older marine invasions have consistently ended in extinction. Simulations show that marine extinctions help drive the pattern of young, depauperate marine clades. This role for extinction is not discernible from molecular phylogenies alone, and not predicted by most previously hypothesised explanations for this gradient. Our results have important implications for the marine-terrestrial biodiversity gradient, and studies of biodiversity gradients in general.
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Biodiversidade , Especiação Genética , Filogenia , Organismos Aquáticos , Ecossistema , Extinção Biológica , Oceanos e Mares , Fatores de TempoRESUMO
OBJECTIVE: The transversus abdominis plane (TAP) block anesthetizes the anterior branches of spinal nerves that innervate the abdominal wall from T6 to L1 dermatomes and provide effective postoperative analgesia after abdominal surgery. Several applications of TAP catheters are described for both acute and chronic abdominal wall pain, but there are no reported cases of TAP catheters used during pregnancy. CASE REPORT: We present the case of a 23-year-old primigravida admitted to the hospital on multiple occasions during her pregnancy for right lower quadrant abdominal "stabbing" pain, with a visual analog scale score intensity of 8/10 abdominal pain. The diagnosis was unclear despite the presence of prominent ileocolic lymph nodes visualized on magnetic resonance imaging. The abdominal pain persisted despite escalating doses of intravenous opioids (up to 30 mg of intravenous hydromorphone daily). At 34 weeks' gestation, a right-sided TAP block with 0.25% bupivacaine was inserted under ultrasound guidance, and this relieved all of her pain (visual analog scale score, 0). The patient was managed with repeated boluses of 0.5% ropivacaine via a TAP catheter, which resulted in complete resolution of her pain within 3 days and allowed for complete discontinuation of opioid medications. The patient was discharged home, with no recurrence of pain. She had an uneventful cesarean delivery at term. CONCLUSIONS: A TAP catheter inserted under ultrasound guidance can be effective for the treatment of chronic abdominal pain during pregnancy and may provide an alternative analgesic modality when intravenous opioids are not providing relief or when neuraxial analgesia techniques are not feasible or contraindicated.
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Dor Abdominal/cirurgia , Parede Abdominal/cirurgia , Bloqueio Nervoso/métodos , Complicações na Gravidez/cirurgia , Dor Abdominal/diagnóstico por imagem , Parede Abdominal/diagnóstico por imagem , Analgesia Obstétrica/métodos , Bupivacaína/administração & dosagem , Feminino , Humanos , Gravidez , Complicações na Gravidez/diagnóstico por imagem , Resultado do Tratamento , Ultrassonografia de Intervenção/métodos , Adulto JovemRESUMO
PURPOSE: In 2011, the hysterectomy enhanced recovery (HER) pathway, a multi-disciplinary, evidence-based care plan designed to improve recovery after open gynecologic surgery for non-malignant lesions, was introduced at The Ottawa Hospital (TOH). This before-and-after study examined the impact of the HER pathway on postoperative day (POD) 1 hospital discharge. METHODS: Ethical approval was obtained. This retrospective cohort study included patients who had undergone open abdominal gynecologic surgery for non-malignant lesions at TOH Civic Campus between July 2010 and September 2012 (the year before and year after HER implementation). Patients were analyzed in either a pre-HER or post-HER group depending on their surgery date. Patients with chronic pain and emergent surgery were excluded. Data were obtained via medical chart review. Our primary outcome was the percentage of POD 1 discharges before and after HER implementation. Secondary outcomes included return to hospital within 30 days of discharge, median length of stay (LOS), clinician compliance with HER, and an exploratory analysis with multivariable modelling to evaluate which aspects of the HER independently predicted POD 1 discharge. Variables used included American Society of Anesthesiologists physical status (≥ II), prior abdominal surgery, body mass index, use of transversus abdominis plane blocks, and anesthetic type. RESULTS: Among the 223 patients, significantly more POD 1 discharges occurred for post-HER compared to pre-HER patients (34% vs 7%, respectively; adjusted odds ratio [OR] = 7.33; 95% confidence interval [CI] = 3.05 to 17.62). Rates of return to hospital at 30 days were similar between the groups (10% post-HER and 13% pre-HER; adjusted OR = 0.74; 95% CI = 0.32 to 1.74). The median length of stay was two days in the post-HER group and three days in the pre-HER group (P < 0.0001). Only inhalational general anesthesia was independently associated with decreased odds of POD 1 discharge (adjusted OR = 0.16, 95% CI = 0.04 to 0.65). CONCLUSION: For patients undergoing abdominal hysterectomy, implementation of a HER pathway is associated with a higher POD 1 discharge rate, with no increase in the early return to hospital rate.
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Medicina Baseada em Evidências/métodos , Histerectomia/métodos , Alta do Paciente/estatística & dados numéricos , Readmissão do Paciente/estatística & dados numéricos , Adulto , Anestesia por Inalação/métodos , Estudos de Coortes , Feminino , Humanos , Tempo de Internação , Pessoa de Meia-Idade , Ontário , Período Pós-Operatório , Estudos Retrospectivos , Fatores de TempoRESUMO
To define the role of rare variants in advanced age-related macular degeneration (AMD) risk, we sequenced the exons of 681 genes within all reported AMD loci and related pathways in 2,493 cases and controls. We first tested each gene for increased or decreased burden of rare variants in cases compared to controls. We found that 7.8% of AMD cases compared to 2.3% of controls are carriers of rare missense CFI variants (odds ratio (OR) = 3.6; P = 2 × 10(-8)). There was a predominance of dysfunctional variants in cases compared to controls. We then tested individual variants for association with disease. We observed significant association with rare missense alleles in genes other than CFI. Genotyping in 5,115 independent samples confirmed associations with AMD of an allele in C3 encoding p.Lys155Gln (replication P = 3.5 × 10(-5), OR = 2.8; joint P = 5.2 × 10(-9), OR = 3.8) and an allele in C9 encoding p.Pro167Ser (replication P = 2.4 × 10(-5), OR = 2.2; joint P = 6.5 × 10(-7), OR = 2.2). Finally, we show that the allele of C3 encoding Gln155 results in resistance to proteolytic inactivation by CFH and CFI. These results implicate loss of C3 protein regulation and excessive alternative complement activation in AMD pathogenesis, thus informing both the direction of effect and mechanistic underpinnings of this disorder.
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Complemento C3/genética , Complemento C9/genética , Fator I do Complemento/genética , Degeneração Macular/genética , Envelhecimento , Substituição de Aminoácidos , Sequência de Bases , Ativação do Complemento/genética , Predisposição Genética para Doença , Variação Genética , Genótipo , Humanos , Risco , Análise de Sequência de DNARESUMO
Three new species of the triplefin blenny genus Enneanectes found in the Pacific Ocean off southern Mexico are described. Two, Enneanectes glendae and Enneanectes macrops, are mainland species, while the third, Enneanectes exsul, is endemic to the Islas Revillagigedo. A key to the five species of Enneanectes known from the tropical eastern Pacific is provided.
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Perciformes/classificação , Distribuição Animal , Estruturas Animais/anatomia & histologia , Estruturas Animais/crescimento & desenvolvimento , Animais , Feminino , Masculino , México , Tamanho do Órgão , Oceano Pacífico , Perciformes/anatomia & histologia , Perciformes/crescimento & desenvolvimentoRESUMO
C3 deficiency is a rare disorder that leads to recurrent pyogenic infections. Here we describe a previously healthy 18 y/o Caucasian male with severe meningococcal disease. Total hemolytic activity was zero secondary to an undetectable C3. The C3 gene was normal by sequencing. Mixing the patient's serum with normal human serum led to C3 consumption. An IgG autoantibody in the patient's serum was identified that stabilized the classical pathway C3 and C5 convertases, thus preventing decay of these enzyme complexes. This autoantibody is an example of a C4 nephritic factor, with an additional feature of stabilizing the C5 convertase. Previous patients with C4 nephritic factor had membranoproliferative glomerulonephritis. Two years after presentation, this patient's C3 remains undetectable with no evidence of renal disease. We revisit the role of autoantibodies to classical pathway convertases in disease, review the literature on C4-NeF and comment on its detection in the clinical laboratory.
Assuntos
Autoanticorpos/sangue , C3 Convertase da Via Clássica do Complemento/metabolismo , Complemento C3/deficiência , Infecções Meningocócicas/etiologia , Adolescente , Complemento C3/genética , Complemento C3/imunologia , C3 Convertase da Via Clássica do Complemento/imunologia , C5 Convertase da Via Clássica do Complemento/imunologia , C5 Convertase da Via Clássica do Complemento/metabolismo , Proteínas do Sistema Complemento , Estabilidade Enzimática , Humanos , Imunoglobulina G/sangue , Masculino , Meningite Meningocócica/etiologia , Meningite Meningocócica/imunologia , Infecções Meningocócicas/imunologia , Modelos Imunológicos , Sepse/etiologia , Sepse/imunologia , Análise de Sequência de DNARESUMO
Atypical hemolytic uremic syndrome (aHUS) is a rare renal thrombotic microangiopathy commonly associated with rare genetic variants in complement system genes, unique to each patient/family. Here, we report 14 sporadic aHUS patients carrying the same mutation, R139W, in the complement C3 gene. The clinical presentation was with a rapid progression to end-stage renal disease (6 of 14) and an unusually high frequency of cardiac (8 of 14) and/or neurologic (5 of 14) events. Although resting glomerular endothelial cells (GEnCs) remained unaffected by R139W-C3 sera, the incubation of those sera with GEnC preactivated with pro-inflammatory stimuli led to increased C3 deposition, C5a release, and procoagulant tissue-factor expression. This functional consequence of R139W-C3 resulted from the formation of a hyperactive C3 convertase. Mutant C3 showed an increased affinity for factor B and a reduced binding to membrane cofactor protein (MCP; CD46), but a normal regulation by factor H (FH). In addition, the frequency of at-risk FH and MCP haplotypes was significantly higher in the R139W-aHUS patients, compared with normal donors or to healthy carriers. These genetic background differences could explain the R139W-aHUS incomplete penetrance. These results demonstrate that this C3 mutation, especially when associated with an at-risk FH and/or MCP haplotypes, becomes pathogenic following an inflammatory endothelium-damaging event.
Assuntos
Complemento C3/genética , Síndrome Hemolítico-Urêmica/genética , Mutação de Sentido Incorreto , Mutação Puntual , Adolescente , Adulto , Idoso , Substituição de Aminoácidos , Síndrome Hemolítico-Urêmica Atípica , Células Cultivadas/efeitos dos fármacos , Pré-Escolar , Complemento C3/química , Complemento C3/metabolismo , Fator B do Complemento/metabolismo , Progressão da Doença , Células Endoteliais/efeitos dos fármacos , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Feminino , Haplótipos/genética , Síndrome Hemolítico-Urêmica/sangue , Síndrome Hemolítico-Urêmica/complicações , Síndrome Hemolítico-Urêmica/imunologia , Humanos , Lactente , Falência Renal Crônica/etiologia , Glomérulos Renais/patologia , Masculino , Proteína Cofatora de Membrana/metabolismo , Pessoa de Meia-Idade , Modelos Moleculares , Penetrância , Conformação Proteica , Ressonância de Plasmônio de Superfície , Adulto JovemRESUMO
OBJECTIVES: To determine the factors that put Canadian women at risk for not supplementing with folic acid (FA) in the three months before conception, as recommended for the prevention of infant neural tube defects. METHODS: This study used data from the Canadian Maternity Experiences Survey. We used Poisson regression analysis with a robust variance to determine which factors were associated with women not supplementing with FA in the three months prior to pregnancy as compared with women who did supplement. RESULTS: Of the 6421 women surveyed, 57.7% were supplementing with FA pre-conceptionally. The risk factors associated with a lack of FA supplementation pre-conceptionally were maternal age <19 (prevalence ratio [PR] = 0.50; 95% CI 0.36 to 0.69) or 20 to 24 (PR = 0.75; 95% CI 0.67 to 0.84); education below high school level (PR = 0.73; 95% CI 0.61 to 0.87), at high school level (PR = 0.77; 95% CI 0.71 to 0.83), or at post-secondary level other than university (PR = 0.93; 95% CI 0.88 to 0.97); being at or below the low-income cut-off (PR = 0.74; 95% CI 0.67 to 0.81); smoking before pregnancy (PR = 0.79; 95% CI 0.73 to 0.86); being non-fluent in the language of the health care provider (PR = 0.66; 95% CI 0.49 to 0.88); being obese (BMI ≥ 30) (PR = 0.91; 95% CI 0.85 to 0.98); being unemployed (PR = 0.94; 95% CI 0.89 to 1.00); and being born outside of Canada (PR = 0.79; 95% CI 0.74 to 0.84). CONCLUSION: Young maternal age, low education, low income, smoking, language barriers, obesity, unemployment, and being born outside Canada are risk factors for suboptimal or lack of FA supplementation pre-conceptionally.
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Ácido Fólico/administração & dosagem , Cuidado Pré-Concepcional , Adolescente , Adulto , Canadá , Estudos Transversais , Suplementos Nutricionais , Escolaridade , Feminino , Humanos , Idade Materna , Defeitos do Tubo Neural/prevenção & controle , Obesidade , Pobreza , Cuidado Pré-Concepcional/estatística & dados numéricos , Gravidez , Fatores de RiscoRESUMO
OBJECTIVE: Maternal smoking and preeclampsia independently increase the risk of adverse pregnancy outcomes; however, smoking decreases the risk of preeclampsia. We sought to estimate the risk of adverse pregnancy outcomes among preeclamptic women who smoke and hypothesized that this risk would be increased, compared with nonpreeclamptic women who smoke or preeclamptic women who do not smoke. STUDY DESIGN: With the use of the Niday Perinatal Database and multiple logistic regressions, we estimated the risk of adverse pregnancy outcomes in nonpreeclamptic women who smoke, preeclamptic women who do not smoke, and preeclamptic women who smoke in relation to nonpreeclamptic women who do not smoke. RESULTS: The incidence of adverse pregnancy outcomes was more than twice as high among preeclamptic women who smoke as among nonpreeclamptic women who do not smoke. The following data were observed: small-for-gestational-age infant (odds ratio [OR], 3.40; 95% CI, 2.27-4.89), preterm birth (OR, 5.77; 95% CI, 4.50-7.35), very preterm birth (OR, 5.44; 95% CI, 3.51-8.11), abruption (OR, 6.16; 95% CI, 3.05-11.01), Apgar <4 at 5 minutes (OR, 3.11; 95% CI, 1.48-5.72), and stillbirth (OR, 3.39; 95% CI, 1.33-6.99). CONCLUSION: Smoking decreases the risk of preeclampsia, but smokers with preeclampsia have an increased risk for adverse pregnancy outcomes.
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Pré-Eclâmpsia/epidemiologia , Resultado da Gravidez , Fumar/efeitos adversos , Descolamento Prematuro da Placenta/epidemiologia , Adulto , Índice de Apgar , Feminino , Humanos , Recém-Nascido , Recém-Nascido Pequeno para a Idade Gestacional , Ontário/epidemiologia , Gravidez , Nascimento Prematuro/epidemiologia , Risco , Fumar/epidemiologia , Natimorto/epidemiologiaRESUMO
Atypical hemolytic uremic syndrome (aHUS) is a disease of complement dysregulation. In approximately 50% of patients, mutations have been described in the genes encoding the complement regulators factor H, MCP, and factor I or the activator factor B. We report here mutations in the central component of the complement cascade, C3, in association with aHUS. We describe 9 novel C3 mutations in 14 aHUS patients with a persistently low serum C3 level. We have demonstrated that 5 of these mutations are gain-of-function and 2 are inactivating. This establishes C3 as a susceptibility factor for aHUS.
Assuntos
Complemento C3/genética , Síndrome Hemolítico-Urêmica/genética , Mutação , Adolescente , Adulto , Criança , Pré-Escolar , Códon sem Sentido , Complemento C3/análise , Análise Mutacional de DNA , Predisposição Genética para Doença , Síndrome Hemolítico-Urêmica/etiologia , Síndrome Hemolítico-Urêmica/imunologia , Heterozigoto , Humanos , Lactente , Mutação de Sentido Incorreto , Adulto JovemRESUMO
Human monocytic ehrlichiosis is caused by a tick-transmitted rickettsia, Ehrlichia chaffeensis. We recently reported that E. chaffeensis grown in tick cells expresses different proteins than bacteria grown in macrophages. Therefore, we tested the hypothesis that immune responses against E. chaffeensis would be different if the mice are challenged with bacteria grown in macrophages or tick cells. We assessed the E. chaffeensis clearance from the peritoneum, spleen, and liver by C57BL/6J mice using a TaqMan-based real-time reverse transcription-PCR assay. Macrophage-grown E. chaffeensis was cleared in 2 weeks from the peritoneum, whereas the pathogen from tick cells persisted for nine additional days and included three relapses of increasing bacterial load separated by three-day intervals. Tick cell-grown bacteria also persisted in the livers and spleens with higher bacterial loads compared to macrophage-grown bacteria and fluctuated over a period of 35 days. Three-day periodic cycles were detected in T-cell CD62L/CD44 ratios in the spleen and bone marrow in response to infections with both tick cell- and macrophage-grown bacteria and were accompanied by similar periodic cycles of spleen cell cytokine secretions and nitric oxide and interleukin-6 by peritoneal macrophages. The E. chaffeensis-specific immunoglobulin G response was considerably higher and steadily increased in mice infected with the tick cell-derived E. chaffeensis compared to DH82-grown bacteria. In addition, antigens detected by the immunoglobulins were significantly different between mice infected with the E. chaffeensis originating from tick cells or macrophages. The differences in the immune response to tick cell-grown bacteria compared to macrophage-grown bacteria reflected a delay in the shift of gene expression from the tick cell-specific Omp 14 gene to the macrophage-specific Omp 19 gene. These data suggest that the host response to E. chaffeensis depends on the source of the bacteria and that this experimental model requires the most natural inoculum possible to allow for a realistic understanding of host resistance.
Assuntos
Ehrlichia chaffeensis/crescimento & desenvolvimento , Ehrlichia chaffeensis/imunologia , Ehrlichiose/imunologia , Macrófagos/microbiologia , Carrapatos/microbiologia , Animais , Proteínas da Membrana Bacteriana Externa/genética , Western Blotting , Células Cultivadas , Modelos Animais de Doenças , Cães , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Imunoglobulina G/sangue , Ativação de Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Linfócitos T/microbiologiaRESUMO
Both isoforms of rat phosphatidylinositol transfer protein (PITP) mediate the intermembrane transfer of sphingomyelin (CerPCho). In the plasma membrane, CerPCho often segregates with cholesterol into microdomains such as lipid rafts and caveolae. To test the hypothesis that PITP exhibits a preference for CerPCho- and cholesterol-rich membranes, we prepared unilamellar vesicles containing variable amounts of these two lipids. We also used CerPCho species with different acyl composition and treated vesicles with agents known to sequester and remove cholesterol. We observed that the beta isoform of rat PITP was more sensitive to membrane cholesterol than was the alpha isoform, as shown by increases in specific activities of lipid transfer of 2-6-fold. A relatively high membrane content of cholesterol (mole fraction > 0.4) was required to elicit such enhancements. Treatment of cholesterol-rich membranes with a series of beta cyclodextrins demonstrated that, upon depletion of cholesterol from participating membranes, the PITPbeta activity changes were fully reversible. We finally noted that the mechanism by which cholesterol enhances the activity of PITPbeta appeared to involve a decreased affinity of the protein for the membrane surface, in a manner that was independent of vesicle size and membrane microviscosity. We conclude that PITPbeta interacts transiently but productively with the liquid-ordered phase formed by CerPCho and cholesterol and discuss the possibility of PITP interactions in vivo with sphingolipid- and cholesterol-rich membrane microdomains.