Short-term high fat feeding increases organ injury and mortality after polymicrobial sepsis

The purpose of this study was to examine the effect of short-term high fat feeding on the inflammatory response in polymicrobial sepsis. Male C57BL/6 mice at 6 weeks of age were randomized to a high-fat diet (HFD) (60% kcal fat) or control diet (CD) (16% kcal fat) for 3 weeks. After 3 weeks of feeding, sepsis was induced by cecal ligation and puncture (CLP) and animals were monitored for survival. In a separate experiment, after 3 weeks of feeding mice underwent CLP and were sacrificed at various time points thereafter. Tissue was collected for biochemical studies. Mice fed a HFD gained more weight and had a greater fat mass compared to CD-fed mice. Mice on a HFD had a lower probability of survival and more severe lung injury compared with CD-fed mice following sepsis. Myeloperoxidase (MPO) activity, an indicator of neutrophil infiltration, was increased in the lung and liver after CLP in HFD-fed mice compared with CD (P < 0.05). The plasma cytokines tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were increased in both groups after CLP, however, TNF-α and IL-6 levels were lower in HFD mice at 3 h after CLP compared with CD and consistent with lung, but not liver, messenger RNA (mRNA) expression. Leptin levels were higher in HFD-fed mice at 18 h after sepsis compared to baseline levels (P < 0.05). Polymicrobial sepsis increased hepatic nuclear factor-κB (NF-κB) activation in HFD-fed mice after CLP vs. CD-fed mice. Short duration high fat feeding increases mortality and organ injury following polymicrobial sepsis. These effects correspond to changes in NF-κB.     

An Outbreak of Tularemia in a Colony of Outdoor-Housed Rhesus Macaques (Macaca mulatta)

Since an epizootic and detection of clinical cases of tularemia (Francisella tularensis) in 1996 at the Oregon National Primate Research Center, only 8 cases were identified in the succeeding 13 y. However, within a period of 7 mo, primarily during Winter 2010, 6 rhesus macaques were confirmed positive for Francisella tularensis type B by the Centers for Disease Control and Prevention by culture and fluorescent antibody testing. All cases had similar gross pathologic findings, which included necrotizing splenitis and lymphadenitis. Recent colony management efforts have focused on minimizing nonhuman primate exposure to commonly observed reservoir species and controlling rodent access to corral-style housing. Strategies continue to evolve with regard to managing a large breeding colony of nonhuman primates in the presence of this threat.Abbreviation: ONPRC, Oregon National Primate Research CenterFrancisella tularensis, the causative agent of tularemia, is a small pleomorphic gram-negative coccobacillus.¹¹ Severe disease and potentially death in humans can result from exposure to as few as 10 cfu of this highly infectious organism.^7,10 The disease is also known as rabbit fever and deer-fly fever, reflecting 2 common sources of infection for humans.³ F. tularensis is classified by the United States Department of Health and Human Services as a Category A Select Agent.⁶ It is considered a potential agent of biologic warfare, and in fact, has been weaponized and stockpiled in the past.¹⁰ The 2 biovars that are referenced most frequently in published human and nonhuman primate literature are tularensis (type A) and holarctica (formerly paleartica; type B).^4,12,13,23 An additional biovar, novicida (type C), has been described, but its virulence in humans is decreased due to its lack of a capsule.¹⁰Tularemia is endemic to many parts of the northern hemisphere, which includes the region surrounding the Oregon National Primate Research Center (ONPRC), an AAALAC-accredited facility.²⁰ Tularemia has one of the broadest host ranges of all bacteria, encompassing well over 200 mammalian species primarily, in addition to birds, amphibians, fish, and various arthropods such as fleas, ticks, mosquitoes, and flies.^10,15,19,20 The ONPRC is located in a mixed forest and field environment which is bordered by wetlands and residential neighborhoods outside of Portland. More than 4500 nonhuman primates are housed here, and most live outdoors in breeding groups. Therefore, exposure to this potentially life-threatening and zoonotic pathogen is inevitable, due to its persistence in the environment and the close proximity of several reservoir species. Presumed reservoir species commonly observed at ONPRC include meadow voles (Microtus pennsylvanicus), brown rats (Rattus norvegicus), deer mice (genus Peromyscus), house mice (Mus musculus), and California ground squirrels (Spermophilus beecheyi). Potential arthropod vectors that are monitored regularly at ONPRC include biting flies and mosquitoes. At this time, testing of prospective rodent carriers for tularemia is ongoing; therefore, in the interest of caution, all of the rodent and arthropod species listed are considered potential carriers of the disease.Tularemia was first recognized at the ONPRC in 1996 during an epizootic that resulted in 24 deaths among corral-housed rhesus macaques. Serology results from banked sera and sera collected during and after the outbreak demonstrated a seroconversion rate of approximately 25% in 723 animals. During the succeeding 13 y, only 8 sporadic cases were diagnosed. However, within a period of 3 mo during the winter of 2010, 5 rhesus macaques were diagnosed with Francisella tularensis type B by the Centers for Disease Control and Prevention (Atlanta, GA). Four months later, an additional case was confirmed. All 6 macaques were younger than 1 y and were assigned to a breeding colony protocol approved by the ONPRC Animal Care and Use Committee. The current report describes the clinical signs and gross and histologic findings associated with these cases, as well as methods for prevention and control of future cases of disease.     

Functional analysis of alleged NOGGIN mutation G92E disproves its pathogenic relevance

We identified an amino acid change (p.G92E) in the Bone Morphogenetic Protein antagonist NOGGIN in a 22-month-old boy who presented with a unilateral brachydactyly type B phenotype. Brachydactyly type B is a skeletal malformation that has been associated with increased Bone Morphogenetic Protein pathway activation in other patients. Previously, the amino acid change p.G92E in NOGGIN was described as causing fibrodysplasia ossificans progressiva, a rare genetic disorder characterized by limb malformations and progressive heterotopic bone formation in soft tissues that, like Brachydactyly type B, is caused by increased activation of Bone Morphogenetic Protein signaling. To determine whether G92E-NOGGIN shows impaired antagonism that could lead to increased Bone Morphogenetic Protein signaling, we performed functional assays to evaluate inhibition of BMP signaling. Interestingly, wt-NOGGIN shows different inhibition efficacies towards various Bone Morphogenetic Proteins that are known to be essential in limb development. However, comparing the biological activity of G92E-NOGGIN with wt-NOGGIN, we observed that G92E-NOGGIN inhibits activation of bone morphogenetic protein signaling with equal efficiency as wt-NOGGIN, supporting that G92E-NOGGIN does not cause pathological effects. Genetic testing of the child's parents revealed the same amino acid change in the healthy father, further supporting that p.G92E is a neutral amino acid substitution in NOGGIN. We conclude that p.G92E represents a rare polymorphism of the NOGGIN gene-- causing neither brachydactyly nor fibrodysplasia ossificans progressiva. This study highlights that a given genetic variation should not be considered pathogenic unless supported by functional analyses.     

Tuberculosis incidence rates during 8 years of follow-up of an antiretroviral treatment cohort in South Africa: comparison with rates in the community

Background

Although antiretroviral therapy (ART) is known to be associated with time-dependent reductions in tuberculosis (TB) incidence, the long-term impact of ART on incidence remains imprecisely defined due to limited duration of follow-up and incomplete CD4 cell count recovery in existing studies. We determined TB incidence in a South African ART cohort with up to 8 years of follow-up and stratified rates according to CD4 cell count recovery. We compared these rates with those of HIV-uninfected individuals living in the same community.

Methodology/Principal Findings

Prospectively collected clinical data on patients receiving ART in a community-based cohort in Cape Town were analysed. 1544 patients with a median follow-up of 5.0 years (IQR 2.4–5.8) were included in the analysis. 484 episodes of incident TB (73.6% culture-confirmed) were diagnosed in 424 patients during 6506 person-years (PYs) of follow-up. The TB incidence rate during the first year of ART was 12.4 (95% CI 10.8–14.4) cases/100PYs and decreased to 4.92 (95% CI 3.64–8.62) cases/100PYs between 5 and 8 years of ART. During person-time accrued within CD4 cell strata 0–100, 101–200, 201–300, 301–400, 401–500, 501–700 and ≥700 cells/µL, TB incidence rates (95% CI) were 25.5 (21.6–30.3), 11.2 (9.4–13.5), 7.9 (6.4–9.7), 5.0 (3.9–6.6), 5.1 (3.8–6.8), 4.1 (3.1–5.4) and 2.7 (1.7–4.5) cases/100PYs, respectively. Overall, 75% (95% CI 70.9–78.8) of TB episodes were recurrent cases. Updated CD4 cell count and viral load measurements were independently associated with long-term TB risk. TB rates during person-time accrued in the highest CD4 cell count stratum (>700 cells/µL) were 4.4-fold higher that the rate in HIV uninfected individuals living in the same community (2.7 versus 0.62 cases/100PYs; 95%CI 0.58–0.65).

Conclusions/Significance

TB rates during long-term ART remained substantially greater than rates in the local HIV uninfected populations regardless of duration of ART or attainment of CD4 cell counts exceeding 700 cells/µL.     

Subterranean, herbivore-induced plant volatile increases biological control activity of multiple beneficial nematode species in distinct habitats

While the role of herbivore-induced volatiles in plant-herbivore-natural enemy interactions is well documented aboveground, new evidence suggests that belowground volatile emissions can protect plants by attracting entomopathogenic nematodes (EPNs). However, due to methodological limitations, no study has previously detected belowground herbivore-induced volatiles in the field or quantified their impact on attraction of diverse EPN species. Here we show how a belowground herbivore-induced volatile can enhance mortality of agriculturally significant root pests. First, in real time, we identified pregeijerene (1,5-dimethylcyclodeca-1,5,7-triene) from citrus roots 9-12 hours after initiation of larval Diaprepes abbreviatus feeding. This compound was also detected in the root zone of mature citrus trees in the field. Application of collected volatiles from weevil-damaged citrus roots attracted native EPNs and increased mortality of beetle larvae (D. abbreviatus) compared to controls in a citrus orchard. In addition, field applications of isolated pregeijerene caused similar results. Quantitative real-time PCR revealed that pregeijerene increased pest mortality by attracting four species of naturally occurring EPNs in the field. Finally, we tested the generality of this root-zone signal by application of pregeijerene in blueberry fields; mortality of larvae (Galleria mellonella and Anomala orientalis) again increased by attracting naturally occurring populations of an EPN. Thus, this specific belowground signal attracts natural enemies of widespread root pests in distinct agricultural systems and may have broad potential in biological control of root pests.     

Silk Film Culture System for in vitro Analysis and Biomaterial Design

Silk films are promising protein-based biomaterials that can be fabricated with high fidelity and economically within a research laboratory environment ^1,2 . These materials are desirable because they possess highly controllable dimensional and material characteristics, are biocompatible and promote cell adhesion, can be modified through topographic patterning or by chemically altering the surface, and can be used as a depot for biologically active molecules for drug delivery related applications ^3-8 . In addition, silk films are relatively straightforward to custom design, can be designed to dissolve within minutes or degrade over years in vitro or in vivo, and are produce with the added benefit of being transparent in nature and therefore highly suitable for imaging applications ^9-13. The culture system methodology presented here represents a scalable approach for rapid assessments of cell-silk film surface interactions. Of particular interest is the use of surface patterned silk films to study differences in cell proliferation and responses of cells for alignment ^12,14 . The seeded cultures were cultured on both micro-patterned and flat silk film substrates, and then assessed through time-lapse phase-contrast imaging, scanning electron microscopy, and biochemical assessment of metabolic activity and nucleic acid content. In summary, the silk film in vitro culture system offers a customizable experimental setup suitable to the study of cell-surface interactions on a biomaterial substrate, which can then be optimized and then translated to in vivo models. Observations using the culture system presented here are currently being used to aid in applications ranging from basic cell interactions to medical device design, and thus are relevant to a broad range of biomedical fields.     

烟草种间杂交亲和性

以野生烟草Nicotiana alata、N.rustica、N.repanda、N.stocktonnii与栽培烟草K326、红花大金元、Yun87、Yun97为材料,进行种间正反杂交,研究种间杂交亲和性。田间观察杂交后的坐果情况并统计坐果率,采用显微荧光染色观察授粉后花粉管在雌蕊上的生长情况,并结合杂交后代萌发检测的方法。结果表明：N.rustica、N.repanda、N.stocktonnii与栽培烟草杂交不亲和。N.rustica花粉能够穿过K326花柱,N.repanda和N.stocktonnii花粉在K326柱头上很少萌发生长。N.alata花粉可以穿过K326的花柱,并得到果实,但是萌发实验显示其种子无活力。N.alata作为母本与栽培烟草杂交不亲和。     

中国外来入侵植物研究现状及存在的问题

生物入侵是当今各国发展的一大挑战,特别是像中国这样的发展中国家。然而,我们对外来入侵种的本底资料还很不清楚,更缺乏相关的深入调查;对生物入侵所造成的生态和经济影响还没有引起足够的重视,同时也忽视了盲目引种可能带来的各种危害。本文在文献调研的基础上,从外来植物的入侵历史和文化、中国外来入侵植物的调查现状、外来植物入侵对生态安全及经济的影响等方面进行了全面系统的总结,并提出了中国外来入侵植物研究领域存在的问题及应对措施。     

The effects of land use and climate change on the carbon cycle of Europe over the past 500 years

The long residence time of carbon in forests and soils means that both the current state and future behavior of the terrestrial biosphere are influenced by past variability in climate and anthropogenic land use. Over the last half‐millennium, European terrestrial ecosystems were affected by the cool temperatures of the Little Ice Age, rising CO₂ concentrations, and human induced deforestation and land abandonment. To quantify the importance of these processes, we performed a series of simulations with the LPJ dynamic vegetation model driven by reconstructed climate, land use, and CO₂ concentrations. Although land use change was the major control on the carbon inventory of Europe over the last 500 years, the current state of the terrestrial biosphere is largely controlled by land use change during the past century. Between 1500 and 2000, climate variability led to temporary sequestration events of up to 3 Pg, whereas increasing atmospheric CO₂ concentrations during the 20th century led to an increase in carbon storage of up to 15 Pg. Anthropogenic land use caused between 25 Pg of carbon emissions and 5 Pg of uptake over the same time period, depending on the historical and spatial pattern of past land use and the timing of the reversal from deforestation to afforestation during the last two centuries. None of the currently existing anthropogenic land use change datasets adequately capture the timing of the forest transition in most European countries as recorded in historical observations. Despite considerable uncertainty, our scenarios indicate that with limited management, extant European forests have the potential to absorb between 5 and 12 Pg of carbon at the present day.