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1.
Barrie J. Anthony Kylie R. James Geoffrey N. Gobert Grant A. Ramm Donald P. McManus 《PloS one》2013,8(6)
Hepatic fibrosis induced by egg deposition is the most serious pathology associated with chronic schistosomiasis, in which the hepatic stellate cell (HSC) plays a central role. While the effect of Schistosoma mansoni eggs on the fibrogenic phenotype of HSCs has been investigated, studies determining the effect of eggs of
S
. japonicum
on HSCs are lacking. Disease caused by
S
. japonicum
is much more severe than that resulting from S. mansoni infection so it is important to compare the pathologies caused by these two parasites, to determine whether this phenotype is due to the species interacting differently with the mammalian host. Accordingly, we investigated the effect of
S
. japonicum
eggs on the human HSC cell line, LX-2, with and without TGF-β (Transforming Growth Factor beta) co-treatment, so as to determine the impact on genes associated with fibrogenesis, inflammation and matrix re-organisation. Activation status of HSCs was assessed by αSMA (Alpha Smooth Muscle Actin) immunofluorescence, accumulation of Oil Red O-stained lipid droplets and the relative expression of selected genes associated with activation. The fibrogenic phenotype of HSCs was inhibited by the presence of eggs both with or without TGF-β treatment, as evidenced by a lack of αSMA staining and reduced gene expression of αSMA and Col1A1 (Collagen 1A1). Unlike S. mansoni-treated cells, however, expression of the quiescent HSC marker PPAR-γ (Peroxisome Proliferator-Activated Receptor gamma) was not increased, nor was there accumulation of lipid droplets. In contrast,
S
. japonicum
eggs induced the mRNA expression of MMP-9 (Matrix Metalloproteinase 9), CCL2 (Chemokine (C-C motif) Ligand 2) and IL-6 (Interleukin 6) in HSCs indicating that rather than inducing complete HSC quiescence, the eggs induced a proinflammatory phenotype. These results suggest HSCs in close proximity to
S
. japonicum
eggs in the liver may play a role in the proinflammatory regulation of hepatic granuloma formation. 相似文献
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Tina R. Clark Amanda M. Lackey Betsy Kleba Lonnie O. Driskell Erika I. Lutter Craig Martens David O. Wood Ted Hackstadt 《Journal of bacteriology》2011,193(18):4993-4995
Transformation frequencies of a mariner-based transposon system in Rickettsia rickettsii were determined using a plaque assay system for enumeration and isolation of mutants. Sequence analysis of insertion sites in both R. rickettsii and R. prowazekii indicated that insertions were random. Transposon mutagenesis provides a useful tool for rickettsial research. 相似文献
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Tasnime N. Akbaraly Mark Hamer Jane E. Ferrie Gordon Lowe G. David Batty Gareth Hagger-Johnson Archana Singh-Manoux Martin J. Shipley Mika Kivim?ki 《CMAJ》2013,185(16):E763-E770
Background:
The importance of chronic inflammation as a determinant of aging phenotypes may have been underestimated in previous studies that used a single measurement of inflammatory markers. We assessed inflammatory markers twice over a 5-year exposure period to examine the association between chronic inflammation and future aging phenotypes in a large population of men and women.Methods:
We obtained data for 3044 middle-aged adults (28.2% women) who were participating in the Whitehall II study and had no history of stroke, myocardial infarction or cancer at our study’s baseline (1997–1999). Interleukin-6 was measured at baseline and 5 years earlier. Cause-specific mortality, chronic disease and functioning were ascertained from hospital data, register linkage and clinical examinations. We used these data to create 4 aging phenotypes at the 10-year follow-up (2007–2009): successful aging (free of major chronic disease and with optimal physical, mental and cognitive functioning), incident fatal or nonfatal cardiovascular disease, death from noncardiovascular causes and normal aging (all other participants).Results:
Of the 3044 participants, 721 (23.7%) met the criteria for successful aging at the 10-year follow-up, 321 (10.6%) had cardiovascular disease events, 147 (4.8%) died from noncardiovascular causes, and the remaining 1855 (60.9%) were included in the normal aging phenotype. After adjustment for potential confounders, having a high interleukin-6 level (> 2.0 ng/L) twice over the 5-year exposure period nearly halved the odds of successful aging at the 10-year follow-up (odds ratio [OR] 0.53, 95% confidence interval [CI] 0.38–0.74) and increased the risk of future cardiovascular events (OR 1.64, 95% CI 1.15–2.33) and noncardiovascular death (OR 2.43, 95% CI 1.58–3.80).Interpretation:
Chronic inflammation, as ascertained by repeat measurements, was associated with a range of unhealthy aging phenotypes and a decreased likelihood of successful aging. Our results suggest that assessing long-term chronic inflammation by repeat measurement of interleukin-6 has the potential to guide clinical practice. Chronic inflammation has been implicated in the pathogenesis of age-related conditions, 1 such as type 2 diabetes, 2 , 3 cardiovascular disease, 4 cognitive impairment 5 and brain atrophy. 6 Chronic inflammation may result from or be a cause of age-related disease processes (illustrated in Appendix 1, available at www.cmaj.ca/lookup/suppl/doi:10.1503/cmaj.122072/-/DC1 ). For example, obesity increases inflammation, and chronic inflammation, in turn, contributes to the development of type 2 diabetes by inducing insulin resistance, 7 , 8 and to coronary artery disease by promoting atherogenesis. 9 Thus, raised levels of inflammation appear to be implicated in various pathological processes leading to diseases in older age. Of the various markers of systemic inflammation, interleukin-6 is particularly relevant to aging outcomes. There is increasing evidence that interleukin-6 is the pro-inflammatory cytokine that “drives” downstream inflammatory markers, such as C-reactive protein and fibrinogen. 10 , 11 Interleukin-6, in contrast to C-reactive protein and fibrinogen, is also likely to play a causal role in aging owing to its direct effects on the brain and skeletal muscles. 12 , 13 In addition, results of Mendelian randomization studies of interleukin-6 and studies of antagonists are consistent with a causal role for interleukin-6 in relation to coronary artery disease, again in contrast to C-reactive protein and fibrinogen. 14 However, current understanding of the link between chronic inflammation and aging phenotypes is hampered by the methodologic limitations of many existing studies. Most studies reported an assessment of inflammation based on a single measurement, precluding a distinction between the short-term (acute) and longer-term (chronic) impact of the inflammatory process on disease outcomes. 7 We conducted a study using 2 measurements of interleukin-6 obtained about 5 years apart to examine the association between chronic inflammation and aging phenotypes assessed 10 years later in a large population of men and women. Because inflammation characterizes a wide range of pathological processes, we considered several aging phenotypes, including cardiovascular disease (fatal and nonfatal), death from noncardiovascular causes and successful aging (optimal functioning across different physical, mental and cognitive domains). 相似文献10.
Lucien Rufener Nicola Bedoni Roland Baur Samantha Rey Dominique A. Glauser Jacques Bouvier Robin Beech Erwin Sigel Alessandro Puoti 《PLoS pathogens》2013,9(8)
Monepantel is a member of the recently identified class of anthelmintics known as the amino-acetonitrile derivatives (AADs). Monepantel controls all major gastro-intestinal nematodes in sheep including those that are resistant to the classical anthelmintics. Previous studies have shown that the Caenorhabditis elegans acr-23 and the Haemonchus contortus Hco-mptl-1 genes may be prominent targets of monepantel. With this discovery it became possible to investigate the mode of action of monepantel in nematodes at the molecular level. In the present study, we show that a C. elegans mutant acr-23 strain is fully rescued by expressing the wild-type acr-23 gene. Moreover, we present a new mutant allele, and characterize acr-23 alleles genetically. We also show that acr-23 is expressed in body wall muscle cells, and provide therefore a possible explanation for the paralysis caused by monepantel. Furthermore, genetic evidence suggests that the chaperone RIC-3 is required for expression of full monepantel resistance. Finally, we present reconstitution of the C. elegans ACR-23 receptor in Xenopus laevis oocytes and provide direct evidence of its modulation by monepantel. Conversely, co-injection of the chaperone RIC-3 had no impact for channel reconstitution in X. laevis oocytes. These results reinforce the involvement of the ACR-23 family in the mode of action of monepantel and advance our understanding of this new class of anthelmintics. 相似文献
11.
Heme and chlorophyll accumulate to
high levels in legume root nodules and in photosynthetic tissues,
respectively, and they are both derived from the universal tetrapyrrole
precursor δ-aminolevulinic acid (ALA). The first committed step in
ALA and tetrapyrrole synthesis is catalyzed by glutamyl-tRNA reductase
(GTR) in plants. A soybean (Glycine max) root-nodule
cDNA encoding GTR was isolated by complementation of an
Escherichia coli GTR-defective mutant for restoration of
ALA prototrophy. Gtr mRNA was very low in uninfected
roots but accumulated to high levels in root nodules. The induction of
Gtr mRNA in developing nodules was subsequent to that of
the gene Enod2 (early nodule)
and coincided with leghemoglobin mRNA accumulation. Genomic analysis
revealed two Gtr genes, Gtr1 and a 3′
portion of Gtr2, which were isolated from the soybean
genome. RNase-protection analysis using probes specific to
Gtr1 and Gtr2 showed that both genes were
expressed, but Gtr1 mRNA accumulated to significantly
higher levels. In addition, the qualitative patterns of expression of
Gtr1 and Gtr2 were similar to each other
and to total Gtr mRNA in leaves and nodules of mature
plants and etiolated plantlets. The data indicate that
Gtr1 is universal for tetrapyrrole synthesis and that a
Gtr gene specific for a tissue or tetrapyrrole is
unlikely. We suggest that ALA synthesis in specialized root nodules
involves an altered spatial expression of genes that are otherwise
induced strongly only in photosynthetic tissues of uninfected plants.Soybean (Glycine max) and numerous other legumes can
establish a symbiosis with rhizobia, resulting in the formation of root
nodules comprising specialized plant and bacterial cells (for review,
see Mylona et al., 1995). Rhizobia reduce atmospheric nitrogen to
ammonia within nodules, which is assimilated by the plant host to
fulfill its nutritional nitrogen requirement. The high energy
requirement for nitrogen fixation necessitates efficient respiration by
the prokaryote within the microaerobic milieu of the nodule. The plant
host synthesizes a nodule-specific hemoglobin (leghemoglobin) that
serves to facilitate oxygen diffusion to the bacterial endosymbiont and
to buffer the free oxygen concentration at a low
tension (for review, see Appleby, 1992). Both of these functions
require that the hemoglobin concentration be high, and, indeed, it
exceeds 1 mm in soybean nodules (Appleby, 1984)
and is the predominant plant protein in that organ. Once thought to be
confined to legume nodules, hemoglobins are found throughout the plant
kingdom, and leghemoglobin likely represents a specialization of a
general plant phenomenon (for review, see Hardison, 1996). A gene
encoding a nonsymbiotic hemoglobin has been identified in soybean and
other legumes (Andersson et al., 1996); therefore, expression in
nodules involves the specific activation of a subset of genes within a
gene family. Leghemoglobin genes may have arisen from gene duplication,
followed by specialization (Andersson et al., 1996).Hemes and chlorophyll are tetrapyrroles synthesized
from common precursors; chlorophyll is quantitatively the major
tetrapyrrole in plants, with heme and other tetrapyrroles being present
in minor amounts. Legume root nodules represent an exception, in which
heme is synthesized in high quantity in the absence of chlorophyll,
thus requiring the activity of enzymes not normally expressed highly in
nonphotosynthetic tissues. Heme is synthesized from the universal
tetrapyrrole precursor ALA by seven successive enzymatic steps;
chlorophyll formation diverges after the synthesis of protoporphyrin,
the immediate heme precursor (for review, see O''Brian, 1996).
Biochemical and genetic evidence shows that soybean heme biosynthesis
genes are strongly induced in root nodules (Sangwan and O''Brian, 1991,
1992, 1993; Madsen et al., 1993; Kaczor et al., 1994; Frustaci et al.,
1995; Santana et al., 1998), and immunohistochemical studies
demonstrate that induction is concentrated in infected nodule cells
(Santana et al., 1998).ALA is synthesized from Glu in plants by a three-step mechanism called
the C5 pathway (Fig.
(Fig.1);1); the latter two steps are committed to
ALA synthesis and are catalyzed by GTR and GSAT, respectively (for
review, see Beale and Weinstein, 1990; Jahn et al., 1991). Plant cDNA
or genes encoding GTR (Gtr, also called HemA) and
GSAT (Gsa) have been identified in several plant species
(Grimm, 1990; Sangwan and O''Brian, 1993; Hofgen et al., 1994; Ilag et
al., 1994; Frustaci et al., 1995; Wenzlau and Berry-Lowe, 1995; Bougri
and Grimm, 1996; Kumar et al., 1996; Tanaka et al., 1996). Two genes
for each enzyme have been described, and some genes are reported to be
specific to a tissue, tetrapyrrole, or light regimen (Bougri and Grimm,
1996; Kumar et al., 1996; Tanaka et al., 1996). However, soybean
Gsa1 is highly expressed in both leaves and nodules and
contains a cis-acting element in its promoter that binds to
a nuclear factor found in both tissues. (Frustaci et al., 1995). In
this study we isolated soybean Gtr1 and characterized the
genetic basis of GTR expression in root nodules.
Figure 1C5 pathway for ALA synthesis. The
committed steps for ALA synthesis catalyzed by GTR and GSAT are boxed.
Glutamyl-tRNA synthetase (GluRS) and glutamyl-tRNAGlu also
participate in protein synthesis. The gene designations in plants are
shown in parentheses ... 相似文献
12.
Myxobacteria are predatory and are prolific producers of secondary metabolites. Here, we tested a hypothesized role that secondary metabolite antibiotics function as weapons in predation. To test this, a Myxococcus xanthus Δta1 mutant, blocked in antibiotic TA (myxovirescin) production, was constructed. This TA− mutant was defective in producing a zone of inhibition (ZOI) against Escherichia coli. This shows that TA is the major M. xanthus-diffusible antibacterial agent against E. coli. Correspondingly, the TA− mutant was defective in E. coli killing. Separately, an engineered E. coli strain resistant to TA was shown to be resistant toward predation. Exogenous addition of spectinomycin, a bacteriostatic antibiotic, rescued the predation defect of the TA− mutant. In contrast, against Micrococcus luteus the TA− mutant exhibited no defect in ZOI or killing. Thus, TA plays a selective role on prey species. To extend these studies to other myxobacteria, the role of antibiotic corallopyronin production in predation was tested and also found to be required for Corallococcus coralloides killing on E. coli. Next, a role of TA production in myxobacterial fitness was assessed by measuring swarm expansion. Here, the TA− mutant had a specific swarm rate reduction on prey lawns, and thus reduced fitness, compared to an isogenic TA+ strain. Based on these observations, we conclude that myxobacterial antibiotic production can function as a predatory weapon. To our knowledge, this is the first report to directly show a link between secondary metabolite production and predation. 相似文献
13.
Model organisms such as budding yeast, worms and flies have proven instrumental
in the discovery of genetic determinants of aging, and the fission yeast
Schizosaccharomyces
pombe is a promising new system for these
studies. We devised an approach to directly select for long-lived
S.
pombe mutants from a random DNA insertion
library. Each insertion mutation bears a unique sequence tag called a bar code
that allows one to determine the proportion of an individual mutant in a culture
containing thousands of different mutants. Aging these mutants in culture
allowed identification of a long-lived mutant bearing an insertion mutation in
the cyclin gene clg1
+. Clg1p, like
Pas1p, physically associates with the cyclin-dependent kinase Pef1p. We
identified a third Pef1p cyclin, Psl1p, and found that only loss of Clg1p or
Pef1p extended lifespan. Genetic and co-immunoprecipitation results indicate
that Pef1p controls lifespan through the downstream protein kinase Cek1p. While
Pef1p is conserved as Pho85p in Saccharomyces
cerevisiae, and as cdk5 in humans, genome-wide
searches for lifespan regulators in S. cerevisiae have
never identified Pho85p. Thus, the S. pombe system
can be used to identify novel, evolutionarily conserved lifespan extending
mutations, and our results suggest a potential role for mammalian cdk5 as a
lifespan regulator. 相似文献
14.
Joao P. C. Pinto Araz Zeyniyev Harma Karsens Hein Trip Juke S. Lolkema Oscar P. Kuipers Jan Kok 《Applied and environmental microbiology》2011,77(18):6687-6690
Plasmid pSEUDO and derivatives were used to show that llmg_pseudo_10 in Lactococcus lactis MG1363 and its homologous locus in L. lactis IL1403 are suitable for chromosomal integrations. L. lactis MG1363 and IL1403 nisin-induced controlled expression (NICE) system derivatives (JP9000 and IL9000) and two general stress reporter strains (NZ9000::PhrcA-GFP and NZ9000::PgroES-GFP) enabling in vivo noninvasive monitoring of cellular fitness were constructed. 相似文献
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Metabolism of
d-Glycero-d-Manno-Heptitol,
Volemitol, in Polyanthus. Discovery of a Novel Ketose
Reductase 下载免费PDF全文
Volemitol
(d-glycero-d-manno-heptitol,
α-sedoheptitol) is an unusual seven-carbon sugar alcohol that
fulfills several important physiological functions in certain species
of the genus Primula. Using the horticultural hybrid
polyanthus (Primula × polyantha) as
our model plant, we found that volemitol is the major nonstructural
carbohydrate in leaves of all stages of development, with
concentrations of up to 50 mg/g fresh weight in source leaves (about
25% of the dry weight), followed by sedoheptulose
(d-altro-2-heptulose, 36 mg/g fresh weight),
and sucrose (4 mg/g fresh weight). Volemitol was shown by the
ethylenediaminetetraacetate-exudation technique to be a prominent
phloem-mobile carbohydrate. It accounted for about 24% (mol/mol) of
the phloem sap carbohydrates, surpassed only by sucrose (63%).
Preliminary 14CO2 pulse-chase radiolabeling
experiments showed that volemitol was a major photosynthetic product,
preceded by the structurally related ketose sedoheptulose. Finally, we
present evidence for a novel NADPH-dependent ketose reductase,
tentatively called sedoheptulose reductase, in volemitol-containing
Primula species, and propose it as responsible for the
biosynthesis of volemitol in planta. Using enzyme extracts from
polyanthus leaves, we determined that sedoheptulose reductase has a pH
optimum between 7.0 and 8.0, a very high substrate specificity, and
displays saturable concentration dependence for both sedoheptulose
(apparent Km = 21 mm) and NADPH
(apparent Km = 0.4 mm). Our
results suggest that volemitol is important in certain
Primula species as a photosynthetic product, phloem
translocate, and storage carbohydrate.Alditols (sugar alcohols or acyclic polyols) may be chemically
described as reduction products of aldose or ketose sugars. The most
prevalent plant alditols are the hexitols sorbitol, mannitol, and
galactitol. However, as many as 17 different alditols occur naturally
in higher plants (for review, see Bieleski, 1982; Lewis, 1984; Loescher
and Everard, 1996). The lesser-known alditols are often restricted in
their occurrence but still fulfill important functions in those plants
where they do occur. Volemitol (Fig. (Fig.1)
1)
is a good example of a less common but important alditol. This
seven-carbon sugar alcohol seems to be confined to certain sections of
the genus Primula, so much so that it has been suggested as
a useful chemotaxonomical marker (Kremer, 1978). Very little is known
about the physiology and metabolism of volemitol in primulas, except
that it was an early photosynthetic product in cowslip (Primula
veris) and oxslip (Primula elatior) (Kremer, 1978).
Figure 1Fischer projections of volemitol and its four
structurally related seven-carbon sugars. Nomenclature follows that of
Collins (1987); trivial names are underlined.The physiological roles of alditols are manifold and largely resemble
those of disaccharides and oligosaccharides. They include
photosynthetic assimilation, translocation and storage of carbon, and
reducing power, as well as protection against different types of
stresses (for review, see Bieleski, 1982; Lewis, 1984; Loescher and
Everard, 1996; Stoop et al., 1996). The biosynthetic pathways of the
hexitols sorbitol (glucitol), mannitol, galactitol (dulcitol), and the
pentitol ribitol have been established in higher plants. They generally
use NADPH as a hydrogen donor and aldose phosphate as a hydrogen
acceptor, in concert with the corresponding phosphatases. One exception
might be galactitol, which was suggested to be formed directly from
unphosphorylated Gal (and NADPH) (Negm, 1986). Although all foliar
alditols are thought to be phloem-mobile (Lewis, 1984), this has only
been demonstrated for sorbitol, mannitol, and galactitol (Zimmermann
and Ziegler, 1975; Davis and Loescher, 1990; Moing et al., 1992; Flora
and Madore, 1993).To expand our knowledge of alditol metabolism in higher plants beyond
that of hexitols, we studied the carbohydrate metabolism of polyanthus
(Primula × polyantha). This popular
horticultural hybrid of primrose (Primula
vulgaris), oxlip, and cowslip (Mabberley, 1997) was
chosen because preliminary experiments showed that its volemitol
content is very high, similar to that of the wild-type species, and
because it may be easily grown both outdoors and indoors.We give a general overview on volemitol metabolism in polyanthus with
special emphasis on the role of volemitol in plant development and
phloem transport. We also report on a novel enzyme, a NADPH-dependent
ketose reductase, which forms volemitol by the reduction
of sedoheptulose. 相似文献
19.
Tracheary Element Differentiation Uses a Novel Mechanism
Coordinating Programmed Cell Death and Secondary
Cell Wall
Synthesis 总被引:25,自引:2,他引:23
Tracheary element differentiation requires strict coordination of secondary cell wall synthesis and programmed cell death (PCD) to produce a functional cell corpse. The execution of cell death involves an influx of Ca2+ into the cell and is manifested by rapid collapse of the large hydrolytic vacuole and cessation of cytoplasmic streaming. This precise means of effecting cell death is a prerequisite for postmortem developmental events, including autolysis and chromatin degradation. A 40-kD serine protease is secreted during secondary cell wall synthesis, which may be the coordinating factor between secondary cell wall synthesis and PCD. Specific proteolysis of the extracellular matrix is necessary and sufficient to trigger Ca2+ influx, vacuole collapse, cell death, and chromatin degradation, suggesting that extracellular proteolysis plays a key regulatory role during PCD. We propose a model in which secondary cell wall synthesis and cell death are coordinated by the concomitant secretion of the 40-kD protease and secondary cell wall precursors. Subsequent cell death is triggered by a critical activity of protease or the arrival of substrate signal precursor corresponding with the completion of a functional secondary cell wall. 相似文献
20.
The Glycosylphosphatidylinositol-Anchored Phosphatase from
Spirodela oligorrhiza Is a Purple Acid
Phosphatase 下载免费PDF全文
Hiroshi Nakazato Takashi Okamoto Miwa Nishikoori Kenji Washio Naoki Morita Kensaku Haraguchi Guy A. Thompson Jr. Hidetoshi Okuyama 《Plant physiology》1998,118(3):1015-1020
We recently presented clear evidence that the major low-phosphate-inducible phosphatase of the duckweed Spirodela oligorrhiza is a glycosylphosphatidylinositol (GPI)-anchored protein, and, to our knowledge, is the first described from higher plants (N. Morita, H. Nakazato, H. Okuyama, Y. Kim, G.A. Thompson, Jr. [1996] Biochim Biophys Acta 1290: 53–62). In this report the purified 57-kD phosphatase is shown to be a purple metalloenzyme containing Fe and Mn atoms and having an absorption maximum at 556 nm. The phosphatase activity was only slightly inhibited by tartrate, as expected for a purple acid phosphatase (PAP). Furthermore, the protein cross-reacted with an anti-Arabidopsis PAP antibody on immunoblots. The N-terminal amino acid sequence of the phosphatase was very similar to those of Arabidopsis, red kidney bean (Phaseolus vulgaris), and soybean (Glycine max) PAP. Extracts of S. oligorrhiza plants incubated with the GPI-specific precursor [3H]ethanolamine were treated with antibodies raised against the purified S. oligorrhiza phosphatase. Radioactivity from the resulting immunoprecipitates was specifically associated with a 57-kD band on sodium dodecyl sulfate-polyacrylamide gels. These results, together with previous findings, strongly indicate that the GPI-anchored phosphatase of S. oligorrhiza is a PAP. 相似文献