text:
Give me a start - Ambrose
Published: 8:00 AM February 3, 2003
EXCLUSIVEDARREN Ambrose has called on Blues boss Joe Royle to let him off the leash earlier and give him the chance to show what he can do for the full 90 minutes.
DARREN Ambrose has called on Blues boss Joe Royle to let him off the leash earlier and give him the chance to show what he can do for the full 90 minutes.
Teenager Ambrose is not criticising his manager and is trying hard to stay patient but believes he, and fellow teenage sensation Darren Bent, can make even more of an impact by starting games.
Sir Bobby Robson and his Newcastle United chairman Freddie Shepherd were at Valley Parade as Ipswich slumped to a 2-0 defeat, and there will be no prizes for guessing who they were checking on once again.
Although the duo were unable to make a big impact after the Blues were reduced to 10 men following Hermann Hreidarsson's dismissal, Ambrose is convinced their time has come.
He said: "I always want to start. I still trust the manager's decision and I'm ready to come off the bench if needed or to start.
"We would love the opportunity to start and hopefully that will happen soon and when it does we will be ready and hopefully do enough so we can then keep our places.
"It is good to come off the bench because you are fresh against tired defenders. But I'd like to start because you have a lot more of the game to show what you can do and a lot more of the game to score and set up a few.
"But having said that I still trust the manager and respect his decision and will come off the bench when he wants me to."
Darren Bent replaced Fabian Wilnis at half time at Bradford as Town switched to a more attacking 4-3-3 formation after dominating the first half without seizing the advantage.
Even when Hreidarsson was sent off, Royle showed his attacking intentions by replacing Pablo Counago with Ambrose and then Martijn Reuser for midfielder Jim Magilton and going to a 3-4-2 formation.
While Marcus Bent hit a post in the first half, Darren Bent crashed an effort against the crossbar and Ambrose might have done better with a ball rolled to him by the younger Bent.
Ambrose admitted: "We had a couple of chances to make a difference and perhaps should have scored but overall we are disappointed not to get the victory.
"It wasn't the best of performances by us , but credit to Bradford, they kept at us although we shouldn't have allowed them to score either of the differences.
"The sending off killed us really as we were about to make another substitution and attack them even more, so it was very disappointing."
Buoyed by the red card for Hreidarsson after he caught Simon Francis, Bradford snatched the lead through Claus Jorgensen before 18-year-old debutant Danny Forrest clinched the game.
Although way below par, Town were still clearly the better side and defender Chris Makin had no doubt two decisions swung the game, the sending off and a penalty denied when Mark Bower clearly tripped Marcus Bent.
Makin said: " We were not at our best anyhow but the sending off made all the difference. Bradford did come out brightly in the second half in fairness, but the sending off had a big bearing on the result.
"We dominated the first half but we didn't create chances ,which was disappointing because we are known for creating chances.
"The referee has not given us anything. It was a definite penalty. Everyone in the stadium knew it was except him.
"We have to pick ourselves up for the Sheffield United game, which is massive for us. Luckily enough in football you do get the chance to put things right and pick yourself up.
"Hermann will be a big miss but there are people waiting in the wings for a chance and they will now get that opportunity, it is up to them to take it."
That could mean more playing time for another impressive 18-year-old as Matt Richards looks to fill the defensive gap, or Ambrose getting his much-desired starting spot.
ONLY an outstanding piece of manouvering by Paul the Ipswich Town coach driver ensured the team arrived safely to Bradford.
During their journey to Yorkshire on Friday during the worst of the snow storms, a car spun on the ice and it looked as if the coach would not be able to avoid crushing the vehicle with a man and woman inside, before Paul took evasive action.
A Town insider said: "It was an excellent piece of driving, things could have been far worse but for Paul."
The car struck the coach a glancing blow and both vehicles suffered some bodywork damage but no-one was hurt.


Confaab, a global provider of comprehensive Hong Kong Business Executives and Business Owners List offers prepackaged & Customized Hong Kong Business Executives email list, Telemarketing lists and Direct mailing lists . We customize Hong Kong Business Executives Email Lists and Hong Kong Mailing List according to your requirements and offer a highly targeted Opt-in Emails and mails to maximize your ROI and improve conversions. Customers can download prepackaged lists instantly.
Hong Kong is one of the largest national economy in Asia. Most of the country's products are in engineering, especially in automobiles, machinery, metals, and chemical goods. Hong Kong is the leading producer of wind turbines and solar power technology in the world.


Thank you! We'll follow up shortly.
Congrats! Your loan has been approved.
To fund your loan, verify your coverage or get quotes from our partner at Newfront Insurance.
Help your customers get General Liability insurance quick and easy. Plus get notified automatically when they purchase.
Cyber Liability insurance can help protect your customers from leaks and hacks. Just another way you can help protect your customer.
Have another idea? Let us know by requesting access!


text:
All issues Volume 41 / No 5 (September–October 2010) Vet. Res., 41 5 (2010) 68 Full HTML
Volume 41, Number 5, September–October 2010
https://doi.org/10.1051/vetres/2010040
Vet. Res. (2010) 41:68
6-hydroxydopamine-mediated release of norepinephrine increases faecal excretion of Salmonella enterica serovar Typhimurium in pigs
Gillian D. Pullinger1, Pauline M. van Diemen1, Sonya C. Carnell1,2, Holly Davies1, Mark Lyte3 and Mark P. Stevens1*
1 Enteric Bacterial Pathogens Laboratory, Institute for Animal Health, Compton, Berkshire RG20 7NN, United Kingdom
2 Present address: Institute for Cell and Molecular Biosciences & School of Biomedical Sciences, University of Newcastle, Framlington Place, Newcastle NE2 4HH, United Kingdom
3 School of Pharmacy, Texas Tech University Health Sciences Center, Lubbock, TX 79430-8162, USA
* Corresponding author: mark-p.stevens@bbsrc.ac.uk
Accepted: 5 July 2010
Salmonella enterica serovar Typhimurium is an animal and zoonotic pathogen of worldwide importance. In pigs, transport and social stress are associated with reactivation and spread of Salmonella Typhimurium infection. The stress-related catecholamine norepinephrine (NE) has been reported to activate growth and virulence factor expression in Salmonella; however the extent to which NE contributes to stress-associated salmonellosis is unclear. We studied the impact of releasing NE from endogenous stores during Salmonella Typhimurium infection of pigs by administration of 6-hydroxydopamine (6-OHDA), which selectively destroys noradrenergic nerve terminals. Treatment of pigs with 6-OHDA 7 or 16 days post-oral inoculation with Salmonella Typhimurium produced elevated plasma NE levels and transiently, but significantly, increased faecal excretion of the challenge strain. Oral administration of NE to Salmonella Typhimurium-infected pigs also transiently and significantly increased shedding; however pre-culture of the bacteria with NE did not alter the outcome of infection. Salmonella has been proposed to sense and respond to NE via a homologue of the adrenergic sensor kinase QseC. A ΔqseC mutant of Salmonella Typhimurium was consistently excreted in lower numbers than the parent strain post-oral inoculation of pigs, though not significantly so. 6-OHDA treatment of pigs infected with the ΔqseC mutant also increased faecal excretion of the mutant strain, albeit to a lesser extent than observed upon 6-OHDA treatment of pigs infected with the parent strain. Our data support the notion that stress-related catecholamines modulate the interaction of enteric bacterial pathogens with their hosts.
Key words: Salmonella / stress / norepinephrine / virulence / colonisation
© The authors, published by INRA/EDP Sciences, 2010
This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted use, distribution, and reproduction in any noncommercial medium, provided the original work is properly cited.
Salmonella enterica serovar Typhimurium is responsible for over 60% of acute porcine salmonellosis in the United Kingdom [2]. In a UK-wide survey of pigs at slaughter, Salmonella Typhimurium was detected in 11.1% of caeca and 2.1% of carcases [10], indicating significant potential for entry of the pathogen into the human food chain and environment. It is evident that stress influences susceptibility of pigs to Salmonella and the risk of zoonosis. Transportation has been correlated with reactivation of subacute Salmonella Typhimurium infections in pigs and intestinal and carcass contamination increases with time spent in lairage [7, 20, 21, 29]. Moreover, social stress caused by mixing increases faecal excretion and translocation of Salmonella Typhimurium to intestinal lymph nodes in early-weaned pigs [8]. Such phenomena do not appear restricted to Salmonella, as mixing of litters, isolation from the sow or short-lived cold stress has been reported to increase faecal excretion of enterotoxigenic Escherichia coli relative to control piglets [22]. Even mild physical handling of pigs involving a daily weight measurement increased the faecal excretion of E. coli and total coliforms relative to control animals [12]. A plausible explanation for such links is that activation of adrenal axes under stress leads to the release of stress-related catecholamines that may impair innate and adaptive immunity. In recent years however, it has become clear that many bacteria are able to sense mediators of the host stress response and respond by activating growth and the expression of virulence factors (reviewed in [17, 41]).
A key response of the enteric nervous system to stress is the release of norepinephrine (NE) from sympathetic nerve fibres that originate in the prevertebral ganglia and innervate the gut mucosa. Elevated NE levels could be found in the intestines of mice subject to partial hepatectomy compared to control animals and were associated with increased susceptibility to gut-derived sepsis and virulence gene expression by Pseudomonas aeruginosa [1, 24]. In relation to Salmonella, NE has been reported to resuscitate Salmonella Typhimurium from a viable non-culturable state [35] and to enhance its growth from low inocula in serum-rich nutrient-limited medium [15]. The ability of NE to activate growth in such media has been associated with binding of NE to ferric iron complexed with lactoferrin and transferrin and its reduction to Fe(II), for which the proteins have a lower affinity [36]. NE has also been reported to activate the expression of virulence-associated factors in Salmonella Typhimurium, including flagella-mediated motility [5, 28] and Type III protein secretion [28, 32], though others have not reproduced such effects [31]. Exogenous iron can activate both motility and Type III secretion system (T3SS)-1 [6, 13], indicating that the effect could be a consequence of NE-mediated iron supply. An alternative explanation is that Salmonella senses and responds to stress-related catecholamines via homologues of adrenergic receptors identified in E. coli O157:H7 called QseC [9] and QseE [34]. In E. coli O157:H7 these proteins autophosphorylate on binding of epinephrine (Epi) and transfer the phosphate moiety to cognate transcriptional regulators, modulating the expression of genes under their control [9, 34]. QseC has been reported to be required for full virulence of Salmonella Typhimurium in pigs [5, 6] and mice [28]. Moreover, an inhibitor of E. coli O157:H7 QseC signalling (LED209) controls systemic salmonellosis in a murine model [32]. However, two-component sensory systems are known to integrate multiple signals and the extent to which such sensors influence virulence by sensing stress-related catecholamines is unclear.
The ability of NE to alter the outcome of Salmonella infection has been suggested in several reports. NE augments Salmonella-induced enteritis in a bovine ligated ileal loop model of infection in a manner independent of QseC and QseE [31]. Analysis of plasmid partitioning in NE-stimulated and unstimulated bacteria in this model indicated that the effect was associated with enhanced net replication of Salmonella Typhimurium [31]. Though useful for evaluating multiple strains, neurochemicals and doses for their effects on local secretory and inflammatory responses, ligated intestinal loop models may be considered artificial in the context of stressed animals in that high inocula and hormone concentrations are held statically over surgically-manipulated mucosa for protracted periods. Oral dosing of mice or chickens with NE enhances intestinal and systemic Salmonella infection [27, 46], albeit that high hormone doses were again used. In calves, intramuscular dosing with NE enhances translocation of various S. enterica serovars from the intestines and promotes encephalopathy [26]. Pre-culture of Salmonella Typhimurium with NE has been reported to result in higher levels of colonisation of the porcine intestinal tract [43]. However, NE-treated and untreated bacteria were cultured in distinct media and it is likely that the physiological status of the bacteria differed substantially, making it difficult to separate effects of NE from growth or media-related effects [16]. In transgenic mice unable to produce Epi or NE owing to mutation of dopamine beta-hydroxylase (dbh −/−), Salmonella Typhimurium transcribed T3SS-1 genes at a lower level than in wild-type mice, consistent with a role for stress-related catecholamines in control of virulence gene expression in vivo [28]. However, dbh −/− mice lacking NE/Epi were more sensitive to Salmonella Typhimurium infection than dbh +/− mice owing to immune deficiency and it is therefore difficult to separate direct effects of NE/Epi on Salmonella from the immune status of the animals [28].
Here, we sought to mimic the effect of acute stress during recovery from experimental oral Salmonella infection in pigs. The selective neurotoxin 6-hydroxydopamine (6-OHDA) was used to invoke the release of NE by destruction of noradrenergic nerve terminals in the periphery without crossing the blood-brain barrier or affecting cholinergic neurons [11, 25]. The effect is transient and damaged noradrenergic neurons heal and recover, enabling the course of Salmonella infection to be monitored in the days post-treatment. Intraperitoneal administration of 6-OHDA to mice invokes the outgrowth of coliforms relative to controls, though coliform numbers returned to normal as damage to noradrenergic neurons was repaired over a two week period [25]. Having established such a model in pigs, we evaluated the role of the QseC sensor kinase in both intestinal colonisation and 6-OHDA-induced responses. Moreover, we examined the impact of treating the inoculum, or infected animals, directly with NE. Our data provide valuable insights into the role of stress-related catecholamines in the outcome of Salmonella infection in reservoir hosts.
2.1. Bacterial strains and media
A spontaneous nalidixic acid resistant mutant of Salmonella Typhimurium strain ST4/74 of well-defined virulence in pigs was used [30, 45]. ST4/74 is the parent of strain SL1344, a hisG auxotroph derived by transduction [19], that has previously been reported to respond to NE [15, 28, 35]. A ST4/74 nalR deletion mutant lacking qseC (also known as preB or STM3178) was created by lambda Red recombinase-mediated integration of a linear PCR amplicon comprising a kanamycin resistance gene flanked by the sequences 5′ and 3′ of qseC, followed by P22int transduction of the mutated allele into the archived strain and flippase-mediated excision of the kanamycin resistance cassette as described [31]. Bacteria were cultured in Luria-Bertani medium containing 20 μg/mL nalidixic acid.
2.2. Experimental animals
Animal experiments were conducted according to the Animal (Scientific Procedures) Act 1986 (license 30/2485) with the approval of the local Ethical Review Committee. Large-White × Landrace pigs of mixed sex and aged ca. 6 weeks old were obtained from a commercial supplier with no history of acute salmonellosis and housed in containment level 2 accommodation with access to antibiotic-free irradiated weaner pellets and water ad libitum. Pigs were confirmed to be culture-negative for Salmonella prior to infection by overnight enrichment of rectal swabs in Rappaport broth (at 37 °C) and selenite brilliant green broth (at 42 °C) followed by plating to modified brilliant green agar (Oxoid, Basingstoke, UK).
2.3. Salmonella infection
ST4/74 nalR or its isogenic ΔqseC mutant were grown to stationary phase in Luria-Bertani broth at 37 °C with shaking. In studies to evaluate the impact of pre-culture with NE, ST4/74 nalR was cultured as above, then subcultured 1:10 into fresh LB broth containing L-norepinephrine bitartrate (Sigma, Poole, UK, reference A9512) at a final concentration of 5 mM or diluent for 2 h at 37 °C with shaking. For inoculation, approximately 1 × 108 colony-forming units (CFU) were suspended in 10 mL antacid (5% (w/v) Mg(SiO3)3, 5% (w/v) NaHCO3 and 5% (w/v) MgCO3 in H2O) and administered by oral gavage using a 10FG catheter. The number of bacteria delivered was determined by retrospective plating of serial ten-fold dilutions of inocula. Rectal temperatures were recorded and animals monitored for clinical signs of disease twice daily. The magnitude and duration of faecal excretion of the challenge strain was followed by plating of serial ten-fold dilutions of fresh faeces collected by rectal palpation to MacConkey agar supplemented with 20 μg/mL nalidixic acid. Data presented are the least square mean viable count (expressed as log10 CFU/g) and the limit of detection by direct plating was 2.0 log10 CFU/g.
2.4. 6-hydroxydopamine and L-norepinephrine treatment
Seven or 16 days post-inoculation with Salmonella Typhimurium groups of pigs were dosed by the intravenous route with 6-hydroxydopamine or placebo. A 240 mg/mL (0.96 M) solution of 6-OHDA (Fluka Chemicals, Gillingham, UK, reference 55238) was freshly prepared in 0.9% (w/v) saline containing 1 mM ascorbic acid and sterilised by passing through a 0.22 μm filter. In the first study, three pigs were given 6-OHDA 7 days post-inoculation with ST4/74 nalR in a series of rising doses of 1 mg/kg, 4 mg/kg then 35 mg/kg at intervals of 20 min via an ear vein. The total dose of 6-OHDA was similar to that used to produce chemical sympathectomy in adult miniature swine (50 mg/kg intravenously) [42]. At the same time, 3 age-matched control pigs infected with ST4/74 nalR were given three injections of an equal volume of diluent at the same intervals and site. All reasonable precautions were taken to minimise handling effects between treated and control groups. In the second experiment, 4 pigs were given a single dose of 40 mg/kg and 2 pigs were given diluent via the vena cava at 16 days post-inoculation with ST4/74 nalR. The latter regime was also used to dose groups of 3 pigs with 6-OHDA or diluent at 16 days post-inoculation with ST4/74 nalR ΔqseC. In all cases, a sample of venous blood was withdrawn from the vena cava using a needle into heparin-coated tubes under vacuum immediately before and 1 h after 6-OHDA treatment, and plasma stored at −80 °C for quantification of circulating NE. Animals were observed continuously for 4 h post-treatment and at 8 h intervals thereafter. Faecal samples were collected immediately before and twice daily after dosing with 6-OHDA or diluent and viable nalidixic acid resistant bacteria were enumerated as above.
To determine the effect of oral dosing of Salmonella Typhimurium-infected pigs with NE, 15 mL of a 167 mg/mL freshly prepared solution of L-norepinephrine or diluent (PBS) was given to duplicate pigs using a 10FG catheter 17 days after oral inoculation with ST4/74 nalR. The final estimated dose was ca. 200 mg/kg.
2.5. NE enzyme-linked immunosorbent assay (ELISA)
Plasma NE levels in pigs pre- and post-treatment with 6-OHDA or diluent were quantified using a sandwich ELISA designed for human plasma and urine according to the manufacturer's instructions (IBL International GMBH, Hamburg, Germany). NE standards of known concentration were used to convert absorbance readings to NE concentrations in ng/mL.
Measured traits were analysed for the effect of strain or treatment by two-way analysis of variance (Statistical Analysis System 1995, SAS Institute, Cary, USA). The magnitude and duration of faecal excretion of strains was analysed after a log10 transformation of the number of excreted bacteria, with daily values taken as repeated measurements (Proc Mixed). Where strain or treatment was identified as a significant variable, pair-wise comparisons of least square means were performed. p values ≤ 0.05 were considered significant.
3.1. 6-OHDA-mediated release of NE increases faecal excretion of Salmonella Typhimurium in pigs
To mimic the impact of acute stress during Salmonella Typhimurium infection, two groups of six 6-week-old pigs were inoculated via the oral route with ST4/74 nalR then treated at 7 or 16 days post-inoculation with 6-OHDA or diluent. In both sets of pigs, ST4/74 nalR induced rapid but short-lived pyrexia and diarrhoea, and recoveries of viable bacteria in the faeces over time were comparable to those recorded previously [30, 45]. By 3–4 days post-inoculation rectal temperatures and clinical signs returned to normal levels. In the first study, 3 pigs were dosed at 7 days post-inoculation with 3 doses of 6-OHDA 20 min apart (1, 4 and 35 mg/kg) via an ear vein. Three control pigs were given an equivalent volume of diluent via an ear vein at the same intervals. Only upon administration of 35 mg/kg did pigs exhibit an overt response to 6-OHDA, characterised by pyrexia, rapid breathing, increased salivation and slight ataxia. Control pigs appeared normal throughout. 6-OHDA treatment was confirmed to elicit a significant increase in plasma NE levels 1 h post-exposure compared to control pigs (p = 0.0027) and pre-treatment levels (p = 0.003; Fig. 1A). Analysis of the course of faecal excretion revealed that 6-OHDA treatment produced a transient increase in faecal excretion of ST4/74 nalR of ca. 1 log10 CFU/g 1 day post-treatment relative to control pigs (Fig. 1B). The difference in the number of Salmonella Typhimurium excreted 1 day after 6-OHDA treatment was not significantly different to the diluent-treated group by a narrow margin (p = 0.057). However, there was a highly significant rise in the mean log10 CFU/g Salmonella Typhimurium excreted 1 day after 6-OHDA treatment compared to faecal samples from the same animals immediately prior to treatment (p = 0.0003; mean log10 CFU/g 1.43 fold higher, equivalent to a rise from 1.14 × 103 CFU/g to 1.53 × 104 CFU/g). By 2 days post-treatment, a comparable number of viable ST4/74 nalR were excreted by 6-OHDA- and diluent-treated pigs.
Intravenous administration of 6-OHDA to pigs increases plasma NE levels and faecal excretion of Salmonella Typhimurium. Plasma NE (ng/mL) was quantified by ELISA immediately before and 1 h after treatment with a total of 40 mg/kg 6-OHDA or diluent 7 days post-inoculation with ST4/74 nalR (panel A) or 16 days post-inoculation (panel C). The impact of treatment on the course of faecal excretion is shown for pigs treated at 7 and 16 days post-inoculation in panels B and D, respectively. Values shown are the least square means (LSM) ± standard error (SE) of the LSM. p values ≤ 0.05 are marked with an asterisk. † denotes significant differences in the 6-OHDA group relative to pre-treatment samples.
In the second trial, a single dose of 40 mg/kg 6-OHDA was given via the vena cava to 4 pigs 16 days post-oral inoculation with ST4/74 nalR. The 6-OHDA-treated pigs showed significantly elevated plasma NE levels 1 h post-treatment compared to 2 control pigs given an equivalent volume of diluent via the same route (Fig. 1C; p = 0.0041). Plasma NE levels after 6-OHDA treatment were also significantly higher than the corresponding pre-treatment samples (p = 0.0004). In 6-OHDA-treated animals, the number of ST4/74 nalR excreted in the faeces was again elevated after 6-OHDA treatment relative to the control group, this time by ca. 2 log10 CFU/g 1 day post-inoculation, but the difference between the two groups was not significant (Fig. 1D; p = 0.1). The mean log10 CFU/g Salmonella Typhimurium excreted was significantly higher after 6-OHDA treatment than in faeces samples from the same animals immediately prior to treatment (p = 0.0047; mean log10 CFU/g 2.68 fold higher, equivalent to a rise from 1.19 × 101 CFU/g to 7.60 × 102 CFU/g). The effect was sustained for longer than observed upon 6-OHDA treatment 7 days after ST4/74 nalR challenge, albeit not significantly so relative to control pigs at 2 days post-treatment.
3.2. Oral administration of NE, but not pre-culture with NE, alters the course of Salmonella Typhimurium infection in pigs
Consistent with the effect of releasing NE from endogenous stores, we observed that oral dosing of Salmonella Typhimurium-infected pigs with NE transiently increases faecal excretion of Salmonella. Groups of two pigs inoculated with ST4/74 nalR as above were allowed to recover then given a single oral bolus of ca. 200 mg/kg NE or diluent 17 days after Salmonella Typhimurium infection by oral gavage with a catheter. Owing to inter-animal variation in shedding levels, data are presented as the fold-change in mean log10 CFU of the challenge strain excreted per gram of faeces relative to the pre-treatment sample. In the NE-treated animals, the mean log10 CFU/g ST4/74 nalR excreted rose by c. 1.4 fold 1 and 2 days after NE treatment, with significant differences to the pre-treatment levels in the same animals being detected (Fig. 2A; p ≤ 0.007). No significant change in the course of excretion of ST4/74 nalR was seen in the group given diluent (Fig. 2A). Clinical signs were similar in both groups.
Oral administration of NE, but not pre-culture with NE, alters the course of Salmonella Typhimurium infection in pigs. Panel A shows the impact of oral administration of c. 200 mg/kg NE or diluent to groups of 2 pigs 17 days post-inoculation with ST4/74 nalR. Values are the fold change in mean log10 CFU Salmonella/g relative to samples collected immediately prior to treatment. Panel B shows the magnitude of faecal excretion of Salmonella Typhimurium after oral dosing of groups of 3 pigs with c. 1 × 108 CFU ST4/74 nalR amplified in LB medium for 2 h in the presence of 5 mM NE or diluent. Values shown are the least square means (LSM) ± standard error (SE) of the LSM. p values ≤ 0.05 are marked with an asterisk.
It has been reported that cultivation of Salmonella Typhimurium in serum-rich nutrient-limited medium containing 2 mM NE enhances the number and tissue distribution of bacteria in the porcine intestines after oral dosing relative to bacteria grown in LB medium [43]. However, the use of different media raises the possibility that the effect is due to distinct physiological states of the bacteria rather than direct effects of NE, as discussed elsewhere [16]. Given the assertion that 50 μM NE activates motility and Type III secretion [5, 28] and evidence that the transcriptome of Salmonella Typhimurium during growth in LB broth alters in response to Epi [23] and NE [38], we examined if pre-culture of ST4/74 nalR in LB medium with or without 5 mM L-norepinephrine for 2 h altered the outcome of infection of pigs. This NE concentration was selected as it augments enteritis induced by LB-grown Salmonella Typhimurium in calf ileal loops [31]. Groups of 3 pigs were inoculated with c. 1 × 108 CFU NE-treated or untreated LB culture and faecal excretion monitored twice daily for 3 days. No statistically significant differences were detected in the magnitude of faecal excretion (Fig. 2B), except at 72 h when the number of NE-pretreated bacteria excreted was lower than in the pigs given untreated bacteria. Similarly, there were no statistical differences in pyrexial responses or colonisation of intestinal sites at post mortem examination between the two groups (data not shown).
3.3. Role of the putative adrenergic sensor kinase QseC in intestinal colonisation of pigs by Salmonella Typhimurium and the response to 6-OHDA
A qseC mutant of Salmonella Typhimurium has been reported to be significantly impaired in intestinal colonisation at 1–7 days after co-infection of 13-week-old pigs with the parent strain [5] or 2–7 days post-inoculation of 7-week-old pigs when given separately [6]. However, only modest attenuation of a Salmonella Typhimurium qseC mutant was detected by co-infection of mice [28] and no significant defect in colonisation of most intestinal sites was observed in calves [31]. As QseC has been proposed as a novel drug target for Salmonella [32, 33], we evaluated its role in colonisation of pigs by separately dosing 6-week-old pigs by the oral route with ST4/74 nalR (n = 5) or its isogenic ΔqseC mutant (n = 6). The magnitude and duration of faecal excretion of the challenge strains was followed daily for 16 days. ST4/74 nalR ΔqseC was consistently shed in lower numbers than the parent strain, though not statistically significantly so (Fig. 3). Values at days 14 and 15 post-inoculation showed a tendency toward significance (p ≤ 0.1). No significant differences in rectal temperature or diarrhoea severity were recorded between the wild-type and ΔqseC mutant groups.
The putative adrenergic sensor kinase QseC does not significantly influence intestinal colonisation of pigs. 6-week-old pigs were inoculated with 1.50 × 109 CFU ST4/74 nalR (n = 5) or 1.53 × 109 CFU ST4/74 nalR ΔqseC (n = 6) and the course of faecal excretion of the bacteria followed daily for 16 days. Values shown are the least square means (LSM) ± standard error (SE) of the LSM. † denotes p values ≤ 0.1.
To determine if QseC is required for the response of Salmonella Typhimurium to 6-OHDA treatment, 16 days after inoculation of pigs with ST4/74 nalR ΔqseC 3 pigs were given 40 mg/kg 6-OHDA into the vena cava and 3 control pigs were given an equivalent volume of diluent. 6-OHDA elicited a significant increase in plasma NE levels relative to the control group (p = 0.008) and pre-treatment samples (p = 0.01; Fig. 4A). The mean log10 CFU/g of the ST4/74 nalR ΔqseC mutant excreted in the faeces rose in response to 6-OHDA treatment compared to animals given diluent, but not significantly so (Fig. 4B). The peak fold change in mean log10 CFU/g ST4/74 nalR ΔqseC excreted after 6-OHDA treatment relative to samples collected from the same animals immediately prior to treatment was 1.09 (equivalent to a rise from 7.67 × 103 CFU/g to 1.75 × 104 CFU/g) and was not statistically significant. The data suggest that QseC contributes to the 6-OHDA-mediated increase in faecal excretion of Salmonella Typhimurium.
QseC influences, but is not essential for, increased faecal excretion of Salmonella Typhimurium in response to 6-OHDA treatment. Plasma NE (ng/mL) was quantified by ELISA immediately before and 1 h after treatment with 40 mg/kg 6-OHDA or diluent 16 days post-inoculation with CFU ST4/74 nalR ΔqseC (panel A). The impact of 6-OHDA treatment on the course of faecal excretion of ST4/74 nalR ΔqseC is shown in panel B. Values shown are the least square means (LSM) ± standard error (SE) of the LSM. p values ≤ 0.05 are marked with an asterisk.
Epidemiological and experimental studies support a role for stress in the development of porcine salmonellosis. Stress modulates multiple processes, and the contribution of stress-related catecholamines to observed phenomena is unclear. Our studies show that release of NE from endogenous stores by administration of the selective neurotoxin 6-hydroxydopamine can enhance faecal excretion of S. enterica serovar Typhimurium during recovery from acute infection. Intravenous delivery of 6-OHDA markedly elevated plasma NE levels, and though NE levels in the intestines are difficult to quantify for reasons we have discussed [44], the enteric nervous system is densely innervated by noradrenergic neurons and it is likely that significant spill over of NE into the intestinal lumen occurred. The ability of 6-OHDA to invoke responses that are specifically due to NE has previously been demonstrated using the catecholamine uptake blocker desipramine hydrochloride [25], and we propose that the model may be suitable for studies with other pig pathogens. Previous studies using a glucose analogue, 2-deoxy-D-glucose, indicated that it could induce many of the hallmark parameters of physiological stress in pigs [39], yet it failed to reactivate Salmonella Choleraesuis at enteric or systemic sites when given to carrier animals [40]. However, catecholamine levels in treated and control animals were not measured and further studies are required to determine how intestinal levels of stress-related catecholamines in experimental animal models compare to those of stressed animals in the field.
Though Salmonella Typhimurium was excreted in the faeces in significantly greater numbers in 6-OHDA-treated pigs relative to pre-treatment samples, it is noteworthy that the effect was relatively modest and transient even though there was clear evidence of physiological shock. Furthermore, the reactivation of Salmonella Typhimurium infection was less pronounced when 6-OHDA was given at day 7 post-inoculation (Fig. 1B) than when 6-OHDA was given at day 16 post-inoculation (Fig. 1D). In both cases, an explanation for the short-lived effect may be that priming of innate immunity during acute infection allows pigs to more effectively control rising Salmonella levels. It is known for example that prior infection of gnotobiotic piglets with avirulent S. enterica serovar Infantis can protect the animals from subsequent challenge with virulent Salmonella Typhimurium in a manner associated with neutrophil recruitment to the intestines [14]. The elevated faecal excretion of Salmonella Typhimurium upon 6-OHDA treatment at day 16 compared to day 7 post-challenge may therefore be associated with a waning protective innate response.
The ability of NE to alter the magnitude of Salmonella Typhimurium excretion by pigs was confirmed by oral administration of NE to recovering animals. This invoked a substantial rise in the number of excreted bacteria compared to controls, consistent with the ability of orally-administered NE to promote intestinal colonisation and systemic translocation by Salmonella Enteritidis in chicks [27] and Salmonella Typhimurium in mice [27, 46]. The concentrations required to promote this effect were very high in a physiological context, however the survival and transit times of orally administered NE through anatomical compartments are unknown. It is conceivable that elevated excretion of Salmonella in response to 6-OHDA or NE treatment may reflect changes in intestinal motility.
To investigate the mechanism(s) by which exogenous or endogenous NE alter the outcome of Salmonella infection in pigs, we examined the role of the putative adrenergic sensor kinase QseC. A Salmonella Typhimurium ΔqseC null mutant was consistently shed in lower numbers than the parent strain, though not significantly so. The data are consistent with the absence of significant attenuation of the same mutant in calves [31] and the modest attenuation of an independent ΔqseC mutant in mice [28], but differ markedly from other studies with a Salmonella Typhimurium qseC mutant in pigs [5, 6]. The disparities between pig studies may be due to use of distinct mutants (a qseC::cat insertion mutant was used by Bearson et al.), inoculation routes (intranasal vs. intragastric), or other parameters. Given the assertion that QseC signalling represents a novel drug target [32, 33], further studies are required to evaluate the role of QseC and inhibitors thereof in reservoir hosts.
Faecal excretion of the Salmonella Typhiumurium ΔqseC mutant could be increased by 6-OHDA treatment, indicating that QseC is not absolutely essential for the effect. However, the magnitude of the increase in faecal excretion of ST4/74 nalR ΔqseC was lower than observed with the parent strain when pigs were treated with 6-OHDA at the same time post-inoculation and when Salmonella levels in the faeces were similar (1.09 fold increase in mean log10 CFU/g ST4/74 nalR ΔqseC compared to a 2.68 fold increase for the parent strain upon 6-OHDA treatment 16 days post-infection), and was not statistically significant. This suggests that QseC plays a role in the response to 6-OHDA, but that other factors may be involved. We caution that the effect of 6-OHDA on faecal excretion of the ∆qseC mutant and parent strains was tested in separate experiments and direct comparisons within a single cohort of treated pigs are required. The potential for QseC-independent phenomena is consistent with the existence of at least one other adrenergic sensor kinase in E. coli O157:H7 (QseE: [34]), and with the ability of NE to modulate other processes such as growth, iron supply and virulence in a manner that is independent of QseCE [31]. Another possible explanation for the reduced response of the ∆qseC mutant to 6-OHDA treatment (Fig. 4B) when compared to wild-type bacteria tested separately but in the same way (Fig. 1D), may be that the mutant is subtly attenuated and unable to replicate as efficiently upon release of NE. In this respect, the lower response of the ∆qseC mutant to 6-OHDA treatment could be due to impaired fitness rather than absence of adrenergic sensing, and studies with other attenuated mutants may support this.
It is also plausible that release of NE mediated by 6-OHDA or exogenous NE may promote Salmonella infection via changes in the microflora. It is known that 6-OHDA profoundly enhances the number of coliforms, particularly E. coli, in the murine intestinal tract [25]. Furthermore, deep-sequencing of intestinal microflora in a restraint-based mouse model of chronic stress was recently reported to alter the composition of the microflora in a manner associated with elevated susceptibility to the murine enteric pathogen Citrobacter rodentium [3]. It is established that the microflora plays an important role in determining the outcome of Salmonella infection, since streptomycin-mediated depletion of the microflora allows Salmonella Typhimurium to produce colitis in inbred mice, whereas mice with intact gut flora exhibit minimal enteric pathology [4]. The bacterial families and genera perturbed by antibiotics that alter susceptibility to salmonellosis have been defined at low resolution [18, 37], and it will be of interest to determine if the same groups are affected by stressors. Further studies are also required to examine the relative importance of growth activation, putative adrenergic sensor kinases, and the modulation of host processes in the context of stress and susceptibility to microbial infection.
In conclusion, we report that the selective neurotoxin 6-OHDA can be used to elicit the release of NE in pigs and enhances the faecal excretion of Salmonella Typhimurium. Our findings are consistent with the ability of NE to augment Salmonella pathogenesis in other animal models [26, 27, 31, 46] and may provide a partial explanation for the elevated excretion of zoonotic pathogens by livestock subject to transport or social stress.
The authors gratefully acknowledge the support of the Biotechnology & Biological Sciences Research Council, United Kingdom (grant number BB/C500822/1).
Alverdy J., Holbrook C., Rocha F., Seiden L., Wu R.L., Musch M., et al., Gut-derived sepsis occurs when the right pathogen with the right virulence genes meets the right host: evidence for in vivo virulence expression in Pseudomonas aeruginosa, Ann. Surg. (2000) 232:480–489. [CrossRef] [PubMed] [Google Scholar]
Anonymous, Zoonoses Report, United Kingdom 2008, Department for Environment, Food and Rural Affairs Publications, London, 2010. [Google Scholar]
Bailey M.T., Dowd S.E., Parry N.M., Galley J.D., Schauer D.B., Lyte M., Stressor exposure disrupts commensal microbial populations in the intestines and leads to increased colonization by Citrobacter rodentium, Infect. Immun. (2010) 78:1509–1519. [CrossRef] [PubMed] [Google Scholar]
Barthel M., Hapfelmeier S., Quintanilla-Martínez L., Kremer M., Rohde M., Hogardt M., et al., Pretreatment of mice with streptomycin provides a Salmonella enterica serovar Typhimurium colitis model that allows analysis of both pathogen and host, Infect. Immun. (2003) 71:2839–2858. [CrossRef] [PubMed] [Google Scholar]
Bearson B.L., Bearson S.M., The role of the QseC quorum-sensing sensor kinase in colonization and norepinephrine-enhanced motility of Salmonella enterica serovar Typhimurium, Microb. Pathog. (2008) 44:271–278. [CrossRef] [PubMed] [Google Scholar]
Bearson B.L., Bearson S.M., Lee I.S., Brunelle B.W., The Salmonella enterica serovar Typhimurium QseB response regulator negatively regulates bacterial motility and swine colonization in the absence of the QseC sensor kinase, Microb. Pathog. (2010) 48:214–219. [CrossRef] [PubMed] [Google Scholar]
Berends B.R., Urlings H.A., Snijders J.M., Van Knapen F., Identification and quantification of risk factors in animal management and transport regarding Salmonella spp. in pigs, Int. J. Food Microbiol. (1996) 30:37–53. [CrossRef] [PubMed] [Google Scholar]
Callaway T.R., Morrow J.L., Edrington T.S., Genovese K.J., Dowd S., Carroll J., et al., Social stress increases fecal shedding of Salmonella typhimurium by early weaned piglets, Curr. Issues Intest. Microbiol. (2006) 7:65–71. [PubMed] [Google Scholar]
Clarke M.B., Hughes D.T., Zhu C., Boedeker E.C., Sperandio V., The QseC sensor kinase: a bacterial adrenergic receptor, Proc. Natl. Acad. Sci. USA (2006) 103:10420–10425. [CrossRef] [Google Scholar]
Davies R.H., Dalziel R., Gibbens J.C., Wilesmith J.W., Ryan J.M., Evans S.J., et al., National survey for Salmonella in pigs, cattle and sheep at slaughter in Great Britain, (1999–2000), J. Appl. Microbiol. (2004) 96:750–760. [CrossRef] [PubMed] [Google Scholar]
de Champlain J., Degeneration and regrowth of adrenergic nerve fibers in the rat peripheral tissues after 6-hydroxydopamine, Can. J. Physiol. Pharmacol. (1971) 4:345–355. [Google Scholar]
Dowd S.E., Callaway T.R., Morrow-Tesch J., Handling may cause increased shedding of Escherichia coli and total coliforms in pigs, Foodborne Pathog. Dis. (2007) 4:99–102. [CrossRef] [PubMed] [Google Scholar]
Ellermeier J.R., Slauch J.M., Fur regulates expression of the Salmonella pathogenicity island 1 type III secretion system through HilD, J. Bacteriol. (2008) 190:476–486. [CrossRef] [PubMed] [Google Scholar]
Foster N., Lovell M.A., Marston K.L., Hulme S.D., Frost A.J., Bland P., Barrow P.A., Rapid protection of gnotobiotic pigs against experimental salmonellosis following induction of polymorphonuclear leukocytes by avirulent Salmonella enterica, Infect. Immun. (2003) 71:2182–2191. [CrossRef] [PubMed] [Google Scholar]
Freestone P.P., Haigh R.D., Lyte M., Specificity of catecholamine-induced growth in Escherichia coli O157:H7, Salmonella enterica and Yersinia enterocolitica, FEMS Microbiol. Lett. (2007) 269:221–228. [CrossRef] [PubMed] [Google Scholar]
Freestone P.P., Lyte M., Microbial endocrinology: experimental design issues in the study of interkingdom signalling in infectious disease, Adv. Appl. Microbiol. (2008) 64:75–105. [CrossRef] [PubMed] [Google Scholar]
Freestone P.P., Sandrini S.M., Haigh R.D., Lyte M., Microbial endocrinology: how stress influences susceptibility to infection, Trends Microbiol. (2008) 16:55–64. [CrossRef] [PubMed] [Google Scholar]
Garner C.D., Antonopoulos D.A., Wagner B., Duhamel G.E., Keresztes I., Ross D.A., et al., Perturbation of the small intestine microbial ecology by streptomycin alters pathology in a Salmonella enterica serovar Typhimurium murine model of infection, Infect. Immun. (2009) 77:2691–2702. [CrossRef] [PubMed] [Google Scholar]
Hoiseth S.K., Stocker B.A., Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines, Nature (1981) 291:238–239. [CrossRef] [PubMed] [Google Scholar]
Hurd H.S., McKean J.D., Griffith R.W., Wesley I.V., Rostagno M.H., Salmonella enterica infections in market swine with and without transport and holding, Appl. Environ. Microbiol. (2002) 68:2376–2381. [CrossRef] [PubMed] [Google Scholar]
Isaacson R.E., Firkins L.D., Weigel R.M., Zuckermann F.A., DiPietro J.A., Effect of transportation and feed withdrawal on shedding of Salmonella typhimurium among experimentally infected pigs, Am. J. Vet. Res. (1999) 60:1155–1158. [PubMed] [Google Scholar]
Jones P.H., Roe J.M., Miller B.G., Effects of stressors on immune parameters and on the faecal shedding of enterotoxigenic Escherichia coli in piglets following experimental inoculation, Res. Vet. Sci. (2001) 70:9–17. [CrossRef] [PubMed] [Google Scholar]
Karavolos M.H., Spencer H., Bulmer D.M., Thompson A., Winzer K., Williams P., et al., Adrenaline modulates the global transcriptional profile of Salmonella revealing a role in the antimicrobial peptide and oxidative stress resistance responses, BMC Genomics (2008) 9:458. [CrossRef] [PubMed] [Google Scholar]
Laughlin R.S., Musch M.W., Hollbrook C.J., Rocha F.M., Chang E.B., Alverdy J.C., The key role of Pseudomonas aeruginosa PA-I lectin on experimental gut-derived sepsis, Ann. Surg. (2000) 232:133–142. [CrossRef] [PubMed] [Google Scholar]
Lyte M., Bailey M.T., Neuroendocrine-bacterial interactions in a neurotoxin-induced model of trauma, J. Surg. Res. (1997) 70:195–201. [CrossRef] [PubMed] [Google Scholar]
McCuddin Z.P., Carlson S.A., Sharma V.K., Experimental reproduction of bovine Salmonella encephalopathy using a norepinephrine-based stress model, Vet. J. (2008) 175:82–88. [CrossRef] [PubMed] [Google Scholar]
Methner U., Rabsch W., Reissbrodt R., Williams P.H., Effect of norepinephrine on colonisation and systemic spread of Salmonella enterica in infected animals: role of catecholate siderophore precursors and degradation products, Int. J. Med. Microbiol. (2008) 298:429–439. [CrossRef] [PubMed] [Google Scholar]
Moreira C.G., Weinshenker D., Sperandio V., QseC mediates Salmonella enterica serovar Typhimurium virulence in vitro and in vivo, Infect. Immun. (2010) 78:914–926. [CrossRef] [PubMed] [Google Scholar]
Morgan I.R., Krautil F.L., Craven J.A., Effect of time in lairage on caecal and carcass Salmonella contamination of slaughter pigs, Epidemiol. Infect. (1987) 98:323–330. [CrossRef] [PubMed] [Google Scholar]
Paulin S.M., Jagannathan A., Campbell J., Wallis T.S., Stevens M.P., Net replication of Salmonella enterica serovars Typhimurium and Choleraesuis in porcine intestinal mucosa and nodes is associated with their differential virulence, Infect. Immun. (2007) 75:3950–3960. [CrossRef] [PubMed] [Google Scholar]
Pullinger G.D., Carnell S.C., Sharaff F.F., van Diemen P.M., Dziva F., Morgan E., et al., Norepinephrine augments Salmonella enterica-induced enteritis in a manner associated with increased net replication but independent of the putative adrenergic sensor kinases QseC and QseE, Infect. Immun. (2010) 78:372–380. [CrossRef] [PubMed] [Google Scholar]
Rasko D.A., Moreira C.G., Li de R., Reading N.C., Ritchie J.M., Waldor M.K., et al., Targeting QseC signaling and virulence for antibiotic development, Science (2008) 321:1078–1080. [CrossRef] [PubMed] [Google Scholar]
Rasko D.A., Sperandio V., Anti-virulence strategies to combat bacteria-mediated disease, Nat. Rev. Drug Discov. (2010) 9:117–128. [CrossRef] [PubMed] [Google Scholar]
Reading N.C., Rasko D.A., Torres A.G., Sperandio V., The two-component system QseEF and the membrane protein QseG link adrenergic and stress sensing to bacterial pathogenesis, Proc. Natl. Acad. Sci. USA (2009) 106:5889–5894. [CrossRef] [Google Scholar]
Reissbrodt R., Rienaecker I., Romanova J.M., Freestone P.P., Haigh R.D., Lyte M., et al., Resuscitation of Salmonella enterica serovar Typhimurium and enterohemorrhagic Escherichia coli from the viable but nonculturable state by heat-stable enterobacterial autoinducer, Appl. Environ. Microbiol. (2002) 68:4788–4794. [CrossRef] [PubMed] [Google Scholar]
Sandrini S.M., Shergill R., Woodward J., Muralikuttan R., Haigh R.D., Lyte M., Freestone P.P., Elucidation of the mechanism by which catecholamine stress hormones liberate iron from the innate immune defense proteins transferrin and lactoferrin, J. Bacteriol. (2010) 192:587–594. [CrossRef] [PubMed] [Google Scholar]
Sekirov I., Tam N.M., Jogova M., Robertson M.L., Li Y., Lupp C., Finlay B.B., Antibiotic-induced perturbations of the intestinal microbiota alter host susceptibility to enteric infection, Infect. Immun. (2008) 76:4726–4736. [CrossRef] [PubMed] [Google Scholar]
Spencer H., Karavolos M.H., Bulmer D.M., Aldridge P., Chhabra S.R., Winzer K., et al., Genome-wide transposon mutagenesis identifies a role for host neuroendocrine stress hormones in regulating the expression of virulence genes in Salmonella, J. Bacteriol. (2010) 192:714–724. [CrossRef] [PubMed] [Google Scholar]
Stabel T.J., Evaluation of 2-deoxy-D-glucose for induction of a stress response in pigs, Am. J. Vet. Res. (1999) 60:708–713. [PubMed] [Google Scholar]
Stabel T.J., Fedorka-Cray P.J., Effect of 2-deoxy-D-glucose induced stress on Salmonella choleraesuis shedding and persistence in swine, Res. Vet. Sci. (2004) 76:187–194. [CrossRef] [PubMed] [Google Scholar]
Stevens M.P., Modulation of the interaction of enteric bacteria with intestinal mucosa by stress-related catecholamines, in: Lyte M., Freestone P. (Eds.), Microbial Endocrinology, Springer Press, London, 2010, pp. 111–134. [CrossRef] [Google Scholar]
Thomas G.D., O'Hagan K.P., Zambraski E.J., Effects of 6-hydroxydopamine in hypertensive adult miniature swine, Clin. Exp. Hypertens. A (1990) 12:647–661. [CrossRef] [PubMed] [Google Scholar]
Toscano M.J., Stabel T.J., Bearson S.M.D., Bearson B.L., Lay D.C., Cultivation of Salmonella enterica serovar Typhimurium in a norepinephrine-containing medium alters in vivo tissue prevalence in swine, J. Exp. Anim. Sci. (2007) 43:329–338. [CrossRef] [Google Scholar]
Vlisidou I., Lyte M., van Diemen P.M., Hawes P., Monaghan P., Wallis T.S., Stevens M.P., The neuroendocrine stress hormone norepinephrine augments Escherichia coli O157:H7-induced enteritis and adherence in a bovine ligated ileal loop model of infection, Infect. Immun. (2004) 72:5446–5451. [CrossRef] [PubMed] [Google Scholar]
Watson P.R., Paulin S.M., Jones P.W., Wallis T.S., Interaction of Salmonella serotypes with porcine macrophages in vitro does not correlate with virulence, Microbiology (2000) 146:1639–1649. [PubMed] [Google Scholar]
Williams P.H., Rabsch W., Methner U., Voigt W., Tschäpe H., Reissbrodt R., Catecholate receptor proteins in Salmonella enterica: role in virulence and implications for vaccine development, Vaccine (2006) 24:3840–3844. [CrossRef] [PubMed] [Google Scholar]
Mutations in the Salmonella enterica serovar Choleraesuis cAMP-receptor protein gene lead to functional defects in the SPI-1 Type III secretion system
Vet. Res. 41, 1-14 (2010)
Contamination of pigs by nose-to-nose contact or airborne transmission of Salmonella Typhimurium
Physiology, pathogenicity and immunogenicity of lon and/or cpxR deleted mutants of Salmonella Gallinarum as vaccine candidates for fowl typhoid
The mycotoxin deoxynivalenol promotes uptake of Salmonella Typhimurium in porcine macrophages, associated with ERK1/2 induced cytoskeleton reorganization
Vet. Res. 40, 1- (2009)
Examination of animal and zoonotic pathogens using microarrays

redpajama_set_name: RedPajamaCommonCrawl


Fast forward 30 years and guess what, global warming projections for the last three decades have turned out to be just as wrong as the global cooling projections of the 70s. The Wall Street Journal has just released a review of the predictions, and they have never even come close to being correct. It's almost like they just made it up from whole cloth.
According to the primary source material from the predictions, actual surface temperatures were increasing between the years 1960 and 1990. But in 1990, the projections look like a rocket while actual surface temperatures increase at a much slower rate. Temperature predictions for the current year are double what they actually are.
Climate models developed by the United Nations around the same time are equally faulty. But how could they be so wrong? The projections were based on a sharp increase in temperatures between the years 1984 and 1989. Surface temperatures dipped sharply in the previous four years, and then began to rise. The projections in the Global Warming Forecast of 1988 was based on a four-year trend.
It's almost as if these experts took a crash course in global climatology in 1983 and then started looking at current data and panicked. That's the most generous interpretation of the facts we could muster. But the fact that they wanted to hijack the entire government and rig the economy transforms what might have been a huge mistake into an attempted power grab of epic proportions.
The models for global warming are based on very brief temperature trends and do not take even decades-long temperature records into account. On top of this, they want us to believe that 97% of climate scientists agree with projections based on a four year period. That would mean 97% of them are wrong, and the next time they claim a huge consensus, we should simply ignore it outright.
All of this may be laughable. Yes, it makes people like Al Gore- who predicted that the ice caps would be liquefied by now- look like standard issue con men. But don't think for a moment that they didn't get everything they wanted. They did.
Huge environmental agencies with lots of political power have been created, and over the years, more than a billion dollars has been shoved into the pockets of the people crying wolf.
If you want a prediction you can count on, try this one on for size: The climate change scare will fade into obscurity over the next ten years. The public will forget about it. Then, sometime around 2088, global cooling will make headlines… and with a high degree of confidence, we predict Iron Maiden will stage a comeback tour.


text:
package org.fintx.http;

import okhttp3.Call;
import okhttp3.Callback;
import okhttp3.FormBody;
import okhttp3.Headers;
import okhttp3.OkHttpClient;
import okhttp3.Request;
import okhttp3.RequestBody;
import okhttp3.Response;
//import okhttp3.logging.HttpLoggingInterceptor;

import java.io.IOException;
import java.net.URL;
import java.security.KeyManagementException;
import java.security.NoSuchAlgorithmException;
import java.security.SecureRandom;
import java.security.cert.CertificateException;
import java.security.cert.X509Certificate;
import java.util.Map;
import java.util.concurrent.TimeUnit;

import javax.net.ssl.HostnameVerifier;
import javax.net.ssl.SSLContext;
import javax.net.ssl.SSLSession;
import javax.net.ssl.SSLSocketFactory;
import javax.net.ssl.TrustManager;
import javax.net.ssl.X509TrustManager;

/**
 * @author bluecreator(qiang.x.wang@gmail.com)
 *
 */
public final class AsyncHttpClient {
    private AsyncHttpClient() {
        throw new AssertionError("No HttpClient instances for you!");
    }

    static private OkHttpClient client;

    static {
        // HttpLoggingInterceptor interceptor=new HttpLoggingInterceptor(new HttpLogger());
        // interceptor.setLevel(HttpLoggingInterceptor.Level.BODY);
        client = new OkHttpClient().newBuilder().sslSocketFactory(createSSLSocketFactory()).hostnameVerifier(new TrustAllHostnameVerifier())
                .retryOnConnectionFailure(true).connectTimeout(20, TimeUnit.SECONDS).writeTimeout(40, TimeUnit.SECONDS).readTimeout(60, TimeUnit.SECONDS)
                /* .addNetworkInterceptor(interceptor) */.build();
    }

    /**
     * 默认信任所有的证书 TODO 最好加上证书认证,主流App都有自己的证书.
     *
     * @return
     */
    // @SuppressLint("TrulyRandom")
    private static SSLSocketFactory createSSLSocketFactory() {

        SSLSocketFactory sslSocketFactory = null;

        try {
            SSLContext sc = SSLContext.getInstance("TLS");
            sc.init(null, new TrustManager[] {new TrustAllManager() }, new SecureRandom());
            sslSocketFactory = sc.getSocketFactory();
        } catch (IllegalStateException | KeyManagementException | NoSuchAlgorithmException e) {
            e.printStackTrace();
        }

        return sslSocketFactory;
    }

    private static class TrustAllManager implements X509TrustManager {
        @Override
        public void checkClientTrusted(X509Certificate[] chain, String authType) throws CertificateException {
        }

        @Override
        public void checkServerTrusted(X509Certificate[] chain, String authType)

                throws CertificateException {
        }

        @Override
        public X509Certificate[] getAcceptedIssuers() {
            return new X509Certificate[0];
        }
    }

    private static class TrustAllHostnameVerifier implements HostnameVerifier {
        @Override
        public boolean verify(String hostname, SSLSession session) {
            return true;
        }
    }

    public static void getAsync(URL url) throws IOException, IllegalStateException {
        Request request = new Request.Builder().url(url).build();

        client.newCall(request).enqueue(new Callback() {
            @Override
            public void onFailure(Call call, IOException e) {
                e.printStackTrace();
            }

            @Override
            public void onResponse(Call call, Response response) throws IOException {
                if (!response.isSuccessful())
                    throw new IllegalStateException("Unexpected code " + response);

                Headers responseHeaders = response.headers();
                for (int i = 0, size = responseHeaders.size(); i < size; i++) {
                    System.out.println(responseHeaders.name(i) + ": " + responseHeaders.value(i));
                }

                System.out.println(response.body().string());
            }
        });
    }

}

redpajama_set_name: RedPajamaGithub


Having a fence around your pool is more than a luxury. It's also the law.
It's important to understand the state and county laws that protect you, your family, and everyone else that may find themselves in or around your pool area.
Laws in Maryland and Virginia differ from state to state. Within the states themselves, however, the county laws differ from county to county. It's imperative that you're absolutely sure of the requirements your state and county have in place for your pool fencing. A local fencing professional is the best person to contact when you have any questions or concerns regarding your pool fencing.
Wooden fencing requires proper care and maintenance. If you own a wooden fence, take note on how to properly take care for and maintain it.
Do you have a wrought iron fence? They look beautiful, but you want to make sure the continue looking beautiful for a long time!
Selling your home? Here are some facts you should know about your fence before you put your home on the market.
Vines look beautiful and add privacy to your yard, but you must be sure you're planting the right type of vine.
Can You Install a Fence in the Winter?
Find out why winter is actually a great time to install your new fence!
If you want to enhance the curb appeal of your home, don't forget about your fence!
When Do I Need a New Fence?
It can be difficult to know when it's time to replace your fence.
should consider investing in a new pool fence?
A pool fence isn't just a beautiful addition to your yard — it can save lives!


text:
Optimal maximum time from referral to diagnosis and treatment
Clinical practice guidelines for the prevention, early detection and management of colorectal cancer > Clinical question
Guideline contents > SPT1-2 (b): What signs/symptoms alone or in combination are most predictive of CRC and what is the optimal maximum time from referral to diagnosis and treatment (diagnostic interval)?
Professor Jon Emery MA, MBBCh, FRACGP, MRCGP, DPhil — Author
A/Prof Gregor Brown — Co-author
Professor Finlay Macrae AO; MBBS; MD; FRACP; FRCP; AGAF — Co-author
Dr Cameron Bell MD FRACP — Co-author
A/Prof Justin Tse — Co-author
Dr Iain Skinner — Co-author
Emery, J, Ben Lee-Bates, Brown, G, Macrae, F, Bell, C, A/Prof Justin Tse, Dr Iain Skinner, Cancer Council Australia Colorectal Cancer Guidelines Working Party. Clinical question:Diagnosis interval in colorectal cancer . In: Clinical practice guidelines for the prevention, early detection and management of colorectal cancer. Sydney: Cancer Council Australia. [Version URL: https://wiki.cancer.org.au/australiawiki/index.php?oldid=173048, cited 2023 Jan 27]. Available from: https://wiki.cancer.org.au/australia/Guidelines:Colorectal_cancer.
1.1 The diagnostic pathway
1.2 Methodological issues
2 Systematic review evidence
2.1 Mortality
2.2 Colorectal cancer-specific mortality
2.3 Tumour stage at diagnosis
3 Evidence summary and recommendations
3.1 Considerations in making these recommendations
4 Benefits and harms
5 Health system implications
5.1 Clinical practice
5.2 Resourcing
5.3 Barriers to implementation
6.1 Unresolved issues
6.2 Studies currently underway
6.3 Future research priorities
8 Appendices
Intuitively, it would be expected that diagnosing cancer quickly would be beneficial, as tumours grow and are more likely to metastasise with time. Indeed, perception of a 'delayed diagnosis' of cancer is a leading cause of medicolegal complaints in primary and ambulatory care, on the assumption that harm occurred as a result of late diagnosis.[1]
The diagnostic pathway[edit source]
So-called delays in cancer diagnosis can occur at various points along the diagnostic pathway.[2] Patients may take time appraising their symptoms before seeking healthcare, they may experience multiple visits to their GP about their symptoms before referral for specialist diagnostic tests,[3] and there may be long waiting times to access these diagnostic tests. This latter point along the diagnostic pathway, from GP referral to diagnosis, is the focus of this section.
Access to timely colonoscopy is an important contributor to the overall diagnostic interval for colorectal cancer (defined usually as the time a patient first presents to healthcare until the time of diagnosis).[4]
Methodological issues[edit source]
Proving that earlier detection of symptomatic cancer matters is epidemiologically challenging, the 'waiting time paradox' describes the phenomenon in which patients with late stage cancers present with severe symptoms and are therefore often diagnosed promptly, but have poorer outcomes.[5] This type of confounding by indication is an important source of bias in studies examining the effect of time to diagnosis on outcomes in symptomatic cancer populations. Many studies that have examined associations between the diagnostic interval and clinical outcomes have assumed a linear relationship between time to diagnosis and mortality. Their analyses, therefore, have not accounted for potential effects of the waiting time paradox. More recent studies have introduced the use of spline regression to allow for flexible associations between the diagnostic interval and clinical outcome.[6][7] These important methodological considerations must be taken into account when interpreting the evidence, which includes apparently inconsistent findings. When making recommendations, we applied greater weight to studies that attempted to account for the waiting time paradox.
Systematic review evidence[edit source]
In symptomatic patients without a colorectal cancer diagnosis, what is the optimal maximum diagnostic interval that achieves better than or equivalent outcomes in terms of survival, mortality, and diagnosis of metastatic disease? (SPT1-2b)
Nine studies[8][9][10][7][6][11][12][13][14] examined the effect of the diagnostic interval on colorectal cancer related outcomes including mortality, cancer specific survival and mortality, and stage of tumour at diagnosis. Seven studies[8][9][10][6][7][11][12] had a moderate risk of bias and two had a high risk of bias.[13][14]
The search strategy, inclusion and exclusion criteria, and quality assessment are described in detail in the Technical report.
Mortality[edit source]
A Canadian retrospective cohort study[10] found that diagnostic interval had no significant effect of diagnostic interval length on colorectal cancer mortality with 1–6 years follow-up.
Danish prospective population-based cohort studies in primary care,[7][11] a UK retrospective cohort study,[9] and a study that included one retrospective and two prospective primary care cohort studies conducted in Denmark and the UK[6] reported significantly higher 3- and 5-year mortality rates associated with shorter waiting periods (all < 1 month). These findings are consistent with the 'waiting time paradox' where patients with severe symptoms associated with later stage disease are diagnosed promptly.
Three Danish and UK primary care cohort studies[6] reported U-shaped associations between diagnostic interval and overall mortality (at 3 or 5 years) using spline regression analyses. Analysis of combined datasets found that higher 5-year mortality was associated with diagnostic intervals greater than 130 days (HR=1.28 95% CI 1.28-1.55).
A large US retrospective study of > 9,000 patients diagnosed with colorectal cancer between 1998 and 2005[8] found that, for patients with colon cancer only, diagnostic intervals of ≥ 8 months compared with 14–59 days showed a significant effect on overall mortality (OR 1.31, 95% CI 1.08 to 1.58). For local stage rectal cancer, mortality was higher for diagnostic intervals < 2 weeks and 2–4 months, compared with 14–59 days, consistent with the U-shaped associations demonstrated in UK and Danish populations.[6][7][11]
Colorectal cancer-specific mortality[edit source]
In an analysis of a large US dataset of medical records for adults aged ≥ 66 years with invasive colon or rectal cancer, colorectal cancer-specific mortality was reported separately for patients diagnosed with either colon cancer or rectal cancer.[8] For those diagnosed with colon cancer, in unadjusted analysis, higher mortality was reported for shorter diagnostic delay (< 2 weeks), compared with 14–59 days (OR 1.27, p < 0.05). Significantly higher mortality was reported when comparing short diagnostic interval (14–59 days) with longer diagnostic intervals of 4–8 months and ≥ 8 months (OR 0.76, p < 0.05, and OR 0.82, p < 0.05, respectively), thus failing to demonstrate any evidence of a U-shaped association between interval and colorectal cancer-specific mortality.
A cohort study comparing outcomes in patients with early and late diagnosis[14] reported significantly higher 5-year cancer-specific survival for a diagnostic interval ≥ 50 days compared with < 50 days when all participants were included in the analysis (94% versus 73%, respectively, p = 0.007).[14] No attempt was made to account for the waiting time paradox in this study.
Tumour stage at diagnosis[edit source]
Four studies[12][13][14][10] examined associations between diagnostic intervals and tumour stage but only one[10] conducted analyses to account for a potential waiting time paradox.
A retrospective cohort study[12] compared stages for three interval cut-offs (> 41 days, > 60 days, > 90 days), assuming a linear effect of time. Shorter intervals were associated with more advanced stage disease.[12]
Another retrospective cohort study[13] reported shorter diagnostic intervals were associated with earlier stages of cancer, however this effect was non-significant.[13]
A cohort study comparing outcomes in patients with early and late diagnosis[14] reported greater rates of Dukes' stage A cancer in participants with a diagnostic interval ≥ 50 days (57.1%) compared with < 50 days (15.2%, p = 0.006).[14]
A large Canadian retrospective cohort study[10] reported higher rates of stage III/IV colorectal cancer for participants with a diagnostic interval < 15 days compared with 51 to < 116 days or ≥ 116 days (OR 0.59, CI 0.39 to 0.89 and OR 0.50, CI 0.33 to 0.75, respectively) but not 15 to < 51 days, consistent with a U-shaped association between diagnostic interval and clinical outcome.[10]
Summary[edit source]
The studies that performed analyses to account for the waiting time paradox found potentially important U-shaped associations between diagnostic intervals and (1) overall mortality[6][7][11][8] and (2) late-stage disease at diagnosis,[10] but not colorectal cancer-specific mortality[8]
The following cut-off intervals for first presentation to healthcare to diagnosis were associated with poorer outcomes:
130 days in the largest study combining three datasets from Danish and UK primary care cohorts[6]
8 months (approximately 243 days) in a large US retrospective study[8]
116 days in a Canadian retrospective study from population-based cancer registry and administrative database.[10]
In the Australian setting, the presentation–diagnosis interval would most commonly represent the time from GP consultation to diagnostic colonoscopy (or other diagnostic procedure) in specialist care.
Evidence summary and recommendations[edit source]
Evidence summary
Analyses of cohort data have reported U-shaped associations between diagnostic interval and (1) overall mortality and (2) late-stage disease at diagnosis, but not colorectal cancer-specific mortality. III-2 [11], [7], [6], [8], [10]
Diagnostic interval cut-off points associated with poorer outcomes range between 116 days and 8 months. III-2 [6], [8], [10]
Evidence-based recommendation
For patients with symptoms suggestive of colorectal cancer, the total time from first healthcare presentation† to diagnostic colonoscopy should be no more than 120 days. Diagnostic intervals greater than 120 days are associated with poorer clinical outcomes.
† First healthcare presentation is defined as the date of presentation in general practice with symptoms suggestive of colorectal cancer or positive iFOBT for screening.
A diagnostic interval of 120 days should be the maximum time from first healthcare presentation† to diagnostic colonoscopy for triage Categories 1 and 2, whether it is for a patient with symptoms or after a positive iFOBT used for colorectal cancer screening. Diagnostic intervals greater than 120 days are associated with poorer clinical outcomes.
Consensus-based recommendation
Triage category 1 patients, whether due to symptoms or positive iFOBT, should continue to be considered most urgent and prioritised for diagnostic colonoscopy, in any model of care at any jurisdictional level.
Colonoscopy for symptomatic patients should be performed as promptly as possible after referral from general practice, especially for those meeting triage Category 1 criteria. If cancer is present, there is no evidence that prognosis is worsened within 120 days from first presentation to diagnostic colonoscopy. However, performing colonoscopy as promptly as possible after referral from general practice is to minimise the risk of psychological harm in symptomatic or iFOBT-positive patients who are potentially anxious while awaiting investigation. Prompt scheduling will also help to ensure that any unexpected delays between general practice referral and colonoscopy triaging do not flow on to exceed the 120-day threshold after which prognosis can worsen if cancer is present.
Considerations in making these recommendations[edit source]
These recommendations are based on the consensus of the guideline development group and interpretation of the best available evidence. There will of course never be Level I evidence to inform these recommendations as RCTs of different diagnostic intervals would be deemed unethical. A maximum diagnostic interval of 120 days from first presentation to healthcare (first healthcare presentation is defined as the date of presentation in general practice with symptoms suggestive of colorectal cancer or positive iFOBT for screening) to diagnosis should be the target to prevent poorer outcomes in those with colorectal cancer. We noted the current recommendation in the Optimal care pathway[15] for colorectal cancer of a maximum of four weeks from referral to colonoscopy for people with symptoms suggestive of colorectal cancer. Recognising that there will be a small proportion of people with colorectal cancer in triage Category 2 (approximately 1-2%), we recommend that all Category 1 and Category 2 colonoscopies (screen positive iFOBT or symptomatic patients) should be performed no later than 120 days from first presentationi to healthcare. Ideally colonoscopy should be performed sooner than this to reduce the risk of psychological harm to patients.[16][17]
The Working Party and subcommittee members had robust discussion regarding the maximum optimal time from first healthcare presentation to diagnostic colonoscopy and treatment. Although the group was in agreement about the interpretation of the systematic review evidence, there was concern about de-emphasising the need for prompt evaluation. The Working Party acknowledges that the guideline may be read with the expectation that it will assist in triage of colonoscopy patients. The authors resolved it was appropriate to maintain the evidence-based recommendations, acknowledging the grade and limitations of the available evidence, but also add the practice point about the ideal interval for symptomatic patients. Given the unavoidable delays along the pathway, all people with a positive iFOBT or with symptoms suggestive of colorectal cancer should have a colonoscopy as promptly as possible.
i Date of first presentation is defined as the positive screening iFOBT.
Benefits and harms[edit source]
There is evidence to suggest that a greater proportion of the diagnostic interval occurs from the point of referral to colonoscopy, rather than in primary care, especially where there is poorer access to colonoscopy. While recognising the current challenges of meeting demand in public health endoscopy services, the guideline development group recommended a target diagnostic interval of a maximum of 120 days from first presentation to healthcare (first healthcare presentation is defined as the date of presentation in general practice with symptoms suggestive of colorectal cancer or positive iFOBT for screening) for all patients meeting either Category 1 or Category 2 criteria.
Health system implications[edit source]
Clinical practice[edit source]
GPs will need to remain alert to the possibility of colorectal cancer as a possible cause of a patient's symptoms and investigate and refer promptly based on the evidence summarised in the previous section.
Resourcing[edit source]
Endoscopy services will need to establish clear diagnostic pathways for patients with suspected colorectal cancer and establish systems to apply the triage criteria and organise timely colonoscopy. Meeting a 120 day target from first presentation for all Category 1 and Category 2 will have significant resource implications for some public hospital endoscopy services.
Barriers to implementation[edit source]
These recommendations are made in the context of the roll-out of the NBCSP, due to be fully implemented by 2020 which will place additional demand for colonoscopy. We acknowledge the challenges of measuring this target given that the evidence is based on the diagnostic interval commencing at the time of first presentation to healthcare. In order to monitor the 120 day diagnostic interval target, referrals will need to record the date of first presentation to healthcare with symptoms suggestive of colorectal cancer. We recognise that this may be logistically challenging to collect and recommend that this information is collected within the standardised GP referral proforma[18] (see Resourcing).
Discussion[edit source]
Unresolved issues[edit source]
Timely diagnosis of colorectal cancer is important for improving survival. While there are inevitable limitations in defining the optimal maximum time to diagnose someone with suspected colorectal cancer, we have applied the current best evidence to make our recommendations. The triage criteria and associated maximum intervals for colonoscopy in Category 1 and 2 patients are designed to improve the efficiency of the referral and triage processes for people with symptoms suggestive of colorectal cancer.
Studies currently underway[edit source]
The authors are not aware of any studies underway that may provide more information on this topic.
Future research priorities[edit source]
Further well-designed research, which accounts for the waiting time paradox, is needed to confirm the estimates of minimum diagnostic intervals associated with poorer colorectal cancer outcomes. In addition, studies should monitor the impact of the implementation of colonoscopy triage categories on waiting times, diagnostic intervals and colorectal cancer outcomes.
↑ Wallace, E. Lowry, J. Smith, S. M. Fahey, T.. The epidemiology of malpractice claims in primary care: a systematic review. BMJ Open 2013;3(7).
↑ Walter F, Webster A, Scott S, Emery J. The Andersen Model of Total Patient Delay: a systematic review of its application in cancer diagnosis. J Health Serv Res Policy 2012 Apr;17(2):110-8 Available from: http://www.ncbi.nlm.nih.gov/pubmed/22008712.
↑ Lyratzopoulos G, Neal RD, Barbiere JM, Rubin GP, Abel GA. Variation in number of general practitioner consultations before hospital referral for cancer: findings from the 2010 National Cancer Patient Experience Survey in England. Lancet Oncol 2012 Apr;13(4):353-65 Available from: http://www.ncbi.nlm.nih.gov/pubmed/22365494.
↑ Weller D, Vedsted P, Rubin G, Walter FM, Emery J, Scott S, et al. The Aarhus statement: improving design and reporting of studies on early cancer diagnosis. Br J Cancer 2012 Mar 27;106(7):1262-7 Available from: http://www.ncbi.nlm.nih.gov/pubmed/22415239.
↑ Neal RD, Tharmanathan P, France B, Din NU, Cotton S, Fallon-Ferguson J, et al. Is increased time to diagnosis and treatment in symptomatic cancer associated with poorer outcomes? Systematic review. Br J Cancer 2015 Mar 31;112 Suppl 1:S92-107 Available from: http://www.ncbi.nlm.nih.gov/pubmed/25734382.
↑ 6.0 6.1 6.2 6.3 6.4 6.5 6.6 6.7 6.8 6.9 Tørring ML, Frydenberg M, Hamilton W, Hansen RP, Lautrup MD, Vedsted P. Diagnostic interval and mortality in colorectal cancer: U-shaped association demonstrated for three different datasets. J Clin Epidemiol 2012 Jun;65(6):669-78 Available from: http://www.ncbi.nlm.nih.gov/pubmed/22459430.
↑ 7.0 7.1 7.2 7.3 7.4 7.5 7.6 Tørring ML, Frydenberg M, Hansen RP, Olesen F, Hamilton W, Vedsted P. Time to diagnosis and mortality in colorectal cancer: a cohort study in primary care. Br J Cancer 2011 Mar 15;104(6):934-40 Available from: http://www.ncbi.nlm.nih.gov/pubmed/21364593.
↑ 8.0 8.1 8.2 8.3 8.4 8.5 8.6 8.7 8.8 Pruitt SL, Harzke AJ, Davidson NO, Schootman M. Do diagnostic and treatment delays for colorectal cancer increase risk of death? Cancer Causes Control 2013 May;24(5):961-77 Available from: http://www.ncbi.nlm.nih.gov/pubmed/23446843.
↑ 9.0 9.1 9.2 Redaniel MT, Martin RM, Ridd MJ, Wade J, Jeffreys M. Diagnostic intervals and its association with breast, prostate, lung and colorectal cancer survival in England: historical cohort study using the Clinical Practice Research Datalink. PLoS One 2015;10(5):e0126608 Available from: http://www.ncbi.nlm.nih.gov/pubmed/25933397.
↑ 10.00 10.01 10.02 10.03 10.04 10.05 10.06 10.07 10.08 10.09 10.10 Singh H, Shu E, Demers A, Bernstein CN, Griffith J, Fradette K. Trends in time to diagnosis of colon cancer and impact on clinical outcomes. Can J Gastroenterol 2012 Dec;26(12):877-80 Available from: http://www.ncbi.nlm.nih.gov/pubmed/23248786.
↑ 11.0 11.1 11.2 11.3 11.4 11.5 Tørring ML, Frydenberg M, Hansen RP, Olesen F, Vedsted P. Evidence of increasing mortality with longer diagnostic intervals for five common cancers: a cohort study in primary care. Eur J Cancer 2013 Jun;49(9):2187-98 Available from: http://www.ncbi.nlm.nih.gov/pubmed/23453935.
↑ 12.0 12.1 12.2 12.3 12.4 Wattacheril J, Kramer JR, Richardson P, Havemann BD, Green LK, Le A, et al. Lagtimes in diagnosis and treatment of colorectal cancer: determinants and association with cancer stage and survival. Aliment Pharmacol Ther 2008 Nov 1;28(9):1166-74 Available from: http://www.ncbi.nlm.nih.gov/pubmed/18691351.
↑ 13.0 13.1 13.2 13.3 13.4 Gómez-Domínguez E, Trapero-Marugán M, del Pozo AJ, Cantero J, Gisbert JP, Maté J. The colorectal carcinoma prognosis factors. Significance of diagnosis delay. Rev Esp Enferm Dig 2006 May;98(5):322-9 Available from: http://www.ncbi.nlm.nih.gov/pubmed/16944992.
↑ 14.0 14.1 14.2 14.3 14.4 14.5 14.6 Rupassara KS, Ponnusamy S, Withanage N, Milewski PJ. A paradox explained? Patients with delayed diagnosis of symptomatic colorectal cancer have good prognosis. Colorectal Dis 2006 Jun;8(5):423-9 Available from: http://www.ncbi.nlm.nih.gov/pubmed/16684087.
↑ Cancer Council Victoria. Optimal care pathway for people with colorectal cancer.; 2014 Available from: www.cancer.org.au/ocp.
↑ Lindholm E, Berglund B, Kewenter J, Haglind E. Worry associated with screening for colorectal carcinomas. Scand J Gastroenterol 1997 Mar;32(3):238-45 Available from: http://www.ncbi.nlm.nih.gov/pubmed/9085461.
↑ Parker MA, Robinson MH, Scholefield JH, Hardcastle JD. Psychiatric morbidity and screening for colorectal cancer. J Med Screen 2002;9(1):7-10 Available from: http://www.ncbi.nlm.nih.gov/pubmed/11943790.
↑ Akbari A, Mayhew A, Al-Alawi MA, Grimshaw J, Winkens R, Glidewell E, et al. Interventions to improve outpatient referrals from primary care to secondary care. Cochrane Database Syst Rev 2008 Oct 8;(4):CD005471 Available from: http://www.ncbi.nlm.nih.gov/pubmed/18843691.
Appendices[edit source]
View recommendation components
View pending evidence
View body of evidence
View literature search
View PICO
NHMRC Evidence statement form SPT1-2b
Systematic review report SPT1-2b
A medical professional who treats acute and chronic illnesses and provides preventive care and health education to a wide range of patients.
A recommendation formulated after a systematic review of the evidence, indicating supporting references.
A test that can detect microscopic amounts of blood in stools. Types of FOBT include immunochemical FOBTs (iFOBTs), which directly detect haemoglobin using antibodies specific for the globin moiety of human haemoglobin, and guaiac FOBTs (gFOBTs), which detect peroxidase activity, an indirect method for identification of haemoglobin.
A recommendation formulated in the absence of quality evidence, after a systematic review of the evidence was conducted and failed to identify admissible evidence on the clinical question.
An examination of the large bowel using a camera on a flexible tube, which is passed through the anus.
The National Bowel Cancer Screening Program. An Australian screening program that aims to reduce illness and death from bowel cancer through early detection or prevention of the disease.
Retrieved from "https://wiki.cancer.org.au/australiawiki/index.php?title=Clinical_question:Diagnosis_interval_in_colorectal_cancer&oldid=173048"


text:
Articles in Sweden ( 2,408 )
2,408 Articles of interest in Sweden
Click on them to get its location and coordinates
Karlevi Runestone
The Karlevi Runestone, designated as Öl 1 by Rundata, is commonly dated to the late 10th century and located near the Kalmarsund straight in Karlevi on the island of Öland, Sweden. It is one of the most notable and prominent runestones and constitut…
 Show coordinates
Kalmar Cathedral
Kalmar Cathedral (Swedish: Kalmar domkyrka) is in the city of Kalmar in Småland in southeast Sweden, situated on the Baltic Sea.
Hovgården
Hovgården is an archaeological site on the Lake Mälaren island of Adelsö in Ekerö Municipality in central-eastern Sweden. During the Viking Age, the centre of the prospering Mälaren Valley was the settlement Birka, founded in the mid-8th century and…
Helgeandsholmen
Helgeandsholmen is a small island in central Stockholm, Sweden. It is located north of Stadsholmen, and east of Strömsborg, with which, together with Riddarholmen, it forms Gamla stan, the old town of Stockholm.
Halmstad University, Sweden
Halmstad University or Halmstad University College (Swedish: Högskolan i Halmstad) (HH) is a University college (Swedish: högskola) in Halmstad, Sweden. It was established in 1983. Halmstad University is a public higher education institution offerin…
Gustavianum
Gustavianum is the former main building of Uppsala University, built 1622–1625. The name Gustavianum comes from Gustavus Adolphus who in the 1620s donated money for its construction. The Gustavianum was Uppsala University's previous main building, u…
Gothenburg Cathedral
Gothenburg Cathedral (Swedish: Gustavi domkyrka / Göteborgs domkyrka) is a cathedral in Gothenburg, the second largest city in Sweden.
Fittja
Fittja is a part of Botkyrka Municipality and the name of the Stockholm metro station in the area. It was settled during the 1970s and consists mostly of rental apartments.
Diplomatstaden
Diplomatstaden (Swedish for "The diplomat city") is a neighbourhood in the Östermalm district in central Stockholm, Sweden.
Saab Arena
Saab Arena, formerly named Cloetta Center between 2004–2014, is an arena in Linköping, Sweden. It opened in 2004 and holds 8,500 people during sport events and 11,500 during concerts.
Cirkus (Stockholm)
Cirkus (Cirkus Arena och Restaurang AB) is an arena in Djurgården, Stockholm, that holds 1,650 people.
Battle of Gestilren
The Battle of Gestilren took place on July 17, 1210. The battle was fought between the exiled King of Sweden Sverker and the ruling King Eric. Sverker had been beaten in the previous Battle of Lena, but had returned with new forces. Sverker was kill…
Arjeplog Municipality
Arjeplog Municipality (Swedish: Arjeplogs kommun, Northern Sami: Arjepluovvi gielda) is a municipality in Norrbotten County in northern Sweden.
ABB Arena is the common name for the two biggest indoor arenas in Västerås, Sweden.[1] "ABB Arena Nord" is the renovated arena, which was earlier called Rocklundahallen. ABB Arena Nord is the ice hockey arena and VIK Västerås HK's home arena. It was…
Örebro Airport
Örebro Airport (IATA: ORB, ICAO: ESOE) is located 10 kilometers southwest of Örebro and is Sweden's 23rd largest passenger airport and the 4th largest cargo airport.
Åre Municipality
Åre Municipality (Swedish: Åre kommun) is a municipality in Jämtland County in northern Sweden.
Visingsö is an island in the southern half of the lake Vättern in Sweden.
Vadstena Abbey
The Abbey of Our Lady and of St. Bridget (Latin: Monasterium sanctarum Mariæ Virgìnis et Brigidæ in Vatzstena), more commonly referred to as Vadstena Abbey, situated on Lake Vättern, in the Diocese of Linköping, Sweden, was the motherhouse of the Br…
Uppåkra
Uppåkra is a village located five kilometres south of Lund in Scania in southernmost Sweden.
Torneträsk
Torneträsk or Torne träsk (Saami: Duortnosjávri, Finnish/Meänkieli: Tornio or Torniojärvi) is a lake in Kiruna Municipality, Lapland, Norrbotten County in Sweden, in the Scandinavian mountain range. Träsk is the local word for lake (in Standard Swed…
Telenor Arena Karlskrona
Telenor Arena Karlskrona (formally Arena Rosenholm) is a multi-purpose sporting arena for indoors and outdoors, located in Karlskrona, Sweden.
Stora Karlsö
Stora Karlsö is a small Swedish island in the Baltic Sea, situated about 6 km west of Gotland. It has an area of about 2.5 km² and is up to 52 meters high. Most of the island consists of a limestone plateau, bordered by steep cliffs along the shore.…
Sofiero Castle
Sofiero Castle or just Sofiero (Swedish: Sofiero slott or just Sofiero) in Helsingborg Municipality, Scania, was one of the Swedish royal family's country mansions, located 5 kilometres (3.1 mi) north of Helsingborg. It was originally a Scanian farm…
Partille Municipality
Partille Municipality (Partille kommun) is a municipality in Västra Götaland County in western Sweden.
Olympia (Helsingborg)
Olympia, sometimes referred to as Olympiastadion (English: Olympic Stadium), is a football stadium in Helsingborg, Sweden. It was opened in 1898, but has been rebuilt both 1993 and 1997, and has a capacity of 17,200 (9,673 seated and about 7,500 gro…
Nils Ericson Terminal
The Nils Ericson Terminal is a major bus terminus in Gothenburg, Sweden. It is placed in the city centre just next to Gothenburg Central Station (Centralstationen), and across the street from the main Nordstan shopping centre. The main street and mo…
Museum of Medieval Stockholm
The Museum of Medieval Stockholm (Swedish: Stockholms medeltidsmuseum), centrally located north of the Royal Palace, was constructed around old monuments excavated in an extensive archaeological dig (dubbed Riksgropen, "National/State Pit") in the l…
Läckö Castle
Läckö Castle (in Swedish: Läckö Slott ) is a medieval castle in Sweden.
Linnaean Garden
The Linnaean Garden or Linnaeus' Garden (Swedish: Linnéträdgården) is the oldest of the botanical gardens belonging to Uppsala University in Sweden.
Kungliga begravningsplatsen
Kungliga begravningsplatsen (translates as the Royal Cemetery) was first used in 1922 and has been the only official burial place of the Swedish Royal Family since 1950, replacing Riddarholm Church.
Kristianstad Airport
Kristianstad Airport (IATA: KID, ICAO: ESMK) is situated outside Kristianstad in Scania, Sweden.
Kopparvallen
Kopparvallen is a multi-use stadium in Åtvidaberg, Sweden. It is used mostly for football matches and is the home stadium of Åtvidabergs FF.
Karlskoga Motorstadion, also known as Gelleråsen is the oldest permanent motorsport race track in Sweden. The circuit is located 6 km north of Karlskoga.
Jokkmokk Municipality
Jokkmokk Municipality (Swedish: Jokkmokks kommun, Finnish: Jokimukan kunta, Northern Sami: Johkamohkki gielda, Meänkieli: Jokinmukka) is a municipality in Norrbotten County in northern Sweden.
Hötorget buildings
The Hötorget buildings (Swedish: Hötorgshusen or Hötorgsskraporna, "-scrapers") are five high-rise office buildings in Stockholm, Sweden.
Hägersten-Liljeholmen is a borough (stadsdelsområde) in Stockholm, Sweden.
Page 12 of 67 « 1 … 7 8 9 10 11 12 13 14 15 16 17 18 …67 »


text:
Home Innovation Computing Processors
X-Box: The big picture
An overall look at the controversy and glitz surrounding the X-Box launch
Written by ZDNet UK, Contributor on March 13, 2000
Microsoft turned its attention from business and home desktop computer systems to the less familiar territory of high-end gaming on Friday with the announcement of its much anticipated X-Box console.
The event was not without controversy, however. At the last minute, Microsoft mysteriously decided to shun expected partner AMD in favour of its traditional microprocessor pal Intel. Question marks also remain over Microsoft's ability to capture gamers' attention from the likes of Sony and Sega, as well as deliver the console as planned.
Here is a round-up of all the news surrounding one of Microsoft's biggest ever gambles.
AMD licks its wounds Did Intel win the X-Box deal at the 11th hour? Mon, 13 March, 2000,
X-Box delays and extra costs for Europe? As per usual, UK gamers may have to wait longer than Japanese and American customers for X-Box. Fri, 10 Mar 2000
Opinion: X-Box dumps AMD Observers say Microsoft's decision to dump AMD in favour of an Intel processor is not surprising. Fri, 10 Mar 2000
Official: X-Box will use Intel New Microsoft games console won't use AMD's Athlon chip as expected. Fri, 10 Mar 2000
Microsoft rips the veil off X-Box Bill Gates and a bevy of third-party developers talk about Microsoft's long-rumoured entry into the game console market, slated for release in autumn 2001. Fri, 10 Mar 2000
The battle for your living room Microsoft readies itself to enter the gaming market and go head-to-head in the living room with Sony, Nintendo and Sega. Wed, 8 Mar 2000
Bill Gates is like a pushy salesman. If he can get one foot in the door, he'll barge on through. Go to AnchorDesk UK with Jesse Berst for the news comment.
I love games, so take me to Gamespot immediately.
Go to the AnchorDesk UK Talkback forum on the Microsoft X-Box.
What do you think? Tell the Mailroom and read what others have to say.


text:
Q: How can I browse OSX shared resources from Ubuntu server? What I want is to check what resources are shared on OSX from my Ubuntu server, then I can mount particular resource using mount.cifs. From Windows 7 I can browse and see what resources are available (root level shares), however I'm not sure how to do that from headless Ubuntu.

A: Use smbclient -L server to list shares on a particular SMB server.

redpajama_set_name: RedPajamaStackExchange


Once you've taken control of Connor ride your horse to the marked place on the map and speak with Washington. He will give you the task of finding the person responsible for stealing the supplies.
Head to the abandoned church where you will meet your father.
After having spoken to him search for traces of the thief. The first clue can be found on the road in front of the building.
The second is on the same road but slightly to the left.
Examine them, head to the green zone marked on the map and start chasing the cart.
After you�re done interrogating the driver, head to the marked spot and begin following the convoy passing by there.
Begin following the people whilst staying inside the evesdrop circle. You can jump inside the moving haybale but this will cost you full synchronisation.
Jump out of the haybale and head into the bushes on the right and quickly kill the soldier on the left.
Once he's dead continue following the gang members while staying within the circle.
It is advised to stick to the right side of the passage. From the bushes there you will be able to kill the second enemy.
The this is waiting in the hut by which the pair of thieves will stop.
As you reach the end of the camp, you will note that your father has been captured by the mercenaries. Free him and kill all thirteen enemies to end the mission.


The Q-Sense E4 is a real-time analytical instrument for studies of molecular events occurring on surfaces. The E4 measures mass and viscoelastic properties of molecular layers as they build up or change on the sensor surface. Areas of interest include materials, protein and surfactant research.
Everyone must first be trained on the instrument before using it. Please fill out the Training Request Form and indicate which instrument(s) you would like to do; please check all that apply.


text:
The Complete English Poems (Paperback)
By John Donne, A. J. Smith (Editor)
On Our Shelves Now (Please wait for us to confirm and & contact you when it's ready)
Spacecraft Voyager 1: New and Selected Poems (Paperback)
By Alice Oswald
Fresh Out of the Sky (Paperback)
By George Szirtes
Blake: Collected Poems (Routledge Classics) (Paperback)
By William Blake, W. B. Yeats (Editor)
The Poems of Dorothy Molloy (Paperback)
By Dorothy Molloy
Bloom for Yourself (Paperback)
By April Green
David Whyte Essentials (Paperback)
Paradise Lost (Paperback)
By John Milton, John Leonard (Editor), John Leonard (Introduction by), John Leonard (Notes by)
Old Possum's Book Of Practical Cats, Illustrated Edition (Hardcover)
Sir Gawain and the Green Knight (Paperback)
By Simon Armitage
The Complete Works of William Shakespeare (Leather-bound Classics) (Leather / fine binding)
By William Shakespeare, Michael A. Cramer, PhD (Introduction by)
The House of Belonging (Paperback)


3. Fraud: By becoming a member, you confirm that the information provided in this form is true and that you agree to abide by the Terms and Conditions of use of this site. Please note that your membership can be cancelled without notice if it is determined that false or misleading information has been provided, the Terms and Conditions of use have been violated, or other abuses have occurred as determined by "Arx Valdex Systems" in its sole discretion. If membership has been revoked, "Arx Valdex Systems" reserves the right to refuse application or readmission to the membership program.
18. Return Policy. Due to the nature of our online site, and the products listed, we "Arx Valdex Systems" have a strict NO RETURN policy for any product. No product shall be able to be returned for a like item, or for a cash refund.