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Panasonic Lumix DMC-GX1 Review
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17 Movie Mode
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Panasonic has been including video capability in its Micro Four Thirds lineup since the introduction of the Panasonic Lumix DMC-GH1 back in 2009. The mirrorless design of the G-series cameras makes them particularly suited to movie shooting since unlike DSLRs, the sensor is unimpaired by a mirror in the light-path, and is used full-time for image composition and focussing.
Video specification
The GX1 has a video capture rate of 30fps and can be set to capture video in AVCHD format. It also offers the ability to record in MP4 format, rather than the Motion JPEG option in previous G-series models. In AVCHD mode, when set to its highest quality setting (FSH), the GX1 offers 60i (interlaced) HD video capture at 1920 x 1080 pixels. In SH mode, 60p (progressive) video is captured at 1280 x 720 pixels. MP4 video can be captured in a choice of either 1920 x 1080, 1280 x 720 or 640 x 480 pixel resolution.
The GX1 has stereo microphones located in front of its hot shoe and a small monoaural speaker located on the camera's left body plate for video playback.
FSH: 1920 x 1080, (60i*, 17 Mbps)
SH: 1280 x 720, (60p*, 17 Mbps)
1920 x 1080, 30fps
1280 x 720, 30fps
640 x 480, 30fps
* Sensor output = 30fps
|Dolby Digital Stereo, wind-cut feature
|File size
|Max 17 MB/sec. (1080i AVCHD)
|Max file size per clip
|4 GB for MP4, card capacity for AVCHD
|Running time
|Up to 29 minutes, 59 seconds for MP4, up to capacity of card for AVCHD (Limited to 29 minutes on models sold in Europe).
Handling in video mode
Capturing video on the GX1 is no more complicated than pressing the dedicated motion picture button on the top of the camera. Yet the lack of a separate movie mode does have some limitations. While you can capture video in any of the 'Photo Styles' or 'Creative Control' modes, your selection is shared between both still image and motion picture modes. This means you cannot, for example, configure a Monochrome Photo Style for still images while simultaneously designating a color mode for video.
Live view, by default is set to the 4:3 still image format. The lack of a selectable movie mode means that you will not actually see a 16:9 format live view until after you have pressed the movie record button, making precise framing difficult at the start of a video. You can manually switch live view to a 16:9 format via the Q. Menu, but then encounter the same issue should you want to capture a still image when live view is set for a video format.
The GX1 allows you to shoot still images while recording uninterrupted video. The camera's stereo microphones will pick up faint, but audible clicks of the shutter button being pressed. And the resulting JPEG-only images are captured in a 16:9 format and at a reduced size (2 MP).
As we've seen with previous G-series cameras (excluding the video-oriented GH2), exposure control options when shooting video are rather limited. User-defined ISO settings are overridden by the camera. In PASM modes both shutter speed and aperture are set by the camera, with no correlation between the values that appear on the screen before you start recording and those actually used during the recording. Existing exposure compensation, white balance, focus and metering values are honored when shooting video, but cannot be altered during the recording.
|This screen shows video capture with key shooting information overlaid and grid lines enabled.
|In iA+ mode, the 'defocus' slider allows you to change aperture during the video and the camera honors existing exposure compensation values.
Interestingly, the only method of manual aperture control is found by activating the automated intelligent Auto (iA) mode. By dragging a 'defocus' slider while filming, you can control the depth of field beyond your area of focus. Should you enable iAuto, however, you give up more extensive white balance and Photo Style options. It should also be noted that while the exposure compensation range for still images is +/-5 stops EV, the act of recording video automatically reduces this range to +/- 3 stops EV.
During video recording you can focus manually or use continuous AF so that the camera will automatically lock focus on a subject within the AF area. As with the Panasonic Lumix DMC-G3 and GF3, the GX1 is capable of rack focus during videos. This feature allows you to choose a specific area on which to set focus simply by touching the screen. The camera will then shift focus from its current location to the one you have just designated.
You can change the amount of information shown on the screen during video capture by pressing the DISP. button. The GX1 has a fixed rear LCD, making the optional LVF-2 electronic viewfinder the only remedy for eliminating glare on bright sunny days.
Video image quality
The GX1 produces videos which, like the camera's still images, exhibit reasonably accurate white balance and show the effect of metering that errs if anything on the conservative side, placing high priority on avoiding highlight clipping, at the expense of a slightly darker scene exposure. The AF system can be a bit slow to acquire focus initially and we've generally found it quicker to set focus via the touchscreen before pressing the record button. The camera's AF is set to continuous focus by default and we do find a noticeable amount of focus hunt when panning the camera across a scene with objects at different distances.
Audio recording, while certainly acceptable tends to be a bit muddy with the close proximity of the GX1's stereo microphones perhaps responsible for less separation between channels than we've heard from competitors like the Nikon 1 V1 and the Sony NEX-7. The camera's wind-cut filter, although offering no options other than Auto and Off, does a passable job of minimizing wind noise in semi-exposed outdoor settings.
Panasonic's optical image stabilization system can be set to counter movement in all directions or just vertical shifts when you are panning the camera. The effect of stabilization is quite noticeable in hand-held video, producing much steadier results than when it is disabled.
Video samplesThese videos are representative of our day-to-day shooting experiences with the GX1. Video performance is essentially the same as we found in our G3 review video samples.
Sample Video 1This video clip of a tennis rally highlights the GX1's lack of direct manual control over shutter speed. While the camera does a fine job of metering for a pleasing exposure under indoor lighting, the relatively slow shutter speed means that the tennis ball - as well as players' arms and rackets during strokes - succumbs to motion blur. This can be most easily seen when you advance the clip frame by frame. Typically a shutter speed of 1/500 is required to freeze this type of action and the GX1 has chosen a value that is significantly slower.
|1920 x 1080i, AVCHD, .MTS file, 12 sec. 28.46 MB Click here to download original .MTS file
Sample video 2
This hand-held video highlights the audio capability of the GX1's stereo microphones. From a camera to subject distance of less than 10 feet, the audio is a bit muddy and undefined, although background conversation emanating from behind the camera is kept to a minimum. Even with the wind-cut filter enabled, the semi-exposed conditions produce pockets of noticeable wind noise. The ability to adjust exposure compensation is demonstrated. The performers were in a fully shaded interior directly in front of open sky. An exposure compensation of 1 1/3 stops was used to achieve a 'normal' exposure for their faces.
|1920 x 1080i, AVCHD, .MTS file, 29 sec. 64.83 MB Click here to download original .MTS file
Sample video 3
This clip shows a very obvious rolling shutter effect. The GX1's relatively slow refresh rate means that as either subjects or the camera move horizontally across the frame, vertical lines appear slanted. The trolley car in the clip below was moving a relatively slow speed, yet it appears distorted as it travels past the camera. The panning of the camera that follows show the same effect in the buildings, which appear to be leaning towards the right.
|1920 x 1080i, AVCHD, .MTS file, 15 sec. 34.83 MB Click here to download original .MTS file
Sample video 4
This hand-held video highlights the power zoom capability of the Lumix G X Vario PZ 14-42mm F3.5-5.6 ASPH OIS lens. Image stabilization is set to the default 'normal' mode. The lens itself can be set to either a 'high', 'medium' or 'low' maximum zoom speed via either Q.Menu operation or a Fn button designation. The zoom operation is smooth and and the use of a lever versus a lens ring makes it easy to hold the camera steady while zooming. We find, however, that the short throw of the lever requires a deft touch to zoom reliably at a speed slower than the maximum. Here you can see the camera's AF system hunt for focus towards the long end of the zoom.
|1920 x 1080i, AVCHD, .MTS file, 16 sec. 38.22 MB Click here to download original .MTS file
I own it
I want it
I had it
Discuss in the forums
Bought one new a couple of years ago on closeout for a frighteningly low price. Yes, the design is nearly five years old, but every time I think about replacing it, I find that the potential replacement is either larger, missing features, or not measurably better. It feels great in my hands, I know the controls inside and out (having previously owned a DMC-LX3, it was easy to learn the GX1's menu system) and it's small and light enough to take anywhere.
Long live the GX1!
I was impressed with the video quality of the GX1 after buying this camera and works with autofocus which quicker than some DSLRs in video AF (except a few of the new dslrs with on sensor AF). Recommended for that alone, over my earlier model Pen. Panasonic knows video. Maybe Olympus is better now. I also have the Olympus 12-50mm and it records fine detail capture on the GX1. I recommend upgrade from the kit. I also have the original Panasonic 14-45mm, sharp lens with OIS which helps stabilize with video in a smaller lens than the Panasonic 14-140mm if carrying size if a consideration. Great lens also, and still can be found online.
Iam really like the design of the GX1, though I do have a few quibbles. It's a little bigger than the rest of the field, but none of these is sufficiently small with a lens attached to be considered significantly smaller than the others. It's got a nice-size grip and thumb rest, which makes it comfortable to hold and shoot single-handed, and it feels well constructed.


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Enterohaemorrhagic Escherichia coli (EHEC) comprise a group of zoonotic diarrhoeal pathogens of worldwide importance. Cattle are a key reservoir; however the molecular mechanisms that promote persistent colonization of the bovine intestines by EHEC are ill-defined. The large plasmid of EHEC O157:H7 encodes several putative virulence factors. Here, it is reported that the pO157-encoded Type V-secreted serine protease EspP influences the intestinal colonization of calves. To dissect the basis of attenuation, a bovine primary rectal epithelial cell line was developed. Adherence of E. coli O157:H7 to such cells was significantly impaired by espP mutation but restored upon addition of highly purified exogenous EspP. Data of this study add to the growing body of evidence that cytotoxins facilitate intestinal colonization by EHEC.
Enterohaemorrhagic Escherichia coli (EHEC) pose a significant public health threat worldwide. Ruminants are a key reservoir and humans are frequently infected following direct or indirect exposure to ruminant faeces. Strategies to reduce the carriage of EHEC in ruminants are expected to lower the incidence of human disease. However, cross-protective vaccines do not yet exist and the molecular mechanisms that influence the persistence of EHEC in the bovine intestines are incompletely understood. By allelic exchange and signature-tagged mutagenesis (STM), portfolios of bacterial genes that influence colonization of calves by EHEC O157:H7 and O26:H-have been identified, including conserved and serotype-specific factors (Dziva et al., 2004; van Diemen et al., 2005). Among the EHEC O26:H- genes putatively required for the intestinal colonization of calves is pssA (van Diemen et al., 2005). PssA (protein secreted by Shiga toxin-producing E. coli) was first described in EHEC O26 strains and is highly related to a family of serine protease autotransporters of Enterobacteriaceae (SPATEs; Djafari et al., 1997). Parallel studies on EHEC O157:H7 identified a pO157-encoded secreted serine protease (EspP) which cleaves human coagulation factor V (Brunder et al., 1997). Through hybridization and immunoblot assays, EspP was shown to be widespread in EHEC O157:H7 strains and to exist in EHEC O26 strains (Brunder et al., 1997) and it is now clear that PssA and EspP are 99% identical at the amino acid level. Despite this close homology, PssA has been reported to cleave casein and exhibit cytopathic activity for Vero cells, phenotypes that have not been reported with EspP (Djafari et al., 1997; Dutta et al., 2002).
Several SPATEs have been extensively characterized in vitro, including EspC of enteropathogenic E. coli (EPEC; Stein et al., 1996; Navarro-Garcia et al., 2004), Pet of enteroaggregative E. coli (EAEC; Eslava et al., 1998), EatA of enterotoxigenic E. coli (ETEC; Patel et al., 2004), SigA of Shigella flexneri (Al-Hasani et al., 2000), and the uropathogenic E. coli (UPEC) proteins Sat (Guyer et al., 2000) and PicU (Parham et al., 2004). In rabbit ileal loop models, mutation of sat (of diffusely adhering E. coli), ETEC eatA and Shigella SigA impaired the induction of intestinal inflammatory and secretory responses (Al-Hasani et al., 2000; Patel et al., 2004; Taddei et al., 2005). Furthermore, EAEC Pet causes damage to human intestinal explants cultured ex vivo (Henderson et al., 1999) and EPEC EspC is toxic to rat jenunal explants (Mellies et al., 2001). Taken together, these observations indicate that SPATEs may play key roles in host–pathogen interactions at epithelial surfaces (reviewed in Henderson et al., 2004).
The majority of SPATEs are encoded on large plasmids. In addition to EspP, the large plasmid of EHEC O157:H7 (pO157) encodes enterohaemolysin (EhxA), catalase-peroxidase (KatP), a large clostridial toxin homologue (ToxB), a metalloprotease that cleaves C1 esterase inhibitor (StcE, initially identified as TagA) and the ecf operon (Burland et al., 1998; Makino et al., 1998). It has been reported that pO157 is required for full adherence to epithelial cells (Toth et al., 1990) and ToxB and StcE have since been implicated in this process (Tastuno et al., 2001; Stevens et al., 2004; Grys et al., 2005). Although it was initially reported that pO157 does not influence E. coli O157:H7-induced enteritis and adherence in gnotobiotic piglets (Tzipori et al., 1987), it was recently reported that pO157 influences colonization of the bovine terminal rectum (Sheng et al., 2006) and that the ecf operon is required for persistence in the bovine intestines (Yoon et al., 2005). With the exception of toxB, which is not required for colonization of young calves (Stevens et al., 2004), the role of other pO157-encoded genes in vivo has not been defined. Evidence for expression during E. coli O157:H7 infection of humans has been obtained for EspP (Brunder et al., 1997), EhxA (Schmidt et al., 1995), StcE (Paton & Paton, 2002) and a pO157-encoded inner membrane acyltransferase (MsbB2; John et al., 2005).
Here, it is shown that EspP contributes to adherence to bovine primary rectal cells and colonization of the bovine intestines by the predominant EHEC serotype affecting humans in Europe and North America.
Materials and methods
Bacterial strains and media
Strain 85-170nalR is a nalidixic acid-resistant nontoxigenic E. coli O157:H7 strain that reliably colonizes the intestines of calves and induces AE lesions in vivo (Stevens et al., 2004; Vlisidou et al., 2006a). The laboratory E. coli strain HB101 was transformed with plasmid pB9-5espP (kindly supplied by Anthony Scott-Tucker, University of Birmingham) for expression and purification of EspP. Bacterial strains were grown in Luria–Bertani (LB) broth or agar supplemented with ampicillin (Amp, 100 µg mL−1), kanamycin (Kan, 50 µg mL−1) and nalidixic acid (Nal, 25 µg mL−1) as appropriate. For adherence assays, bacteria were grown in minimum essential medium buffered with HEPES (MEM-HEPES; Sigma, St Louis).
Construction and characterization of an E. coli O157:H7 ΔespP::kanR mutant
The espP gene was replaced by a kanamycin resistance cassette by integration of a linear PCR product upon transient expression of λRed recombinase, essentially as described (Datsenko & Wanner, 2000). The amplicon was generated using Vent™ proofreading DNA polymerase (New England Biolabs, Hitchin, UK) and primers espP1F-FRT (5′-GATAAACATACTGTGTTTGTTATCCGTATGATA ACAAACACATAAAGGAGGTGTAGGCTGGAGCTGCTTC-3′), and espP2-FRT (5′-CGGCAGGCACTGAGGGTAAAGGGCCCGCAGGCCCTTTTGAATACGGAGTACATATGAATATCCTCCTTAG-3′) which contain 50 nucleotide homology extensions corresponding to the regions immediately 5′ and 3′ of espP and priming sequences for the kanR cassette of pKD4 (bold; Datsenko & Wanner, 2000). The PCR product was DpnI digested to remove residual template, gel purified and electroporated into 85-170nalR carrying the temperature-sensitive λRed helper plasmid pKD46 following induction of λRed recombinase with 10 mM l-arabinose at 30°C. A recombinant (85-170nalRΔespP::kanR) was selected on LB agar containing nalidixic acid and kanamycin and cured of pKD46 by growth at 42°C. The insertion was confirmed by PCR using the espP flanking primers espP-F (5′-CTCCTTTACTGTATCCCTCAT-3′) and espP-R (5′-AGGCACTGAGGGTAAAGG-3′).
To verify the mutant, 85-170nalR and 85-170nalRΔespP::kanR were cultivated in Dulbecco's Modified Eagles Medium (DMEM) to induce protein secretion and secreted proteins were prepared by trichloroacetic acid precipitation (Stevens et al., 2004). Secreted proteins were analysed by 4–15% gradient sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and visualized following staining with GelCode Blue reagent (Perbio Science UK Ltd, Cheshire, UK).
Bovine primary cell culture
Bovine primary epithelial cells were derived from the region 0 to 5 cm proximal to the rectal-anal junction of cattle up to 30 months old using a modification of the method described for colonic epithelial cultures (Hoey et al., 2003). Unless stated otherwise, reagents for the procedure were obtained from Sigma-Aldrich. Briefly, mucosal scrapings from the terminal rectum were digested in DMEM containing 1% (v/v) foetal calf serum (FCS), 100 U mL−1 penicillin, 30 µg mL−1 streptomycin, 25 µg mL−1 gentamicin, 75 U mL−1 collagenase and 20 µg mL−1 dispase (Roche, Rockford) with gentle shaking at 37°C until isolated crypts could be observed microscopically. A series of differential centrifugation steps with DMEM containing 2% (w/v) sorbitol was used to enrich the isolated crypts from undigested material, endogenous microbial communities and single cells including fibroblasts (Booth et al., 1995). The crypt cell pellet was resuspended in cell culture medium [DMEM, 2.5% (v/v) FCS, 0.25 U mL−1 insulin, 10 ng mL−1 epidermal growth factor (EGF) and 30 µg mL−1 gentamicin]. Approximately 400–600 crypts were seeded per well into 24-well Costar culture plates (Corning, Cambridge) precoated with collagen (Vitrogen Collagen, Nutacon, Netherlands). To inhibit fibroblast growth, the medium was replaced with MEM d-Valine medium containing 10% (v/v) batch-tested FCS, 0.25 U mL−1 insulin, 10 ng mL−1 EGF and 30 µg mL−1 gentamicin after 24 h as described previously (Hoey et al., 2003). The cells were grown to confluence (c. 3 × 105 cells per well, typically achieved 10–14 days after isolation). The cultured cells were confirmed by immunocytochemistry to possess epithelial cell-specific cytokeratins (CK4, 5, 6, 8, 10, 13 and 18) and lack a marker for fibroblasts as described (Hoey et al., 2003).
Purification of exogenous EspP
Purified recombinant EspP was prepared essentially as described (Navarro-Garcia et al., 2004). Briefly, the EspP-overproducing strain HB101(pB9-5 espP) was grown in LB overnight at 37°C and the supernatant harvested following centrifugation. The culture supernatant was filtered through a 0.22 µm pore-size low-protein-binding membrane, then concentrated by centrifugation in against a Centricon filter with a 100 kDa cut-off (Millipore, Bedford). The retentate was reconstituted in MEM-HEPES, checked for purity by SDS-PAGE and confirmed to contain EspP by Western blotting. The amount of protein was estimated using the standard BCA method (Pierce Biotechnology Inc, Rockford) and stored at −20°C until required.
Adherence assays
Stationary-phase LB cultures of wild-type and the mutant bacteria grown in MEM-HEPES with appropriate antibiotics were diluted 1 : 10 in MEM-HEPES and incubated at 37°C with shaking (200 r.p.m.) to an OD at 600 nm of 0.3–0.4. Confluent bovine rectal primary epithelial cells (above) were separately infected in duplicate with each of the strains at a multiplicity of infection of c. 100 in MEM-HEPES and incubated for 1 h at 37°C in a humidified 5% CO2 atmosphere. Purified recombinant EspP was supplemented to a final concentration of 178.8 ng mL−1 of protein where appropriate. Nonadherent bacteria were removed by washing three times with phosphate-buffered saline (PBS) and adherent viable bacteria were enumerated following dispersion with PBS-0.1% (v/v) Triton X-100, serial 10-fold dilution and plating onto selective media. The assays were carried out independently on three occasions and the data analysed with a generalized linear mixed model (GLMM) assuming a Poisson response variable (McCullagh & Nelder, 1989). The canonical link function was used. The experimental replicate was fitted as a random effect. Strain and the presence/absence of exogenous EspP were fitted as fixed effects. Evaluation of the significance of fixed effects was carried out using the Chi-squared approximation for the Wald test.
Oral infection of calves with E. coli O157:H7
Procedures for oral inoculation of calves with E. coli O157:H7 have been described (Stevens et al., 2004) and comply with the Animals (Scientific Procedures) Act 1986 (licence 30/2009). Three 12-day-old Friesian bull calves were coinfected with c. 1 × 1010 CFU each of 85-170nalR and 85-170nalRΔespP::kanR grown to stationary phase in brain heart infusion broth in a 1 : 1 ratio. The magnitude and duration of faecal excretion of the bacteria were followed at least once daily for 12 days by plating of triplicate serial dilutions of fresh faeces collected by rectal palpation to sorbitol MacConkey agar (Oxoid, Basingstoke, UK) supplemented with 2.5 µg mL−1 potassium tellurite and 20 µg mL−1 nalidixic acid (T–SMAC–Nal) and T–SMAC–Nal containing 50 µg mL−1 kanamycin (T–SMAC–Nal–Kan). The number of wild-type bacteria was calculated by subtracting the viable count on T–SMAC–Nal–Kan from that obtained using T–SMAC–Nal as previously described (Dziva et al., 2004). Recoveries of wild-type and the mutant bacteria were confirmed by PCR from selected colonies using espP-flanking primers. The faecal shedding data were analysed for the effect of mutation by means of an F-test, with the data taken as repeated measurements and the animal as a covariant (Proc Mixed, Statistical Analysis System 1995, SAS Institute, Cary). P values <0.05 were taken to be significant.
Results and discussion
Characterization of an E. coli O157:H7 espP::kanR mutant
A espP::kanR mutant of E. coli O157:H7 strain 85-170nalR was constructed by λRed-mediated linear recombination and verified by PCR with espP-flanking primers (data not shown) and SDS-PAGE analysis of proteins secreted by the wild-type and mutant strains. While both the mutant and parent strains secreted comparable quantities of the Type III secreted proteins EspA, -B, -D and Tir, a protein of the expected size of EspP was absent in the secreted fraction of the mutant (Fig. 1). The protein present in the secreted proteome of the wild-type at this position has been independently confirmed to be EspP by tandem MS (Roe et al., 2007). The growth rate of the 85-170nalRespP::kanR mutant was confirmed to be identical to that of the parent strain in both rich and minimal media by real-time spectrophotometry (data not shown).
EspP plays a role in the intestinal colonization of calves by E. coli O157:H7
The role of EspP in the intestinal colonization of calves was determined using coinfection experiments, using a model that has reliably identified attenuating defects previously (Stevens et al., 2002; Dziva et al., 2004; Vlisidou et al., 2006a, b). From day 5 postoral inoculation, the 85-170nalRΔespP::kanR mutant was shed in the faeces in lower numbers than the parent strain and the reduction became statistically significant from day 9 onwards (P values <0.05; Fig. 2). Although the role of secreted serine protease in persistence of the bovine intestines was first suggested by the isolation of two independent signature-tagged mini-Tn5Km2 mutants of EHEC O26:H- with insertions in pssA (van Diemen et al., 2005), it has been observed that some genes identified by STM are not required for persistence when defined mutants are constructed and screened in isolation or by coinfection in the same model (map; Dziva et al., 2004; nleD; Marchés et al., 2005). Thus, the data herein provide novel independent validation of the role of EspP in intestinal colonization. The extent of attenuation caused by the espP mutation was slight compared with the effect of intimin and Tir mutations in the same strain in calves of the same age (Vlisidou et al., 2006a), but is significant when compared with an 85-170nalR mutant lacking the putative cytotoxin/adhesin ToxB (Stevens et al., 2004).
Although it has been shown that EspP is required for intestinal colonization of cattle by E. coli O157:H7, it is noteworthy that sorbitol-fermenting EHEC O157:H-strains isolated from cattle lack espP (Brunder et al., 1999; Karch & Bielaszewska, 2001) and a mutated form of espP was reported in five of 30 bovine E. coli O157:H7 isolates (McNally et al., 2001). However, the possibility of redundancy in the repertoire of secreted proteases in these strains cannot be precluded. Data of this study add to the growing number of secreted cytotoxins that play a role in EHEC-intestinal interactions. Shiga toxin was recently observed to influence intestinal colonization of mice by E. coli O157:H7, possibly by modulating the surface expression of intimin coreceptors (Robinson et al., 2006). Furthermore, an inhibitor of lymphocyte proliferation and cytokine synthesis (LifA, lymphostatin) influences intestinal colonisation in calves by EHEC O5:H- and O111:H- (Stevens et al., 2002) and persistence and the induction of colonic hyperplasia by Citrobacter rodentium in mice (Klapproth et al., 2005). However, lifA mutations also impair adherence (Stevens et al., 2002) and the contribution of direct cytotoxicity to colonization is not fully understood. Enterohaemolysin was also suggested to be important in colonization of calves by EHEC O26:H- by STM and the gene is highly conserved among EHEC (van Diemen et al., 2005). It is possible that such cytotoxins act in concert to promote intestinal colonization.
EspP influences adherence of E. coli O157:H7 to bovine primary rectal epithelial cells
To assess the basis of attenuation, bovine primary rectal epithelial cells were isolated for in vitro adherence assays. Epithelial cells were obtained from the terminal rectum of cattle c. 0–5 cm proximal to the rectal–anal junction. The relevance of cells derived from this site is suggested by the finding that colonization of cattle can be established by the direct administration of E. coli O157:H7 to terminal rectal mucosa (Sheng et al., 2004) in a manner dependent on pO157 (Sheng et al., 2006). The cultured cells were polarized and differentiated, expressed the epithelial cell-specific cytokeratins and resembled those seen previously with bovine colonic primary epithelial cell cultures (Hoey et al., 2003).
To examine the impact of secreted EspP on adherence, EspP were overexpressed in E. coli strain HB101 and the protein from the supernatant was purified. The purity of the protein preparation was confirmed by SDS-PAGE (Fig. 3a) and the identity of the protein species verified by Western blotting (data not shown). The purification method has previously been used to prepare homogenous catalytically active EspC from EPEC (Mellies et al., 2001; Navarro-Garcia et al., 2004).
Mutant and parent strains were allowed to interact with primary rectal epithelial cells in the presence or absence of purified exogenous EspP. The mean adherence data indicated that the 85-170nalRΔespP::kanR mutant was significantly (P<0.001) impaired in its ability to adhere to bovine rectal primary epithelial cells compared with the parent strain (Fig. 3b). The cells showed no evidence on cytotoxicity or cytoskeletal changes (data not shown). The adherence defect of the mutant was rescued by addition of highly purified recombinant EspP, suggesting that the phenotype of the mutant was due to loss of EspP rather than due to second-site or polar effects. Adherence of the wild-type strain was increased by the addition of exogenous EspP, but not significantly so (P=0.03).
Although the mechanism by which EspP affects intestinal colonization and adherence to cultured cells is unknown, a previous study indicated that it cleaves pepsin A and human coagulation factor V (Brunder et al., 1997). Degradation of factor V is a shared feature among SPATES from other E. coli pathotypes (Dutta et al., 2002) and this has been suggested to contribute to mucosal haemorrhage in humans (Brunder et al., 1997). Some SPATEs possess mucinase activity (reviewed in Henderson et al., 2004); however, it has been reported that EspP is unable to cleave bovine submaxilliary mucus in vitro (Dutta et al., 2002). Type V-secreted proteins typically possess a β-domain that remains anchored on the bacterial outer membrane and a passenger domain that is often cleaved and released in the milieu. Recently, it has been reported that some autotransported proteins are capable of mediating adhesion directly (Fink et al., 2003) or by acting as scaffolds for bacterial adhesins (Veiga et al., 2003) and it remains possible that surface-anchored EspP may act in this way. The contribution of the secreted and surface-associated portions of EspP and other EHEC cytotoxins in colonization and pathogenesis merits further investigation.
The authors gratefully acknowledge the support of the Department for the Environment, Food and Rural Affairs (project no. OZ0707), the support from DEFRA VTRI (project no. VT0102) and the Scottish Executive Environment and Rural Affairs Department (WP2.2-22050).


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Bone erosions in rheumatoid arthritis can be repaired through reduction in disease activity with conventional disease-modifying antirheumatic drugs
DOI: 10.1186/ar1943
© Ideguchi et al.; licensee BioMed Central Ltd. 2006
Received: 24 February 2005
Accepted: 21 March 2006
Published: 28 April 2006
We conducted the present study to determine whether repair of erosions occurs in patients with rheumatoid arthritis (RA) treated with conventional disease-modifying anti-rheumatic drugs (DMARDs) and to compare clinical characteristics between patients exhibiting and not exhibiting erosion repair. We included in the study a total of 122 RA patients who fulfilled the 1987 American College of Rheumatology criteria for RA; all patients had paired sequential radiographs of both hands and wrists showing erosive changes at baseline. Patients were classified into two groups according to the presence of repair of erosions at follow up, namely the 'repair observed' and 'repair not observed' groups. Clinical characteristics, disease activity, radiographic scores and treatment in the two groups were compared. Forty-four repairs were observed in 13 patients (10.7%). Compared with the repair not observed group, the functional class of the patients in the repair observed group was lower at baseline (P < 0.01) and the mean disease activity was lower at follow up (P < 0.005). The changes in radiographic scores per year (total radiographic score and erosion score) were lower (P < 0.05 and P < 0.01, respectively) in the repair observed group. No difference in treatment was observed. Repair of erosions was detected in 10.7% of RA patients treated with conventional DMARDs. Repairs were associated with low functional class at baseline and low disease activity at follow up. These observations support the importance of reduction in disease activity in RA patients. Because repair of erosions was detected in a substantial number of patients, assessment of erosion repair should be incorporated into the radiographic evaluation and scoring of RA.
Rheumatoid arthritis (RA) is a chronic, destructive autoimmune inflammatory disorder of unknown aetiology that occurs in about 1% of the adult population [1].
The radiograph has evolved into the 'gold standard' for evaluation of RA progression because it best demonstrates the anatomical destruction of joint structures [2]. However, repair of erosions or reparative changes in RA have rarely been reported [3–10]. There are several possible reasons for this. First, radiographs are rarely obtained in patients who appear to be experiencing remission, in whom repair phenomena may be observed. Second, most clinical trials are conducted in patients with longstanding destructive RA with high disease activity, and in such cases it is difficult to define repair clearly. Third, there is an interval between clinical findings and corresponding radiographic phenomena in clinical trials, and most clinical trials have insufficient follow up to identify repair phenomena. Fourth, the most commonly used scoring methods, namely those of Sharp and Larsen and their groups, are not designed to describe reparative changes. The Sharp/van der Heijde method includes 16 areas for erosions and 15 for joint space narrowing in each hand. The erosion score per joint can range from 0 to 5. Joint space narrowing is scored with range from 0 to 4. The maximum erosion score of all joints in both hands is 160 and the maximum score for joint space narrowing in all joints of both hands is 120. Finally, serial high-quality radiographs must be obtained to permit detection of repair.
In recent years considerable advances have been made in the pharmacological management of RA [11]. Previously, the mainstays of treatment for RA were disease-modifying antirheumatic drugs (DMARDs), which suppress the inflammatory process and slow disease progression. Today's biological agents are specifically designed to block key mediators of the RA inflammatory process.
Recent trials with tumour necrosis factor-α blocking drugs in RA have demonstrated improvements in radiographic scores, suggesting drug-induced repair of erosions [12–14]. These findings have renewed interest in the radiographic detection of repair, and Sharp and coworkers [15] instituted a subcommittee of the OMERACT (Outcome Measures in Rheumatology) Imaging Committee in a first attempt to confirm whether repair occurs in RA and, if so, to determine how repair should be assessed. They provided evidence that repair of bone damage does occur in RA, resulting in some degree of improvement, which was recognized by the majority of a panel of experts. A consensus on definitions for the morphological features of repair was achieved for the following: sclerosis, cortication, filling-in, remodeling and restoration [15].
We wondered whether repair of erosions can be detected in the real world setting among RA patients who have longstanding arthritis, who do not achieve remission easily. To examine whether repair of erosions in RA patients treated with conventional DMARDs can be detected, we examined 122 RA patients with paired sequential radiographs of both hands and wrists. We also assessed the clinical characteristics of these patients.
Materials and methods
All patients were diagnosed with RA according to the 1987 American College of Rheumatology criteria for RA. From among RA patients seen in the rheumatology clinic by two of the authors (HI and SO) at Yokohama City University Medical Center between January 2001 and March 2004, patients with paired sequential radiographs of both hands and wrists were enrolled in this study.
Clinical evaluation
Clinical characteristics, including patient's sex, age, disease duration, rheumatoid factor status, prescription (dose of prednisone, bisphosphonates and DMARDs such as methotrexate and sulfasalazine), functional class (determined using American College of Rheumatology criteria) and RA stage (determined according to Steinbrocker criteria), were recorded.
Disease activity was assessed using the Disease Activity Score (DAS) in 28 joints, DAS28-3. Modifications to the original DAS index have been applied, yielding the DAS28 (which is a composite index that includes variables such as the number of tender and swollen joints using 28 joint counts, erythrocyte sedimentation rate (ESR), and the patients' assessment of disease activity) and DAS28-3 (excluding patients' assessment of disease activity from DAS28) indices; these indices have been and validated and, for reasons of simplicity, are preferred over the original index in clinical practice [16–18]. The DAS28-3 value was calculated as follows:
Radiographic evaluation
Detection of repair of erosions
Standard radiographs of the hands and wrists were obtained in two planes (anteroposterior and oblique projections). Radiographic examinations were interpreted independently by two board-certified rheumatologists with musculoskeletal reading experience (HI and SO). The films were reviewed to identify erosion repair specifically. Bone erosion was defined as a discrete interruption of the cortical surface, based on standard plain film radiograph criteria [19, 20]. The films were evaluated with known sequences in order to ensure maximum sensitivity in detecting erosion repair [21, 22]. The evaluators were blinded to patient identity. Repair was defined as follows [10]: category 1, reappearance of the cortical plate at a bone site where it had been destroyed; category 2, partial or complete filling in of an erosion; and category 3, subchondral bone sclerosis and osteophyte formation (secondary osteoarthritis). Where there was a difference in opinion regarding whether repair was present, these cases were discussed until agreement was reached. To confirm the existence of repair, these films were re-examined in random order, without knowledge of the sequence, by two other rheumatologists (HH and AS) as well as by the initial ones.
Radiographic scoring method
Radiographic joint damage of the hands and wrists was assessed using the van der Heijde modification to the total Sharp scoring system (vdH-S) [23]. Two rheumatologists (HI and SO) scored the films of each patient independently, with knowledge of the order of the radiographs. Erosion scores can range from 0 to 160. Joint space narrowing (JSN) scores can range from 0 to 120. Total radiographic scores can range from 0 to 280. Readers were allowed to record improvement in scores. For each set of radiographs, the mean score by the two readers was used in the analysis. In the present study the smallest detectable change was 12.8, which reflects that component of a measurement that was statistically attributable to error from the measurement process [24–26]. Because the time interval between pairs of radiographs varied among patients, radiographic progression was determined by dividing the difference between scores of each pair of radiographs by the years that had elapsed between radiographs; the yearly adjusted progression rate for each patient was expressed as changes in score (total radiographic, erosion, or JSN) per year.
Statistical analysis
Statistical analysis was conducted using SPSS 11.0 statistical analysis software (SPSS Inc., Chicago, Illinois, USA). We analyzed group differences with respect to sex, rheumatoid factor and prescribed bisphosphonates using Fisher's exact probability test. Then, we examined whether the data exhibited a normal distribution using F test, and in cases of normal distribution we used parametric examination. If equal variance was gained, we used the Student's t test. We analyzed group differences in age and DAS28-3 score using Student's t test. If equal variance was not gained, then we used the Mann-Whitney U test. We also analyzed group differences in disease duration, dosage of prednisone, class, radiographic score and prescribed DMARDs using the Mann-Whitney U test. Statistical significance was defined as a P value less than 0.05. Significant variables in univariate analysis were entered simultaneously into multivariate logistic regression to identify variables with independent predictive value for repair.
Informed consent was obtained from each patient, and the Ethics Committee of the Yokohama City Medical Center approved the study protocol.
Characteristics of the study population
P value
Repair observed
Repair not observed
Patients (n)
Female (n [%])
10 (76.9%)
93 (85.3%)
Age (mean [range]; years)
64.2 ± 9.8 (39–78)
60.6 ± 12.6 (22–85)
Disease duration (mean [range]; years)
12.5 ± 9.6 (2.7–38)
14.3 ± 10.8 (1–45)
RF positivity (n [%])
Dosage of prednisone (mean [range]; mg)
2.0 ± 2.1 (0.0–6.0)
3.7 ± 3.3 (0.0–14.5)
Prescribed bisphosphonate (n [%])
8 (61.5%)
79 (72.5%)
Functional class
RA stage
DAS28-3 score at baseline (mean [range])
4.4 ± 1.2 (2.7–6.0)
4.0 ± 1.1 (1.0–7.0)
DAS28-3 score at follow up (mean [range])
2.7 ± 1.1 (1.5–5.4)
3.7 ± 1.2 (0.2–8.1)
ΔDAS28-3 (mean [range])
-1.7 ± 1.2 (-3.5 to +0.1)
-0.3 ± 1.0 (-2.4 to +2.6)
Time interval between pairs of radiographs, years (mean [range])
2.3 ± 1.0 (1.4–3.1)
1.6 ± 1.8 (1.0–1.6)
ΔDAS28-3/year (mean [range])
-0.9 ± 0.8 (-2.6 to +0.1)
-0.2 ± 0.9 (-2.8 to +3.4)
Forty-four repairs were detected in 13 patients (10.7%). Of these repairs, initial difference in opinion between HI and SO existed for only one repair. Subsequently, to confirm the existence of repair, 44 pairs of images were independently presented to two other readers as well as HI and SO, without knowledge of the sequence, and they were asked to indicate which image was worse on global evaluation of erosion and which erosion was larger in size. Complete agreement for both evaluations among the four readers was achieved in 95.5% of repairs. Based on this high level of interobserver agreement, these 44 repairs in 13 patients were included in the following analysis. According to the definition of repair given above, the 44 repairs were classified as follows: five repairs were deemed to be in category 1 (reappearance of the cortical plate at a bone site where it had been destroyed); 39 repairs were in category 2 (partial or complete filling in of an erosion); and no repair was in category 3 (subchondral bone sclerosis and osteophyte formation). In other words, none of the erosions was judged as exhibiting 'repair' as a result of osteophyte development.
Radiographic joint damage scores at baseline
Repair observed
Repair not observed
Total radiographic scorea
Mean ± SD
117.1 ± 39.2
106.1 ± 66.3
Median (range)
119.0 (47.0–188.5)
100.5 (9.0–242.0)
Interquartile range
Erosion scorec
Mean ± SD
54.7 ± 26.7
42.6 ± 38.3
Median (range)
57.5 (9.5, 101.0)
28.8 (0.5, 135.0)
Interquartile range
30.1, 69.0
11.5, 66.8
JSN scored
Mean ± SD
62.3 ± 13.9
63.6 ± 30.9
Median (range)
62.5 (37.5, 87.5)
67.3 (8.5, 115.0)
Interquartile range
55.5, 72.0
34.0, 92.8
Prescriptions of DMARDs, prednisone and bisphosphonates
Repair observed
Repair not observed
7 (53.8%)
59 (54.1%)
4 (30.8%)
35 (32.1%)
Methotrexate and sulfasalazine
1 (7.7%)
18 (16.5%)
1 (7.7%)
11 (10.1%)
2 (15.4%)
22 (20.2%)
7 (53.8%)
75 (68.8%)
8 (61.5%)
79 (72.5%)
Univariate analysis revealed that class status at baseline (P < 0.01), DAS28-3 at follow up (P < 0.005), ΔDAS28-3 (P < 0.001), ΔDAS28-3/year (P < 0.01), change in total radiographic score/year (P < 0.05) and change in erosion score/year (P < 0.01) were significant predictors of repair. These factors were entered into a multivariate logistic regression model to identify variables with independent predictive value for repair. ΔDAS28-3 was found to be a significant independent predictor of repair (P < 0.007).
Bone sites where repairs of erosions were observed in individual patients
Patient number
Bone site and number of erosions
Number of repairs
rB2 × 1
rA3 × 2, rB3 × 2
rA4 × 1, lA3 × 1, lB3 × 1
rC2 × 1, lI × 1
lE5 × 1, lF × 1, lG × 1, rH × 1, lH × 1
rD2 × 1, rI × 1
lA4 × 1, lB4 × 1, rE4 × 1, rE5 × 1, rG × 1
lC3 × 1, rE5 × 1
rD2 × 1, lH × 1, rI × 1, lI × 1, lJ × 1
rD2 × 1, rI × 1
rA5 × 2, rB5 × 2, lI × 1
rE5 × 1, rF × 1
rA4 × 1, lA5 × 2, rB4 × 1, lB5 × 2
Most of the scoring systems of radiographic data and their multiple modifications are designed to quantify the speed of progressive destruction over time in selected joints of the hands, wrists and feet. However, they are not designed to document improvement. If readers were allowed to score improvements in scores, these scoring methods – with appropriate statistical analysis – may accurately reflect repair in serial films. However, in reality the majority of healing phenomena do not result in changes to radiographic scores [10].
Several investigations have reported that magnetic resonance imaging (MRI) and ultrasonography are highly sensitive in detecting erosions in the hands and wrists of patients with RA [27–29]. Indeed, erosions shown by MRI may be visible 6–12 months or more before they are observed on radiographs, providing a means of early detection and immediate therapeutic intervention [28, 30]. Stewart and coworkers [31] examined 41 patients with RA. MRI scans of the dominant wrist in 31 patients obtained at 1 year and 6 years were compared. Twenty-two patients had an increase in erosion score in the interval and three patients exhibited a decrease in erosion score, suggesting erosion repair.
Based on these findings, we investigated whether repair of erosions can be detected in RA patients treated with conventional DMARDs at our rheumatology clinic, and we compared clinical characteristics between patients exhibiting and not exhibiting erosion repair. To ensure optimal sensitivity in detecting erosion repair, the films were evaluated with knowledge of the sequence. Although this is an issue that still attracts much debate, one study [32] suggested that films should preferably be read in chronological order because this leads to an increase in the detection of clinically relevant changes, without serious overestimation of irrelevant changes. In another study [33] reading films in chronological order was shown to yield a better signal-to-noise ratio when compared with paired reading. We chose the chronological approach in the present study because we believe it to be the most sensitive scoring method; however, more data are needed before we may arrive at a definitive conclusion regarding the optimal approach in observational studies. Furthermore, interobserver agreement was tested independently by a total of four rheumatologists without knowledge of sequence, resulting in high agreement.
We assessed radiographic joint damage using vdH-S. There were no significant differences between groups in total radiographic score, erosion score, or JSN score at baseline. The change in total radiographic score/year (P < 0.05) and change in erosion score/year (P < 0.01) were significantly lower in the repair observed group. To our surprise, patients with erosion repair at any bone site of the hands exhibited lower overall radiographic progression rates, as evaluated using vdH-S, which does not include scoring of healing phenomena. The change in erosion score/year of 1.1 ± 2.2 in the repair observed group means that erosive progression does occur simultaneously with repair in other joints in the same patient. In fact, new erosions were observed in four patients (30.8%) in the repair observed group. Another interesting finding from our study was that erosion repair was observed in patients who were not taking any DMARDs. This suggests the existence of a physiological process of self-repair in RA patients with low disease activity.
The great majority of repairs (20 repairs [45.5%]) were detected in PIP joints. We believe we can partially explain this by the fact that more erosions were observed at PIP joints than at other bone sites at baseline. The number of 'repairs' becomes higher with greater numbers of erosions at baseline.
The process of bone destruction in RA is considered to be correlated with arthritis activity [34], although evidence is accumulating that bone destruction can occur independent of arthritis activity [35–38]. Molenaar and coworkers [35] followed up 187 patients with RA who were in remission clinically and radiologically for 2 years. After 2 years of follow up, remission persisted in 52% of patients. Clinically relevant progression of damage was more frequent in patients with exacerbation than in those with persistent remission.
However, in 15% of patients with persistent remission, erosions developed in previously unaffected joints. Those investigators concluded that the DAS area under the curve was a stronger predictor of radiographic progression than were baseline variables and absence of persistent remission. In the present study there were eight patients in persistent remission, defined as DAS28-3 score below 2.6. Erosion repair was detected in none of these patients; indeed, three patients (37.5%) even exhibited radiographic progression exceeding the smallest detectable change (total radiographic score increase in vdH-S >12.8). On the other hand, although 12 patients (92.3%) in the repair observed group exhibited reduced DAS28-3 score at follow up, one patient actually had a slight increase of disease activity (0.1 increases in ΔDAS28-3). In the repair observed group, 10 patients were in remission or had low disease activity at follow up (DAS28-3 score <3.2), two patients had intermediate activity (DAS28-3 score greater than 3.2 but less than 5.1) and one patient had high disease activity (DAS28-3 score >5.1). Our data confirm the finding of Molenaar and coworkers [35] that bone destruction can occur independently of clinical activity.
The present study has several limitations resulting from its retrospective design. The number of patients with erosion repair was relatively small, and the radiographic evaluations were limited to hands because the number of radiographs of the feet taken at routine clinical setting was small. However, these limitations do not diminish the importance of the study, which, to our knowledge, is the first to explore the clinical characteristics of RA patients exhibiting erosion repair.
Our study suggests that structural repair can be achieved by reducing disease activity in RA, and this phenomenon is not limited to patients with early disease or treatment with biological agents. The importance of targeting remission with aggressive therapy in all patients with RA is highlighted. A reliable method for evaluating and scoring of erosion repair must be established and factors predictive of bone repair identified in the near future.
Disease Activity Score
disease-modifying antirheumatic drugs
joint space narrowing
magnetic resonance imaging
proximal interphalangeal
rheumatoid arthritis
- vdH-S:
the van der Heijde modification of the total Sharp scoring system.
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- van der Heijde DM, van Leeuwen MA, van Riel PL, Koster AM, van 't Hof MA, van Rijswijk MH, van de Putte LB: Biannual radiographic assessments of hands and feet in a three-year prospective followup of patients with early rheumatoid arthritis. Arthritis Rheum. 1992, 35: 26-34.View ArticlePubMedGoogle Scholar
- Hulsmans HM, Jacobs JW, van der Heijde DM, van Albada-Kuipers GA, Schenk Y, Bijlsma JW: The course of radiologic damage during the first six years of rheumatoid arthritis. Arthritis Rheum. 2000, 43: 1927-1940. 10.1002/1529-0131(200009)43:9<1927::AID-ANR3>3.0.CO;2-B.View ArticlePubMedGoogle Scholar
- Bruynesteyn K, van der Heijde D, Boers M, Lassere M, Boonen A, Edmonds J, Houben H, Paulus H, Peloso P, Saudan A, et al: Minimal clinically important difference in radiological progression of joint damage over 1 year in rheumatoid arthritis: preliminary results of a validation study with clinical experts. J Rheumatol. 2001, 28: 904-910.PubMedGoogle Scholar
- van der Heijde DM: Assessment of radiographs in longitudinal observational studies. J Rheumatol Suppl. 2004, 69: 46-47.PubMedGoogle Scholar
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- Bruynesteyn K, Boers M, Kostense P, van der Linden S, van der Heijde D: Deciding on progression of joint damage in paired films of individual patients: smallest detectable difference or change. Ann Rheum Dis. 2005, 64: 179-182. 10.1136/ard.2003.018457.PubMed CentralView ArticlePubMedGoogle Scholar
- van der Heijde D, Lassere M, Edmonds J, Kirwan J, Strand V, Boers M: Minimal clinically important difference in plain films in RA: group discussions, conclusions, and recommendations. OMERACT Imaging Task Force. J Rheumatol. 2001, 28: 914-917.PubMedGoogle Scholar
- Lassere MN, van der Heijde D, Johnson KR, Boers M, Edmonds J: Reliability of measures of disease activity and disease damage in rheumatoid arthritis: implications for smallest detectable difference, minimal clinically important difference, and analysis of treatment effects in randomized controlled trials. J Rheumatol. 2001, 28: 892-903.PubMedGoogle Scholar
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- Bruynesteyn K, Van Der Heijde D, Boers M, Saudan A, Peloso P, Paulus H, Houben H, Griffiths B, Edmonds J, Bresnihan B, et al: Detecting radiological changes in rheumatoid arthritis that are considered important by clinical experts: influence of reading with or without known sequence. J Rheumatol. 2002, 29: 2306-2312.PubMedGoogle Scholar
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This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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Colorado Art Ranch Residency Program [1]
The Colorado Art Ranch offers a residency at Carpenter Ranch in Routt County, Colorado [3], from September 1 to September 30 for one poet, fiction writer, or creative nonfiction writer whose work relates to land and water issues. The resident will be provided with housing and work space, but is responsible for their own food, travel, and other expenses. Submit up to 20 pages of poetry or prose with a $50 application fee by May 15. Visit the website for an application and complete guidelines.
Colorado Art Ranch Residency Program, 269 Snowcliff Road, Livermore, CO 90536. (303) 279-5198. Grant Pound, Executive Director.


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The Horse Forum (http://www.horseforum.com/forumindex.php)
- Horse Grooming (/horse-grooming/)
- - Plaiting (http://www.horseforum.com/horse-grooming/plaiting-122325/)
My and my boy have an upcoming show, just a simple schooling show. But just for fun, I'd really like to do a hunter braid on him.
I don't know how to braid. o.o.. Sad, yes. I was never taught. Is it just folding each strand of hair over the other? Is there a certain type of braid I'm going to be using for the hunter braid?
I would really appreciate the help.
Easy peasy, 3 strands (left, middle, right) fold left over middle (left then becomes middle) then fold right over middle (right then becomes middle) then left over middle, right over middle, and keep on going.
like this:
just practise :)
I always think of it like this
break into three strands - then twist two strands (One of the outside strands and the inside one) then twist the middle and the OTHER outside strand... and just think twist twist twist twist....
practice as said above - get a barbie or wool or something to practice on
Oh my gosh, thank you two soooo much! You just like... Saved my pride.
Another question, that video used a french braid for the forelock. Do you HAVE to use the french braid or can you use just a normal braid? If you do have to use a french braid to make it look decent, how do you do a french braid..? xD
French braiding is easy it's just a normal plait that you add bits into as you go, still keeping 3 strands.
Your actually supposed to do an under french plait (you put the strands under instead of over) but you don't have to do it under, you can do it normally (I find it's easier)
this should help
How to Create a French Braid - Hair
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text: Flight Attendant Logs 53 Years In-FlightSadie Stein12/29/10 12:53pmFiled to: Fly girlsNorma heapair travelContinental AirlinesFlyingStewardessesflight attendantsAir hostesses31EditPromoteShare to KinjaToggle Conversation toolsGo to permalink Norma Heap started air-hostessing in 1957; 26 million miles later, she's Continental's longest-serving flight-attendant. Says Norma, 74, "I'm just fortunate that I chose something that I enjoy doing...and I've never lost the love for flying or traveling, or serving people."Reply31 repliesLeave a reply


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Hotfrog Canada provides information regarding North York Community Care Access Centre in North York ON. North York Community Care Access Centre is located at 45 Sheppard Ave E Suite 700, North York ON M2N 5W9 and provides Community And Government services. Contact them on (416) 222-2241.
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The epitome of polished, masculine styling, this tie clip from BOSS will complete your sartorial look to perfection. In a sleek silver design, this tie clip features engraved branded detailing to complete a stylish piece.
Thank you for visiting BOSS Teodor Tie Clip, Silver on shoppingfind.co.uk
We bring the products from many websites to one website for you to search, save and compare.


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The Manic Street Preachers are avid students and past masters of rock mythology. It enabled them to survive the likely fatal disappearance of their talismanic lyricist and friend Richey Edwards in 1995, an enduring personal loss they fully understood in terms of the band's legend. It has also let them find a way past mid-career shakiness.
Postcards From A Young Man (2010) and Rewind the Film (2013) added middle-aged ruefulness at the appropriate moment to their initial flaming, short-fused punk spirit. Most importantly, the Manics are still a passionate rock band. And they insist on an old, not quite broken idea of what rock'n'roll is for: to steel you to resist injustice throughout your life, even as middle-aged compromise creeps in.
Brighton was always a core town for Manics disciples. They're treated to a high-class support act, Scritti Politti, essentially Green Gartside.
Having only returned to performing after 26 years' stage-frightened absence in 2006, Green's legs stay tensely braced, but he does tentatively dip his honeyed voice into his stock of polished hits. "Wood Beez (Pray Like Aretha Franklin)" and "Absolute" are the sort of intellectually rigorous 1980s pop the Manics grew up on, making their fellow Welshman a fitting guest.
The Manics make their own entrance with one of their biggest hits, "A Design for Life", and its undebeatable opening line: "Libraries gave us power," But 2010's "(It's Not War) Just the End of Love" is where they come into focus for me tonight. It's both epic in sound, and as intimately close in spirit as an idealistic friend. It's fuelled by singer James Dean Bradfield's individual take on the rock guitar's classic wail,
"Rewind the Film", a low-down, Lee Hazlewood-style Welsh valleys ballad from last year, is helped by video footage of rust-red collieries, abandoned rugby goalposts, and miners' strike newspaper cuttings. "I want the world to see it all," Bradfield sighs. With these images, the song becomes an elegy to Welsh communities buried by Thatcher, and laid out straight here. Bradfield's mournfully slow, solo acoustic version of "From Despair to Where" is similarly affecting.
The feather boa draped on a mic-stand left eternally ready for Richey Edwards' return has been quietly retired. But he isn't forgotten. "This one's for you, Richey!" Nicky Wire yells, introducing "Bored Out Of My Mind"'s Zepellin-heavy rock. The crowd are inevitably older and more passive than any Edwards witnessed in his time in the band, when it was impossible to be indifferent to them.
The massed, raised fists when they burn through that era's anthem, "Motorcycle Emptiness", shows, though, that these fans do still have faith in the Manics. Last year's Motown-mainlining "Show Me the Wonder" is just as thrilling. "If You Tolerate This Your Children Will Be Next", a 1998 number one hit based on a Spanish Civil War slogan, is a fine way to finish. We still love them, and they still love rock, and the rebel visions it allows them.Reuse content


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Welcome to Pop Warner Football!
Dear Scripps Ranch Pop Warner Parents:
Welcome to Scripps Ranch Pop Warner! We are very excited that your child will be playing football this season! Whether your son is a first time player or a seasoned veteran, we hope you will enjoy your experience with our organization.
Here is everything you need to know about participating in the 2016 season:
TWO-WEEK MANDATORY CONDITIONING: Begins Monday, August 1st for 2 weeks and includes 20 hours of mandatory conditioning. The first 10 hours of conditioning are just in helmets and league issued short and t-shirts. The second 10 hours are in full pads. We strongly recommend scheduling family vacations before August 1st and after mid-November. The first few weeks are very important for conditioning, player safety (via USA Football's - Heads Up Football), offensive and defensive positions and plays. Drills involving contact start after the first 10 hours of conditioning. Scrimmages start after the second 10 hours of conditioning. If you are concerned about any conflicts, please let us know so we can let the coaches know and develop a plan to work around them.
CONDITIONING (Tackle): First 10 hours (Helmets, Shorts/Shirts)
- Monday | August 1 | 5:00-7:30 p.m.
- Tuesday | August 2 | 5:00-7:30 p.m.
- Wednesday | August 3 | 5:00-7:30 p.m.
- Thursday | August 4 | 5:00-7:30 p.m.
CONDITIONING (Flag) : First 8 hours
- Monday | August 1 | 5:15-7:15 p.m.
- Tuesday | August 2 | 5:15-7:15 p.m.
- Wednesday | August 3 | 5:15-7:15 p.m.
- Thursday | August 4 | 5:15-7:15 p.m.
CONDITIONING (Tackle) : Second 10 hours (Helmets, Full pads)
- Monday | August 8 | 5:00-7:30 p.m.
- Tuesday | August 9 | 5:00-7:30 p.m.
- Wednesday | August 10 | 5:00-7:30 p.m.
- Thursday | August 11 | 5:00-7:30 p.m.
CONDITIONING (Flag) : Second 8 hours
- Monday | August 8 | 5:15-7:15 p.m.
- Tuesday | August 9 | 5:15-7:15 p.m.
- Wednesday | August 10 | 5:15-7:15 p.m.
- Thursday | August 11 | 5:15-7:15 p.m.
EARLY SEASON PRACTICE (Tackle) : May be modified based upon scrimmage dates
- Week 3 - Monday - Thursday | August 15 - 18 | 5:00-7:30 p.m.
- Week 4 - Monday - Thursday | August 22 - 25 | 5:00-7:30 p.m.
EARLY SEASON PRACTICE (Flag) : May be modified based upon scrimmage dates
- Week 3 - Monday - Thursday | August 15 - 18 | 5:15-7:15 p.m.
- Week 4 - Monday - Thursday | August 22 - 25 | 5:15-7:15 p.m.
REGULAR PRACTICES: Three days per week beginning the week of August 22. Each team may have a slightly different schedule. You will be informed by your team head coach or business manager.
- Lower teams: Dingeman Elementary (Includes Flag, Jr. Mitey Mite and Mitey Mite)
- Upper teams: SRHS Soccer Field (Includes: Jr. Pee Wee, Pee Wee and Jr. Midget)
- Tackle teams at Dingeman: Tues, Wed and Fridays from 5:15 - 7:30 p.m
- Tackle teams at SRHS Soccer Field: Tues, Wed and Thurs or Fridays from 5:15 to 7:30 p.m.
- Flag teams at Dingeman: Tues, Wed and Fridays from 5:15 to 6:45 p.m.
Games are on Saturdays beginning August 27th. Home tackle football games are held at the Scripps Ranch High school football stadium. Parents are needed to help in the snack bar, help with chains and in the announcer booth on home game days.
Flag home games are held at the Scripps Ranch High school soccer field.
Away games are held at the other 17 Association's fields in the Palomar Conference which include playing teams from Poway, Rancho Bernardo, Oceanside, La Costa all the way up to Temecula and Murrietta. For a complete list of associations, please visit the Palomar Pop Warner Conference at www.palomarpopwarner.org.
Flag plays a total of 8 games with a final Flag Fest at the end of the season. Jr. Mitey Mite through Midget teams play 10 games and with additional possible playoff and championship games. The season ends at the end of October for Flag, the beginning of November for Jr. Mitey Mite and Mite and usually mid to late November for all Jr. Pee Wee-Midget teams. Teams can chose to continue playing after the season is over at Invitational and Bowl games hosted by other Associations.
Flag: $225 | Tackle: $375
- $50 Sibling discount
- $50 Jersey re-use Discount
Your fees include:
- Complete uniform provided by SRPW, with a jersey and spirit pack* to keep (*tackle teams only)
- Individual and Team picture package
- A minimum of 8 games for Flag and 10 games for Tackle teams as well as possible playoff game(s)
- Fees pay for Pop Warner insurance and Palomar conference team fees as well as all high school field, lights, custodial and referee fees
- Registration fees also cover all coaches uniform costs, training, practice equipment and coaches bags
UNIFORM COST: There is no cost for the helmets, practice and game uniforms, however, all equipment (excluding jersey) must be turned in at end of season or the family will be charged. Players must supply their own cleats, under-armor and mouth guard.


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fdcontinental20 Feedback reviews, 95% positive
We are a family run business with over 30 years experience. We offer a reliable ,honest and fairly priced approach to work and all of our work is quality assured.
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Brett Jones Contracting33 Feedback reviews, 100% positive
Welcome to my page. I offer a friendly service specialising in all types of fencing and gate installation. I started in the fencing trade and over the last few years I have developed my skills to also include decking and shed building. Based in Newmarket I cover all...
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pca home&garden services22 Feedback reviews, 95% positive
PCA is based in St Ives, and chatteris .we serve all surrounding area .Are main trades are hard landcaping i;e fencing ;patios decking and all aspects of landscaping work .we are fully insured .we also offer fully itemised free quotations.we have been in the building trade...
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jobrite landscaping1 Feedback review, 100% positive
i will always offer a 'can do' service, if we do not know how to, then we will know who can. my attitude to work is always positive and reliable as i want our family business to continue to grow with my children joining the team as they leave school or college.
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Fence Installers35 Feedback reviews, 100% positive
My name is David Blacker, I am a family man who enjoys outside work and having something to be proud of at the end of each day. I have built up a considerable portfolio of domestic fencing as you can see from my pictures. Please put as much detail as you can when you...
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K Thomas Property Maintenance2 Feedback reviews, 100% positive
I'm a responsible reliable family man whose aim is to do every job to the best of my ability at an affordable price. All my previous jobs have had 100% feedback
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Premier Stone Pavers13 Feedback reviews, 92% positive
We are a father and son business with over 30 years experience in driveways patios fencing and landscaping services.All of our pavers are time served and qualified to undertake all aspects of work to the best standards ensuring the workmanship lasts.we Are Marshall paving...
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Borderline Fencing & Gates7 Feedback reviews, 86% positive
I have worked in the fencing profession for over 16 years, specialising in assembling and repairing fences and gates. I am a reliable, trustworthy and hardworking person who works in collaboration with the client to complete work to a high standard.
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NJ Contractors6 Feedback reviews, 100% positive
We are a trustworthy family run business with over 145 years joint experience in general building. Our team of professional friendly tradesmen offer an array of different trades from groundwork to carpentry. We have completed many job over the years from large jobs including...
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e-timber Products1 Feedback review, 100% positive
E-timber products has been established for over six years and specialise in the manufacture and installation of garden furniture, fencing and decking. Over the last five years we have grown substantially by providing fist class pricing, quality and service.
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INL Landscapes18 Feedback reviews, 100% positive
INL Landscapes is a family orientated company was started in January 2008 by Ian Roberts. Ian had spent the previous 8 years learning his trade with a local landscaping company. INL Landscapes specialises in block paving, patio's and fencing, planting and maintenance and...
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Gryall Landscapes1 Feedback review, 100% positive
Hello. Thank you for looking at my profile. My name is Gary ryall . I'm 47 years old . I have been in the landscaping and building trade for 30 yrs .. serving apprenticeship for 3 yrs at beginning of my career. Gaining full honours in landscaping. I have a big client base...
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Evergreen Landscapes97 Feedback reviews, 99% positive
Hi I am a reliable trustworthy and punctual guy and my aim is too provide a excellent services to all of my customers. All my work is done to a high standard so you homeowner will have the confidence that you have the right guy for the job. Portfolio and sites to view on...
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MJL Services8 Feedback reviews, 88% positive
Why us? Friendly, honest and reliable. Locally based in Bedfordshire, I pride myself on 100% customer satisfaction. A small family run business with the high specification finish and service you would expect from a larger firm. Quotations and site visits undertaken with no...
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Grand Outdoors5 Feedback reviews, 100% positive
Grand Outdoors Landscaping are East Anglia's foremost professional service for patio and drive way installers, fencing contractors, tree care and general landscape design and installation. We are affordable, professional and reliable. We do not require any deposits for...
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By Randy Dotinga |
Felicia Shaw has a history of setting the stage for something unusual.
Years ago, she convinced her then-husband, Chargers football player Pete Shaw, to perform in a ballet production with a couple of his teammates. It was a nerve-wracking moment for the men on stage, but they survived and were thrilled to see the athleticism of the ballet artists at work, Felicia Shaw recalls.
Pete Shaw was so stunned that he pondered their power afterward: "With all they can do with their bodies, could you imagine what they could do on the field?"


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The St. Louis Cardinals added two starting pitchers in the last two days, bringing in John Lackey and Justin Masterson.
Numbers Game looks at two trades that improve the Cardinals' chances of returning to the postseason for the fourth straight season.
The Cardinals Get: RHP John Lackey and RHP Justin Masterson.
Lackey, 35, has been a solid starter for the Red Sox, particularly in the past two seasons since undergoing Tommy John surgery in 2012. After a year off for recovery, Lackey has gone 21-20 with a 3.55 ERA and 1.19 WHIP in 50 starts. Not dominant, but good enough to rank 24th among starting pitchers in Fan Graphs WAR over the past two seasons and rank 49th among starting pitchers in the TSN.ca Player Rankings this year.
He's relying more on his fastball, throwing it 65.6% of the time, his highest percentage since 2003, and his average velocity (92.1 MPH) is the best of his career.
The Cardinals add a steady veteran pitcher, with postseason experience (3.03 ERA, 1.30 WHIP in 19 postseason appearances), one that can easily fit in the middle of the rotation for a St. Louis team that, one game out of a wild card spot, justifiably has playoff aspirations.
While those numbers paint a picture of a nice addition to the Cardinals' rotation, there is a definite cherry on top of the sundae and that is that Lackey's contract. While it pays him $15.25-million this year, there is a 2015 club option at league minimum salary because Lackey missed the entire 2012 season (a clause in the contract acknowledging Lackey's previous elbow troubles). Provided that Lackey still intends to pitch next season, he would provide exceptional value to the Cardinals.
Masterson is a 29-year-old who has had some good seasons with the Indians, but is undeniably struggling this year, posting a career-worst 5.51 ERA and 1.65 WHIP in 19 starts. He's recently spent time on the DL with a knee injury, but Masterson's bigger issue is that he's lost some velocity. In 2011, his fastball was clocked, on average, at 92.7 MPH. This year, it's down to 89.1 MPH, and for a guy who throws such a high percentage of fastballs, that's a killer.
According to Fan Graphs, Masterson's fastball has been the least effective in the majors this season. He has been somewhat unlucky with a career-worst .350 batting average on balls in play, but some of that is simply a reflection of Masterson getting hit hard this season.
However, if Masterson fills the fifth spot in the Cardinals' rotation --behind Adam Wainwright, Lance Lynn, Lackey and Shelby Miller -- then that's an adequate contribution for the last couple months of this season. He would have to dramatically improve his performance to qualify as a starter for the postseason, but he may be a viable bullpen arm to face right-handed hitters (getting rocked to the tune of a .935 OPS by left-handed batters this season).
Earning $9.7625-million this season, Masterson will be eligible for arbitration at season's end and can be a free agent after the 2015 season. This is an opportunity for the Cardinals to buy low and see if they can get Masterson back on track, while hopefully helping their chances to get into the playoffs this season.
The Red Sox Get: RHP Joe Kelly and RF/1B Allen Craig.
A 26-year-old righthander, Kelly has made 38 starts (and 30 relief appearances) for the Cardinals over the past three seasons. As a starter, he's 15-11, with a 3.25 ERA and 1.39 WHIP, with 130 strikeouts in 213 1/3 IP. His ERA is the same, but Kelly's strikeout rate is dramatically higher coming out of the bullpen (49 K in 52 2/3 IP).
Kelly throws hard (average fastball speed of 94.7 MPH) and has cut back on his ineffective slider, having more success in this season's small sample, with his curve ball. He should have a great opportunity to start regularly in Boston over the next couple months, in an effort to establish that he can be a rotation regular next season.
This early in his career, Kelly is an inexpensive option, making $523,000 this year, and not arbitration eligible until 2016 at the earliest.
Craig is a 30-year-old who was a productive hitter before suffering a foot injury last season. He still has an .803 OPS for his career, but has struggled this year, posting seven homers and a .237 batting average, with a .638 OPS.
There has to be some expectation on the Red Sox part that they can get Craig back on track. He simply hasn't made as much solid contact this season, with a career-high 56.5% groundball rate. Presuming that Craig can get healthy and bounce back to his previous levels, he's a potential middle of the order bat for the Red Sox who is signed through 2017. Craig is making $2.75-million this year and his contract will pay $26.5-million over the next three seasons, with a club option for 2018.
Craig should get a look in left field with the Red Sox, effectively taking over for Jonny Gomes, who was traded to Oakland earlier in the day. Of Craig's 117 career starts in the outfield, 35 have been in left.
With Craig leaving St. Louis, that leaves an everyday job for top Cardinals prospect Oscar Taveras. The 22-year-old has struggled (hitting .208 with a .508 OPS in 32 games) this season, but he's the top prospect in the organization, so he'll get a crack at the everyday job. If he can't hack it, Jon Jay can handle right field for the Cardinals, with Peter Bourjos in centre, if need be.
Lackey's contract situation obviously helped the Red Sox secure real value in return and they got two pieces that could be part of a contending club in 2015.
The Indians Get: OF James Ramsey.
A first-round pick by the Cardinals in 2012, 24-year-old James Ramsey is an outfielder that is having a pretty strong showing at Double-A, hitting 13 home runs, and hitting .300 with a .916 OPS in 67 games. Ranked by Baseball America as the Cardinals' No. 8 prospect in the offseason, Ramsey is likely not too far away from getting his shot in the big leagues.
Considering how the season was going for Masterson, that's not a bad move for the Tribe. They get a prospect who will have a chance to play in their outfield in the not-too-distant future, and they can keep young righthanders Trevor Bauer and Danny Salazar in the rotation for the rest of the season.
Scott Cullen can be reached at email@example.com and followed on Twitter at http://twitter.com/tsnscottcullen. For more, check out TSN Fantasy on Facebook.
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