Loa22 is an outer membrane protein encoded

Loa22 is an outer membrane protein encoded buy MK-1775 within all Leptospira genomes sequenced to date. It has been observed to be upregulated in vivo[27] and it is one of very few leptospiral proteins so far that has been shown to be necessary for virulence [3]. Additional studies are needed to define the precise context of NulO expression on L. interrogans and understand its potential significance in virulence. Conclusions Based on a combination of experimentation and in silico genomic analysis, we have demonstrated the function of NulO biosynthetic gene clusters in pathogenic and intermediately pathogenic species of Leptospira, several of which are capable of synthesizing di-N-acetylated

NulO species, as well as the true sialic acid, N-acetyneuraminic acid, a finding of considerable consequence for the leptospirosis field. This finding expands the number of important human pathogens that utilize endogenous biosynthetic pathways to elaborate surface structures containing sialic acids and related NulO

molecules [16]. Sialic acids have proven roles in complement LY2874455 chemical structure evasion, intracellular survival, and biofilm formation [29], and evidence is emerging that some human pathogens with Neu5Ac on their surfaces can engage sialic acid-binding receptors (Siglecs) on leukocyte cell surfaces, resulting in phagocytosis or dampening of bactericidal activities [30–32]. The roles of other NulO molecules such as legionaminic and pseudaminic acids are less well defined, but these molecules have been shown to play roles in behaviors such as autoagglutination, motility, and host colonization [33–37]. Curiously, disease caused by L. interrogans includes bacteremia and meningitis as components of the clinical disease spectrum, similar to the well-characterized Neu5Ac-expressing human bacterial pathogens Group B Streptococcus Neisseria meningitidis E. coli K1, and Haemophilus influenzae. As genetic tools and small animal infection systems for study of Leptospira are further refined, analysis Transferase inhibitor of the

contribution of NulO biosynthesis to the virulence of this neglected disease can be further elucidated. Methods Strains and STA-9090 culture conditions Intermediately pathogenic strains L. licerasiae serovar Varillal strains MMD3731, MMD4847 and CEH008 (isolated from rodents in Peru), L. fainei serovar Hurstbridge strain BUT 6T and the saprophyte L. biflexa serovar Patoc were used for these experiments. Pathogenic Leptospira used in this study included L. interrogans serovar Copenhageni strain L1-130, L. interrogans serovar Lai strain 55601, and L. interrogans serovar Icterohaemorrhagiae wild rodent isolate MMD 3731 that were passaged fewer than 5 times in vitro after re-isolation from hamster liver to maintain virulence. L. santarosai and L. alexanderi serovar Manhao were originally isolated from clinical cases of leptospirosis and now serve as reference laboratory strains.

1989) All raters have followed a trainings program and are certi

1989). All raters have followed a trainings program and are certified, and have to attend a refresher course twice a year. The Ergo-Kit lifting tests were found to be reliable in subjects both with and without SBI-0206965 chemical structure musculoskeletal complaints with respect to the lifting tests (Gouttebarge et al. 2005,

2006). There is no information known to us from international literature about the reliability of the other tests of the Ergo-Kit FCE, neither is there information available about the predictive validity of this FCE. Claimants with a medical contra-indication for FCE, e.g., recent myocardial infarct, heart failure or recent surgery, were excluded from the test. Outcomes The questionnaire presented to all IPs https://www.selleckchem.com/products/VX-765.html contained three questions: 1. The IP was asked whether the FCE assessment had complementary value for the assessment of the physical work ability of the patient. The response choices were dichotomous: yes or no. With regard to the sub-question, characteristics of IPs and claimants that were believed to influence the answer of IPs about the complementary value of FCE information were classified. The characteristics selected for the IP group were work experience and familiarity with FCE. Work experience was found to be a factor that influences the way IPs come to their judgment about work ability (Razenberg 1992; Kerstholt et al. 2002). Familiarity with FCE was

judged HSP inhibitor to be another reason why IPs might think differently about the complementary value. It was deemed possible that earlier contact with FCE information led to a negative opinion, as shown in the study about the utility of FCE information (Wind et al. 2006). The characteristics registered in the claimant group were the location of the disorder, their working situation, and functional disability. Location of disorders could be a factor for differences in judgment of the complementary value

of FCE information. It is possible that FCE information could be judged as more valuable in assessments of claimants with general disorders than specifically localized disorders. Work status is another characteristic of the claimants that could lead to a difference between the group of IPs that considers FCE information to be of complementary value versus those that do not. The information Carteolol HCl that a claimant is currently working might make the information from an FCE assessment appear less valuable, and thus influence the IP’s perception of the complementary value of FCE information. Functional disability was also assessed with the revised Oswestry questionnaire. The revised Oswestry questionnaire is derived from the Oswestry questionnaire (Fairbank et al. 1980) and is a 10-item instrument designed to measure the effects of pain on functional disability. Results of the revised Oswestry questionnaire were noted in numbers of claimants according to the five classes outlined by the revised Oswestry questionnaire: 0–20, 20–40, 40–60, 60–80, 80–100%.

Heaney RP (2003) M

Heaney RP (2003) Normalizing calcium intake: projected population effects for body weight. J Nutr 133:268S–270SPubMed 23. Parikh SJ, Yanovski JA (2003) Calcium intake and adiposity. Am J Clin Nutr 77:281–287PubMed 24. Barr SI (2003) Increased dairy product or calcium intake: is body weight or composition affected in humans? The Journal of nutrition 133:245S–248SPubMed 25. Trowman R, Dumville VX-680 JC, Hahn S, Torgerson DJ (2006) A systematic review of the effects of calcium supplementation on body weight. Br J Nutr 95:1033–1038PubMed 26. Lanou

AJ, Barnard ND (2008) Dairy and weight loss hypothesis: an evaluation of the clinical trials. Nutr Rev 66:272–279PubMed 27. Bolland MJ, Avenell A, Baron JA, Grey A, MacLennan GS, Gamble GD, Reid IR (2010) Effect of calcium supplements on risk of myocardial infarction and cardiovascular events: meta-analysis. BMJ 341:c3691PubMed 28. Bolland MJ, Barber PA, Doughty RN, Mason B, Horne A, Ames R, Gamble GD, Grey A, Reid IR (2008) Vascular Selleck TGFbeta inhibitor events in healthy older women receiving calcium supplementation: randomised controlled trial. BMJ 336:262–266PubMed 29. Reid IR, Schooler BA, Hannan SF, Ibbertson HK (1986) The acute biochemical effects of four proprietary calcium preparations. Aust N Z J Med 16:193–197PubMed 30. Foley RN, Collins AJ, Ishani A, Kalra PA (2008) Calcium-phosphate levels and cardiovascular disease in community-dwelling adults: the Atherosclerosis Risk in Communities

(ARIC) Study. Am Heart J 156:556–563PubMed 31. Vestergaard P, Mollerup CL, Frokjaer VG, Christiansen P, Blichert-Toft M, Mosekilde L (2003) Cardiovascular events before and after surgery for primary hyperparathyroidism. buy Erismodegib World J Surg 27:216–222PubMed 32. Jackson RD, LaCroix AZ, Gass M et al (2006) Calcium

plus vitamin D supplementation and the risk of fractures. N Engl J Med 354:669–683PubMed ADP ribosylation factor 33. Hsia J, Heiss G, Ren H et al (2007) Calcium/vitamin D supplementation and cardiovascular events. Circulation 115:846–854PubMed 34. Wang TJ, Pencina MJ, Booth SL, Jacques PF, Ingelsson E, Lanier K, Benjamin EJ, D’Agostino RB, Wolf M, Vasan RS (2008) Vitamin D deficiency and risk of cardiovascular disease. Circulation 117:503–511PubMed 35. Autier P, Gandini S (2007) Vitamin D supplementation and total mortality: a meta-analysis of randomized controlled trials. Arch Intern Med 167:1730–1737PubMed 36. Lewis JR, Calver J, Zhu K, Flicker L, Prince RL (2011) Calcium supplementation and the risks of atherosclerotic vascular disease in older women: results of a 5-year RCT and a 4.5-year follow-up. J Bone Miner Res 26:35–41PubMed 37. Park Y, Leitzmann MF, Subar AF, Hollenbeck A, Schatzkin A (2009) Dairy food, calcium, and risk of cancer in the NIH-AARP Diet and Health Study. Arch Intern Med 169:391–401PubMed 38. Martinez ME, Willett WC (1998) Calcium, vitamin D, and colorectal cancer: a review of the epidemiologic evidence. Cancer Epidemiol Biomarkers Prev 7:163–168PubMed 39.

Authors’ contributions XZ and JM participated in the study design

Authors’ contributions XZ and JM participated in the study design, constructed lentiviral plasmid vector

,conducted the real-time PCR assays and drafted the manuscript; YJW and YL statisticsed the patient information and conducted immunohistochemical staining; HZL carried out the western bolt assay; QL and XJL carried out the proliferation and cell migration assay; PM conduced the trials in vivo. ; HYL conceived of the study, and participated in its design and coordination, and reviewed the manuscript. All authors read and approved the final manuscript.”
“Introduction Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the third leading cause of cancer-related death [1]. Although significant advances in surgical techniques and perioperative care over the last two decades, the long-term prognosis of HCC remains dismal largely due to the high frequency of metastasis or recurrence. Recently, more evidences suggest that #Mocetinostat randurls[1|1|,|CHEM1|]# HCC metastasis involves a complex cascade of signal events between tumor cells and host stroma microenvironment. AZD5363 supplier These crosstalking might modulate or determine

the process of HCC invasion and metastasis. Thus, exclusive reliance on tumor cell itself for research cannot enable insight into the diverse pathological changes occurring in HCC metastasis. Generally, the microenvironment of HCC is composed of stromal cells (e.g., hepatic stellate cells, fibroblasts, invading inflammatory/immune cells, and endothelial cells) and non-cellular components (e.g., growth factors, proteolytic enzymes, inflammatory cytokines, and extensive extracellular matrix proteins). A lot of studies on HCC have validated the important roles of stromal cells in HCC progression [2]. Hepatic stellate

cells (HSCs) increase HCC growth and invasion both in vitro and in vivo. Conditioned media derived from HSCs induce HCC cell proliferation and migration. Moreover, on a three-dimensional spheroid co-culture system as well as an in vivo implantation of a mixture of HSCs and HCC cells, HSCs obviously accelerate HCC growth and diminish the extent of central necrosis [3, 4]. Activated HSCs also enhance HCC progression by other means such as regulating T cells that create Sclareol an immunosuppressive microenvironment and stimulating angiogenesis [5]. Through the release of different factors like cytokines, chemokines, or enzymes, tumor-associated macrophages (TAMs) can regulate tumor growth, angiogenesis, invasion, and metastasis [6]. Particularly, some secreted factors from TAMs also induce cancer cell motility, thereby enhancing tumor cell invasion capacity [7]. These data demonstrate that stromal cells can actively modulate the malignant characteristics of HCC cells and further determine the outcome of HCC. Given that tumors have abundant blood vessels for supplying oxygen and nutrition, endothelial cells (ECs) are ubiquitous within solid tumors.

The filtered single cell suspensions were stained with Trypan Blu

The filtered single cell suspensions were stained with AZD4547 ic50 Trypan Blue. The living cells were counted, and primary culture was completed within 2 h, followed by inoculation in simplified serum-free medium (DMEM/F12, containing 2% B27, 20 μg/L EGF and 20 μg/L bFGF), and then culture at 37°C in 5% CO2 saturated humidity incubator. The medium was changed every 3~4 days. The cells were passaged by 1:2 subculture every 7 days and observed under the inverted phase contrast microscope. The cells were passaged three times.

After the cell spheres became regularly shaped, they were dissociated into single cells with 0.25% trypsin + mechanical Caspase pathway method, and inoculated into a 96-well plate at 1 living cell/well, with each well added with 100 μL simplified serum-free medium. The wells containing only one cell were labeled under the inverted microscope, and supplemented with 100 μL simplified serum-free medium for further culture.

The formation of single cell colonies was recorded by dynamic observation. The cells were observed under the inverted microscope CT99021 nmr after culture for about one week, and the proliferated cells were collected and transferred into a culture flask for further culture and proliferation. The purified BTSCs after colony screening were used in the following experiments.   (2) Immunofluorescent identification of BTSCs: On the 5th day of passage, BTSs that grew well were re-suspended in culture medium containing a small amount

of serum (DMEM/F12 containing 10%FBS), and dropped onto a poly-L-lysine-coated coverslip. After standing still for about 4 h until the solution adhered to the coverslip, the coverslip was fixed in 4% paraformaldehyde for 30 min, blocked with normal goat serum for 20 min, incubated with rabbit anti-human CD133 antibody overnight at 4°C, and then incubated with Cy3-labeled sheep anti-rabbit IgG at 37°C for 60 min, followed by DAPI counterstaining of the nuclei and coverslipping with buffered glycerol. Following each step, the coverslip was rinsed with 0.01 mol/L PBS three times, each for 5 minutes. The coverslip was observed after mounting and pictures were taken.   (3) Assessment of the effect of ATRA on proliferation of BTSCs: The BTSCs were collected and divided into groups as described below, put into the corresponding see more culture medium, disaggregated into single cell suspensions by mechanical dissociation, and inoculated into a 96-well plate at the density of 1000 living cells/well, with 100 Ml in each well. According to the different treatments, the BTSCs were divided into: (1) control group: basic medium (DMEM/F12 with 2% B27) containing the same amount of anhydrous ethanol as in the ATRA group (the final concentration < 0.1%); (2) ATRA group: containing 1 μmol/L ATRA; (3) ATRA/growth factor group: containing 1 μmol/L ATRA, and 20 μg/L EGF and 20 μg/L bFGF; (4) growth factor group: containing 20 μg/L EGF and 20 μg/L bFGF.

In this work, we study the case, in which the distances between a

In this work, we study the case, in which the distances between atoms are quite large, so that the average distances between atoms are greater or in the same order than the

‘resonant transition’ wavelength. Therefore, we prepare an ensemble of N two-level atoms initially in ground state, Proteasome inhibitors in cancer therapy and a single mode of the radiation field is excited in a ‘Fock’ state (so called one-photon state). This is the case of a purely monochromatic wave with zero line width under the consideration. A laser output in single mode operation can approximate this situation due to its high degree of monochromaticity (small line width) for instance. The mode of electromagnetic field is specified completely by giving its wave vectors k 0 with atomic transition frequency ω = c|k 0| and its polarization j (j = 1, 2). The main feature, differentiating our research from others in this domain, is the developed direct and consistent solution to the N-particle equations, describing the time evolution of the N atomic probability state amplitudes. Besides, in certain sense, we explained the nature of the widely used Weisskopf-Wigner approximation that was not found in the reviewed by us scientific

literature. The goal of this paper can be formulated as an attempt to propose an adapted and simple in practical use theory, for example in the highly applied nanoscale physics. The proposed theoretical material requires corresponding RG-7388 molecular weight experimental verification. As an idea of an application, the model system can be realized on atomic (developing the method proposed in [1] for the nuclei of 57Fe in certain composites, but this time for a visible region), chains of trapped ions (like in [8]), and molecular structures for further developing such techniques like FRET (described for instance in [12]), atomic chains like carbyne loops (for example, [13]), and microhole array synthesized by femtosecond laser radiation (see [14], for an instance). Let us first provide below some general theoretical premises. More detailed derivations of the corresponding

mathematical model Adenosine triphosphate can be found in [11]. Methods The equations of motion for the state GDC-0068 clinical trial amplitudes We have assumed that the atomic energy levels have no linewidth, so that, only if , the atoms can be able to absorb a photon. Obviously, this is an unrealistic case since it is impossible to have a completely monochromatic wave. In addition, for the case of the Fock initial state, in which we measured the energy precisely of the mode, the average electric field will be zero. In the forth of the law of energy conservation, an emitted photon will correspond to the same frequency (we can say it will occur with a high probability after a quite long time interval if the system has a damping). Therefore, consider a collection of N identical atoms, at positions r 1,…,r α ,…,r N , coupled to a one mode electromagnetic (EM) field. Each atom α = 1..

In this work, we found that both the F- and V-type ATPases are ex

In this work, we found that both the F- and V-type ATPases are expressed C. themocellum. Co-presence of V- and F-type ATPases in a bacterium is uncommon. Previously, only Enterococcus hirae was reported to utilize both types of ATPases [18]. The E. hirae

V-type ATPase differs from typical V-type ATPase in preferentially transporting Na+ [19, 20] instead of H+. In the thermophilic Clostridium fervidus, a second example of Na+-pumping V-type ATPase was reported [21]. It is reasonable to speculate that the V-type ATPase in C. thermocellum is a Na+-pumping ATPase. Most bacteria contain either F-type or V-type ATPase, among those that contain PI3K Inhibitor Library both types of ATPases, new functional variants of ATPases could be identified and their roles in bacterial physiology could be investigated. Bifunctional acetaldehyde/alcohol Mocetinostat dehydrogenase (ALDH-ADH, Cthe_0423, 96 kDa) was detected at over 880 kDa. ADHs could be classified into 3 classes based on their length: short chain ADH (approximately 250 residues) and medium chain ADH (approximately 370 residues) exist in a homotetramer form [22], but a structure of long chain ADH (over 380 amino acids and often as many as 900 amino acid residues) was not reported. The ALDH-ADH of C. thermocellum appears to be a long chain ADH and forms a homo-multimer like the ADH in Entamoeba histolytica [23]. Alcohol dehydrogenases were reported to be membrane-bound protein complexes

[24–26], it is reasonable to PXD101 supplier observe ADH in C. thermocellum membrane fraction. Complexes in lipid transport and metabolism Carboxyl transferase (CT, Cthe_0699, 56 kDa) was identified at ~220 kDa. In eubacteria, CT is part of acetyl coenzyme A carboxylase (ACC) complex, which normally consists

of biotin carboxylase (BC), biotin carboxyl carrier protein (BCCP), and CT. Typically, CT contains two subunits in a stable α2β2 form [27, 28]. But, in Streptomyces coelicolor, the ACC enzyme has Vildagliptin a subunit (590 residues) with fused BC and BCCP domains, and another subunit (530 residues) that contains the fused CT domains [29]. In archaea, ACC is a multi-subunit enzyme, with BC, BCCP and CT subunits. The archael CT subunit is also a single protein (520 residues) in a CT4 form, rather than two separate subunits, which is similar to the β subunit (CT) of the ACC from Streptomyces [30]. In C. thermocellum, CT is a 56 kDa protein, which contains two domains of carboxyl transferase, and we did not detect other ACC subunits on BN/SDS-PAGE. So the CT appears to be a sub complex of CT4 not associated with BC and BCCP. CT was also detected at over 880 kDa, which maybe due to precipitation during electrophoresis or CT formed a large complex with other subunits of ACC. Previous studies also suggested ACC may form a membrane-associated protein complex [31, 32]. Complexes in amino acid transport and metabolism Serine-Acetyl-Transferase (SAT, Cthe_1840, 33.

5 km/h Therefore, only in EAH-C-R4 we can assume that race speed

5 km/h. Therefore, only in EAH-C-R4 we can assume that race speed was one of the factors which influenced EAH in our tested group. Fluid intake and race performance An important finding was the fact that in the ultra-MTBers (R1,R2), fluid intake was positively related to the number of kilometers achieved during 24-hour MTB race, which is in agreement with previous studies [3, 15, 25, 30, 47]. The

ultra-MTBers in the 24-hour MTB races R1 and R2 who drank more finished ahead of those who drank less. Furthermore, the ultra-MTBers in 24-hour MTB R2 with greater body mass losses achieved more kilometers in the race than those with lower body mass losses. In a recent study, Knechtle et al. showed similar findings in 24-hour ultra-runners [30]. In contrast to the ultra-MTBers in R1 and R2, in the ultra-runners in R3 fluid intake was not 4EGI-1 nmr related to race performance. We assume that the ultra-MTBers in R1 and R2 with a better race performance who did not develop EAH drank more than the others, however, still in accordance with IMMDA. In 219 runners in a 100-km ultra-marathon,

the faster runners had a support crew to provide drinks in contrast to the slower runners with no support crew [15]. Presumably, also our faster ultra-MTBers used this possibility of an additional fluid intake. In Knechtle et al. [15], PI3K Inhibitor Library order the faster athletes who probably had a higher sweating rate lost more fluids and consequently drank more fluids. The finding that fluid intake was positively correlated with race performance suggests that athletes in R1 and R2 were drinking appropriately. Faster athletes were working harder and required more water than slower athletes. We hypothesised that in cases of fluid overload, fluid intake would be related to Selleckchem Daporinad post-race body mass, Δ body mass, post-race plasma [Na+], and Δ plasma [Na+], respectively. In none of the races was fluid intake associated with post-race body mass, Δ body mass, Δ plasma [Na+], Δ plasma

volume, or Δ urine specific gravity. Another finding was that the finishers with a better race performance had lower post-race plasma [Na+] in R2 and R3, and a higher body mass loss in R2. Also in Hoffman et al. [11], Knechtle et al. [15] and Noakes [63] faster runners tended to lose more body mass. Likewise, fluid intake Flucloronide was negatively associated with Δ body mass in a recent study [25]. In a 24-hour running race Δ body mass showed no association with post-race plasma [Na+], however, no subject developed EAH [31]. Moreover, fluid intake correlated negatively to average running speed [31]. However, it is difficult to explain the decrease in body mass despite the increased fluid intake and the lower post-race plasma [Na+]. In a recent study, faster runners lost more body mass, and faster runners drank more fluid than slower runners [65]. Also, faster ultra-MTBers in R2 lost more body mass although they drank more.

The reactivity of PCV2-positive serum and mAb 8E4 to these mutant

The reactivity of PCV2-positive serum and mAb 8E4 to these mutants in the IPMA is summarized in Figure 1c. PCV2-positive serum produced strong signals with the two mutants, whereas there was no reactivity with mAb 8E4 (Figure 5j and 5k). Another mutant was then generated in the PCV2/YJ-ORF2 background that contained a single mutation of R to A at position 59 of capsid protein (Figure 1c, rYJ-CL-1-59). The PCV2-positive serum produced a strong signal with the mutant, however, mAb 8E4 did not produce a positive reaction RXDX-101 molecular weight (Figure 5l).

Discussion Several studies have suggested that genetic differences in PCV2 are associated with the geographical region from which the isolates originated, and a classification system that has been proposed divides PCV2 into three genotypes (a, b and c); a and b are the two major genotypes of PCV2 [8,

22–26], but c is only isolated in Demark [9]. Therefore, PCV2c was not used in the present study. Until now, only one serotype has been identified among strains of PCV2. However, mAbs directed against PCV2 (except PCV2c) have shown some differences in reactivity with different PCV2 strains [7, 14]. MAb 8E4 AZD5363 mouse generated in the present study reacted with PCV2a (LG, CL and JF2), by the IPMA and capture ELISA, and had the capacity to neutralize PCV2a (LG, CL and JF2). Therefore, using mAb 8E4, three strains of PCV2a could be differentiated

learn more from three PCV2b strains. However, mAb 8E4 did not give a positive reaction by western blot analysis. Thus, the above results suggest that mAb 8E4 recognizes a conformational epitope in the capsid protein of PCV2. There were several regions of diversity identified by alignment of the amino acid sequences of the capsid protein between PCV2a and PCV2b strains in the present study. The first 46 residues at the N terminus of the capsid protein are probably not involved in the formation of conformational epitopes. This region contains residues rich in basic amino acids and thus may be involved in the formation of the interior surface of the virion, and may interact with the negative charges of genomic DNA during virus assembly, as reported for many icosahedral viruses [27–29]. Amino acids from residue 47 to the C terminus within the capsid protein may be important for formation of PCV2 capsid protein. Several AZD6244 concentration epitopes in the PCV2 capsid protein that are involved in reactions with antibodies are also within this range [6, 7, 30]. Therefore, five regions (aa 47-72, 80-94, 110-154, 190-211 and 230-235) were chosen for construction of PCV2-ORF2-CL/YJ chimeras that included amino acids that differed between PCV2a and PCV2b.

These values are close to those found when other genes have been

These values are close to those found when other genes have been examined; a similar 7-fold increase in adherence to INT-407 cells was found with a cj1461 (methyltransferase) mutant versus the wild-type strain [8]. Mutants in waaF showed a 14-fold reduction in invasion of INT407 cells compared with the wild type strain [9]. Disruption mutants of adhesin-encoding genes cadF and

flpA exhibited a 72% and 62% reduction in adherence, respectively [10]. Insertion SYN-117 chemical structure mutagenesis of cj0588 encoding the TlyA product caused a significant reduction in adherence to Caco-2 cells in culture of C. jejuni strains 81–176 (decreased to 59% compared with wild type) and 81116 (reduced to 48% compared with wild mTOR activation type) [11]. Results

from our assays were quite similar to these studies, showing a 0.5 to 1.0 log reduction in adherence of the isolate without the CJIE1-family prophage (Table 2). The presence of the prophage therefore makes a substantial contribution to the adherence of the lysogenized bacterium. Though the trend to much higher adherence by isolates carrying the prophage was clear in all experiments, the differences in the adherence of isolates with and without the prophage did not reach statistical significance. This was likely partly due to the inter-experimental variability in the adherence and invasion assays, which has been noted before [12] and appears to be a characteristic of the assay. Differences in adherence in vivo can be very significant even when cell culture assays demonstrate no difference between strains [13]. It is critically important that the role of the prophage be assessed in a relevant animal model and with functional mutagenesis

studies. Invasion of Caco-2 cells was reduced in tlyA mutants to 56% and 31% of wild-type in C. jejuni strains 81–176 and 81116, respectively [11]. The 16- to 21-fold difference in invasion detected in the isolates with and without the CJIE1-family prophage was similar to this but much less than the 50-fold reduction in invasion of INT-407 cells resulting ADP ribosylation factor from an insertion mutation of cj1461 [8]. However, the cj1461 mutant also resulted in a motility defect, which is known to have profound effects on invasion [14, 15]. In contrast, no gross alterations in motility were seen in C. jejuni isolates with and without the prophage in the present study. The relative numbers of invaded bacteria expressed as a percentage of those adherent at 30 min post-inoculation was higher than seen by Christensen et al. [16]. However, the differences between adherence and invasion of bacteria with and without the CJIE1-family prophage were consistent in all experiments, suggesting that whatever technical differences resulted in the higher %I/A values were also consistent. The measurable differences in adherence and invasion associated with prophage carriage found in this study appear to be STI571 mouse substantiated.