It has been reported that German cockroach extract is capable of activating protease-activated receptor

(PAR)-2 and provoking IL-8 secretion from bronchial epithelial cells [7], indicating that cockroach allergen may affect the expression of PARs and hypersecretion of cytokines. Indeed, we recently demonstrated that recombinant Per a (rPer a) seven can upregulate the expression of PARs and provoke Th2 cytokine, IL-4 and IL-13, production in P815 cells [8]. As Per a 1s are major allergens in American cockroach and their functions in provoking allergic reactions remain obscure and mast cells play a key role in allergic reactions, we generated rPer a 1.0101 and rPer a 1.0104 and investigated their influence on the expression of PARs and cytokine production in P815 cells in the current study. Patients and samples.  A total of 21 allergic rhinitis patients with positive skin prick to allergen extracts selleck screening library and four healthy controls (HC) were recruited in the study. check details Among the allergic patients, 15 of them were allergic to American cockroach and six of them to ragweed. The informed consent from each volunteer

according to the declaration of Helsinki and agreement with the ethical committee of the First Affiliated Hospital of Nanjing Medical University was obtained. Serum (2 ml) from peripheral venous blood was collected from each patient and HC for Western blot analysis. Expression of Per a 1.0101 and 1.0104 proteins in E. coli.  The procedures were mainly adopted from the one described previously for Per a 7 [8]. Briefly, pMD-Per a 1.0101 and pMD-Per a 1.0104 plasmids were digested and then ligated into unique Nde I and Hind III sites next in a pET-28a expression vector, respectively. The resulting plasmids were transformed into E. coli BL21 (DE3) for the expression of proteins. The final expression condition, under which the proteins were expressed mostly in soluble form, was at 25 °C for 12 h in the presence of 0.6 mm of IPTG. rPer a 1.0101 and rPer a 1.0104 proteins

were purified using BugBuster Ni-NTA His bind purification kit according to manufacturer’s protocol as described previously [8]. Endotoxin contamination was examined with the LAL assay according to the manufacturer’s instructions. The endotoxin levels detected with limulus amebocyte lysate chromogenic endpoint assay for endotoxin (Hycult Biotech, Uden BV, The Netherlands) were very low, being <0.01 EU/mg in rPer a 1.0101 and rPer a 1.0104 proteins. Evaluation of solubility of American cockroach allergens.  In order to express American cockroach allergens in a soluble form in E. coli, a statistical model for prediction of solubility of protein expression in E. coli was used [9]. A composite parameter canonical variable (CV), which is dependent on the contribution of each of the individual amino acid, was calculated as follows: CV = 15.43 (N + G + P + S)/n−29.

A hemodynamic Selleckchem CDK inhibitor sensitivity analysis showed that DM2 networks were predicted to be less robust in their ability to maintain perfused network surface area in the event of upstream terminal arteriole constriction. Conclusions:  This study illustrates that capillary network connectivity is altered by DM2 and this negatively impacts microvascular hemodynamics. This work can serve as a basis for a

more quantitative approach to evaluating DM2 microvascular networks and their potential use as an early diagnostic aid and/or method for identifying therapeutic targets. “
“Please cite this paper as: Cheung and Daanen (2012). Dynamic Adaptation of the Peripheral Circulation to Cold Exposure. Microcirculation 19(1), 65–77. Humans residing or working in cold environments exhibit a stronger cold-induced vasodilation (CIVD) reaction in the peripheral microvasculature than those living in warm regions of the world, leading Ivacaftor in vitro to a general assumption that thermal responses to local

cold exposure can be systematically improved by natural acclimatization or specific acclimation. However, it remains unclear whether this improved tolerance is actually due to systematic acclimatization, or alternately due to the genetic pre-disposition or self-selection for such occupations. Longitudinal studies of repeated extremity exposure to cold demonstrate only ambiguous adaptive responses. In field studies, general cold acclimation may lead to increased sympathetic activity that results in reduced finger blood flow. Laboratory studies offer more control over confounding parameters, but in most studies, no consistent changes in peripheral blood flow occur even after repeated exposure for several weeks. Most studies are performed Unoprostone on a limited amount of subjects only, and the variability of the CIVD response demands more subjects to obtain significant results. This review systematically surveys the trainability of CIVD, concluding that repeated

local cold exposure does not alter circulatory dynamics in the peripheries, and that humans remain at risk of cold injuries even after extended stays in cold environments. Circulatory flow in the extremities adjusts rapidly and dynamically to cold exposure and also to the thermal state of the body [26]. Shortly upon exposure to cold environments, a sympathetically mediated vasoconstriction results in reduced blood flow to the peripheries in favor of a central pooling of blood in the torso and deep body core. Due to the vasoconstriction of the peripheral microvasculature and the high surface area-to-volume ratio, the skin temperature of the fingers and toes tends to rapidly and exponentially decrease to a level approaching that of the ambient environment.

Mannan C. albicans serotype B-specific sera antibodies levels induced by immunization Etoposide cell line with M5-BSA conjugate did no correlate with specific antibody-secreting cells counts. Alteration of mannan C. albicans serotype A-specific IgM and IgG antibody production induced by immunization with M6-BSA conjugate distinctively revealed

IgM/IgG isotype switch (Fig. 4). Purified cell wall mannan does not always maintain their native conformation. To maintain mannan native conformation intact for the analysis of antibodies generated during the immunization with conjugates, we used intact yeast and hyphal cells of C. albicans serotype A in whole cells ELISA assays to determine natural cell wall mannan-specific antibodies levels (Fig. 5). We observed higher yeast and hyphae-specific IgM sera levels following M5-BSA immunization in comparison with control although without significant alteration throughout immunization. M5-BSA conjugate immunization induced significantly higher

levels of yeast and hyphae-specific IgG antibody levels in comparison with IgG levels in sera of controls only after the primary sc injection of conjugate (Fig. 5). IgG levels induced by subsequent M5-BSA conjugate injections were comparable or lower than IgG levels Selleck Dactolisib in sera of controls for both morphological forms of C. albicans serotype A. Yeast and hyphae-specific IgA levels significantly increased after primary M5-BSA conjugate injection and decreased after the primary sc booster administration to the levels comparable with IgA levels in sera of controls (Fig. 5). For M6-BSA conjugate and whole cell–specific IgM levels, we obtained similar results as for M5-BSA conjugate immunization. Etomidate Hyphae-specific IgM levels in immune sera were slightly higher than or comparable with control (yeast form, secondary ip booster injection, 3rd ip) but without significant

alteration throughout immunization (Fig. 5). Immunization with M6-BSA conjugate induced C. albicans serotype A yeast form specific IgG levels comparable with yeast that form specific IgG levels in sera of controls. For hyphal form of C. albicans serotype A, primary sc booster injection and secondary booster injections (both routes of administration, 3rd ip and 3rd sc) induced significantly higher IgG levels in comparison with sera of controls with maximal peak after secondary ip booster injection (Fig. 5). Only for hyphal form of C. albicans, whole cell–specific IgA levels significantly increased after primary M6-BSA conjugate injection. The alterations in C. albicans serotype A whole cell–specific IgG levels after individual administrations of conjugates reveal differences between conjugates. Different changes of the whole cell–specific IgG levels in the course of immunization with conjugates indicated different specificity of induced IgG antibodies and indicate M6-BSA conjugate as preferable for induction of potentially opsonizing antibodies.

We observed no significant change in this measurement (Fig. 2c, P = 0·4691). Plasma TGF-β levels were relatively stable over time in both groups (Fig. 2d). We next measured plasma cytokine and chemokine levels in both groups using multiplex assays. Twenty-seven analytes were measured, and no significant differences were found LY2157299 in the change from baseline between the placebo and sitagliptin groups at any of the time-points. The levels of cytokines and chemokines in both the drug and placebo groups at day 3 are shown in Fig. 2e. Similar results were obtained at other time-points (data not shown). The

percentage of lymphocyte subsets in PBMCs were measured by flow cytometry. The frequency of major lymphocyte subsets (B cells, T cells: both CD4+ and CD8+, NK, NKT cells and monocytes) was measured, and no significant differences were found between groups (data not shown). Vismodegib solubility dmso In addition, numbers of regulatory T cells (CD4+CD25+FoxP3+) were assessed. In mice, increases in regulatory

T cells with DPP-4 inhibition have been reported [18]. However, we observed no significant changes in the percentage of regulatory T cells with sitagliptin treatment (Fig. 3a,b). A small increase in neutrophil and total white blood cell count after sitagliptin treatment was reported to the Federal Drug Administration. In our study, CBC values were also measured, and no significant differences were found between groups in any measure, including white blood cells (WBC) and neutrophils (data not shown and Supporting information, Fig. S1). CD26/DPP-4

is expressed differentially on naive (CD45RA+) and memory (CD45RO+) T cells [25]. Therefore, we measured the percentage of naive and memory T cells in both the CD4+ and CD8+ T cell compartment. The percentage of CD8+CD45RO+ cells increased significantly on day 3 in the sitagliptin group compared to the placebo (P = 0·0104) and was also higher on day 14 (P = 0·0351) (Table 1). We also measured CD26 expression, gating on three populations: CD26– cells defined by fluorescence-1 controls, CD26lo cells, corresponding to the low level found on most naive CD45RA+ cells and CD26hi cells found primarily among the memory CD45RO+ population Glutamate dehydrogenase (Fig. 3c). We observed changes consistent with an increase in CD26 expression early after sitagliptin treatment compared to placebo treatment, including increases in the percentages of CD4+CD45RO+CD26hi and CD8+CD26hi cells, and in the fluorescence levels of CD26 on CD4+CD26hi, CD3+CD26hi and CD3+CD45RO+CD26hi populations (Fig. 3d and Table 1). A significant decrease in the percentage of CD8+CD26lo cells was also observed in sitagliptin-treated individuals compared to placebo, which is consistent with increased CD26, as these cells probably shifted to the CD8+CD26hi population.

The use of force in response to peers’ taking over toys was evident before the first birthday, but more common thereafter, although

only a minority of children in each sample used force. Analysis of a combined data set revealed that force was deployed more often by 2-year-olds than younger infants, and was significantly associated with verbal references to people’s possession of objects. These observations show that toddlers do deploy force intentionally to defend their possessions. “
“We examined the relation between 6- and 7-month-old infants’ (N = 60) manual activity with objects during free play and their perception of the learn more features of dynamic, multimodal events. Infants were habituated to a single event in which a hand reached for and manipulated a colorful, multifeatured object, and a sound was heard (e.g., a hand squeezed a purple round object, causing a whistling sound) and then their response to events that involved a change in the appearance of the object, the action, or the sound was assessed. Infants responded least to changes CHIR-99021 chemical structure in the appearance of the objects, and their

sensitivity to this feature was related to their manual activity with objects during free play. Infants’ responding to changes in the sound or action was unrelated to motor activity, suggesting that at this age motor achievements related to object exploration are associated with infants’ perception of some, but not all, object features. “
“Little research hitherto has examined how individual differences in attention, as assessed using standard experimental paradigms, relate to individual differences in how attention is spontaneously allocated in more naturalistic contexts. Here, we analyzed the time intervals between refoveating eye movements (fixation durations) while typically developing 11-month-old infants viewed a 90-min battery ranging from complex dynamic

to noncomplex static materials. The same infants also completed experimental assessments of cognitive control, psychomotor reaction times (RT), processing speed (indexed via peak look during habituation), and arousal (indexed via tonic pupil size). High test–retest reliability was found for fixation duration, across testing sessions and across types of viewing material. Increased cognitive control and increased arousal were associated with reduced this website variability in fixation duration. For fixations to dynamic stimuli, in which a large proportion of saccades may be exogenously cued, we found that psychomotor RT measures were most predictive of mean fixation duration; for fixations to static stimuli, in contrast, in which there is less exogenous attentional capture, we found that psychomotor RT did not predict performance, but that measures of cognitive control and arousal did. The implications of these findings for understanding the development of attentional control in naturalistic settings are discussed.

Most patients with type 1 diabetes (T1DM) and reduced eGFR have classic glomerular changes of DN regardless Bortezomib clinical trial of albuminuria status. Typical renal structural changes of DN are usually also observed in patients with T2DM, reduced eGFR and albuminuria. However, predominantly interstitial, tubular or vascular damage or near normal renal structure have also been reported in biopsies obtained from patients with T2DM, regardless of eGFR or albuminuria status, in the absence of any other known cause for renal dysfunction. Despite the above, in people with diabetes and proteinuria, non-diabetic kidney disease (NDKD) alone or superimposed on DN changes

is not an uncommon finding.[6] It is important that NDKD is diagnosed. Despite the attention to strict metabolic control and blockade of the renin–angiotensin-aldosterone system,

proteinuric DKD is usually progressive, whereas NDKD is potentially treatable, depending on aetiology. Therefore, we have briefly reviewed the contemporary spectrum of DKD, the histology and clinical predictors of NDKD and present several clinical vignettes (Box 1) to illustrate the variability of renal disease in diabetic patients that have presented to one of our hospitals. Case 1. DKD in T1DM A 47-year-old man was diagnosed with T1DM since childhood, BMS-354825 solubility dmso with multiple complications including proliferative retinopathy, peripheral neuropathy and cerebrovascular disease. Other medical history included obesity and hypertension; there was no family history of renal disease. He presented with worsening nephrotic-range proteinuria (24 h urinary protein 6.5 g) and rapid deterioration in renal function; HbA1C was 9.8%. Renal biopsy confirmed Class IV DN (Fig. 1). Case 2. DKD in T2DM A 38-year-old obese woman presented with rapid weight gain (12 kg in one week) associated with bilateral oedema to her upper thighs. She had significant proteinuria (urinary protein/creatinine Rebamipide ratio 378 mg/mol) with impaired renal function (serum creatinine 122 μmol/L). Past history was notable for gestational diabetes. She was diagnosed with T2DM (HbA1c 13.4%) and renal biopsy confirmed Class III DN with nodular glomerulosclerosis

(Fig. 2). Case 3. FSGS causing nephrotic syndrome A 43-year-old obese woman with 11 year history of T2DM, presented with nephrotic syndrome (gross peripheral oedema, urinary protein/creatinine 913 mg/mol, serum albumin 26 g/L) and preserved renal function (eGFR 77 mL/min). Her HbA1c was 7% with no known diabetic complications. Renal biopsy demonstrated FSGS with mild chronic tubulointerstitial damage (Fig. 4). Case 4. Hypertensive kidney disease A 74-year-old man with T2DM for 7 years was referred with gradually worsening renal impairment (eGFR 21 mL/min). His HbA1C was 6.3% on oral agents with no vascular complications. Other medical history included hypertension and obstructive sleep apnoea. Urine sediment did not show any proteinuria; kidneys were small-sized on ultrasonography.

The see more authors thank Dr. Yusaku Nakamura, the director of Tsuda Hospital, for collection of patients’ serum and urine samples. The authors thank

Dr Makito Ito, Department of Parasitology, Aichi Medical University for valuable technical advice concerning the immune reactions of urine samples and Yasuko Nishimura and Mariko Kuroda for valuable technical assistance. This work was supported by research grant D from Kansai Medical University, by a Grant-in-Aid for Scientific Research (C) 2 from the Japanese Ministry of Education, Culture, Sports, Science and Technology (No. 14570365). The authors declare no conflicts of interest associated with this study. “
“Murine polyomavirus is used in various models of persistent virus infection. This study was undertaken to assess the spatial and temporal patterns of MPyV infection in the brains of immunocompetent (BALB/c) and immunocompromised (KSN nude) mice. MPyV was stereotaxically microinfused into the brain parenchyma, and the kinetics of infection were examined by quantitative PCR. In BALB/c mice, the amount of viral DNA

in the brain peaked at 4 days p.i. and then rapidly diminished. In contrast, MPyV DNA levels increased up to 4 days and then gradually decreased over the 30-day observation period in the brain of KSN mice. In both mouse strains, viral DNA was readily detected around the sites of inoculation from 2 to 6 days p.i., and continued to be detected for up to 30 days p.i. In addition, MPyV infection did not lead to a drastic MLN0128 price induction of innate immune response in the brains, nor did MPyV-inoculated mice show any signs of disease. These results indicate that MPyV establishes an asymptomatic long-term infection in the mouse brain. Members of the family Polyomaviridae (polyomaviruses) are small non-enveloped viruses with a circular

double-stranded DNA genome of approximately 5 kbp (1). Polyomaviruses are widely distributed among vertebrates including birds, rodents Adenosine and primates (1). Mammalian polyomaviruses show narrow host specificities and frequently establish subclinical and persistent infections in their natural hosts (2). The major sites of persistence for mammalian polyomaviruses are the cells of peripheral organs, such as the kidney, urinary tract and spleen (3, 4). In addition, many studies have suggested that the low amounts of JCPyV, a human polyomavirus, are asymptomatically present in the human brain (5). It has also been revealed that the frequency of JCPyV DNA detection in the brain without obvious disease is increased in patients with immunodeficiency disorders (6–8); however, due to its narrow host range in vivo, experimental animals, such as small rodents and non-human primates, do not permit productive replication by JCPyV (9). Thus, the study of JCPyV infection of the brain has been hampered by the lack of suitable animal models.

Amplicons were then purified and cloned into a pGEM-T Easy Vector (Promega, Madison, WI, USA). Two Cys-to-Ser substitution mutants (C213S and C178,213S) were generated by PCR-based site-directed mutagenesis. The primer sets were as follows: for substitution of Cys at 213, 5′-GTACTGGGTGACGCTCATCTGCTC-3′ and 5′-GAGCAGATGAGCGTCACCCAGTAC-3′, and for substitution of Cys at 178, 5′-GTGATATTGACGCTGTCGTGCACG-3′, and 5′-TTCGTGCACGACAGCGTCAATATCAC-3′. PCR to amplify the 5′ and 3′ portions of mutants was performed using the 5′ forward and 3′ reverse primers in combination with the primers above and plasmid MK-8669 manufacturer cloning MoPrP as a template. MoPrP, C213S, and C178, 213S were re-cloned from the pGEM-T Easy Vector into

pET15b (Novagen, Madison, WI, USA) at NdeI and AZD3965 in vitro BamHI sites, and the vectors carrying PrP were transformed into E. coli BL21 (DE3) (Novagen). Expression was carried out according to the manufacturer’s instructions. After solubilization of inclusion bodies in binding buffer (0.5M NaCl, 20 mM imidazole, 8 M urea in 20 mM phosphate buffer, pH 7.4), recombinant

PrPs were purified under denaturing conditions using a HisTrap HP Kit (Amersham, Arlington Heights, IL, USA) according to the manufacturer’s instructions. Purified recombinant PrPs were then dialyzed against 2 M Gdn-HCl and 1 mM EDTA in 50 mM Tris-HCl (pH 8.0). The purities of each PrP were estimated to be >90% by SDS-PAGE and CBB staining. Recombinant PrPs were analyzed by Western blotting with the 3F4 antibody to distinguish recombinant PrP from PrPSc used as seed, and signal intensities were evaluated using a Chemi imager. The scrapie isoform of prion protein was prepared from brain tissue collected from affected animals as described previously (11). Prion-infected mouse brains were homogenized in 10% sarkosyl in 10 mM Tris-HCl (pH 7.4) and then centrifuged at

22,000 g for 10 min. The supernatant was then decanted and centrifuged at 540,000 g for 30 min. The pellets were suspended in TSN with the aid of brief sonication and centrifuged again under the same conditions. The pellets suspended in TSN were treated with 50 μg/mL of PK at 37°C for 60 min. The pellets obtained by centrifugation at 22,000 g for 10 min were washed twice with TSN by centrifugation under the same conditions. The purity of the seed PrPSc fraction NADPH-cytochrome-c2 reductase was examined by SDS-PAGE and silver staining (Wako, Osaka, Japan). All prion strain PrPSc fractions were adjusted to 200 μg/mL by comparing their signal intensities after Western blotting with that of MoPrP. Ten micrograms of MoPrP or C213S, and 5 μg of PrPSc derived from the Chandler strain, were incubated in reaction buffer containing DTT or 2ME at 37°C for 24 hr. After incubation, all PrPs were methanol-precipitated and dissolved in 6 M urea in 50 mM Tris-HCl (pH8.0). mBBr was added to a final concentration of 4 mM, and the solutions incubated for 20 min at 25°C to label sulfhydryl groups.

Grace et al. in a review of ANZDATA listed patients starting dialysis between 2000 and 2010 found only 7% of postcodes outside of major Fludarabine in vivo cities were in the most advantaged quartile, compared with 54% of postcodes within major cities[9] Gray et al. in a similar review of non-indigenous patients on dialysis on found significant differences in disease burden between major capitals (MC), inner remote (IM), outer remote (OM) and very remote (VR) areas – Figure 2.[10] Patients want to be treated close to where they reside to avoid the cost of travel and dislocation involved in visiting metropolitan based clinics.

The implementation of renal palliative/supportive care services in rural areas requires a different model to metropolitan areas if these patients are to have the same standard of care as those in metropolitan areas. General practitioners and renal physicians tend to refer on the basis of previous personal exposure. Providing specialist renal palliative/supportive

care services will need to involve some on the ground outreach services to gain the trust and respect of the local physicians. Any model will need to enhance contact between palliative care services and local physicians. A ‘move aside while we show you how it is done in the city’ approach is unlikely to be successful. The knowledge base for renal palliative Selleckchem Selumetinib care will need to be outsourced to the local physicians, GPs, and palliative care nurses to enhance patient care. Given that it is unlikely that rural units will have specialist renal palliative /supportive expertise on site the DNT committee supports the concept of a hub and spoke model of care to provide equity of service in all rural and remote areas.

This implies that metropolitan palliative care services will have a responsibility to provide outreach services and will need adequate resources. The same model is used to provide transplant services successfully in rural areas and not only allows rural patients to access these services locally but provides up skilling of the local workforce. Developments in information technology such as telemedicine are possible solutions to some of the problems associated with distance and isolation. The current Medicare Sodium butyrate rebate for consultations by videoconferencing should promote and compensate set up costs. This can be easily performed with currently available technology including Skype. There is a potential role for web based on going education for rural renal physicians and palliative care physicians in renal supportive care. This could potentially involve cased based scenarios in a chat room environment. A model currently working in the New England Area involves having a local supportive care nurse who is experienced in dialysis assess all patients referred to the service. Referrals can be from nursing colleagues, GPs, allied health workers and renal physicians.

Total melanoma tumor counts were obtained on day 22 by adding

the number of foci counted in the superior, middle, inferior, and postcaval lobes of the right lung to the number of foci counted in the left lung. The endpoint of the study was originally defined as 100 metastases per lung set. All procedures and analyses were performed blind, without knowledge of the test samples. Differences in sCTLA-4 levels between treatments were analyzed using the Wilcoxon Matched-Pairs Signed-Ranks Test, and differences in metastatic melanoma tumor load by Mann–Whitney U test. This work was funded by an endowment grant (04/50) from NHS Grampian, UK, and a Knowledge Transfer Grant from the University of Aberdeen. Dr. Lekh N. Dahal was supported by a studentship from the University of Aberdeen and by Arthritis Research UK (Grant no. 19282). The authors are grateful to Professors this website John Todd and Linda Wicker (University of Cambridge, UK) for helpful discussions and provision of reagents. The authors thank Teva Pharmaceuticals, Tikva, Israel, for their collaborative support in the murine melanoma model. The authors also thank Drs Jennifer Niven and Isabel Crane for their help with the IRBP model of experimental autoimmune uveitis. The authors

(FJW, LND, and RNB) have filed a patent covering the use of the monoclonal Ab JMW-3B3 as a therapeutic. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or before typeset. Technical www.selleckchem.com/screening/pi3k-signaling-inhibitor-library.html support issues arising from supporting information (other than missing files) should be addressed to the authors. “
“Intra-amniotic pathogens and by-products activate innate immune responses encompassing multitudes of signaling molecules and pathways that can result in spontaneous preterm birth (PTB). This study investigates fetal membrane response to bacterial stimulation using a bioinformatics approach. Dysregulated biomarker (IL1-β, IL-2, IL-8, IL-10, and TNF-α) data from fetal membranes at term stimulated with Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma

hominis, E. coli, Group B Streptococci, Polyporhans gingivalis, or Gardnerella vaginalis with 50% (v/v) amniotic fluid (AF) were analyzed by Ingenuity Pathway Analysis. In racially stratified analysis, networks representing late-stage immune inflammation were seen in African-Americans in AF absence. Inflammation was dominant in AF presence as well. In Caucasians, late-stage immune response was dominant with AF, but not in its absence. Fetal membrane biofunctions in response to bacteria reflect early- and late-stage innate immune defenses that vary based on the presence of AF and subject race. “
“Here construction of an attenuated mutant of an avian pathogenic Escherichia coli serovar O78 using an allelic exchange procedure is described.