In this step, opportunities can be provided to patients for addressing misinformation about their diseases and helping them realize unrealistic goals, because they might misunderstand their condition and have unreasonable or unrealistic goals for treatment. However, physicians should not modify or manipulate the goals. After the detailed conversation, patients decide their treatment goals. When patients have multiple goals, they need to rank the importance of the goals during the conversation. Goals

might be related to symptoms (e.g. frequency, urgency, or nocturnal), physical impact (e.g. ability to work, travel, or perform activities), emotion learn more (e.g. worry about leaking urine), sexual function (e.g. decrease in sexual desire), social relationships (e.g. embarrassment in public, avoidance of social activities), Romidepsin clinical trial coping strategies (e.g. wearing pad or changing underwear), or quality of life (e.g. sleep quality). The next step is to identify patient expectations for treatment benefit. Goals are typically stated in terms of lifestyle events that are

affected by the health problem. For example, a patient may say that his or her treatment goal is to “be able to sleep at night without going to the toilet”, or “travel without worry of going to the toilet”. However, patient expectations are generally stated in terms of symptom relief. Additionally, the expectations might include the entire treatment experience, including physician personality, waiting times, hospital facilities, and complications. As in setting goals, physician should Immune system not modify or manipulate patients’ expectations. The final step is to assess goal achievement after treatment. At that time, patients review their pretreatment goals and rate their perceptions of goal achievement compared with the initial expectations. This can be measured using

a visual analog scale or Likert scale. The efficacy of antimuscarinics has been demonstrated in the treatment of overactive bladder (OAB) through well-designed, randomized controlled trials; however, the clinical significance of these findings is in doubt.5–7 Poor compliance and persistence with medication suggest that many patients perceive little ongoing benefit and have unmet expectations from the treatment.8,9 One of the reasons for the discrepancy between investigational and clinical points of view is the lack of patient-driven criteria in outcome assessment. Thus, investigators who are working on outcome research have been testing patient-reported goal achievement in the treatment of OAB.10–12 Choo et al.10 first reported the efficacy of antimuscarinics in terms of goal achievement in OAB patients. After a 12-week treatment with tolterodine, the median rates of goal achievement for each OAB symptom were 60% for frequency, 60% for urgency episodes, and 80% for urgency incontinence compared with the initial expectation of symptom improvement.

2B). Overall, these data suggest that TREM-1 expression in H-iDCs is dependent at least in part on HIF-1. TREM-1 is endowed with proinflammatory and immunoregulatory

potential upon cross-linking [29, 30]. To investigate TREM-1 function in H-iDCs, 4-day H-iDCs were plated on Panobinostat a plastic surface coated with a specific anti-TREM-1 agonist mAb or an isotype-matched control anti-HLA-I mAb for 24 h under hypoxia, and the expression of surface antigens was assessed by flow cytometry. As shown in Figure 3A, surface expression of the T-cell costimulatory molecule, CD86, the CD83 maturation marker, and the CCR7 and CXCR4 chemokine receptors was strongly enhanced in response to TREM-1 compared with that from HLA-I triggering, both in terms of mean fluorescence intensity and/or percentage of positive cells, while no modulation of CD40 costimulatory molecule selleck chemicals llc was observed. We analyzed in parallel supernatants for cytokine and chemokine content by ELISA. Enhanced secretion of

several proinflammatory, Th1/Th17 cell-priming cytokines and chemokines, such as TNF-α, IL-1β, IL-12, CXCL8, CCL5, CCL17, and osteopontin (OPN), was measured in response to TREM-1 engagement compared with that in cells triggered with anti-HLA-I mAb (Fig. 3B). No substantial differences in phenotype and cytokine secretion were observed in HLA-I-stimulated H-iDCs relative to that of unstimulated cells or cells stimulated with an irrelevant isotype-matched mAb (data not shown), confirming that H-iDC activation by anti-TREM-1 mAb was specific. To investigate the functional relevance of TREM-1

engagement on H-iDCs, we compared the ability of anti-TREM-1- and anti-HLA-I-stimulated H-iDCs to activate allogeneic T cells in a 5 day MLR assay. As shown in Figure 4A, T-cell proliferation was significantly higher after culture with allogeneic H-iDCs previously cross-linked with anti-TREM-1 mAb than with anti-HLA-I-stimulated H-iDCs. Moreover, T cells alloactivated with TREM-1-triggered H-iDCs showed an increased ability to produce the Th1 and Th17 cytokines, IFN-γ, and IL-17, compared with those cultured with H-iDCs stimulated with anti-HLA-I (Fig. 4B) or unstimulated (data not shown). No significant differences were observed in the secretion of the typical Th2 Thalidomide cytokines, IL-4 and IL-10, by T cells recovered from coculture with TREM-1- and HLA-I-triggered H-iDCs. Overall, these data suggest that TREM-1 engagement on H-iDCs induces phenotypic and functional changes typical of maturation, stimulating their Th1/Th17-polarizing proinflammatory activity. DCs immunostimulatory properties are acquired during a complex differentiation and maturation process tightly regulated by a network of inhibitory and activating signals transduced by multiple families of cell surface receptors [3, 8, 9, 25-27].

These data show

that like other lymphocyte populations, including NK cells, iNKT cells are sensitive to the immunosuppressive effects of adenosine. Adenosine-related compounds cause the simultaneous engagement of Gs- and Gi-coupled adenosine receptors. We therefore asked whether ligation of the predominant high-affinity A2aR during TCR-mediated stimulation would modulate activation or effector functions, i.e. Fulvestrant datasheet cytokine production, of iNKT cells. To exclude a lack of costimulatory molecules accounting for a lack of IFN-γ secretion, we next used BMDC at day of culture, typically consisting of both immature and mature cells. To exclude responses of the BMDC to A2aR modulation, cells were fixed upon α-galactosylceramide (α-GalCer)-loading. Enriched iNKT cell preparations were thus stimulated in the presence of a specific A2aR agonist or antagonist. Comparable to the effects of the stable adenosine analogue, the exposure to A2aR agonist CGS21680 during the stimulation period inhibited the production of IFN-γ by iNKT cells. In striking contrast, CGS21680 led to a significant increase in IL-4 production. Conversely, the A2aR antagonist ZM241485 inhibited the iNKT cell-mediated mTOR inhibitor secretion of IL-4 and concomitantly increased the production of IFN-γ (Fig. 2B), markedly skewing the Th1/Th2 ratio of cytokines produced by iNKT cells toward IFN-γ. These data were corroborated

by a similar analysis of a human iNKT cell line (Fig. 2C). The requirement of A2aR signaling for IL-4 production clearly is in opposition to the effects of A2aR ligation on conventional T cells, which are inhibited non-selectively 24. These data also provide an explanation for the phenomenon Beldi et al. recently described 17, in which iNKT cells lacking the ecto-enzyme CD39, and hence unable to generate adenosine, were not able to produce IL-4 upon CD1d-mediated activation. To determine

the physiological in vivo significance of these findings, we asked whether iNKT cells in mice lacking the predominant A2aR would be functionally altered. We injected A2aR KO mice or WT mice with α-GalCer and tested the cytokine production 90 min and 5 h later, reflecting the time of appearance Methamphetamine in serum. The production of IL-4 and IL-10 upon α-GalCer administration can be observed early after activation, whereas IFN-γ secretion by iNKT cells requires IL-12 produced by DC upon maturation and hence are detectable later after injection. We detected increases in all four tested cytokines (IL-4, IL-10, TGF-β and IFN-γ) in the serum of α-GalCer-injected WT mice compared to un-injected mice (data not shown). Comparable with the in vitro results, iNKT cells in the absence of A2aR produced significantly lower levels of IL-4 upon α-GalCer injection (Fig. 3A). The expression of another Th2 cytokine IL-10 was also markedly decreased in the A2aR KO mice. In marked contrast, but also comparable with the in vitro results, IFN-γ was increased in the A2aR-deficient mice.

“
“Pretangles are cytoplasmic tau immunoreactivity in neurons without apparent formation of fibrillary structures. In Alzheimer disease, such tau deposition is considered to represent a premature state prior to fibril formation (AD-pretangles), later to form neurofibrillary

BMS 354825 tangles and finally ghost tangles. This morphological evolution from pretangles to ghost tangles is in parallel with their profile shift from four repeat (4R) tau-positive pretangles to three repeat (3R) tau-positive ghost tangles with both positive neurofibrillary tangles in between. This complementary shift of tau profile from 4R to 3R suggests that these tau epitopes are represented interchangeably along tangle evolution. Similar tau immunoreactivity without fibril formation is also observed in corticobasal degeneration (CBD-pretangles). CBD-pretangles and AD-pretangles share: (i) selective 4R tau immunoreactivity without involvement of 3R tau; and (ii) argyrophilia with Gallyas silver impregnation. However, CBD-pretangles neither evolve into ghost tangles nor exhibit 3R tau

immunoreactivity even at the advanced stage. Because electron microscopic studies on these pretangles are selleckchem quite limited, it remains to be clarified whether such differences in later evolution are related to their primary ultrastructures, potentially distinct Rebamipide between AD and CBD. As double staining for 3R and 4R tau clarified complementary shift from 4R to 3R tau along evolution from pretangles to ghost tangles, double immunoelectron microscopy, if possible, may

clarify similar profile shifts in relation to each tau fibril at the ultrastructural dimension. This will provide a unique viewpoint on how molecular (epitope) representations are related to pathogenesis of fibrillary components. “
“This chapter contains sections titled: Introduction Anatomy and Physiology of the Innerear Access of Ototoxicants to the Inner Ear Methods for Studying the Inner Ear Effects and Actions of Ototoxic Drugs Classes of Ototoxic Agents Ototoxic Interactions Summary References “
“Nasu-Hakola disease (NHD) is a rare autosomal recessive disorder, characterized by progressive presenile dementia and formation of multifocal bone cysts, caused by genetic mutations of DNAX-activation protein 12 (DAP12) or triggering receptor expressed on myeloid cells 2 (TREM2). TREM2 and DAP12 constitute a receptor/adapter signaling complex expressed on osteoclasts, dendritic cells (DC), macrophages and microglia. Previous studies using knockout mice and mouse brain cell cultures suggest that a loss-of-function of DAP12/TREM2 in microglia plays a central role in the neuropathological manifestation of NHD. However, there exist no immunohistochemical studies that focus attention on microglia in NHD brains.

We then employed H5N1 infection as

a model to study the antiviral activity of α-defensin-induced MxA. The viral plaque assay in Fig. 4A shows that, similar to IFN-α-pretreated HGECs, α-defensin-1, -2, and -3-pretreated cells significantly inhibited H5N1 replication, suggesting a functional MxA protein. On the other hand, β-defensin-1, -2, -3, and LL-37-pretreated HGECs poorly inhibited viral replication. These findings AZD9291 ic50 were confirmed by microscopically observed cytopathic effects (data not shown). To confirm the antiviral activity of MxA against H5N1, we transfected HGECs with MxA-targeted siRNA, treated the cells with α-defensin-1 overnight, and then infected them with H5N1 virus. MxA-targeted siRNA greatly reduced levels of MxA mRNA expression by 95%, (Fig. 4B) and effectively abolished inhibition of viral replication by 93% in H5N1-infected HGECs (Fig. 4C). These findings were supported by microscopically observed cytopathic effects (Fig. 4D). α-defensins are known as major proteins secreted by PMNs [[32]]. In the physiological condition of healthy gingiva, PMNs and their products are present in the tissue and the crevicular fluid in the gingival sulcus [[33, 34]]. In vitro culture of PMNs (5 × 106 cells/mL)

for 6 h led to secretion of α-defensins in supernatants (which ranged from 90 479 to 98 714 pg/mL). To investigate the role of the PMN-derived α-defensins Ku0059436 in MxA expression, we cultured HGECs with 6 h PMN supernatants. Under this condition, expression of MxA at both mRNA and protein levels in HGEC was observed after 6 h and 24 h treatment, Avelestat (AZD9668) respectively (Fig 5A and B). The MxA-inducing activity was diminished when neutralizing antibody against

α-defensins was added to the culture, whereas neutralizing antibodies against type I IFN (IFN-α and IFN-β) had no effect (Fig. 5B). These data suggest that PMN-derived α-defensins were responsible for the observed MxA expression. The immunostaining results to detect epithelial MxA were obtained using the oral, but not the sulcus, side of periodontal tissue (Fig. 2) because the epithelium at the sulcus side, especially for the junctional epithelium, is generally lost or torn during the surgical procedure. Fig. 6A depicts anatomic landmarks of the gingival sulcus. In this study, we were able to obtain two specimens of gingival sulcus area from healthy periodontal tissue. We then investigated localization of MxA protein in the healthy sulcus and also in relation to α-defensin. Fig. 6C shows that MxA protein was consistently expressed throughout epithelial cells of periodontal tissues. MxA staining was especially intense in the junctional epithelium (Fig. 6C). α-defensins were identified in small round cells with PMN morphology, most of which were found in the connective tissue layer (Fig. 6E). Migratory PMNs in junctional epi-thelium were also observed and highlighted in Fig. 6D.

In this study, the secretion of pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, IL-8 and IL-1β; Th1 cytokines interferon-gamma (IFN-γ), IL-2 and tumor necrosis factor-beta (TNF-β); and Th2 cytokines IL-4, IL-5 and Selleckchem Maraviroc IL-10 by the peripheral blood mononuclear cells (PBMCs) of pulmonary tuberculosis patients was studied. PBMCs were cultured in vitro in the absence and presence of complex mycobacterial antigens and peptides corresponding to 11 regions of difference (RD) of Mycobacterium tuberculosis that are deleted/absent in all vaccine strains of Mycobacterium

bovis bacillus Calmette-Guérin (BCG). The culture supernatants were tested for secreted cytokines by FlowCytomix assay. PBMCs from the majority of patients (53–100%) spontaneously secreted detectable concentrations of all cytokines tested, except for IL2 (29%) and IL-10 (41%). The profiles of proinflammatory cytokines were largely similar for various complex antigens or

RD peptides. However, with respect to Th1 and Th2 cytokines, the antigens could be divided into three groups; the first with Th1-bias (culture filtrate of M. tuberculosis, RD1, RD5, RD7, RD9 and RD10), the second with Th2-bias (whole cells and cell walls of M. tuberculosis, RD12, RD13 and RD15), and the third without Th1/Th2-bias (M. bovis BCG, RD4, RD6 and RD11). Complex mycobacterial antigens and RD proteins with Th1- and Th2-biases may have roles in protection and pathogenesis CHIR-99021 manufacturer of tuberculosis, respectively. Tuberculosis is a major global health problem, about one third of the world’s population being infected with M. tuberculosis, 9 million people developing active disease and 2 million people dying of this disease each year (1). In TB, both protection and pathogenesis are mediated by cellular responses, which primarily Forskolin mouse involve interactions of lymphocytes (mainly T cells) and phagocytes of the monocyte/macrophage lineage (2, 3). These interactions are mostly dependent on the interplay of cytokines produced by these

cells. Although, many cytokines contribute to protective immunity, Th1 responses, dominated by the secretion of cytokines IL-2, TNF-β and IFN-γ, are considered the major players in protective immunity against M. tuberculosis (2–9). In contrast, Th2 responses, characterized by the secretion of cytokines IL-4, IL-5 and IL-10, correlate with lack of protection and increased severity of TB (10–13). In particular, IL-10 is strongly associated with reduced resistance and chronic progressive TB (14). In addition, IL-10 deactivates macrophages and inhibits secretion of the protective Th1 cytokines (14). Furthermore, innate immune response-related pro-inflammatory cytokines with chemotactic activity, namely IL-1β, IL-6, IL-8 and TNF-α, initiate events that curb mycobacterial growth by recruiting monocytes into the lesions and activating them to kill the pathogens (15).

[69] Paradoxically, many of these in vitro approaches to this website prion disease research have been developed using materials from high-titer rodent models of sheep scrapie. The challenge for human prion disease research is to apply these emerging techniques to the study of human prions in humans. Molecular strain typing in the form of classifying the mobility and glycoform ratio of protease-resistant prion protein by Western blotting is a remarkably useful adjunct to neuropathological assessment during the post-mortem diagnosis of human prion diseases

(Fig. 1). The glycoform ratio difference between vCJD and all forms of sCJD is a remarkably robust phenomenon, although the mechanism underlying it remains obscure. All cases of vCJD examined show type 2B PrPres, irrespective of brain region assayed and the PrPres type is also found in lymphoreticular tissues, albeit with presumably tissue-specific minor modification of mobility and an accentuation of the glycoform ratio. Similarly sCJD cases are characterized by a narrow range of glycoform ratios, distinct from vCJD, and the presence of either type 1 or type 2 PrPres (type 1A and type 2A). The PrPres types found in the brain in iCJD and kuru resemble those found in sCJD (type 1A and type 2A), from which they were presumably derived. Individual cases of gCJD, GSS and FFI usually check details have type

1 or type 2 PrPres, but with a glycoform ratio in which the non-glycosylated component is under-represented (which we have termed A/B). However, this is not always true and a broad spectrum of glycoform ratios can be found in genetic prion diseases. Moreover, some cases of GSS are characterized by an approximately 8 kDa (N- and C-terminally truncated) PrPres fragment, and some cases of FFI have little detectable PrPres at all. Despite the diagnostic utility, a simple

one-to-one correspondence between PrPres type and disease phenotype (and by implication to agent strain) seems unlikely in principle and is complicated by the facts. First, the choice of analyzing only that fraction of PrPSc which ID-8 survives a particular concentration of protease may seem arbitrary. Second, the interpretation of a molecular population variable, such as glycosylation site occupancy, as conforming to two or three discrete types, could be seen as simplistic. Lastly, protease digestion may be considered to be a somewhat blunt instrument to distinguish secondary and higher-order conformational differences in PrPSc. Even when genotype (mutations and polymorphisms) is taken into account, three major types (1, 2, 8 kDa) and three wild-type genotypes (MM, MV and VV) provide insufficient molecular variation to account for all the phenotypic variations observed. For example, two forms of sCJD share methione homozygosity and type 2A PrPres but one form closely resembles FFI (without the causative mutation) and the other is CJD-like.

After Rucaparib cell line transplantation the quantity of TFH-cells was the highest in patients with pre-existent DSA. In kidney biopsies taken during rejection, TFH-cells co-localized with B cells and immunoglobulins in follicular-like structures. Our data on TFH-cells in kidney-transplantation demonstrate that TFH-cells may mediate humoral alloreactivity, which is also seen in the immunosuppressed milieu. “
“Compared with HLA-DR molecules, the specificities of HLA-DP and HLA-DQ molecules have only been studied to a limited extent. The description of the binding motifs has been mostly anecdotal and does not provide a quantitative

measure of the importance of each position in the binding core and the relative weight of different amino acids at a given position. The recent publication of larger data sets of peptide-binding to

DP and DQ molecules opens the possibility of using data-driven bioinformatics methods to accurately define the binding motifs of these molecules. Using the CX-5461 research buy neural network-based method NNAlign, we characterized the binding specificities of five HLA-DP and six HLA-DQ among the most frequent in the human population. The identified binding motifs showed an overall concurrence with earlier studies but revealed subtle differences. The DP molecules revealed a large overlap in the pattern of amino acid preferences at core positions, with conserved hydrophobic/aromatic anchors at P1 and P6, and an additional hydrophobic anchor at P9 in some variants. These results confirm the existence of a previously hypothesized supertype encompassing the most common DP alleles. Conversely, the binding motifs for DQ molecules appear more divergent, displaying unconventional anchor positions and in some cases rather unspecific amino acid preferences. The MHC performs an essential role in the cellular immune system, and regulates

immune responses through presentation of processed antigens to T lymphocytes. The MHC is also widely studied because of its association with many autoimmune and inflammatory diseases, including type I diabetes, rheumatoid arthritis, multiple sclerosis and Crohn’s disease, why and certain MHC alleles have been linked to susceptibility to infectious diseases such as malaria and HIV (reviewed in ref. 1). Unlike MHC class I, which samples peptides from cytosolic proteins, MHC class II molecules present short peptide sequences derived from extracellular proteins. Human MHC class II molecules are heterodimers consisting of an α-chain and a β-chain encoded on chromosome 6 in one of three HLA loci: DR, DP and DQ. Compared with DR molecules, the specificities of DP and DQ molecules have only been studied to a limited extent, and their binding motifs are poorly characterized and understood.

A p-value of <0.05 was considered significant. This work was supported by grants R01AI063331 and R01DK091191 from the National Institutes of Health. L. F. was supported by a Research Career Development Award from the Crohn's and Colitis Foundation of America. We would like to thank Randal Kaufman and Yingjie Chen for Pkr−/− mouse femurs. We would also like to thank Peter Kuffa for help in generating anti-Nlrp3 antibody and Sharon Koonse for animal husbandry. Luigi Franchi is an employee of Lycera, a biotechnology

company specializing in the field of inflammation. Raf inhibitor “
“Dengue viruses infect cells by attaching to a surface receptor which remains unknown. The putative receptor molecules of dengue virus type 2 on the surface of mosquito (AP-61) and mammalian (LLC-MK2) cell lines were investigated. The immunochemical detection and structural analysis of carbohydrates demonstrated that the neutral

glycosphingolipids, L-3 (GlcNAcβ1-3Manβ1-4Glcβ1-1’Cer) in AP-61 cells, and nLc4Cer (Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-1’Cer) in LLC-MK2 cells were recognized by the virus. These findings strongly suggest that neutral glycosphingolipids share the key determinant for virus binding and that the β-GlcNAc residue may play an important role in dengue virus binding to the host cell surface. Dengue viruses are the causative agents of dengue fever MAPK inhibitor and its associated complications, dengue hemorrhagic fever and dengue shock syndrome (1).

These lethal conditions may be caused by any of the four virus serotypes (DENV-1, DENV-2, DENV-3 and DENV-4) (2). There is neither effective treatment, nor vaccines currently available for prevention of dengue diseases. A prerequisite for development of antiviral strategies against dengue virus is a better understanding of the infection and replication processes (3). In regard to invasion of the host cells, the virus must attach to the cell Florfenicol surface via cellular receptor(s), but the viral receptor is still unclear. Several studies have demonstrated putative receptor(s) for dengue viruses. By using multiple approaches and different cell lines with different strains of dengue viruses, numerous candidates for dengue virus receptor(s) have been provided. Possible receptors for dengue virus on mammalian cells that have been identified include HS-type GAGs (1, 4–7), C-type lectins dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin and liver/lymph node-specific intercellular adhesion molecule-3-grabbing integrin (8, 9), glucose-regulated protein 78 (10) and the non-integrin receptor, laminin receptor 1 (11, 12). In the case of receptors of mosquito cells, two glycoproteins with molecular masses of 40 and 45 kDa have been identified (13, 14).

E. coli strains were grown in LB medium or TSB (BD Diagnostic Systems, Sparks, MD, USA). Construction of a crp deletion mutant of J29 was performed by the methods of Donnenberg and Kaper (37). In short, the crp gene was amplified by PCR with E. coli J29 as the template. The amplified fragment was cloned into the BamH I and Sal

I sites of pMW119. A 351-base pair internal deletion of crp gene was created by digestion with Hinc II (Toyobo Life Science, Tokyo, Japan) and ligation with T4 DNA ligase (Boehringer Mannheim, Burlington, ON, Canada) according to the manufacture’s recommendations. The internally deleted gene was subcloned into pCVD442 (37), and the resulting Romidepsin mouse plasmid transformed into E. coli SM10λpir (38) by electroporation followed by selection with ampicillin. This recombinant plasmid was transferred from E. coli SM10λpir into a nalidixic resistant clone of E. coli J29 by filter mating followed by selection with nalidixic acid and ampicillin. Plasmid excision events were identified by selection for sucrose resistance followed by screening for ampicillin and kanamycin susceptibility, which is indicative of loss of suicide vector sequences. Deletion of the chromosomal crp gene was confirmed by PCR screening. The primer sets and PCR conditions have been described previously (36). One of the resulting mutant strains was designated AESN1331; the mutant strain was cultured in TSB and stored

as a Napabucasin frozen culture (-80°C) in 50% glycerol. Fertilized eggs and chickens of SPF white leghorns of the

line M were obtained from the Laboratory Animal Research Station, Nippon Institute for Biological Science (Yamanashi, Japan). The eggs were Ribonucleotide reductase incubated at 37–38°C in a relative humidity of approximately 55%. Animal utilization protocols were approved under the guidelines of Nippon Institute for Biological Science on Animal Care. The presence of the O78 surface antigen was established by slide agglutination with the corresponding antiserum (Denka Seiken, Tokyo, Japan). Colony diameter was tested by culturing bacteria on trypticase soy agar (BD Diagnostic Systems) for 24 hrs at 37°C and then measuring the diameters of three separate colonies with a ruler (1 mm resolution). Colony color was assessed following culturing on MacConkey agar (BD Diagnostic Systems for 24 hrs at 37°C. Biotyping was performed with the API20E bacterial identification system (bioMerieux sa, Marcy l’Etoile, France). For assay of hemolytic activity, blood agar plates containing 5% sheep blood in LB medium were streaked with over-night cultures and examined for clear zones of erythrocyte lysis after 20 hrs incubation at 37°C (36). Adsorption of Congo red was tested by the method of Corbett et al. (39). Detection of the following genes was performed by PCR: papC, which encodes P fimbriae; tsh, which encodes temperature-sensitive hemagglutinin; cvaC, which encodes colicin V, and iss, which encodes increased serum survival protein.