An increasing number of studies in haemodialysis (HD) patients show benefits of alternative HD regimens providing more effective treatment and improving surrogate end-points and quality of life.1 There has been growing interest in changes in HD prescription

to facilitate these treatments; and alternative HD schedules have thus become an increasingly popular alternative to conventional thrice-weekly HD (3.5–5 h per session).2–5 Alternative regimens provide greater flexibility and predominantly involve augmentation of the frequency and/or duration of HD. In Australia, longer and more frequent HD is the commonest alternative regimen and is often undertaken in the home environment. This is in contrast to other countries such

as the USA where longer and more frequent HD is predominantly Selleck LY2109761 performed in-centre, and shorter and more frequent dialysis is the most common alternative HD regimen.6 This review outlines dialysis prescriptions for alternative HD regimens, including differences compared with conventional HD with regards to dialysate click here concentrations, blood and dialysate flow rates, ultrafiltration rates, anticoagulation and adequacy of HD. Haemodialysis schedules can vary with respect to duration per session and frequency of sessions per week. HD duration can vary to involve extended hours dialysis referring to 6–12 h performed either during the day or at night (nocturnal). The frequency of HD can also range from three to seven times per week either during the day or nocturnal. ‘Quotidian’ (which literally means ‘daily’) HD has often described any regimen that is undertaken

more than three times weekly and the commonest modalities are short-daily HD (SDHD) and nocturnal HD (NHD) (Table 1).7 SDHD refers to regimens that are delivered between 4 and 6 days almost per week usually <3 h per session. NHD provides extended hours HD overnight and is delivered anywhere from 3 to 7 nights per week, including an alternate-night regimen (3.5 nights per week). The SDHD and NHD may be more ‘physiological’ modes of dialysis than conventional HD with potentially greater solute clearance and more rigorous control of biochemical and physical parameters (Table 2). The rationale for more frequent HD includes a reduction of the interdialytic interval, with less fluid gains and increased haemodynamic stability, and an increase in the efficiency of solute clearance. The rationale for increased duration of HD includes an increase in removal of solutes, especially those cleared in a time-dependent fashion (such as phosphate and β2 microglobulin), and an improvement in haemodynamic stability, with lower pump speeds and slower ultrafiltration rates. Multiple publications report significant improvements with SDHD and NHD for quality of life, anaemia and mineral metabolism management, sleep physiology and cardiovascular end-points including hypertension and cardiac structure and function.

Although falling within the same rhythmic class, Basque and Spanish exhibit significant differences in their distributions of vocalic intervals (within-rhythmic class variation). All infant groups in our study successfully discriminated between the languages, although each group exhibited a different pattern. Monolingual Spanish

infants succeeded only when they heard Basque during habituation, suggesting that they were influenced by native language recognition. The bilingual and the Basque monolingual infants showed no such asymmetries and succeeded irrespective of the language of habituation. Additionally, bilingual infants exhibited longer looking times in the test phase MI-503 as compared with monolinguals, reflecting that bilingual infants attend to their native languages differently than monolinguals. Overall, results suggest that bilingual infants are sensitive to within-rhythm acoustic regularities of their native language(s) facilitating language

discrimination and hence supporting early bilingual acquisition. “
“Recent work has suggested the value of electroencephalographic (EEG) measures in the study of infants’ processing of human action. Studies in this area have investigated desynchronization of the sensorimotor mu rhythm during action execution and action observation in infancy. Untested but critical to theory is whether the mu rhythm shows a differential response to actions which share similar goals but have different motor requirements or sensory outcomes. By varying the invisible property of object weight, buy RXDX-106 those we controlled for the abstract goal (reach, grasp, and lift the object), while allowing other aspects of the action to vary. The mu response during 14-month-old infants’ own executed actions showed a differential hemispheric response between acting on heavier and lighter objects. EEG responses also showed sensitivity to “expected object weight” when infants simply observed an experimenter reach for objects

that the infants’ prior experience indicated were heavier vs. lighter. Crucially, this neural reactivity was predictive—during the observation of the other reaching toward the object, before lifting occurred. This suggests that infants’ own self-experience with a particular object’s weight influences their processing of others’ actions on the object, with implications for developmental social-cognitive neuroscience. “
“Hierarchical structures are crucial to many aspects of cognitive processing and especially for language. However, there still is little experimental support for the ability of infants to learn such structures. Here, we show that, with structures simple enough to be processed by various animals, seven-month-old infants seem to learn hierarchical relations.

Spiller et al. similarly reported that AGP reduced neutrophil migration into the peritoneum in mild sepsis derived from the CLP procedure six hours post-CLP [41]. These discordant findings may derive from differences in procedures and the use of different rodent species, and from the investigation of different vascular beds and anatomical locations, which respond differently to inflammatory stimuli. While we did not measure leukocyte levels in Sunitinib nmr the peritoneum, we observed no effect of AGP on the leukopenia associated with either endotoxemia or CLP. In addition, our experiments were completed within four hours of administration

of LPS or perforation of the cecum, whereas the other reports extended the period of observation

post-CLP to between six hours and one week. Enormous quantities of human plasma are currently fractionated to produce purified plasma protein products such as albumin Sorafenib in vivo or immunoglobulin concentrates [5]. AGP is a currently discarded by-product of plasma fractionation. Our results suggest that AGP may have a role to play in ameliorating the early hepatic inflammatory response that, if uncontrolled, contributes to considerable mortality and morbidity in the critically ill. Future studies are warranted to address this possibility, and to explore the possibility that the timing of AGP administration could be critical to its potential benefit. Further mechanistic understanding may require long-term delivery of AGP during the further development of septic shock in animal models, for instance as achieved in previous work from

our laboratories using adenoviral delivery of cytokines to the liver [30]. Fluid resuscitation in sepsis is a hot topic in the clinical literature. With the demise of the hydroxyethyl starches for this Interleukin-3 receptor patient population, investigators are looking to alternative colloids to improve microcirculatory perfusion and systemic hemodynamics. Our work supports other preclinical studies suggesting the natural positive acute phase plasma protein AGP may have therapeutic potential. Canadian Blood Services (CBS) provided a competitive Graduate Fellowship to TRM (award number XH00030) and competitive operating grant support (award number XH00038) to AEF-R and WPS. Some additional funds used in the final stages of this project were derived from funding of CBS research by Health Canada, a Department of the federal government of Canada; in consequence this report must contain the statement, “The views expressed herein do not necessarily represent the views of the federal government. “
“Please cite this paper as: Baum, Suter, Gerber, Tschanz, Buergy, Blank, Hlushchuk and Djonov (2010). VEGF-A Promotes Intussusceptive Angiogenesis in the Developing Chicken Chorioallantoic Membrane. Microcirculation17(6), 447–457. Objective:  To assess the impact of vascular endothelial growth factor (VEGF) on intussusceptive angiogenesis.

To investigate whether the PS-5 mimetic affects the migratory property of T lymphocytes, we analyzed the ability

of T-cell populations to respond to supernatants from IFN-γ-activated keratinocytes in transwell migration assays. As shown in Figure 5A, supernatants from untreated or NC-treated-keratinocytes stimulated AZD0530 nmr with IFN-γ were fourfold more efficient in eliciting migratory responses of circulating PBMCs previously stained for anti-CD3, compared with supernatants from unstimulated strains. On the contrary, the treatment with PS-5, as well as with KIR peptide, significantly reduced the IFN-γ-dependent migration of PBMCs toward the supernatants of activated keratinocytes. Similar effects were observed in migration experiments performed with skin T-cell lines derived from type 1-mediated inflammatory skin diseases, including psoriasis (Fig. 5B) and lichen planus (Fig. 5C). Finally, we investigated the effects of PS-5 peptide on STAT1 activation and the expression of STAT1-dependent inflammatory genes in organ cultures of normal human skin treated with IFN-γ. As shown in Figure 6, the explants of IFN-γ-treated skin preincubated with PS-5, as well as with KIR peptide, showed a faint epidermal immunoreactivity for phosphorylated STAT1, compared with those observed in skin explants treated with NC peptide or its vehicle (Fig. 6). In these skin explants, phospho-STAT1 expression

BMS-777607 manufacturer was comparable with that observed in abundance in lesional skin obtained from psoriatic patients, used as positive control. In contrast, phospho-STAT1 staining was quite absent in untreated skin explants and in uninvolved zones (nonlesional skin) of psoriatic plaques. As direct consequence of the reduced STAT1 phosphorylation and activation, the Depsipeptide supplier epidermal expression of ICAM-1, HLA-DR, CXCL10 was abrogated in IFN-γ-treated explants of human skin incubated with PS-5 or KIR mimetics, compared with that found in organ cultures treated with NC peptide or vehicle (Fig. 7). The decrease of the number of ICAM-1+,

HLA-DR+, or CXCL10+ epidermal cells in PS-5-treated skin organ cultures was highly significant, as demonstrated by counting positive cells/mm2 in four different stained sections obtained from three skin explants for each condition (Fig. 7). Taken together, these results highlighted the efficiency of PS-5 mimetic to dampen the inflammatory responses triggered by JAK2/STAT1 signaling in human skin. Inhibition of JAK2 activity and the consequent inactivation of the downstream STAT1 transcription factor represent a promising strategy for the attenuation of the inflammatory responses elicited by epidermal keratinocytes following massive exposure to IFN-γ in the skin. In recent years, a number of small molecule inhibitors of IFN-γ signaling have been developed, including mimetics sharing the KIR region of SOCS1 protein [12, 22, 23].

Recently, we reported that unrestricted activation of this pathway in NF-κB2/p100-deficient (p100−/−) knock-in mice alters the phenotype of MZ stroma and B cells. Here, we show that lack of the p100 inhibitor resulted in an expansion of both MZ B and peritoneal B-1 cells. However, these cells failed to generate proliferating FG-4592 nmr blasts in response to T-cell-independent type 2 (TI-2) Ags, correlating with dampened IgM and absent IgG3 responses. This phenotype was in part due to increased activity of the NF-κB subunit RelB. Moreover, p100−/−B6

BM chimeras were more susceptible to infection by encapsulated Streptococcus pneumoniae bacteria, pathogens that induce TI-2 responses. In contrast to the TI-2 defect, p100 deficiency did not impair immune responses to the TI-1 Ag LPS and p100−/−

MZ B cells showed normal Ag transportation into B-cell follicles. Furthermore, p100−/− MZ B and B-1 cells failed to respond to TI-2 Ags in the presence of WT accessory cells. Thus, NF-κB2/p100 deficiency caused a predominant B-cell-intrinsic TI-2 defect that could largely be attributed to impaired proliferation of plasmablasts. Importantly, p100 was also necessary for efficient defense against clinically relevant TI-2 pathogens. “
“Hyaluronan is known to accumulate in tissues during inflammatory diseases associated with graft implantation and rejection of organ allografts. The aim was to evaluate whether hyaluronidase treatment affected hyaluronan content and blood perfusion in graft pancreatitis. Syngeneic selleck chemical rat pancreatic-duodenal transplantations were performed. Two days later blood flow measurements were made with a microsphere technique in both grafted and endogenous pancreas in animals treated with daily injections of vehicle or hyaluronidase (20.000 U/kg). Non-transplanted rats served as controls. Also, samples for analysis of hyaluronan and water content were taken. The hyaluronan content of the pancreatic graft was increased after transplantation. Hyaluronidase treatment markedly reduced total pancreatic and islet blood flow in both grafted and endogenous pancreas, whereas duodenum blood flow was unaffected. No blood flow effects were seen in non-transplanted control rats. Hyaluronan

content was increased in the grafted pancreas, but hyaluronidase treatment ever decreased it to levels comparable to those of the endogenous gland. There were no differences in hyaluronan content in the endogenous pancreases of transplanted and non-transplanted rats. Graft pancreatitis after rat pancreas transplantation is associated with an increased hyaluronan content, which can be reduced by treatment with hyaluronidase. Hyaluronidase treatment of the graft recipients effected a 50% reduction in total pancreatic and islet blood flow in the graft, as well as in the endogenous pancreas. The functional importance of this is at present unknown. Hyaluronan (HA) is a ubiquitous glucosaminoglucan in the extracellular matrix of most tissues [1].

“
“We report a rare case of focal cortical dysplasia (FCD) concurring with diffuse astrocytoma and arachnoid cyst, and also re-evaluate the glial component in archival FCD cases for the differential diagnosis of diffuse gliomas. A 7-year-old boy with a 9-month history of psychomotor seizures disclosed

a hyperintense area accompanied by a cystic lesion in the left temporal lobe on MRI. The surgical specimen displayed dyslamination of the cortices and ectopic neurons in the white matter, associated with dysmorphic neurons, indicating FCD type Trichostatin A solubility dmso IIA. Additionally, the lesion showed diffuse proliferation and infiltration of glial cells, immunopositive for infiltrating glioma markers (nestin, doublecortin, MAP-2e) and p53, and MIB-1 index was 2.0%. These findings indicated coexisting diffuse astrocytoma. Coexistence of diffuse glioma with FCD is unusual, but selleck products we often notice increased population of small glial cells in FCD lesions. Re-evaluation of archival FCD cases with diverse markers revealed that reactive microglia significantly proliferate in the white matter lesions. Therefore, a careful pathological assessment has to be made to define a rare case of diffuse glioma occurring in FCD. “
“M. Sie, E. S. J. M. de Bont, F. J. G. Scherpen, E. W. Hoving and W. F. A. den Dunnen (2010) Neuropathology and Applied Neurobiology36,

636–647 Tumour vasculature and angiogenic profile of paediatric pilocytic astrocytoma; is it much different from glioblastoma? Aims: Pilocytic astrocytomas are the most frequent brain tumours in children. Because of their high vascularity, this study aimed to obtain insights into potential angiogenic related therapeutic targets in these tumours by characterization

of the vasculature and the angiogenic profile. In this study 59 paediatric pilocytic astrocytomas were compared with 62 adult glioblastomas, as a prototype of tumour angiogenesis. Methods: Microvessel density, vessel maturity in terms of basement membrane and pericyte coverage, and turnover of both endothelial and tumour cells, and vascular endothelial growth factor (VEGF) expression were evaluated in tumour tissue, MAPK inhibitor immunohistochemically stained with, respectively, CD34, collagen IV, smooth muscle actin, Ki67/CD34, caspase-3/CD34 and VEGF(-A–D). As an indicator for vessel stability the angiopoietin (ANGPT)-1/ANGPT-2 balance was calculated using Real Time RT-PCR. Results: Pilocytic astrocytoma and glioblastoma showed similar fractions of vessels covered with basement membrane and pericytes. Overlapping ANGPT-1/ANGPT-2 balance and VEGF-A expression were found. Pilocytic astrocytoma had fewer but wider vessels compared with glioblastoma. Turnover of endothelial and tumour cells were relatively lower in pilocytic astrocytoma. Within pilocytic astrocytoma, higher ANGPT-1/ANGPT-2 balance was correlated with fewer apoptotic endothelial cells. Lower numbers of vessels were correlated with higher VEGF-A expression.

Molecules similar to aag (molecular mimicry) can also initiate an autoimmune disease [10, 11, 46, 47]. Infectious agents or their products (exo- or endotoxins) acting as adjuvants can incorporate aag and cause the formation of disease inducing pathogenic aabs [48–50]. Autoimmune

diseases can present themselves as short-term or prolonged chronic progressive diseases following unusual presentation of self or self-like ag, the former causing minimal harm, the latter sometimes resulting in ultrastructural changes leading to organ failure.  Presentation of the inciting agent(s). Inciting agents play a major role in the initiation and maintenance of certain autoimmune diseases. These agents (toxins, chemicals,

drugs, etc.) present self ag to the cells of the immune system as hapten protein conjugates with or without adjuvants (e.g. bacterial breakdown products from sites of infection). Altered self ag evoke pathogenic IgG check details aab responses and start a genuine autoimmune disease characterized by morphological and functional changes of an organ, and clinical signs and symptoms. If the inciting agent is removed from the system then no further production of disease causing pathogenic aabs will occur. That is to say, the continuous presence of a modifying agent (the inciter) is necessary in the system to maintain the production of pathogenic aabs. Normal self ag will not initiate and/or maintain pathogenic aab production [51]. Cancer is included in this group of autoimmune disorders. Cancer – because minimal antigenicity of cancer-specific ag on cancer cell membrane surfaces means that such ag click here are not recognized as non-self – generally evokes no pathogenic aab response. IgM aabs assist in the removal of released cellular components from cancer cells damaged by ischaemia, drugs, etc., but will not lyse whole cancer cells themselves [17, 19]. Presence of pathogenic aabs in the circulation is always tissue damaging [51]. Pathogenic aabs are able to react with modified (chemically or otherwise altered) aag present in the circulation or

at any other location in the body and are also able to cross react with native aag residing in tissues [51]. To illustrate this point, in an experimental autoimmune kidney disease in rats called slowly progressive Methane monooxygenase Heymann nephritis (SPHN) we have observed the following. The injected modified tubular nephritogenic ag [12, 21] in various forms initiates the production of pathogenic IgG aabs that are able to react with native nephritogenic ag localized in both (1) the glomeruli and (2) the tubular brush border (BB) region. As a result, a cycle of events begins. Normal renal ag are liberated from the renal proximal convoluted tubules and contribute with circulating pathogenic IgG aabs (in the presence of complement) to glomerular immune complex (IC) formation.

1.IFN-α does not induce α-defensin production from HGECs. Supporting Information Fig. 2. mRNA expression of STAT1, STAT2, IRF3, IRF7, and IRF9 in α-defensin-1-treated HGEC by real-time RT-PCR. Supporting Information Fig. 3. α-defensin-1 does not induce STAT1 activation in HGECs. “
“The immunomodulatory

ability of mesenchymal stem cells (MSCs) may be used to develop therapies for autoimmune diseases. Flk-1+ MSCs are a population of MSCs with defined phenotype and their safety has been evaluated in Phase 1 clinical trials. We designed this study to evaluate whether Flk-1+ MSCs conferred a therapeutic effect on collagen-induced arthritis (CIA), an animal model of rheumatic arthritis, and to explore the underlying mechanisms. Flk-1+ MSCs, 1–2 × 106, were injected

into CIA mice on U0126 solubility dmso either day 0 or day 21. The clinical course of arthritis was monitored. Serum cytokine profile was determined by cytometric bead array kit or enzyme-linked immunosorbent assay. Flk-1+ MSCs and splenocytes co-culture was conducted to explore the underlying mechanisms. Flk-1+ MSCs did not confer therapeutic benefits. Clinical symptom scores and histological evaluation suggested aggravation of arthritis in mice treated with MSCs at day 21. Serum cytokine profile analysis showed marked interleukin (IL)-6 secretion immediately after MSC administration. Results of in vitro culture of splenocytes confirmed that the addition of Flk-1+ MSCs promoted splenocyte proliferation AZD2014 Leukocyte receptor tyrosine kinase and increased IL-6 and IL-17 secretion. Moreover, splenocyte proliferation was also enhanced in mice treated with MSCs at day 21. Accordingly, MSCs at low concentrations

were found to promote lipopolysaccharide-primed splenocytes proliferation in an in vitro co-culture system. We propose that Flk-1+ MSCs aggravate arthritis in CIA model by at least up-regulating secretion of IL-6, which favours Th17 differentiation. When Flk-1+ MSCs are used for patients, we should be cautious about subjects with rheumatoid arthritis. Mesenchymal stem cells (MSCs) are multi-potential cells with extensive proliferative ability. They have been isolated from both bone marrow and other tissues, and are capable of differentiating into chondrocytes, osteocytes, adipocytes, endothelial cells and neural cells under appropriate cues [1,2]. The ability for isolation and expansion of MSCs in vitro without losing their phenotype or multi-lineage potential has granted MSCs a promising role in tissue engineering and regenerative medicine [3,4]. Extensive evidence has shown that MSCs can exert profound immunosuppressive effects, as they can suppress T cell proliferation in culture and prolong skin graft across MHC barriers [5,6]. In addition to T cells, MSCs are also found to suppress proliferation of B cells [7], natural killer cells [8–10] and differentiation, proliferation and maturation of dendritic cells [11–16].

In addition to demonstrating strain independence, experiments were performed to show that Treg-cell control of GC responses was also antigen independent. Figure 3 summarizes the effect of anti-GITR mAb treatment on splenic GC responses induced by i.p challenge of BALB/c mice with IAV. Whereas SRBC induce a Th2-biased response,5 IAV invokes a Th1-polarized reaction.56Figure 3(a) shows that mice immunized i.p. with IAV generate selleck chemicals a robust splenic GC response which peaks at day 12 (Fig. 3b). Similar to Th2 antigens,5,6 the GC reaction induced by

IAV was characterized by a steady ratio of IgM+ to switched GC B cells (Fig. 3c). Importantly, anti-GITR mAb administration resulted in a higher frequency and total number of splenic GC B cells at several time-points (Fig. 3b), and significantly increased the proportion of switched GC B cells throughout the entire reaction (Fig. 3c). As opposed to GCs induced with SRBC immunization, we observed no significant difference EX 527 nmr in the distribution of IgG isotypes within the switched GC B-cell pool at any time-points after IAV challenge (data not shown). The results generated above demonstrated the role of Treg cells in controlling both the size of SRBC-induced and IAV-induced GC responses, and the ratio of IgM+ to switched B cells within the

GC population. In these experiments, however, total splenic GC B cells were enumerated because the B220+ PNAhi B-cell population induced after SRBC or IAV injection was presumed to be specific for the challenge antigen. (Please note that specific pathogen-free mice do not exhibit splenic GCs in the absence of immunization, Fig. 1.) We therefore sought to confirm the role of Treg cells in governing GC reactions by tracking antigen-binding GC B cells, instead of the entire B220+ PNAhi splenic B-cell pool. To perform these studies, PE was used as the challenge antigen,57–59 and PE-binding GC B cells were analysed in anti-GITR mAb or control rIgG-treated mice. As shown in Fig. 4(a), i.p. immunization with PE precipitated in alum induced splenic B220+ PNAhi GC B cells, a new sub-set of which retained the ability to bind native

PE. In control animals, the PE-binding GC B-cell response peaked at day 12 (Fig. 4b) and like other normal splenic GC responses, displayed a relatively steady ratio of IgM+ to switched B cells (Fig. 4c). As expected, disruption of Treg cells with anti-GITR mAb administration resulted in an increased total PE-binding GC response, and a progressive increase in the proportion and total number of switched PE-binding GC B cells. In Figs 1–4, splenic GC responses were dysregulated when anti-GITR mAb was given before and soon after immunization. To assess whether already established GCs can be altered by late-stage Treg-cell disruption, mice were challenged with SRBC at day 0 and treated with either anti-GITR mAb or control rIgG on days 8 and 12, or days 12 and 16 post-immunization. Splenic GCs from both groups were examined on days 18 and 24.

IgG derived

from a SS patient positive for antibodies to the Talazoparib third extracellular loop had no effect on (Ca2+)I, as well as IgG derived from an anti-M3R antibody-negative SS patient (Figs 3e and 4). Recently, anti-M3R antibodies have been the focus of interest in rheumatology because of their potential pathogenic role, use as diagnostic markers and being therapeutic targets in patients with SS [1]. Several methods have been used to detect anti-M3R antibodies in SS patients [1]. In functional assays using smooth muscles, IgG fractions from patients with SS (SS-IgG) inhibited carbachol-evoked or nerve-evoked bladder or colon contractions [8,9]. In salivary gland cells, SS-IgG inhibited the rise in (Ca2+)i induced by carbachol, and also inhibited pilocarpine-induced AQP5 trafficking to the apical membrane from the cytoplasm [2]. The inhibitory actions of SS-IgG on

the rise in (Ca2+)i was acutely reversible [10]. Anti-M3R antibodies from SS patients can be detected by immunofluorescent analysis using rat lacrimal glands [11], and by flow cytometry using the M3R-transfected Chinese hamster ovary (CHO) cell line [12]. Moreover, anti-M3R antibodies in sera of SS patients were detected by ELISA using synthetic peptides or recombinant proteins of the second extracellular loop of M3R [13]. We have reported previously the presence of anti-M3R antibodies in a group of patients with SS, which recognized the second extracellular loop by ELISA using synthetic Venetoclax molecular weight peptides [4,5]. In the present study, we established a standard method to detect anti-M3R antibodies that can be used for screening large patient populations. Functional assays and flow cytometry are too laborious for routine use. Although ELISA is easy, the results from some ELISA systems used for screening anti-M3R antibodies differ Histidine ammonia-lyase widely with regard to the prevalence of anti-M3R antibodies (from 11 to 90%) [4,14]. Furthermore, Cavill et al.[15]

reported failure to detect anti-M3R antibodies by ELISA using synthetic peptides. In the present study, we reported higher frequencies and titres of anti-M3R antibodies against all extracellular domains in SS patients than the control. The prevalence of anti-M3R antibodies against the second extracellular loop in SS (55%) determined in the present study was much higher than that reported in our previous study (11%) [4]. The reason for this difference is probably related to the change in the methodology, such as increased sensitivity resulting from purity of the synthetic peptides, modification of the washing procedure or other factors introduced in the modified ELISA system. In the present study, we also determined the precise B cell epitopes of M3R molecules.