No microbubble coalescence and no increased size were observed. Adhesion of some microbubbles to leukocytes was observed in various microcirculation models. Microbubbles are captured by Kupffer cells in the liver. Targeted microbubbles were shown to adhere specifically BMS-777607 mouse to endothelial receptors without compromising local blood flow. Conclusion:  These results support the safety of both targeted and nontargeted UCAs as no microvascular flow alteration or plugging of microvessels were observed. They confirm that binding

observed with targeted microbubbles are due to the binding of these microbubbles to specific endothelial receptors. “
“Microcirculation (2010) 17, 348–357. doi: 10.1111/j.1549-8719.2010.00036.x Objective:  The canonical Wnt signaling pathway, heavily studied in development and cancer, has recently been implicated in microvascular growth with the use of developmental and in vitro models. To date, however, no study exists showing the effects of perturbing the canonical Wnt pathway in a complete microvascular network undergoing physiological remodeling in vivo. Our objective was to investigate the effects of canonical Wnt inhibition on the microvascular remodeling of adult rats. Methods:  Canonical Wnt inhibitor

DKK-1, Wnt inhibitor sFRP-1, BSA or saline was superfused onto the exteriorized mesenteric windows selleck chemicals llc of 300 g adult female Sprague-Dawley rats for 20 minutes. Three days following surgery, mesenteric windows were imaged intravitally and

harvested for immunofluorescence staining with smooth muscle alpha-actin and BRDU. Results:  We heptaminol observed prominent differences in the response of the mesenteric microvasculature amongst the various treatment groups. Significant increases in hemorrhage area, vascular density, and draining vessel diameter were observed in windows treated with Wnt inhibitors as compared to control-treated windows. Additionally, confocal imaging analysis showed significant increases in proliferating cells as well as evidence of proliferating smooth muscle cells along venules. Conclusions:  Together, our results suggest that canonical Wnt inhibition plays an important role in microvascular remodeling, specifically venular remodeling. “
“Please cite this paper as: Sundd, Gutierrez, Petrich, Ginsberg, Groisman, and Ley (2011). Live Cell Imaging of Paxillin in Rolling Neutrophils by Dual-Color Quantitative Dynamic Footprinting. Microcirculation 18(5), 361–372. Objective:  Neutrophil recruitment to sites of inflammation involves P-selectin-dependent rolling. qDF is a useful tool to visualize the topography of the neutrophil footprint as it interacts with the substrate. However, elucidating the role of specific proteins in addition to topography requires simultaneous visualization of two fluorochromes. Methods:  To validate DqDF, mouse neutrophils were labeled with the membrane dyes DiO and DiI and perfused into microchannels coated with P-selectin–Fc.

Future directions in this field will also be discussed. MiRNAs were first found in the nematode Caenorhabditis elegans in 1993.1 Since then they have also been described widely in plants and mammals.2 MiRNAs are first transcribed in the nucleus as stem-loop primary miRNA, which are then cleaved into shorter precursor miRNA by Drosha, an RNase III, and its essential GSK1120212 datasheet cofactor called DGCR8 (DiGeorge syndrome critical region 8), a double-stranded RNA-binding protein (Fig. 1).3–6 The precursor miRNAs are transported out of the nucleus via Exportin-5 and once in the cytosol are cleaved into their mature form of 20–22 nucleotides by Dicer, another

RNase III.7,8 After cleavage, the miRNA duplex is unwound and the functional strand is loaded onto the RNA-induced silencing complex (RISC) and functions as its guide.9 The mature miRNA guides the RISC complex to a (near) complementary sequence, usually in the 3′ untranslated region (UTR), of a target messenger RNA (mRNA).9 Upon binding, the RISC causes post-transcriptional gene silencing by

either cleaving the target mRNA or by inhibiting its translation, CRM1 inhibitor so that miRNAs are usually negative regulators of gene expression.10 In addition to their role in such post-transcriptional repression, miRNAs have now been implicated in transcriptional gene silencing by targeting the promoter region but have also been reported to have a positive effect on transcription.11–13 Each miRNA can potentially regulate the translation

of a large number of different mRNA and each mRNA can Protein kinase N1 possess multiple binding sites for a single or for many different miRNA because the specificity of miRNA is mainly determined by Watson-Crick base pairing at the 5′ region of the miRNA. Estimates have suggested that the total number of different miRNA sequences in humans may exceed 1000.14 Computational analysis also predicts that over 60% of human genes are potential targets of miRNAs and that there are a large number of other non-coding RNAs of greater nucleotide length than microRNA, which are also likely to have important functions.15 However, direct experimental evidence defining mRNA targets of miRNA regulation has been reported for only a small number of miRNAs and target mRNAs. Assaying the levels of specific microRNA sequences was initially cumbersome; however, advances in technology now allow detection with a sensitivity and specificity that can enable monitoring in a clinical setting. Originally, RNA blot analyses provided both quantitative and qualitative information about the various forms of a miRNA within a total RNA sample.1,16 As the number of miRNAs in the miRBase registry17 has increased, microarray technology has been adapted to enable the parallel screening of thousands of miRNAs in one sample.18 More recently, real time reverse transcription-polymerase chain reaction has been adapted to enable relative quantification and quantitative analysis of miRNA levels.

This work was supported in part by a Grant-in-aid for Scientific Research (C) (16590366) from the Ministry of Education, Science and Culture of Japan,

a Grant (19-SHINKOU-005) from the Ministry of Health, Labour and Welfare of Japan and Tohoku University 21st COE program ‘CRESCENDO’. The authors have no financial conflict of interest. Fig. S1. Distribution of Gr-1dull+ cells in the R2-SSCmoderately high area. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Non-eosinophilic asthma is characterized by infiltration of neutrophils into the lung and variable responsiveness check details to glucocorticoids.

The pathophysiological mechanisms have not been characterized in detail. Here, we present an experimental asthma model in mice associated with non-eosinophilic airway inflammation and airway hyper-responsiveness (AHR). For this, BALB/c mice were sensitized by biolistic DNA immunization with a plasmid encoding the model antigen β-galactosidase (pFascin-βGal mice). For comparison, eosinophilic airway inflammation was induced by subcutaneous injection of βGal protein (βGal mice). Intranasal challenge of mice in both groups induced AHR to a comparable extent as well as recruitment of inflammatory cells into the airways. In contrast to βGal selleck mice, which exhibited extensive eosinophilic infiltration in the lung, goblet cell hyperplasia and polarization of CD4+ T cells into Th2 and Th17 cells, pFascin-βGal mice showed considerable neutrophilia, but no goblet cell hyperplasia and a predominance of Th1 and Tc1 cells in the airways. Depletion studies in pFascin-βGal mice revealed that CD4+ and CD8+ cells cooperated to induce maximum inflammation, but that neutrophilic infiltration was not a prerequisite Teicoplanin for AHR induction. Treatment of pFascin-βGal mice with dexamethasone before intranasal challenge did not affect neutrophilic infiltration, but significantly

reduced AHR, infiltration of monocytes and lymphocytes as well as content of IFN-γ in the bronchoalveolar fluid. Our results suggest that non-eosinophilic asthma associated predominantly with Th1/Tc1 cells is susceptible to glucocorticoid treatment. pFascin-βGal mice might represent a mouse model to study pathophysiological mechanisms proceeding in the subgroup of asthmatics with non-eosinophilic asthma that respond to inhaled steroids. “
“Enteroviral infections go usually unnoticed, even during pregnancy, yet some case histories and mouse experiments indicate that these viruses may be transmitted vertically. More frequently, however, transmission occurs by (fecal) contamination during and shortly after birth.

To reveal bound antibodies we used horseradish peroxidase (HRP)-conjugated secondary antibodies. Blots were developed with enhanced chemiluminescence (ECL) reagent (Pierce; Thermo Scientific, Rockford, IL, USA). To obtain semi-quantitative

estimates for the total tyrosine phosphorylation, it was quantified and densitometry analysis was performed using Tina 2·0 software (Raytest, Straubenhardt, Germany). Values were normalized to the intensity of actin bands. For comparisons of quantitative values we used the unpaired Student’s t-test. The frequency of autoantibodies in HAE patients and control group was compared using Fisher’s exact test. Two-tailed P-values of 0·05 or less were considered statistically significant. Data are expressed as mean values of MFI ± s.d. In 29 of the 61 (47·5%) patients, at least one of the tested autoantibodies was found in the serum, as detailed

in Table 1. We did PLX3397 in vivo not find any difference in gender ratio when HAE patients with autoantibodies were compared with those without autoantibodies [male (12 of 25), female (17 of 36)]. Additionally, we did not find a difference in the average mean of the complement 4 (C4) levels between these two groups of HAE patients [0·095 ± 0·05 versus 0·088 ± 0·05, P = not significant (n.s.)]. In the healthy control group, five of 50 (10%) had serum autoantibodies. This frequency is statistically lower compared to HAE patients [five of 50 (10%) versus 29 of 61 (47·5%), P = 0·0001]. Two had positive anti-nuclear antibodies (4%), two of 50 CHIR-99021 research buy (4%) had anti-cardiolipin antibodies and in one serum we found positive anti-S. cerevisiae antibodies. Seven of 61 HAE patients (11·4%) suffered from the following CHIR99021 immunoregulatory disorders; one patient had systemic lupus erythematosus (SLE), two patients had coeliac disease,

one patient had mixed connective tissue disease, one patient had systemic sclerosis, one patient had Crohn’s disease and one patient multiple sclerosis-like syndrome. Expression of CD69 and CD5 was found to be statistically higher on memory B cells (CD19+CD27+) from HAE patients compared to healthy controls (4·59 ± 4·41 versus 2·06 ± 1·81, P = 0·04, 8·22 ± 7·17 versus 3·65 ± 3·78, P = 0·05, respectively). Expression of CD21 on memory B cells was also significantly higher when compared to that on memory B cells from healthy controls (2·43 ± 0·54 versus 1·92 ± 0·41, P = 0·01). In contrast, we did not find any statistical difference in the expression of MHC-II, CD40 and CD86 on the memory B cells of the two groups. Results are summarized in Table 2. Memory B cells isolated from the HAE group expressed a significantly higher amount of TLR-9 (8·17 ± 4·1 versus 4·56 ± 1·6, P = 0·0027). Furthermore, the expression of TLR-9 in B cells from HAE patients who had autoantibodies was much higher than that of memory B cells from both the control group (10 ± 4·7 versus 4·56 ± 1·6, P = 0·0002) and from HAE patients without autoantibodies (10 ± 4·7 versus 5·8 ± 0·9, P = 0·036).

We find no predilection or predisposition towards an accompanying TDP-43 pathology in patients with FTLD-tau, irrespective of presence or absence of MAPT mutation, or that genetic changes associated Y-27632 molecular weight with FTLD-TDP predispose towards excessive tauopathy. Where the two processes coexist, this is limited and probably causatively independent of each other. “
cases of primary hydrocephalus. Hyh mice, which exhibit either severe or compensated long-lasting forms of hydrocephalus, were examined and compared with wild-type mice. TGFβ1, TNFα and TNFαR1 mRNA levels were quantified using real-time PCR. TNFα and TNFαR1 were immunolocalized in the brain tissues of hyh mice and four hydrocephalic human

foetuses relative to astroglial and microglial reactions. The TGFβ1 mRNA levels were not significantly different between hyh mice exhibiting severe or compensated hydrocephalus and normal mice. In contrast, severely hydrocephalic mice exhibited four- and two-fold increases in the mean

levels of TNFα and TNFαR1, respectively, compared with normal mice. In the hyh mouse, TNFα and TNFαR1 immunoreactivity was preferentially detected in astrocytes that form a particular periventricular reaction characteristic of hydrocephalus. However, these proteins were rarely detected in microglia, which did not appear to be activated. TNFα immunoreactivity was also detected in the glial reaction in the small RO4929097 group of human foetuses exhibiting hydrocephalus that were examined. In the hyh mouse model of congenital hydrocephalus, TNFα and TNFαR1 appear to be associated with the severity of the disease, probably mediating the astrocyte reaction, neurodegenerative processes and ischaemia. “
“Frontotemporal lobar degeneration (FTLD) is classified mainly into FTLD-tau and FTLD-TDP according to the protein present Aldol condensation within inclusion bodies. While such a classification implies only a single type of protein should be present, recent studies have demonstrated dual tau and TDP-43 proteinopathy can occur, particularly in inherited FTLD. We therefore investigated 33 patients with

FTLD-tau (including 9 with MAPT mutation) for TDP-43 pathological changes, and 45 patients with FTLD-TDP (including 12 with hexanucleotide expansion in C9ORF72 and 12 with GRN mutation), and 23 patients with motor neurone disease (3 with hexanucleotide expansion in C9ORF72), for tauopathy. TDP-43 pathological changes, of the kind seen in many elderly individuals with Alzheimer’s disease, were seen in only two FTLD-tau cases – a 70-year-old male with exon 10 + 13 mutation in MAPT, and a 73-year-old female with corticobasal degeneration. Such changes were considered to be secondary and probably reflective of advanced age. Conversely, there was generally only scant tau pathology, usually only within hippocampus and/or entorhinal cortex, in most patients with FTLD-TDP or MND.

In Braak stages 0–I–II cases, UBL immunoreactivity was detected in a dense fiber network in the neuropil, and in the cell cytoplasm and nucleoplasm of neurons in Cornu Ammonis (CA) fields and dentate gyrus granular neurons. In Braak stages III-IV and V-VI cases, UBL immunoreactivity was reduced in the neuropil BKM120 and in the cytoplasm of the majority of CA1 neurons; some CA1 pyramidal neurons and the majority of CA2/3 pyramidal, CA4 multipolar, and dentate granular neurons had markedly increased UBL immunoreactivity in the nucleoplasm. Dual immunofluorescence analysis of UBL and antibody clone AT8 revealed co-localization most frequently

Navitoclax order in CA1 pyramidal neurons in Braak stage III-IV and V-VI cases. Further processing using the pan-amyloid marker X-34 revealed prominent UBL/X-34 dual labeling of extracellular NFT confined to the CA1/subiculum in Braak stage V-VI cases. Our results demonstrate that in AD hippocampus, early NFT changes are associated with

neuronal up-regulation of UBL in nucleoplasm, or its translocation from the cytoplasm to the nucleus. The perseverance of UBL changes in CA2/3, CA4 and dentate gyrus, generally considered as more resistant to NFT pathology, but not in the CA1, may mark a compensatory, potentially protective response to increased tau phosphorylation in hippocampal neurons; the failure of such a response may contribute to neuronal degeneration in end-stage AD. The ubiquitin (Ub)–proteasome system is the major non-lysosomal proteolytic pathway in eukaryotes.[1] Ubiquilin-1 (also referred to as “protein linking integrin-associated protein to cytoskeleton 1”, or Plic-1) is a Ub-like (UBL) protein with functional domains

on its N-terminus (UB) and C-terminus (Ub-associated; UBA). Ubiquilin interacts with polyubiquitylated proteins through its UBA domain and with two subunits of the 19S proteasome through the UB domain.[2] UBL protein is observed in neurofibrillary tangles (NFT) in aminophylline Alzheimer’s disease (AD) brains,[3] facilitates presenilin synthesis[3] and modulates amyloid precursor protein trafficking and amyloid-beta (Aβ) secretion.[4] Previous studies reported that early in AD, UBL-1 protein levels decrease in the frontal cortex;[5] the status of UBL-1 protein levels in the hippocampus in patients with varying degrees of NFT pathology is unknown. In this study, we used immunohistochemical techniques to examine localization and alterations in UBL-1 protein in the hippocampus from cases at different stages of NFT pathology as classified by Braak and Braak.[6] Multiple-label immunofluorescent microscopy analyses examined the relationship of UBL with early and late NFT changes.

38 To date, however, there are insufficient data to determine whether cure rates differ significantly between the repeat retropubic and transobturator routes, and whether complication rates are higher after secondary than primary MUS procedures. Use of a re-adjustable sling for recurrent SUI with sphincteric deficiency is currently under investigation. Use of the Remeex (Neomedic, Barcelona, Spain) re-adjustable sling (Fig. 1) showed that, after

3 years, 109 of 125 (87.2%) women were continent under stress after initial surgery, including 49 of 55 (84%) with recurrent SUI and 60 of 70 (85.7%) with ISD.44 Moreover, 19 of these patients showed additional benefit from ABT-199 concentration a subsequent re-adjustment. The rate of infection of the re-exposed varitensor during adjustment was lower, while the development of de novo overactivity (8%) was similar to results observed with the other sling type. A prospective study of the AMI adjustable suburethral sling (Agency for Medical Innovation GmbH, 6800 Feldkirch, Austria) (Fig. 245) implanted through the retropubic route in 25 patients with recurrent urodynamic SUI showed that 21 of the patients were urodynamically continent after 12 months.46 A recent study described the use of a transobturator

crossover re-adjustable sling as a salvage procedure for failed anti-incontinence click here procedures (Fig. 3).47 This SAFYRE t plus sling (Promedon, Cordoba, Argentina) consists of a monofilament polypropylene mesh between two self-anchoring Inositol monophosphatase 1 columns. The procedure is performed by creating a spiral sling for better circumferential coaptation

of the urethra. Moreover, silicone washers are used in the genitofemoral fold at the level of the clitoris, both to improve fixation and facilitate later adjustments. Re-adjustments were easily performed under local anesthesia by moving the washers until there was no urine leakage during valsalva maneuver. After 12 months, the overall cure rate was 93.7% (15/16), with only one patient requiring re-adjustment. During surgery, however, one patient experienced a urethral perforation, which was resolved by closing the urethral operation. The adjustable continence therapy (ACT) device consists of two adjustable balloons, each attached to an injection port placed subcutaneously in the labia majora (Fig. 4).48 After 6 years, 68% of patients remained dry.49 The pubovaginal sling has shown success rates ranging from 50 to 90% in the treatment of women with persistent or recurrent SUI. A trial of the pubovaginal sling in patients with all types of SUI divided patients into simple and complex groups, with mean numbers of prior incontinence surgeries of 0.78 and 3.1, respectively.50 After 1-year follow-up, SUI was cured in 183 women (73%) and improved in 48 (19%). After a >10-year follow-up in 20 women, the success rate was 95%.

This finding highlights the potential for an autoantibody-independent effect of B cell depletion on MS disease activity. B cells are important antigen-presenting cells. Physical interaction of B cells and T cells [major histocompatibility complex (MHC)/antigen/T cell receptor] occurs in the presence of co-stimulatory molecules such as CD40/CD40ligand, B7/CD28, OX40 ligand/OX40 on the surface of B cells and T cells, respectively [55]. B cell depletion in mice was found to impact on

CD4+ T cell activation and expansion in vivo, which may explain its positive effect on multiple T cell-mediated autoimmune diseases, including MS [56] and type 1 diabetes [57]. It remains to be seen whether B cell-depleting strategies may alter the ratio of CTL : infected targets cells favourably, and thus enable better control of EBV infection. Furthermore, B cells have the ability to regulate T cell function and inflammation through cytokine production. A recent BMN 673 molecular weight study

found that B cells of MS patients had altered cytokine responses, e.g. increased ratio of lymphotoxin (LT) : IL-10 and increased secretion of tumour necrosis factor (TNF)-α and LT when exposed to the proinflammatory cytokine IFN-γ or bacterial cytosine–guanine dinucleotide (CpG)-DNA bound to Toll-like receptor 9 [58]. Interestingly, CD4 and CD8 T cells of MS patients produced significantly fewer proinflammatory Th1/Th17 cytokines after in vivo or ex vivo B cell depletion. B cell depletion may, therefore, be effective in reducing CNS inflammation. However, HIF cancer cAMP B cells also play an important role in immunoregulation. Animal studies highlight the importance of the IL-10-producing B cell subset (B10) in the suppression of autoimmunity and inflammation [59], which may explain why B cell depletion led to the worsening of inflammatory disease in some EAE models, with delayed production of IL-10 and emergence of regulatory T cells [60]. B cell depletion also exacerbated disease in myelin–oligodendrocyte glycoprotein peptide (MOG p35–55)-induced EAE in mice [61]. Hence, the relative

contribution of B cells to EAE and MS may vary depending on the stage of disease progression, highlighting the existence of an intricate cross-talk between T and B cells. Exciting MS treatments are currently in the pipeline, which reflect important roles for B cells as drivers of MS pathogenesis, an area overshadowed by the emphasis on T cell research in the last decade. Furthermore, the eradication of EBV+ B cells by B cell-depleting strategies is another interesting line of investigation. B cell depletion may also impact on the propensity of latent infections to contribute to neuroinflammation in the CNS, and we may want to test anti-viral strategies in MS directly using drugs which can cross the blood–brain barrier; however, treatment success may also depend on the stage of disease progression.

[11] Anaemia is a common problem in Taiwanese CKD patients. Published data indicate that 58.8% of patients with stage 4 CKD in Taiwan are anaemic, and the prevalence mTOR inhibitor of anaemia increases to 92.5% in patients reaching stage 5 CKD.[10] On 1 March 1995, Taiwan’s government launched the national health insurance (NHI) system, which ensures the right to healthcare for all residents and provides free access and total coverage of medical expenses for renal replacement therapy.

At the same time, the NHI implemented a fully bundled payment system for HD expenses including the actual cost of dialysis, the cost of dialysis-related laboratory tests, and the cost of using calcium-containing phosphate binders, active vitamin D, and ESAs. In order to promote

the use of peritoneal dialysis (PD), the NHI executed a partially bundled system in the PD treatment payment in which the reimbursement for ESAs was not included. Because almost everyone with ESRD in Taiwan is entitled to the NHI, the incentive to select healthier patients is greatly reduced in the case of dialysis. Erythropoiesis-stimulating agents soon became one of the largest drug expenditures in the NHI program of Taiwan. In 1996, the NHI applied more restrictive reimbursement criteria for ESA use targeting to a lower haematocrit in patients with CKD. ESAs are to be initiated when non-dialysis CKD patients have a serum creatinine >6 mg/dL Phosphatidylinositol diacylglycerol-lyase and a haematocrit <28%, and buy Talazoparib to maintain a haematocrit level not exceeding 30%. The maximal dose of epoetin-α or β was capped at 5000 U per week, as opposed to 9000 units per week in Japan or 400 000 units per month in the United States. The target haematocrit range and dose limitation for ESAs were the same for dialysis-dependent

CKD patients. We analyzed data from the Taiwan Renal Registry Data System (TWRDS) to examine the national trends of anaemia management in prevalent dialysis patients from 1995 to 2012. The proportion of HD patients with haematocrit <28% declined from 49% to 11%. By contrast, the proportion of those with haematocrit ≥32% rose from 16% to 32% (Fig. 1a). In 1995, mean haemoglobin was 8.9 g/dL (haematocrit 26.8%) in HD patients (Fig. 1b). Mean haemoglobin increased to 10.1 g/dL (haematocrit 30.4%) in 2004, compared with 10.4 g/dL in Japan and 11.7 g/dL in the United States, and rose steadily to 10.5 g/dL (haematocrit 31.6%) in 2012, similar to that in the United States and Japan from the DOPPS study.[12-14] The proportion of HD patients prescribed ESA remained stable at around 80%, compared with 89% in the United States and 91% in Japan. The year trend in haematocrit distribution for PD patients was similar to HD patients (Fig. 1c). However, the proportion of PD patients prescribed ESAs rose from 74.0% in 2006 to 86.2% in 2012 (Fig. 1d).

To date, this has only been achieved with attenuated N. caninum isolates used as live vaccines (10,11). However, application of a live vaccine poses a series of logistic and economical problems, which render inactivated and/or subunit vaccines much more attractive, provided a reasonable degree of protection against infection and disease can be achieved. Several research groups have reported promising results using recombinant antigens for vaccination studies, but others have reported failures or even anti-protective effects (3,9). This shows that the antigen repertoire of N. caninum contains both protective and immunomodulatory or

even immunosuppressive molecules, and these need to be defined and investigated. In addition, the route of antigen delivery and Daporinad concentration the type of adjuvant employed also need further investigation, MAPK inhibitor considering that they can also alter the efficacy of a given vaccine candidate (41,43,44). Infection studies in cattle do not represent a cost-effective system to work with, and only

a limited number of research groups have taken up the enormous task to work with cattle directly (9,12). Accordingly, murine models have been extensively used for proof-of-concept studies on how an immune response against a vaccine could limit parasite dissemination and pathology. The currently used experimental murine models include (i) cerebral infection models with challenge infections of nonpregnant mice leading to cerebral disease and death, (ii) foetal infection models where mice are challenged during pregnancy and (iii) transplacental transmission of N. caninum tachyzoites leading to stillbirth, abortion or birth of infected offspring (9,49). In

the present study, we employed the acute disease model of cerebral infection in nonpregnant animals. For the vaccine, we employed an innovative approach by analysing the relative efficacy of recNcPDI vaccine antigen associated with nanogel vaccine delivery formulations. RecNcPDI has been previously shown to be ineffective when applied i.p. emulsified in SAPs, but highly effective and mediating protection against cerebral infection and disease when applied i.n. in the presence of cholera toxin (19). The purpose of the current work was to use chitosan-based nanogels, combined with different adjuvants (saponin and CT), as carriers for the E. coli Amisulpride expressed recNcPDI antigen. Thereby, the aims were to investigate whether this nanogel association would influence the immunogenic and efficacy characteristics of the vaccine antigen upon i.p. and i.n. vaccination. SDS–PAGE and immunoblotting showed that recNcPDI was efficiently associated with both types of nanogels employed – alginate-coated chitosan nanogels and mannosylated, alginate-coated nanogels. The vaccine antigen was well associated with the nanogels, in terms of no nanogel-free material being detected. It also retained its antigenic reactivity with a polyclonal anti-recNcPDI antiserum.