For both fHbp and NHBA, antigen peptides with high frequency in the sample were associated mostly with one or two ccs, the most diverse cc being cc41/44 for both antigens. In general each peptide had a similar proportion of coverage when found in strains belonging to different ccs, with the exception of the NHBA peptide 21 that was significantly more covered in cc269 than

in cc35, suggesting a bias in the level of antigen expression associated with the genetic diversity between the two ccs. Albeit strains harboring specific combinations of MLST and antigen genotype were consistently covered (e.g. cc32 and fHbp1.1; cc41/44 and fHbp1.4; cc41/44 and NHBA2) the PF-02341066 solubility dmso majority of genetic profiles had both strains covered and not covered, confirming that antigen genotyping, neither alone nor in combination with MLST, would be sufficient to predict vaccine strain coverage for all isolates. While our active population-based sentinel surveillance data provide the most comprehensive measurement BMN 673 cell line of IMD in Canada, several limitations apply. MenB IMD is rare and the numbers in any given age group or province are small; therefore our ability to detect differences among subgroups is limited, and differences in strain coverage among age or geographic groups were not statistically significant. Approximately 20% of MenB cases in our data were confirmed by PCR only with no isolate available for testing. Additionally, IMPACT surveillance includes

primarily urban areas of Canada and may not be representative of remote or rural regions. The MATS provides a conservative estimate of vaccine coverage, which may be an underestimate [15] and [28]. Finally, although the nadA gene was found in 12 isolates (7%) in our study, only two (1%) expressed NadA with a RP above the PBT. Since expression of NadA is repressed in vitro,

but not in vivo, conditions, MATS may underestimate NadA’s contribution to vaccine strain coverage [29] and [30]. Our study characterizes the current MenB molecular epidemiology and provides a good estimate of the potential coverage of 4CMenB. Accurate post-implementation PDK4 surveillance and/or post-implementation effectiveness studies will be necessary to determine the true effectiveness of this new vaccine [31], taking into account the level of vaccine coverage in the population and any herd protection. We gratefully acknowledge the expert assistance provided by the IMPACT Monitor Liaison (Heather Samson), the IMPACT nurse monitors and staff of the IMPACT data center (Kim Marty, Wenli Zhang, Shu Yu Fan and Debbe Heayn), the National Microbiology Laboratory (Averil Henderson), the HPA laboratory Manchester, UK (Jay Lucidarme, Stefanie Gilchrist and Danielle Thompson) and our public health and infectious disease colleagues. We thank the Directors and staff of the provincial and territorial public health laboratories for providing the isolates for this study. Author contributions: J.A.

Reaction tubes were incubated at 37 °C for 10 min and the reaction was stopped by adding 3 ml of a 0.1 M sodium pyrophosphate/10% trichloroacetic acid (TCA) cold solution. Radioactive polymerized filtrate collected on cellulose nitrate

transfer membranes (0.45 μm, Whatman) was dried and immersed in scintillating fluid. Radioactivity was measured in a scintillating counter and was expressed as counts per minute (CPM). Percentage inhibition was calculated as 100 − [(CPM with extract/CPM without extract) × 100]. Reactions were carried out in duplicate for each of two independent determinations. Azidothymidine (AZT) was used as a positive control.12 Binding of gp120 http://www.selleckchem.com/products/ink128.html to CD4 was analysed using a commercially available gp120 Capture ELISA kit (GenxBio Health Science, India). To determine whether extracts could interfere with the binding of CD4 to gp120 by interaction with soluble gp120, each extract (Final conc. 10 mg/ml) was mixed with 25 ng of purified gp120 in a total volume of 100 μl and incubated

at room temperature for 1 h. This mixture was then added to microtiter plate wells coated with CD4 ligand and incubated at room temperature for 1 h. The solutions were aspirated and the wells were washed 3 times with washing buffer. The extent of gp120 binding was assessed by using detector reagent provided in the kit according to Trichostatin A purchase the manufacturer’s instructions. Negative control was set-up in parallel and heparin was included as a positive control.13 The present study, in-vitro antimicrobial activity of C. coromandelicum extract against 5 Gram-positive and Gram negative bacterial strains and 6 fungal strains

showed a broad spectrum of antimicrobial activity Table 1. The antimicrobial activities of plant extract are compared with standard antibiotics such as Ciprofloxacin and Amphotericin-B which were used as positive controls. The plant extract showed the zone of inhibition on Gram negative bacterial strains Escherichiae coli (19 mm), Klebsiella pneumoniae (14 mm), Salmonella typhi (22 mm), Shigella boydi (16 mm), Shigella almost flexneri (17 mm). The Gram positive strains Bacillus subtilis (14 mm), Micrococcus flavum (13 mm), Micrococcus leuteum (14 mm), Staphylococcus aureus (10 mm), Staphylococcus epidermis (10 mm) showed significant sensitivity. Among the both bacterial strain plant extract showed the very good sensitivity on Gram negative bacterial strain (S. typhi 22 mm) Fig. 1. The plant shows antifungal activity against Aspergillus niger (16 mm), Auricularia polytricha (17 mm), Arthrobotrys oligospora (13 mm), Candida albicans (18 mm), Chaetomella raphigera (15 mm), Monilinia fruticola (10 mm) Fig. 1. The agar well diffusion assay is a qualitative, non-standardized method useful only for the screening of large numbers of samples.

found in macaques ( Maunsell et al., 1999). In all three species, M cells respond faster than P cells, suggesting that the division of pathways serves the same function: M cells encode spatial information and P cells encode color information. The only difference that Usrey and Reid found between owl and squirrel

monkeys was that overall, visual responses in owl monkeys were slower, which they speculated may be due to the nocturnal nature of the species. Between owl and squirrel monkeys, the receptive field surrounds were equally strong for M and P neurons. Based on these studies, it appears there are more similarities than differences between primate species in the early visual this website system, although a full, detailed analysis is beyond the scope of the present work. Compared to the CRF, less is known about the presence of an ECRF in the primate LGN. Indirect inhibitory input to the thalamus has been shown by Babadi and colleagues to modulate LGN responses in cats (Babadi et al., 2010). By identifying retinal input through S-potentials, they were able to exclude the retina as the source of the inhibitory modulation they observed, suggesting a non-retinal source as a likely candidate for extra-classical suppression. This agrees with

the findings of Kaplan et al. (Kaplan et al., 1987), who described Depsipeptide cell line nonlinear contrast gain control in both the cat and monkey LGN through simultaneous S-potential and LGN single unit recordings (i.e. the retinal input could not explain the nonlinear pattern in the LGN output). Solomon, White and Martin

(Solomon et al., 2002) looked extensively at the suppressive effects of ECRF stimulation, or extra-classical inhibition (ECI), in the primate LGN and found that more was present in the M and K pathways than the P pathway. Interestingly, while the strength of ECI increased as contrast increased in the ECRF, it also showed a dependence on the contrast of the RF, supporting their speculation that the ECRF might extend through the CRF as well. They suggested LGN interneurons as a likely source Ergoloid of ECI. Webb and colleagues investigated the spatial distribution, both fine and coarse, of the ECRF for M and P cells (Webb et al., 2005). Their findings show that the ECRF is larger than the CRF, consistent with other reports (Alitto and Usrey, 2008 and Solomon et al., 2002), but found that the ECRF is often asymmetric, concluding that there is no systematic spatial distribution to the ECRF. Webb et al. agree with Solomon et al. in the suggestion that the ECRF has different sources than the CRF, e.g. different retinal or thalamic sources, citing the correspondence between varying spatial configurations of LGN interneuron receptive fields and the asymmetric nature of ECI to also hypothesize that thalamic interneurons are involved in the ECRF.

036) and group 3 (treatment-naïve anti-VEGF injections + no planned supplement intervention; P = .014), but not when compared with group 4 (control; P = .215; Figure 2). Both wet AMD groups not taking omega-3 supplementation (groups 2 and 3) had similar levels of vitreous VEGF-A

(P = .758). Group 3 (treatment naïve) had significantly higher vitreous levels of VEGF-A when compared with nonvascular ocular pathologic features group 4 selleck chemicals (controls; P = .039; Figure 2). Seven of 9 patients in group 1 had concentrations of vitreous VEGF-A lower than all but 1 of the patients in group 2 ( Figure 2). Analysis of plasma levels of VEGF-A revealed no significant change between groups (P = .736; Figure 3). Similarly, although values for CFT tended toward improvement,

no significant benefit was noted with omega-3 supplementation in the sample population investigated in this pilot study (P = .211; Figure 4). In this pilot clinical trial, we Afatinib concentration investigated the influence of omega-3 supplementation on VEGF-A levels in the vitreous of patients undergoing anti-VEGF treatment for wet AMD and noted a significant decrease of VEGF-A in patients receiving omega-3. Dietary intake of omega-3 LCPUFAs and its influence on processes implicated in pathologic retinal angiogenesis has been proposed.18 We previously reported on the pronounced anti-angiogenic effects of certain omega-3 LCPUFA metabolites such as 4-hydroxy-docosahexaenoic acid (a metabolite produced via the 5-lipoxygenase pathway and acting through the peroxisome proliferator-activated TCL receptor). We also demonstrated that increased omega-3 LCPUFA

dietary intake reduces pathologic angiogenesis in experimental animal models of neovascular retinopathies.27, 29 and 32 Our previous genetic work in humans extended these findings to support the influence of omega-3 activated pathway on angiogenesis in wet AMD patients via complement and VEGF signaling systems.33 In the time frame of the current human study, the effects of omega-3 supplementation were exclusive to modulating vitreous levels of VEGF-A in proximity of the site of neovascularization, but not on systemic levels as determined by analysis of plasma. Interestingly, despite the significantly lower levels of VEGF-A in the vitreous of group 1, CFT values were similar to those of group 2 (after an average of 7 prior anti-VEGF injections) and of group 3 (Figure 3 and Table). In accordance with recent work in diabetic macular edema by Sonoda and associates, our findings also demonstrated a lack of correlation between CFT values and vitreous levels of VEGF in patients with active wet AMD (data not shown).34 These data agree with the notion that other factors besides VEGF-A may contribute to disease activity in wet AMD and that combination therapy with other agents is likely necessary in many patients to completely stall CNV activity and promote regression.

This was followed by a randomized, double-blind, placebo controlled Phase IIa study which assessed the formulation of 105.2 FFU/serotype in 60 healthy infants. SII BRV-PV/placebo was administered in 1:1 ratio as three doses with at least four weeks interval between doses. The study assessed the safety, GSK-3 cancer immunogenicity and shedding of the vaccine. Close post-vaccination follow-up showed the vaccine to be safe and well tolerated. A summary of the solicited vaccine

reactogenicity is summarized in Table 2. Almost all the events were mild and transient. Two SAEs (urinary tract infections and septicemia) unrelated to study vaccines were reported and both recovered uneventfully. We saw no effect on laboratory parameters. Three doses of the vaccine were found immunogenic. The seroconversion post dose 2 was 36% and 7.14%, in vaccine and placebo arms respectively (p = 0.0160). The corresponding post dose 3 seroconversion were 48% and 21.43% (p = 0.0492) ( Table 3). The post dose 3 GMTs in vaccine and placebo arms were 18.55 U/ml; and 7.31 U/ml. Following these satisfactory results, a randomized, double-blind, placebo controlled Phase IIb study was conducted which assessed the formulation of 105.6 FFU/serotype in 60 healthy infants. SII BRV-PV/placebo was administered in 1:1 ratio as three doses with at least four weeks interval. This formulation of the vaccine was also found safe and

well tolerated. A summary of the solicited vaccine reactogenicity is summarized in Table 2. click here Almost all the events were mild and transient. No SAE was reported and there all was no effect on laboratory parameters. Three doses of the 105.6 FFU/serotype formulation induced a significant immune response (Table 3). The seroconversion post dose 2 was 56.67% and 11.54%, in vaccine and placebo arms respectively (p value <0.05). The corresponding post dose 3 figures were 60% and 7.69% (p < 0.05). The seroconversion rates indicated that the 105.6 FFU/serotype formulation

is immunogenic in infants. These results are similar to those reported for the Rotarix (GSK) in an Indian study where the seroconversion rates were 58.3% [95% CI: 48.7; 67.4] in the Rotarix group and 6.3%; [95% CI: 2.5; 12.5] in the placebo group [20]. Another Indian study on the 116E vaccine showed 89.7% seroconversion in the vaccine arm and 28.1% in the placebo arm [21]. Another Indian study on Rotateq showed 83% 3-fold rise (seroconversion) in serum IgA antibodies; however the study had no placebo arm [22]. In developed countries, the seroresponses to rotavirus vaccines are high. The examples include a Korean study on Rotarix (88.1%) [23], a Korean study on Rotateq (94.7%) [24], a Japanese study on Rotarix (85.3%) [25], an European study on Rotarix (85.5–89.2%) [26], and a Finnish study on Rotarix (83.7–90.5%) [27]. However, for reasons not completely understood, the seroresponses are lower in developing countries. The examples include an African study on Rotateq (73.8–82.

Recent studies have shown that the HIV elite controllers have elevated numbers of high avidity polyfunctional cytotoxic HIV Gag-specific CD8+ T-cells in the mucosae compare to the HIV progressors [11], [12] and [13]. HIV transmits mostly via the genital tract or rectal mucosa and the first CD4 T cell depletion occurs in the gut mucosae [14]. It is now established that HIV is a disease of the mucosae, thus a mucosal vaccine approach may prove more useful in preventing and controlling HIV infection [15] and [16]. Unfortunately, due to the complexities

associated with delivery, safety and evaluation of vaccines efficacy in the mucosae, no mucosal HIV vaccine strategy has yet entered clinical development. Belyakov and find more co-workers have demonstrated that the intra-rectal immunisation induces local mucosal compartmentalisation of CTL of high “functional avidity” and protection of gastrointestinal CD4+ T cells from SHIV viral depletion in rhesus macaques compared to systemic delivery [17] and [18]. Consistent to their finding we have also found that i.m. rDNA/i.n. rFPV can induce

improved protection in macaques [19]. Since then in our laboratory we have studied the immune outcomes induced following mucosal and systemic heterologous prime-boost vaccination of antigenically distinct poxvirus vectors, Avipoxvirus Epigenetic inhibitor purchase fowlpox virus (FPV)-HIVgag/pol prime followed by an attenuated Orthopoxvirus vaccinia virus (VV)-HIVgag/pol booster vaccination [20]. These studies have shown that according to the route of vaccine delivery the quality or avidity of HIV-specific CD8 T cells can be vastly different and specifically, IL-13 and IL-4 have an inhibitory influence upon the development of high avidity CD8+ T cell responses. Our data has demonstrated that (i) mucosal vaccination

about can induce high avidity HIV-specific CD8+ T cells with reduced IL-4/IL-13 activity and better protective efficacy [21], (ii) IL-13 in the cell milieu has a direct negative impact upon CD8+ T cell avidity [22] and (iii) direct neutralisation of endogenous IL-13 activity using a high affinity cytokine receptor, IL-13Rα2 adjuvanted HIV vaccines delivered intranasal/intramuscular strategy can induce high avidity systemic and mucosal HIV-gag specific CD8+ T cell responses, with enhanced cytokine/chemokine expression and greater protective efficacy [23]. Surprisingly, transient inhibition of IL-13 activity at the site of immunisation in wild-type mice generated similar CD8+ T cell responses in regards to avidity and anti-viral protection as IL-13−/− gene knockout mice immunised with control vaccines [23]. Cytokines IL-4 and IL-13 share sequence similarity, cell surface receptor subunits, intracellular signalling and relatively similar functional effects on cells.

The guideline focuses on evidence underpinning four main areas: the diagnosis of JIA, treatment and management of JIA in the early stage, during acute episodes, and the long term management of JIA. It covers issues such as early and accurate diagnosis, care and referral pathways, use of medications, non-pharmacological management including evidence for land and water exercise, patient self-management education, and psychosocial support requirements. Two

detailed algorithms are presented on pages 8 and 9, covering the diagnosis Birinapant clinical trial and early management of JIA, and the management of JIA. A summary of the 21 recommendations is presented on pages 10–11, with more detailed explanation of the recommendation level and

specific evidence contained in pages 12–24. Three pages of resources are provided on pages 35–37 including publications, electronic sources (websites), and a history and clinical examination checklist to assist with examination and differential diagnosis. “
“Latest update: May 2010. Date of next update: 2014. Patient group: Individuals with chronic obstructive pulmonary disease (COPD). Intended audience: Health professionals who manage patients with COPD. Additional versions: This is the first update to the guidelines. The original guidelines were published in the Medical Journal of Australia in 2003. Caspase activity (http://www.mja.com.au/public/issues/178_06_170303/tho10508_all.html). Expert working group: The guidelines were developed by the Australian Lung Foundation and the Thoracic Society of Australia and New Zealand. The guidelines evaluation committee consisted of 8 Australian health professionals

representing medicine, public health, and physiotherapy. A larger group of 27 experts from Australia and New Zealand including physiotherapists ADAMTS5 also contributed. Funded by: Australian Lung Foundation. Consultation with: Draft versions of the guidelines were available on the RACGP website for public consultation and over 200 stakeholder groups were specifically targeted. Approved by: The Royal Australian College of Physicians, The Royal College of Nursing Australia, the Australian Physiotherapy Association, Australian Asthma and Respiratory Educators Association, and the Asthma Foundation. Location: The website (http://www.copdx.org.au/home) contains the guidelines spread over pages on the site, as well as a .pdf version. Description: The .pdf version is a 71-page document that presents recommendations and the underlying evidence to assist with the diagnosis and management of patients with COPD. The key recommendations are summarised on page 10 in the COPD-X plan: Confirm diagnosis, Optimise function, Prevent deterioration, Develop a self-management plan, and manage eXacerbations.

95% and as 47 ± 1.21% by the standard. During the oxidation process, peroxides were gradually decomposed to lower molecular weight compounds, like malonaldehyde, which could be measured by TBA method on the final day of the incubation period. The antioxidant activity of the nanoparticles was high on 7th day of incubation which was compared with the standard and was shown in Fig. 7. While the standard inhibited lipid peroxidation to 49 ± 1.31%, Selumetinib mouse the sample inhibited to 46 ± 1.71%. Absorbance was measured for various dilutions from 1:1 to 1:256 with concentration

of the sample ranging from 1000 μg/ml to 1.953 μg/ml and the corresponding percentage of cell viability was calculated. The cell viability of Human Epithelium cells of Liver cancer was found to be 16.39% at 1 mg/ml concentration of the sample with GI50 (50% Growth inhibition) NVP-BGJ398 nmr at 93.75 μg/ml as shown in the Fig. 8. The cytotoxic effects of the nano samples were depicted in Fig. 9. Scientists are focusing on medicinal plants to discover

natural antioxidants since some synthetic antioxidants have toxic effects. In addition, natural antioxidants play a vital role in protecting human health.23 Many reports have been published about the biogenesis of silver nanoparticles using several plant extracts but their antioxidant and anticancer activities have not yet been revealed. This study is the first report on the antioxidant and anticancer potential of silver nanoparticles synthesized from the leaf extract of M. pubescens. The activities of antioxidants have Rutecarpine been attributed to various mechanisms such as prevention of chain initiation, decomposition of peroxides, reducing capacity and radical scavenging.24 The silver nanoparticles studied exhibited significant radical scavenging activities. The effect of antioxidants on DPPH is thought to be due to their hydrogen donating activity.25 DPPH is considered as a lipophilic radical which makes it to readily accept electron from the antioxidant compound, converting its

color from purple to yellow which is detected at 517 nm. Superoxide anion radical is a weak oxidant but it gives rise to the generation of powerful and dangerous hydroxyl radicals as well as singlet oxygen, both free radicals contribute to oxidative stress.26 Hydroxyl radical is one of the potent reactive oxygen species in the biological system. It reacts with polyunsaturated fatty acid moieties of cell membrane phospholipids and causes damage to cell.2 In the metal chelating activity, Ferrozine can quantitatively chelate with Fe2+ and forms a complex with red color. This reaction is limited in the presence of other chelating agents and results in the decrease of red color of the ferrozine-Fe2+ complex. Measurement of the color reduction estimates the chelating activity of the sample to compete with ferrozine for the ferrous ions.27 Phosphomolybdenum reduction potential of M.

Tolerability and satisfaction were also measured the same way. Adverse events (such as haemoptysis, pharyngitis, and excessive coughing) were recorded after each treatment session. Whether an adverse event was severe enough to lead to intolerance of the trial intervention was also recorded. A blinded investigator questioned participants Cilengitide price specifically regarding these events. Adherence was assessed by counting unused sachets of hypertonic saline, and through documentation of each session of airway clearance techniques and hypertonic saline in the participant’s hospital case records. Furthermore, a physiotherapist attended each airway clearance session, even if the airway clearance techniques were

to be performed independently, to confirm compliance with the allocated timing regimen. At the conclusion of the 3-day study, participants reported their preferred timing regimen. For participants who repeated the 3-day study during the year of follow-up to determine if their preferred timing regimen had changed, perceived effectiveness, tolerability, satisfaction, preferred timing regimen, adherence, and adverse events were measured as previously. FEV1 was chosen as the primary outcome because

it has the potential to reflect both treatment efficacy and airway narrowing. We were unable to find an estimate of the smallest effect on FEV1 that adults with cystic fibrosis would consider makes using a particular timing regimen worthwhile. However, given that the timing regimens typically require SB431542 concentration similar time, effort, and expense, we postulated that even a very small effect would be worthwhile. Therefore we sought a difference of 150 mL between groups for the change in FEV1 across an individual treatment session. Pilot data provided a SD of 173 mL for this change in FEV1 among four adults with cystic fibrosis who met the eligibility criteria. Assuming this SD, 13 participants would provide 80% power, at the 2-sided 5% significance level, to detect a 150 mL difference in FEV1 as statistically significant between two groups in

the study. We increased Rutecarpine this to 32 to allow for multiple between-group comparisons and some loss to follow-up. We also sought to have sufficient statistical power to identify the smallest effect on satisfaction that would make it worthwhile to use one timing regimen instead of another. Again, given no established value and given that the timing regimens require similar time, effort, and expense, we nominated 10 mm on the 100 mm visual analogue scale as the threshold. Assuming a SD of 20 mm (Dentice et al 2006), 34 participants would provide 80% power, at the 2-sided 5% significance level, to detect a 10 mm difference in satisfaction as statistically significant between two groups in the study. We increased this to 50 to allow for multiple between-group comparisons and some loss to follow-up.

The activated OAg was designated OAg-oxNaIO4. For conjugation to CRM197, OAg-oxNaIO4 was added to CRM197 in NaH2PO4 100 mM pH 7.2 to give a final concentration of 10 and 5 mg/mL, respectively. NaBH3CN was added immediately after (OAg-oxNaIO4:NaBH3CN = 1:1 w/w),

and the reaction mixture stirred overnight at 37 °C. After this time, NaBH4 (OAg-oxNaIO4:NaBH4 = 1:1 w/w) was added and the mixture was stirred at 37 °C for 2 h. The conjugate was designated OAg-oxNaIO4-CRM197. OAg-oxTEMPO-CRM197: random activation of the OAg chain with TEMPO and conjugation to CRM197. OAg (3 mg/mL, corresponding to [CH2OH] of 7.69 mM) and NaHCO3 (molar ratio NaHCO3/CH2OH = 30), were added to a stirred solution of TEMPO (molar ratio TEMPO/CH2OH = 0.05) in DMF. The reaction was cooled Selleck PD98059 to 0 °C and TCC (molar ratio TCC/CH2OH = 1.6) was added. The activated sugar was recovered from the reaction mixture by precipitation with EtOH (85 v/v% in the final mixture) after 2 h of stirring at 0 °C. The pellet was washed twice with 100% EtOH (1.5 volumes with respect to the reaction mixture volume) and lyophilized. The activated OAg was designated OAg-oxTEMPO2h. The same procedure was used for the synthesis of OAg-oxTEMPO12h, increasing the reaction time to 12 h. OAg-oxTEMPO2 h

and OAg-oxTEMPO12h were conjugated to CRM197, using the same conditions for OAg-oxNaIO4. The two corresponding conjugates were designated Z-VAD-FMK supplier OAg-oxTEMPO2h-CRM197 and OAg-oxTEMPO12h-CRM197, respectively. OAg-ADH-SIDEA-CRM197: selective

activation of the terminal KDO with ADH, followed by reaction with SIDEA and conjugation to CRM197. The synthesis of this conjugate was performed as previously Florfenicol described [28] and detailed in SI. OAg-NH2-SIDEA-CRM197: selective activation of the terminal KDO with NH4OAc, followed by reaction with SIDEA and conjugation to CRM197. OAg was solubilized in 500 mM NH4OAc pH 7.0 at a concentration of 40 mg/mL. NaBH3CN was added immediately (NaBH3CN:OAg = 2:5 w/w). The solution was mixed at 30 °C for 5 days. The reaction mixture was desalted on a G-25 column and the OAg-NH2 was dried. The following steps of conjugation were performed as for OAg-ADH-SIDEA-CRM197 and the resulting conjugate was designed OAg-NH2-SIDEA-CRM197. All conjugates were purified by hydrophobic interaction chromatography (HIC) on a Phenyl HP column [GE Healthcare], loading 500 μg of protein for mL of resin in 50 mM NaH2PO4 3 M NaCl pH 7.2. The purified conjugate was eluted in water and the collected fractions were dialyzed against 10 mM NaH2PO4 pH 7.2. Total saccharide was quantified by phenol sulfuric assay [29], protein content by micro BCA (using BSA as standard and following manufacturer’s instructions [Thermo Scientifics]) and the ratio of saccharide to protein calculated. OAg-CRM197 conjugates profiles were compared with free CRM197 by HPLC-SEC and SDS-PAGE (see SI).