The HA ectodomain-encoding cDNA was cloned into the pCD5 expression vector for efficient expression in mammalian cells [9]. The pCD5-Cal/04/09 vector had been modified such

that the HA-encoding cDNA was cloned in frame with DNA sequences coding for a signal sequence, a GCN4 isoleucine zipper trimerization motif (KRMKQIEDKIEEIESKQKKIENEIARIKK) [10] and the Strep-tagII (WSHPQFEK; IBA, Germany). The HA ectodomain was expressed in HEK293T as previously described [11]. HA protein expression and secretion was confirmed by sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) followed by western blotting using a mouse anti-Strep-tag antibody (IBA, Germany). Secreted HA proteins were purified SB431542 cost using Strep-tactin sepharose beads according to the manufacturer’s instructions (IBA, Germany). The concentration of purified protein was determined by

using a Nanodrop 1000 spectrophotometer (Isogen Life Sciences) according the manufacturer’s instructions. Oligomeric status of the HA protein was determined by analyzing the elution profile using a Superdex200GL 10–300 column and by blue-native gel-electrophoresis. The vaccine was formulated with Specol [12] and [13] as an adjuvant, at 25 μg HA per dose of 2 ml. Pigs were vaccinated intramuscularly. Influenza virus A/Netherlands/602/2009 (H1N1)v was isolated from the first confirmed case in the Netherlands [14]. The patient was a 3-year old boy, developing a fever and symptoms of HKI 272 respiratory disease after returning from Mexico with his family. A nasal swab was taken before the patient was treated with oseltamivir. Virus was initially grown on embryonated eggs, and subsequently passaged on Madin–Darby canine kidney (MDCK) cells before it was used to inoculate the pigs. This virus differs by 8 amino acids from the A/California/4/2009 Florfenicol (H1N1)v strain [14]. Because it is, however, closer to the consensus sequence, it is considered representative of the circulating H1N1v influenza strains. Pigs were inoculated with a dose of 107.5 TCID50, suspended in 2 ml PBS, of which 1 ml was nebulised within

each nostril. Clinical symptoms and body temperature were recorded daily from day 3 before inoculation until the end of the experiment. At days 1–3 p.i. clinical symptoms and body-temperature were recorded twice per day with a 12 h interval. Serum samples were collected during both times of vaccination, at the time of inoculation, and 7, 10, 14 and 21 days p.i. Oropharyngeal and nasal swabs were collected daily from all animals still alive from day 0 to 11 p.i., and on days 14, 17 and 21 p.i. For oropharyngeal swabs multi-layered gauze dressings in a pair of tweezers were used to scrape the palatine tonsils at the dorsal pharyngeal wall, behind the soft palate. Nasal swabs were collected using sterile rayon swabs (Medical Wire & Equipment, Corsham, United Kingdom).

Individuals with chronic pain often experience significant functional impairment

as well as difficulty in occupational/ social roles. The CPGQ may not provide a comprehensive assessment of how ongoing pain affects the functions and participation in life roles; however it can be utilised as a preliminary assessment tool to ascertain the extent of disablement resulting from chronic pain. Further research is required to determine if the 5 categories of CPGQ allow thorough and consistent selleck compound discrimination of pain severity and disability among individuals with varying degree of pain/ disablement. Hence, CPGQ with further validation can facilitate individualised management tailored according to the clinical subgroup of the patient (high pain versus high disability). Lastly, responsiveness and MCID of the subscales of the CPGQ need to be established in prospective longitudinal studies. “
“Latest update: 2011. Next update: 2015. Patient group: People aged 18 year or older with contracted (frozen) shoulder. Intended audience: Professionals involved in caring for people with contracted (frozen) shoulder – buy Ku-0059436 physiotherapy teachers and practitioners foremost, but also commissioners/providers of healthcare, GPs, orthopaedic surgeons, radiologists and rheumatologists. The guideline has been written in plain English to be accessible to patients

and their representative organisations. Additional versions: Nil. Expert working group: A 10-member

group of physiotherapists from the United Kingdom (UK) with expertise in the shoulder comprised the expert working group. Funded by: This guideline development received no funding support. Consultation with: The expert working group consulted with a 14 member multidisciplinary Delphi panel including medical specialists and patient representatives from the UK. The before guidelines were reviewed by the Good Practice Panel of the Chartered Society of Physiotherapy and five independent expert reviewers. Approved by: The Chartered Society of Physiotherapy, UK. Location: Hanchard N, et al (2011) Evidence-based clinical guidelines for the diagnosis, assessment and physiotherapy management of contracted (frozen) shoulder v.1.6, ‘standard’ physiotherapy. www.csp.org.uk/skipp Description: This guideline is a 170-page document that aims to identify and critically appraise the best available evidence relating to the diagnosis, assessment, and physiotherapy management of contracted (frozen) shoulder. It begins with a description of key concepts and methods in a manner that a clinician with only limited grounding in research should be able to understand. Information on the anatomy, pathology, and terminology linked to frozen shoulder is presented. Factors to consider and evidence underpinning the diagnosis and usual presentation of this pathology are outlined.

Complications from Gc infections are frequent, debilitating, and disproportionately affect women. selleck products Untreated cervical infections commonly progress to the upper reproductive tract, which contributes to pelvic inflammatory disease (PID), infertility, life-threatening ectopic pregnancy, and chronic pain. Infertility rates following PID are high, at >10% following a single episode and >50% following three or more episodes [1]. In men 10–30% of untreated urethritis cases may progress to epididymitis, a common cause of male infertility in some

regions [2]. During pregnancy, Gc causes chorioamnionitis complicated by septic abortion in up to 13% of women, preterm delivery in 23% of women, and premature rupture of membranes in 29% of women [3]. Neonatal conjunctival infections are destructive, leading to corneal scarring and blindness. Gonorrhea also dramatically increases the acquisition and transmission of human immunodeficiency virus (HIV) [4]. An estimated 106 million Gc infections occur annually, worldwide [5]. Diagnostic capabilities and surveillance systems vary between nations, and thus, infection is greatly underreported and prevalence is often highest among economically or socially disadvantaged populations. Microbiologic culture is diagnostic, but syndromic management alone is

standard for many regions of the world. Rapid DNA-based tests have improved sensitivity, especially for asymptomatic disease, but are not available in all countries. In all situations, treatment find more is empiric at the initial point of care to eliminate further transmission. Antimicrobial resistance patterns guide treatment recommendations, the goal of which is to effectively treat ≥95% of infections at first presentation. Antibiotic resistance is widespread and has developed rapidly with each successive treatment regimen. Alarmingly, with the advent of resistance to extended-spectrum Phosphatidylinositol diacylglycerol-lyase cephalosporins, we have now reached the point where untreatable disease can be anticipated in the

near future [6]. Although rapid effective treatment of gonorrhea decreases long-term sequelae and can eliminate the effect on HIV transmission [7], expansion of multi-drug resistant Gc is a global threat to public health and amplifies the urgent need for novel prevention methods. Development of an effective gonorrhea vaccine is likely to have significant benefits given the impact of gonorrhea on human health. Ebrahim et al. estimated 1326 disability-adjusted life years (DALYs) are attributable to 321,300 Gc infections. Applied to WHO global estimates of new Gc infections, this translates to 440,000 DALYs per year [8] and [9]. The benefits of effective treatment to women also have been estimated: treatment of 100 women with gonorrhea, of which 25% are pregnant, would prevent 25 cases of PID, one ectopic pregnancy, 6 cases of infertility, and 7 cases of neonatal ophthalmia.

Linear regression was used to estimate the difference and associated 95% confidence intervals. Because selleck CRP levels did not follow a normal distribution, it was log-transformed in linear regression models. Last, we created case–PT pairs of participants matched on age (± 5 years) and gender and compared their differences in WBC, CRP, LINE-1 methylation and IL-6 methylation using paired-T tests. All statistical analyses were performed using SAS (version 9.1; SAS institute, Cary, NC). There were 79 car drivers and 101 PT users. Car drivers were older, had higher BMI, and included a greater proportion of males and non-Hispanic whites than

PT commuters (Table 1). Car drivers ate more fruits and more meats than PT users (p = 0.02 and 0.04 respectively, Table 2). We identified two dietary patterns in the study population: the prudent dietary pattern was characterized by high intakes of vegetables and fruits; and the western dietary pattern was characterized by high intakes of meats, grains and dairy products. Overall the two groups did not differ in the adherence to the two dietary patterns, either for the prudent or for the western diet (Table 3). Car drivers reported a higher level of light job activities and a lower level of sedentary activities than PT commuters (p = 0.007 and 0.004 respectively). Overall, car drivers had higher adherence to 2005 DGA for physical activity

than PT commuters (78.5% vs. 65.0%). However, after adjusting for age, gender, race/ethnicity and BMI, the difference in adherence to the 2005 DGA for physical activity became statistically insignificant Alectinib chemical structure (difference: − 14.2%, 95%CI: − 29.0, 0.5) (Table 4). In Table 5, there were no differences in median level of CRP (car vs. PT: 0.6 vs. 0.5 mg/dl, difference in log-CRP: 0.2, 95%CI: − 0.2, 0.5) and mean level of WBC (car vs. PT: 6.7 vs. 6.5 cells/mm3, difference: − 0.4, 95%CI: − 0.9, 0.2). In Table 6, there were no differences in mean levels of LINE-1 methylation (car: 78.0%;

PT: 78.3%, difference: 0.2, 95%CI: − 0.5, 1.0), and IL-6 promoter methylation (car: 56.1%; PT: 58.0%, difference: 1.7, 95%CI: − 2.4, 5.8). Missing values in Table 6 are due to low DNA yield following extraction from the buffy or low quality science calls on pyrosequencing LINE-1 methylation or IL-6 promoter methylation. A total of 58 1-to-1, age-gender matched pairs comprising one PT commuter and one car commuter were formed. No statistically significant differences were found for WBC (difference = 0.07 cells/mm3, 95%CI: − 0.64, 0.77), CRP (difference = 0.03 mg/dl, 95%CI: − 0.67, 0.74), LINE-1 methylation (difference = − 0.07%, 95%CI: − 0.91, 0.77) and IL-6 methylation (difference = − 3.81%, 95%CI: − 10.15, 2.52) between pairs. There remained, however, an age difference of about 1 year (difference = 0.98 year, 95% CI: 0.58, 1.39) within pairs.

15 according to Eq. (A.6). The log Ppara, log Pfilter, and log PABL were added as fixed contributions, as log P0 www.selleckchem.com/MEK.html and log Puptake were refined ( Appendix A.5) for the non-inhibitor and added-inhibitor (50 μM PSC833) sets. Both the intrinsic and the uptake permeability values appeared to be affected by efflux ( Table 3). The two sets were

then combined, with the repeated refinement yielding log P0 = −5.28 ± 0.04, log Puptake = −5.73 (kept fixed), and log Pefflux = −5.80 ± 0.04 for the non-inhibitor set and log Pefflux < −8 for the +50 μM PSC833 set. This suggested that efflux was essentially suppressed by the inhibitor. With the log Pefflux of −5.80, it was possible to rationalize the extent to which the individual-set refined log Puptake and see more log P0 in the two sets were different. Fig. 4c and d shows colchicine and digoxin with added efflux inhibitor (checkered circle) and no-inhibitor (black circles). The addition of inhibitors increases the apparent permeability by nearly the same amount in both drugs, consistent with the suppression of efflux

transporter. To assess the ability to predict in vivo BBB permeability of a compound from permeability data measured using the PBEC model, P0 (in vitro) derived from our PBEC model permeability data was plotted against P0in situ (in vivo) derived from in situ brain perfusion data in rodents ( Fig. 5). Published data from other in vitro porcine BBB models were also included in the linear regression analysis. The r2 value all of 0.61 shows a good correlation for the pooled data. The in vitro blood–brain barrier

(BBB) model from primary porcine brain endothelial cells (PBEC) which shows a restrictive paracellular pathway was used for permeability studies of small drug-like compounds of different chemistry: acid, bases, neutrals and zwitterions. Assay at multiple pH was conducted for the ionizable compounds propranolol, acetylsalicylic acid, naloxone and vinblastine to plot permeability vs. pH. The pCEL-X software (Section 2.5 and Appendix A) was used for detailed permeability data analysis, including aqueous boundary layer (ABL) correction. The ABL was found to restrict propranolol permeability, which was also limited by low pore density of the Transwell®-Clear polyester filter membrane. The intrinsic transcellular permeability P0 showed good correlation with in situ data, indicating the predictive power of the in vitro model. Stirring helps to diminish the ABL thickness, but it cannot reduce it entirely. This is because the aqueous medium adjacent to the membrane surface is less mobile due to hydrogen bonds formed at the interface (Loftsson and Brewster, 2008). Hence, even vigorous stirring is unable to remove the ABL totally. Furthermore, excessive stirring is undesirable, since it can compromize tight junction integrity (cf., Zhang et al., 2006: 600 RPM). Application of the pKaFLUX method for ABL correction using pCEL-X proved useful particularly for ionizable compounds.

Student’s t-test was employed to determine the significance of differences between the studied groups. p values <0.05 (*) were

considered to be significant. DNA fragments encoding bfpA (600 bp) and intimin (eae388–667) (840 bp), were amplified by PCR from EPEC (E2348/69) and ligated into the KpnI and BamHI sites of the pMIP12 vector under the control of the pblaF* promoter IGF-1R inhibitor ( Supplementary Figure); the constructs were named pMH12-bfpA and pMH12-intimin, respectively. The plasmids were electroporated into BCG and Smeg, and the resulting strains were examined for BfpA and intimin expression. Expression of both bfpA and intimin (eae) was confirmed by immunoblotting bacterial whole-cell extracts using anti-BfpA or anti-intimin antisera. As observed in Fig. 1A and B, the antisera specifically recognized bands of approximately 19.5 and 34 kDa, corresponding to BfpA and intimin, respectively, from both rBCG and rSmeg strains. No proteins were recognized by the antisera in whole-cell lysates from BCG or Smeg controls without the plasmid vectors ( Fig. 1A and B). C57BL/6 mice were immunized by oral gavage or intraperitoneal injection with 4 doses of 1 × 108 CFU in 200 μL of rBCG-bfpA, rSmeg-bfpA, rBCG-intimin or rSmeg-intimin at two-week intervals. As a mucosal adjuvant, SBA-15 Hydroxychloroquine solubility dmso silica was used. Control mice were immunized with

non-recombinant BCG or Smeg or with PBS following the same immunization schedule. A significantly higher level of anti-BfpA and anti-intimin IgA or IgG antibodies was observed in

both the feces and serum of mice immunized with rBCG or rSmeg as compared with that of serum collected in the groups that received non-recombinant BCG or Smeg or PBS (p < 0.001) ( Fig. 2A and B). Pre-immune sera and feces that were collected and pooled were evaluated, and presented no reactivity to BfpA or intimin (data not shown), suggesting the absence of anti-BfpA or anti-intimin antibodies prior to immunization. Our analysis of serum IgG subclass Rolziracetam responses also revealed that mice subjected to intraperitoneal immunization predominantly developed an IgG2a response, indicating a Th1-type cell response ( Fig. 2C). To evaluate the involvement of Th1-type cells on the immune responses induced by recombinant BCG-bfpA, BCG-intimin, Smeg-bfpA and Smeg-intimin, spleen cells were recovered 15 days after the final immunization and treated in vitro with the corresponding recombinant protein expressed in the vaccine used. We assayed the supernatants for the presence of the cytokines TNF-α, IFN-γ, IL-4 and IL-5. As is shown in Fig. 2A–C, anti-BfpA and anti-intimin, respectively, IgA and IgG antibodies were detected in feces and serum. Immunization with recombinant vaccine expressing BfpA induced higher production of IFN-γ, in vitro, by spleen cells (Fig. 3).

The increase in availability and use of Epigenetic activity rotavirus vaccines in the

future underlines the importance of surveillance networks to investigate the post-vaccine introduction epidemiology of rotavirus in terms of disease burden and effect on strain types. Sudhir Babji was supported by the Global Infectious Disease Research Training Grant (D43TW007392; PI – GK). None of the authors report a conflict of interest. “
“Rotavirus infection, mostly caused by Group A viruses, is prevalent in human populations worldwide. Although the virus infects older individuals, the disease can be severe in immunologically naïve infants and young children. The burden of severe rotavirus illness and deaths falls heavily upon children in low and middle-income countries: more than 80% of rotavirus-related deaths are estimated to occur in lower income countries of Asia and sub-Saharan Africa [1]. India has an especially large population at risk of clinically significant rotavirus gastroenteritis (GE); of the 1.2 billion people, 11% are <5 years old. Worldwide in 2008, diarrhea attributable to rotavirus infection resulted in 453,000 deaths (95% CI 420,000–494,000) in children younger than 5 years representing CHIR-99021 solubility dmso 37% of deaths attributable to

diarrhea and 5% of all deaths in children younger than 5 years. Five countries accounted for more than half of all deaths attributable to rotavirus infection: Democratic Republic of the Congo, Ethiopia, India, Nigeria, and Pakistan with India alone accounting for 22% of deaths (98,621 deaths) [2]. Typical clinical signs of infection include fever, projectile vomiting, and profuse watery diarrhea, which may significantly dehydrate the infected child. Moderate to severe dehydration in young children is more often associated mafosfamide with rotavirus infection than other enteropathogens. There are no specific medications for rotavirus GE, but rehydration with oral rehydration salts (ORS) has long been a standard therapy for acute infantile diarrhea. Severe dehydration can be life threatening and requires treatment in a clinic or hospital where the child

can receive intravenous (IV) fluids and appropriate case management. The purpose of this observational study was to carry out a hospital-based surveillance of rotavirus gastroenteritis in children ≤59 months of age and develop estimates of disease burden in the population under surveillance. A prospective hospital-based surveillance was conducted at 12 medical centers attached to Medical Schools across India. From North India subjects were enrolled from Dayanand Medical College & Hospital, Ludhiana; Chhatrapati Shahuji Maharaj Medical University, Lucknow; Kalawati Saran Children Hospital, New Delhi; Post Graduate Institute of Medical Education and Research, Chandigarh and Sawai Man Singh Medical College, Jaipur.

For relative importance, the most highly rated cluster was Personal Ability (cluster average = 4.21). For feasibility to implement, the most highly rated cluster was Sidewalks and Crosswalks (cluster average = 3.66). The Go-Zone map (Fig. 3) compared statement ratings from low to high for both relative

Epacadostat price importance and feasibility to implement. The top right quadrant is the ‘Go-Zone’ for action and reflects statements rated as both important and feasible. Rating scores placed 18 statements within the Go-Zone for action. Twelve of these eighteen statements arose from the sidewalks/crosswalks (n = 7) and neighborhood features (n = 5) clusters. We used a novel approach, concept mapping, to identify elements of the built and social environments that are perceived to influence older adults’ outdoor walking. Our findings are important

for three reasons: older adults command an increasing proportion of the global population (World Health Organization, 2011); decisions regarding neighborhood attributes have implications for older adult mobility; and we reside within an increasingly constrained fiscal environment of public accountability that must prioritize scarce resources. Therefore, our findings are timely and important check details as they guide decision makers regarding priority areas of investment in the built environment that promote mobility of an increasingly aging population. Our findings also highlight areas of enquiry for further research. What emerged as a clear priority for participants was both the presence Adenylyl cyclase and the characteristics of sidewalks and crosswalks. About half of all statements within this cluster were considered both important

and feasible to implement; and this is consistent with the literature related to walking outdoors and older adults’ pedestrian mobility. Safely navigating sidewalks and streets is vital for older adults’ outdoor mobility; and walking is impeded if sidewalks are absent or poorly maintained (Corseuil et al., 2011) or if pedestrian crossing times are too short to allow older adults sufficient time to cross the street (Grant et al., 2010). We deemed statements considered both important and feasible to implement as particularly relevant targets for new or renewed policy efforts. For example, building sidewalks on at least one side of the street was important to participants and is already required for new developments in many major municipalities. Thus, some of our findings reinforce what is already known, validating existing and new policies, and priority areas for investment by local and provincial government. Public transportation and pedestrian routes were also identified as highly important and feasible to implement; and accessible private vehicle parking fell just outside the ‘go-zone’ cut-off.

13, 14, 15 and 16 In the present study the binding interactions of some of the 3,4-heteroannelated quinolin-2-ones with DNA Gyrase as well as their antibacterial activity has been reported. These compounds are assumed to bind to inhibit the DNA Gyrase 2 of S. aureus in a similar fashion as ciprofloxacin does, since the compounds share structural similarity with Ciprofloxacin. The potential compounds are identified and their antibacterial activity is evaluated against S. aureus and Escherichia coli and

reported here. To study the Selleck MLN8237 extent of interaction of the synthesized 3,4-heteroannelated quinolin-2-ones with the DNA Gyrase of S. aureus, the compounds were docked to the protein using the GOLD 3.2 (Genetic Optimization for Ligand Docking) software. The docked poses of each ligand were analyzed and fitness Scores are calculated with Silver. Screening of antibacterial activity of title compounds was done by adopting disc diffusion method as described by Cruickshank et al (1975)17 using S. aureus

Gram +ve (Oxford strain) and E. coli Gram −ve (NCTC 10148). The synthesized compounds were constructed and prepared for docking using the Ligprep Protocol of Maestro. Ligand minimization was done using OPLS 2005 Force field. The synthesized compounds were constructed and prepared for docking using the Ligprep Protocol of Maestro. A high resolution (2.1 Å) crystal structure of S. aureus DNA Gyrase is selected and docked using GOLD 3.2. The GOLD INCB024360 supplier fitness function is made up of four components: protein–ligand hydrogen bond energy, protein–ligand Vander Waals energy, ligand internal vdw energy and ligand torsional strain energy and the fitness score is taken as the negative of the sum of the component energy terms. The docked poses of each ligand were visualized and the interactions were analyzed with Silver. The best Fitness

Scores for each ligand are tabulated along with the details of H-bonds and other interactions in Table 1. The binding poses of the ligands to the proteins are shown in Fig. 1. Compound 1b has the highest fitness score of 51.23. Compounds 1a, 1c, 2d and 2j also showed good fitness scores of next to 1b. 11 compounds show good fitness score values as compared to Ciprofloxacin. Screening of antibacterial activity of title compounds was done by adopting disc diffusion method as described by Cruickshank et al (1975).17 The compounds were dissolved in appropriate solvents (AR grade) and Whatman No.1 filter paper discs of 6 mm diameter were prepared with various concentrations of test compounds ranging from 200 to 3.125 μg/disc. The test organisms used were S. aureus Gram +ve (Oxford strain) and E. coli Gram −ve (NCTC 10148).

Cell suspensions were obtained using a cell strainer (70 μm, Becton Dickinson). Cells were washed and cultured in 96-well flat bottom plates at a density of 2.0 × 105 cells/well in triplicate Angiogenesis inhibitor and restimulated with 40 μg/ml OVA. ConA (Sigma–Aldrich) 5 μg/ml was used as a positive control. After 3 days the supernatants

were collected and stored at −80 °C until further use. The amount of IFN-γ in the supernatant was determined by ELISA using a commercial kit (Becton Dickinson) according to the manufacturer’s instructions. Statistical analysis was performed with Prism 5 for Windows (Graphpad, San Diego, USA). Statistical significance was determined either by a one way or a two way analysis of variance (ANOVA) with a Bonferroni post-test, depending on the experiment set-up. With the film hydration method and subsequent extrusion, OVA-containing liposomes with an average size of 130 nm and a positive zetapotential could be prepared in a reproducible manner (Table 1). Ultrafiltration showed that nearly 100% OVA was associated with the liposomes. PAM could be easily incorporated into the liposomes (∼85%)

and the incorporation did not affect the (measured) liposome characteristics. The addition check details of CpG did influence the liposome characteristics as the size augmented by two-fold. Furthermore, CpG reduced OVA association with the liposomes, probably due to competition between the antigen and the TLR ligand as both compounds bear a negative charge. The stability and release of the OVA liposomes was studied over time in PBS at 37 °C. Dilution in PBS had an initial effect on the size of the liposomes as their size decreased from 130 nm to 90 nm, due to the influence of PBS on the hydrodynamic diameter of the liposomes [31]. After this initial size decrease, the size remained stable during the following 8 days

(Fig. 1). During this period OVA was released PDK4 from the liposomes. An initial burst release of 25% was observed and after 5 h already 50% of the OVA was no longer associated with the liposomes. During the following 8 days the remaining OVA was slowly released. PAM and CpG are two TLR ligands. The effect of ligand encapsulation in OVA liposomes on their interactions with the TLRs was studied on HEK293 cells transfected with either TLR2 (receptor for PAM) or TLR9 (receptor for CpG). Non-adjuvanted liposomes and a solution of OVA did not induce TLR2 or TLR9 activation (data not shown). PAM in solution was a stronger TLR2 activator compared to the liposome encapsulated PAM (Fig. 2A). A 15-fold higher dose of PAM was necessary to obtain the same level of IL-8 production from the HEK293-CD14/TLR2 cells. Both PAM in solution and OVA/PAM liposomes activated the cells in a concentration dependent manner. CpG activated TRL9-transfected HEK cells in a concentration dependent way as well.