PEDro includes 564 Cochrane reviews, indicating that a tenth of a

PEDro includes 564 Cochrane reviews, indicating that a tenth of all Cochrane reviews are either directly or indirectly relevant to physiotherapy practice. In The Cochrane Library, 254 of the Cochrane reviews (approximately 5%) are directly indexed as ‘Physical Therapy Modalities’. This is of CP-673451 molecular weight particular importance in supporting evidence-based physiotherapy, because Cochrane reviews relevant to physiotherapy have been demonstrated to be of higher quality than physiotherapy reviews published outside of The Cochrane Library. 1 These reviews provide reliable evidence to inform physiotherapy intervention

decisions and guide practice, and demonstrate the credentials of physiotherapy as a research active and informed profession. Further demonstrating the relevance of The Cochrane Library to physiotherapy, interventions delivered by physiotherapists AZD6738 clinical trial or relevant to physiotherapy feature in 10 of the 20 most accessed reviews in The Cochrane Library, as presented in Table 1. In addition to systematic reviews, The Cochrane

Library includes CENTRAL, a database of randomised controlled trials and other studies eligible for inclusion in Cochrane reviews. These studies have been identified through the efforts of Cochrane’s many contributors and volunteers. Importantly, this database includes trials in languages other than English, or published in journals not indexed in MEDLINE, thereby facilitating access to studies that would otherwise be difficult to find. CENTRAL’s coverage of randomised trials of physiotherapy interventions is as good or better than other major bibliographic databases that index such trials. 2 and 3 As well as automatically including reports of randomised trials indexed in MEDLINE, Edoxaban CENTRAL also contains many reports of trials that are unique to EMBASE. We estimate that at least 12 000 reports of trials of physiotherapy

interventions from MEDLINE and EMBASE are included in CENTRAL. Furthermore, the manual searching of journals and conference proceedings was commonplace in the early days of Cochrane and would often result in discovering reports of trials that would never otherwise be identified. For example, hand searching the Australian Journal of Physiotherapy (1955 to 2009) yielded over 250 trial reports, many of which were only reported as conference abstracts, but which are now available in CENTRAL. The influence of Cochrane on physiotherapy research and education in Australia is further demonstrated by the role of the Australian schools of physiotherapy in authoring Cochrane reviews and including the use of The Cochrane Library in their curricula. Of the 14 Australian schools of physiotherapy listed by the Australian Physiotherapy Council, at least 10 have members of staff who are authors of Cochrane reviews.

If a paracellular

marker was used in the assay to define

If a paracellular

marker was used in the assay to define the paracellular limit, deviation of the experimental data from this limitation could suggest presence of uptake mechanism(s) for the charged form of a compound. With pCEL-X analysis, naloxone and vinblastine showed such pH-dependent deviation in the present study. At physiological pH 7.4, both compounds are charged (cationic). Organic cation transport system could be involved in uptake of these compounds. Although it was not possible to detect uptake transport in the case of acetylsalicylic acid (nor was such a process reported in 3-MA research buy the literature for the molecule), a similar molecule, salicylic acid, the primary metabolite of acetylsalicylic acid, was found at high concentration in the

brain (brain-to-blood concentration ratio 1.06) after intraperitoneal injection of acetylsalicylic acid in mice ( Prins et al., 2009). Our finding of concentration-dependent permeation of naloxone is consistent with in vivo studies by Suzuki et al. (2010) reporting concentration-dependent uptake of naloxone in rat brains as measured by the Brain Uptake Index (BUI). The uptake mechanism is proposed to involve a pH-dependent cationic H1-antagonist transporter ( Suzuki et al., 2010). The results provide evidence that the combination of our in vitro BBB model from PBEC with detailed pKaFLUX analysis reaches the same learn more conclusion as in vivo studies, further validating the PBEC model and confirming its ability to predict in vivo BBB function. The intrinsic transcellular permeability P0 derived from measured Papp can reflect a purely transcellular passive permeation PD184352 (CI-1040) or a combination of passive and carrier-mediated mechanism(s). While uptake of charged forms can be clearly revealed, specific

transport of the neutral form is not as easily recognized unless the assay is repeated to include transport inhibitors or unlabelled compounds to provide competition for uptake. A decrease in P0 in the presence of competing substrates suggests uptake mechanism(s) and an increase in P0 in the presence of inhibitors suggests that the compound may be subject to efflux mechanism(s). For ionizable compounds, if the assay is conducted at a single pH, uncertainty may arise in the analysis. The uncertainty derives from difficulty in determining the pKaFLUX or ‘bend in the curve’ when fitting all the parameters to the experimental data. One way to reduce the uncertainty is by defining at least one boundary, i.e., ABL or paracellular permeation, using appropriate markers. The method would be moderately demanding for screening purposes, but its value would be predictive information from pCEL-X before permeability experiments, helping to design experiments better, thus saving time and resources. Also, detailed data analysis in pCEL-X after experiments gives additional information and insights into permeability mechanisms.

I can only talk for me … but I think that generally as therapists

I can only talk for me … but I think that generally as therapists we quite like to problem solve for our client. There were silences and there were pauses, which did throw it back on the client. (Physiotherapist A, 16 years’ experience) The coaching process was seen to have potential value as part of ongoing negotiation throughout the rehabilitation process and not just at the outset. … but often down the track a little

bit it would be good to have something that you kind of put in place because priorities for people change. (Physiotherapist selleck products D, 5 years’ experience) A notable finding was that aspects of the coaching process did cause discomfort to the physiotherapists. At times a sense of emotional tension was expressed especially if the patients were perceived to be complex or unrealistic. It is interesting to note that these fears were primarily about

potential issues rather than actual issues, and were related to the physiotherapist perceptions of the patients’ vulnerability. There was also a sense of discomfort at the possibility of Galunisertib supplier encountering emotional distress and they perceived this as being potentially harmful. I was a bit concerned about how my client would actually respond for the simple reason that he has a lot of social things going on in his life, and I just wondered … whether it unearthed stuff … He said he was okay, so maybe it was more my discomfort as far as knowing what is going on at home. (Physiotherapist A, 16 years’ experience) For the participants, taking part in the process also allowed them to refocus on what was important to them, which was often accompanied by an increase in motivation to continue to address their chosen rehabilitation goals. She seemed to get to the heart of the matter. She seemed to know that I badly wanted to walk and took steps to encourage that. I felt that she was really interested

in achieving my goal. (Patient D) In a similar way to the physiotherapists, taking part in the coaching session meant that the patients in the study were able to be a more active participant. They described being more intentional in pursuing their goals, taking more these responsibility for achieving this, and were able to articulate more coping strategies to address unexpected barriers that occurred. They were also more likely to revisit and reuse strategies that had been helpful in the past, such as the use of diaries and planning when to exercise. And it’s more associated with what I do, rather than what other people do. So I decided what the goal was and I decided everything and then I had to do everything. (Patient F) The patients also identified that the intervention was not long enough, and that on-going support and tracking of progress could make the process more helpful.

15 Hydro-methanolic seed extracts of the plant also showed inhibi

15 Hydro-methanolic seed extracts of the plant also showed inhibition of deoxyribose degradation by OH− ions, inhibition of nitrite formation by competing with O2, degradation of

H2O2 and inhibition of lipid peroxidation, all from the ethyl acetate fraction. 16 Crude aqueous methanolic seed extracts of H. antidysenterica significantly decrease the size of calcium oxalate crystals and convert them from calcium oxalate monohydrates (COM) to calcium oxalate dehydrate (COD) in vitro. The extract suppresses cell toxicity (induced by COM) and production of lactate dehydrogenase. The extract was tested in vivo in male wistar rats, which showed substantial decrease in polyurea, water intake, Ca++ INCB28060 cost excretion and crystal formation. 17 Stem bark extract of H. antidysenterica in the form of “Kutaja tvak churna” showed healing activity in patients suffering from bleeding piles. 18 Aqueous seed extract of H. antidysenterica showed a significant increase in urine

output of wistar rats at dosage range of 30–100 mg/kg. A substantial increase was also observed in the amount of Na+ and K+ ions excreted through urine of treated rats. 19 A daily intake of the bark powder for 15 days completely cured patients suffering from amoebiasis. Another clinical trial investigated the therapeutic efficacy of “Amoebin cap”, a medicine for amoebiasis containing H. antidysenterica as one of its constituents. 20 Inhibition of acetylcholinesterase is desirable when treating neurological problems such as Alzheimer’s find more disease. Since alkaloids from some plants have already been known to inhibit AChE, a study tested some alkaloids of H. antidysenterica for similar action. Out of five isolated alkaloids (conessine, isoconessimine, conessimine, conarrhimine and conimine), conessimine exhibited the most profound effects, with an IC50 value of 4 μM. The study concluded that these alkaloids can be potentially used in drugs for

treating much neurological disorders. 21 A separate study investigated the CNS-stimulating activity of methanolic bark extract on Swiss albino mice. The results showed that regardless of the dosage, the extract significantly decreased and relaxed the gripping capabilities of the muscles and also the spontaneous locomotive activity, thus indicating a depressing effect on the CNS.22 In-vitro activity of aqueous and ethanolic extracts bark on Pheretima posthuma (earthworm) showed significant results. 23 Ethanolic seed extracts showed concentration-dependent zones of inhibition against bacterial cultures of EPEC bacteria. Since EPEC is resistant to many antibiotics, the ethanolic extract is considered as a promising antibacterial agent. 2 In one study, petroleum ether extract of bark inhibited E. coli at 50 μg/ml with a minimum inhibitory concentration (MIC) of 50 μg/ml while methanol and chloroform extracts did so at higher concentrations, thus having higher MIC values. Yet, compared to other plants, H.

microplus varies according

microplus varies according GDC-0199 research buy to characteristics of the tick population targeted and host factors among other things [14] and [15]. Pen trials conducted in the state of Mato Grosso do Sul, Brazil revealed that the efficacy of Bm86-based vaccines against the Campo Grande strain of R. microplus ranged from 31 to 49% [17] and [18]. Efficacy around 99% against R. annulatus obtained with Bm86-based vaccines is an indication of the consistent high level of anti-R. microplus immunoprotection that a novel antigenic and immunogenic

tick molecule, or combinations thereof, could elicit in vaccinated cattle. Such level of efficacy offers the opportunity to incorporate vaccination as a tool for the integrated eradication of cattle fever tick populations [40] and [41]. The search for protective antigens that are highly efficacious against R. microplus continues. Proteinase inhibitors have received attention as a group of molecules found in ticks with potential for use as Sunitinib datasheet immunogens in an anti-tick vaccine. Several trypsin inhibitors that are present in the egg, larval and adult stages of R. microplus have been described [19], [20] and [21].

It has been suggested that the R. microplus serine protease inhibitors may be involved in larval attachment at the bite site and blood feeding [22]. Trypsin inhibitors from R. microplus larvae purified in their native form elicited a protective immune response in vaccinated cattle yielding 72.8% efficacy, and 69.7% reduction in the number of adult female ticks completing the parasitic phase of their life cycle [22]. However, a peptide ADP ribosylation factor designed from one of the R. microplus larval trypsin inhibitors afforded only 18.4% immunoprotection against tick infestation in crossbred cattle [23]. The use of recombinant trypsin inhibitors can circumvent the challenge of having to purify trypsin inhibitors in sufficient quantities to conduct cattle tick vaccination tests

[21] and [22]. An expressed sequence tag originally identified in R. microplus larvae was later reported to correspond to sequence amplified from ovarian tissue coding for the fragment of a Kunitz-BPTI domain protease inhibitor termed rBmTI-6 [21] and [24]. The rBmTI-6 was expressed in the Pichia pastoris system and characterized as a three-headed Kunitz-bovine pancreatic trypsin inhibitor, but its ability to protect immunized cattle against tick infestation remained to be determined [21]. Here, the partial nucleotide sequence of the putative R. microplus larval trypsin inhibitor was used to produce the recombinant polypeptide in the yeast expression system to probe its immunoprotective properties [24]. Results of the cattle immunization trial and other experiments using the recombinant R. microplus larval trypsin inhibitor (rRmLTI) are also reported. Ticks used for this study were obtained from a laboratory colony maintained at EMBRAPA Beef Cattle.

Additionally, we are identifying associations with a relatively s

Additionally, we are identifying associations with a relatively small number of dependent variables (51), across many independent variables that have correlations, and confidence intervals of the coverage estimations were not considered in the regression.

We have kept the best models we found, however, other good models could also exist. Supplementary Table 1 presents a summary of variables highly correlated with those in the children and high-risk models. Our models provide a solid approach on the analysis of factors Alectinib clinical trial related with coverage. However, care should be taken in relying too heavily on any particular variable or finding without considering its interaction with other variables in the model. The distribution and administration of the H1N1 vaccine provided an opportunity to understand how specific approaches may affect vaccine uptake in priority populations in an emergency situation. Results from this analysis complement those examining factors associated with vaccination of overall adults and suggests that supply chain factors may affect vaccine uptake. The analysis also points to opportunities for future research such as further analysis on uptake and the relationship with spatial access to vaccine or access by provider

type, and the role of urban or rural differences in vaccine uptake. These research questions and others can be informed by more detailed mapping of the process and Pifithrin-�� chemical structure system to show details of demand (e.g., by population or providers), supply (e.g.

details on allocations and shipments including the final point of distribution and the category of provider), lead-times across the system, variations within and across states, where vaccine was administered, when, by who and to what subpopulation. Such data would also allow for a robust comparison of potential distribution systems and processes before they are implemented. C. Davila-Payan collected data, performed statistical analysis, and aided in drafting the manuscript. J. Swann designed ALOX15 the study, advised on methodology and logistical factors, and drafted the manuscript. P. Wortley advised on public health and vaccination programs, assisted in acquisition of data, aided in interpretation of results, and editing the manuscript. All authors approved the final manuscript. C. Davila-Payan was partially supported by the ORISE Fellows program during the research. J. Swann was partially supported as the Harold R. and Mary Anne Nash professor, by the Zalesky Family, and by Andrea Laliberte in gifts to the Georgia Institute of Technology, and was partially supported by the Centers for Disease Control and Prevention (CDC) in an Intergovernmental Personnel Act agreement between the CDC and Georgia Tech. The ORISE Fellows program and the donors to Georgia Tech had no role in this research. Participants at the CDC gave feedback on preliminary results including potential interpretations and reviewed the final manuscript for confidentiality and accuracy.

Also study investigators collected stool samples at participant h

Also study investigators collected stool samples at participant houses for each case of diarrhea. Finally, during the second year, the CSCOM fees (usually higher than the traditional healer’s fee) were paid for by the study, as were costs of medicines prescribed at the discretion of the study physician. With these modifications, the surveillance for detection of RVGE cases was greatly strengthened during the second year of surveillance. Moreover, in the second

year of the study monthly meetings were held with all the traditional healers providing services within the study areas to inform them about the study objectives to ask them to refer gastroenteritis cases that they see to the closest CSCOM. The traditional Lapatinib clinical trial healers were reimbursed for transportation expenses that they incurred in coming to the meeting. VX-770 purchase Once a week the most prominent leaders among the traditional healers were visited at the places where they deliver care to remind them about referring suspected gastroenteritis

cases to the CSCOMs. As reported elsewhere [8], the primary study outcome was severe RVGE, regardless of serotype, occurring ≥14 days after the third dose until the end of the study. Gastroenteritis was defined as ≥3 watery or looser than normal stools within a 24-h period and/or forceful vomiting. Data on ongoing symptoms and signs were collected throughout the course of the episode. These data were used to define severity using the 20-point modified Vesikari Clinical Scoring System (VCSS) [11] and [12]; “severe” was defined as a heptaminol score of ≥11. Secondary efficacy endpoints included efficacy against severe RVGE by individual circulating RV serotypes (not reported

in this manuscript), and efficacy against severe RVGE for all infants who received at least one dose of vaccine (intention-to-treat (ITT) analyses). Other efficacy analyses included efficacy against severe RVGE through the first year of life and during the second year of life in Mali. Rotavirus antigen in stool was detected by enzyme immunoassay (EIA) [9] and the RV genotype was confirmed by RT-PCR [10]. Serum anti-rotavirus IgA responses and serum neutralizing antibody (SNA) responses to human RV serotypes G1, G2, G3, G4, and P1A [8] were measured in serum specimens collected before (pre-dose 1 (pD1)) and following the third dose of vaccine (approximately 14 days post-dose 3 (PD3)) in a subset of 150 infants to document immunologic responses [8]. Pre-dose 1 (pD1) and PD3 geometric mean titers (GMTs) of serum anti-RV IgA and RV SNA responses, as well as the seroresponse rates (≥3-fold rise from pD1 to PD3) of serum anti-RV IgA and RV SNA, were measured along with 95% confidence intervals. Efficacy was defined as (1 − Rvaccine/Rplacebo) × 100%, where R represents the incidence for each group. The number of cases in each group was assumed to follow a Poisson distribution.

two-dose boys = $256 million over 70 years of vaccination in a po

two-dose boys = $256 million over 70 years of vaccination in a population of 10 million, results not shown). Compared to no vaccination, all two- and three-dose girls-only and girls & boys HPV vaccination strategies investigated produce cost-effectiveness ratios below the $40,000/QALY-gained cost-effectiveness threshold ( Fig. 2, and see Supplementary Table 3 for detailed results). In the base-case, two-dose girls-only vaccination (vs. no vaccination) consistently produces the lowest incremental cost-effectiveness ratio with cost/QALY-gained varying between $7900 [IQR: 7000;9700] and $10,400 [IQR: 8800;13,400] ( Fig. 2b–f). The only

exception is when two-dose duration of protection is assumed to be 10 years ( Fig. 2a). In the sensitivity analysis, two-dose girls-only vaccination Tofacitinib solubility dmso cost-effectiveness ratios remained below $40,000/QALY-gained ( Fig. 3a). The maximum cost per dose for two-dose girls-only vaccination to remain cost-effective (vs. no vaccination) is predicted to be $128, $218 and $252 assuming two-dose vaccine protection lasts 10, 20 and 30 years, INCB024360 cost respectively (see Supplementary Fig. 4 and Table 4). The incremental cost-effectiveness ratio of

giving the third dose of vaccine to girls (i.e., of three-dose girls-only vs. two-dose girls-only) is estimated to be below $40,000/QALY-gained if: (i) three doses provide longer protection than two doses (i.e., more than 5 years), and ii) two-dose protection is less than 30 years ( Fig. 2 and Fig. 3). Under most scenarios, two-dose girls & boys vaccination (vs. two-dose girls-only) provides fewer or similar QALYs-gained and is more expensive than three-dose girls-only vaccination (i.e., is dominated; Fig. 2 and Fig. 3). The Thymidine kinase only exceptions are: (i) if the third dose provides little or no additional protection to two doses, (ii) when extreme scenarios for burden of HPV-disease among MSM are assumed (e.g., 7% males are MSM, the relative risk of disease among

MSM vs. male heterosexuals is 17, and girls-only vaccination is assumed to have no effect on HPV-related disease incidence in MSM) or (iii) when vaccine cost for boys is 10–40% of the cost for girls ( Fig. 3b, Supplementary Fig. 3). Finally, the incremental cost-effectiveness ratio of three-dose girls & boys vaccination (vs. three-dose girls-only) is greater than $100,000/QALY-gained under all base-case scenarios and most scenarios investigated in sensitivity analysis ( Fig. 2 and Fig. 3). In the sensitivity analysis, three-dose girls & boys vaccination is estimated to be less than $40,000/QALY-gained if the cost per dose for girls and boys is substantially reduced (Supplementary Fig. 4c).

4 (lane 3) showed absence of DNA band and only a smear of degrade

4 (lane 3) showed absence of DNA band and only a smear of degraded DNA was observed. All the extracts except methanol showed observable protection of DNA intactness. Free radicals are known for DNA strand breaking and damage which eventually contributes to carcinogenesis, mutagenesis and cytotoxicity.16 Various researchers have reported the similar results and used plant extracts and fractions for DNA protection against oxidative damage.16 and 28 One of the interesting finding of present study was that ME did not show significant DNA protection activity which can be attributed to its inability to scavenge OH radicals (Fig. 2). It can be postulated from the results depicted in Fig. 5

that AAPH degraded BSA protein (lane 3). However, pre-treatment INCB018424 clinical trial of H. isora fruit extracts effectively protected the protein from AAPH-induced

oxidation, which can be seen in terms of restoration of band intensity in the gel. These results hold significance and may have a positive role in inhibiting several stress or toxicity induced-protein oxidation. 26 All authors have none to declare. Authors thank the Principals of Modern College and Prof. Ramkrishna More College, Pune for encouragement and support to carry out this work. “
“Pyrroles and their derivatives exhibit different important biological activities, like antibacterial, antioxidant, cytotoxic and insecticidal Paclitaxel purchase properties.1, 2 and 3 Several five membered heteroaromatic systems like 1,2,4-triazole, 4-oxadiazole and 4-oxazolidinones having three hetero atoms at symmetrical PDK4 positions have been studied because of their interesting physiological properties.4, 5 and 6 They exhibit board spectrum

of pharmacological activities such as antiinflamatory,7 and 8 antiviral9 and antibacterial10, 11, 12, 13 and 14 activities. In view of the above mentioned pharmacological activities of pyrrole, 1,2,4-triazole, 4-oxidiazole and 4-oxaazolidinones, a number of the 2-substituted 3,5-dimethyl-2,4-diethoxy carbonyl pyrrole derivative have been synthesized containing above moieties. The reaction sequence leading to the formation of desired heterocyclic compounds are outlined in Scheme 1. The starting material 3,5-dimethyl-2,4-diethoxy carbonyl pyrrole (1) was prepared,15 refluxed with hydrazine hydrate to give 2- (3′, 5′ dimethyl-4′-ethoxy carbonyl pyrrole) acid hydrazide (2) was then refluxed with different iso-cyanate16 and 17 in presence of ethanol for 8 h. The isosemi-carbazide (3a–g) was heated with alkaline ethanolic solution for 3 h afforded 5-(3′,5′-dimethyl-4′-ethoxy carbonyl pyrrole)-4-phenyl-3-hydroxy-1, 2, 4-triazole (4a–g). 5-(3′,5′-dimethyl-4′-ethoxy carbonyl pyrrole)-1-phenyl amino-1,3,4-oxadiazole (5a–g) were obtained by cyclization of (3) by stirring it with conc. H2SO4, for 4 h.

At these doses, immunising strains did not induce clinical signs,

At these doses, immunising strains did not induce clinical signs, were completely cleared with all mice surviving the infection. At 13 weeks postimmunisation clearance of the selleck kinase inhibitor bacteria was confirmed by viable counts from spleens and livers. Mice were subsequently re-challenged either intravenously with 104 CFU, or orally with 108 CFU of SL1344. Age-matched unimmunised mice were included for comparison. Viable counts in the target organs were enumerated as detailed

above. All work was licensed by the UK Home Office. For histopathological analysis, a portion of spleen was fixed in 10% buffered formalin then embedded in paraffin wax. Four 3 μm sections were cut approximately 20–30 μM apart then stained with Haematoxylin and

Eosin (H&E). Spleen sections were examined microscopically. Sonicated SL1344 was used as the ELISA capture SNS-032 cell line antigen to assay anti-Salmonella antibodies following vaccination. This was diluted in carbonate coating buffer (1.59 g/l sodium carbonate, 2.93 g/l sodium bicarbonate, pH 8.2) to 1 × 106 bacteria/ml, based on the viable count of the original culture. 100 μl of this antigen solution was used to coat the wells of an ELISA plate (Immunoplates, Nunc, Thermofisher Scientific, Lutterworth, UK) through overnight incubation at 4 °C. Plates were washed with washing buffer (PBS containing 0.05%, w/v, Tween 20) then wells were blocked with 300 μl/well of blocking buffer (PBS containing 1% bovine serum albumin) for 2 h. Serial fivefold dilutions of heat-inactivated mouse serum were prepared in blocking buffer and 100 μl were added to washed plates. Sera from normal

mice and known positive sera were included on each plate as negative and positive from controls. Plates were incubated for 2 h at room temperature. Total antibody was detected using 100 μl/well of biotinylated goat anti-mouse immunoglobulins (Dako, Ely, UK) diluted 1:1000 in blocking buffer. Subtypes IgG1 and IgG2a were detected using 100 μl/well of biotinylated rat anti-mouse IgG1 or IgG2a antibodies (BD Bioscience, Oxford, UK) diluted 1:500 in blocking buffer. Plates were incubated with secondary antibody for 1 h at room temperature and then washed three times in wash buffer. Then 100 μl/well of streptavidin (BD Bioscience, Oxford, UK), diluted 1:100 in blocking buffer, was added and plates were incubated in the dark for 30 minutes. Plates were then washed and developed with 100 μl TMB substrate solution (BD Bioscience, Oxford, UK) and the reaction stopped with the addition of 50 μl/well of 5N sulphuric acid. Absorbance was read at 450 nm. Data presented are from dilutions of 1:12,500 for total Ig and 1:2500 for Ig subclasses. RAW 264.7 cells were seeded into 96 well plates at a density of 2 × 105 cells/well in RPMI medium (Sigma Dorset, UK) supplemented with 10% FCS and 2 mM l-glutamate. Plates were seeded the evening before infection and incubated throughout at 37 °C with 5% CO2.