m.). Mice were acclimatized to the laboratory for at least 1 h before testing. Animals were used according to the guidelines of the Committee on Care and Use of Experimental Animal Resources, the Federal University of Santa Maria, Brazil. Non-spatial long-term memory was investigated using a step-down inhibitory avoidance task according to the method of Sakaguchi et al. (2006), with some modifications. Each mouse was placed on the platform, and the latency to step-down (four paws on the grid) was automatically recorded in training

and test sessions. In the training session, upon stepping down, the mouse received a 0.5 mA scrambled foot shock for 2 s. Test sessions were performed 24 h later, with the same procedure except that no shock was administered after stepping down; an upper cutoff time of 300 s was set. Six to eight animals were used per group. PEBT at the doses of 5 GDC-0973 chemical structure or 10 mg/kg orally (p.o.) (Souza et al., 2009), or vehicle (canola oil 10 ml/kg, p.o.) were given 1 h before Pfizer Licensed Compound Library training (acquisition), immediately post-training (consolidation), or 1 h before test (retrieval). The oral route dominates contemporary drug therapy and is considered to be safe, efficient and easily accessible

with minimal discomfort compared to other routes of administration (Lennernãs, 2007). Spontaneous locomotor activity was measured in the open-field test (Walsh and Cummins, 1976). The open-field was made of plywood and surrounded by walls 30 cm in height. The floor of the open-field, 45 cm in length and 45 cm in width, was divided by masking tape markers into 9 squares (3 rows of 3). Each animal was placed individually at the center of the apparatus and observed for 4 min to record the locomotor (number of segments crossed with the four paws) and exploratory activities (expressed by the number of time rearing on the hind limbs). Six to eight animals

were used per group. The locomotor and exploratory activities were evaluated after the test session of the step-down inhibitory avoidance task. In order to investigate the possible mechanisms involved in the effect ADP ribosylation factor of PEBT on memory, glutamate uptake and release assays were carried out 1 h (training) or 24 h (test of memory) after oral administration of PEBT (10 mg/kg). Glutamate uptake was performed according to Thomazi et al. (2004). One and 24 h after oral administration of PEBT, mice were killed by cervical dislocation and the brains were immediately removed. Slices (0.4 mm) were obtained by transversally cuts of cortex and hippocampus using a McIlwain chopper. Experiments were made in triplicates. Slices were pre-incubated for 15 min at 37 °C in a Hank’s balanced salt solution (HBSS) containing (in mM): 137 NaCl, 0.63 Na2HPO4, 4.17 NaHCO3, 5.36 KCl, 0.44 KH2PO4, 1.26 CaCl2, 0.41 MgSO4, 0.49 MgCl2 and 1.11 glucose, adjusted to pH 7.2. Then, 0.66 and 0.

(n = 15), or an unknown reason (n = 34). Refusals were included and coded as limited health literacy, as these people are likely to perform with limited health literacy skills in real-life settings (e.g. at the doctor’s office) because of their difficulties. Therefore, they were included to maintain the population-representativeness

of the sample and capture a more accurate range of the health literacy skills of the English population. The present analysis thus included 3087 men and women aged 60–75 years (Fig. 1). Health literacy was assessed using a four-item comprehension test based on a fictitious medicine label from the International Adult Literacy Survey (Thorn, this website 2009) (Appendix A). Health literacy was categorised as ‘adequate’ (4/4 questions answered correctly) or ‘limited’ (< 4/4 answered correctly) to capture the point at which adults begin to have difficulty with everyday health tasks. Although whether and how health literacy skills may change over time are uncertain, health literacy scores among our sample

are expected to be stable between data collection and the times of reported CRC screenings (within one year of wave 5 data collection for 59% of those reporting screening and within two years for 96%). Health literacy was also measured at ELSA wave 2 (2004–5) and the scores did not change between waves 2 and 5 within individuals who remained in the study for both waves. Health literacy scores measured PI3K Inhibitor Library at wave 2 were not used for this analysis, as study attrition between waves was differential by health literacy score. Participants were asked if they had ever used a bowel testing kit (i.e. an FOBT kit) and whether the kit was part of the NHS Bowel Cancer Screening Programme. Only 49 out of the 1709 participants (< 3%) who reported having completed an FOBT kit responded that the kit was not part of the NHS programme

and 3 (< 1%) responded that they did not know whether it was part of the programme; hence for this analysis we assume that completion of a Cytidine deaminase FOBT kit equates with participation in the NHS programme. For convenience, the terms “completion of an FOBT kit” and “CRC screening” will hereupon be used synonymously. Sociodemographic covariates were: age, sex (male; female); educational attainment (no qualification; up to degree level; degree level or equivalent); net non-pension wealth (quintiles stratified at age 65 to account for changes in wealth following retirement) (Bostock and Steptoe, 2012); occupational class according to the 2010 National Statistics Socio-economic Classification (routine; intermediate; managerial or professional) (Office for National Statistics, 2010); and ethnic minority status (non-white; white).

The remaining two countries (India and Sri Lanka) have no formal policy. The consequences to committee members when they report a conflict of interest vary by country. For example, depending on the level of conflict, members of the Australian NITAG might participate and vote, participate but not vote, attend the meeting but remain silent, or be barred from the meeting altogether. The United Kingdom as well report a relatively nuanced policy, based on whether a conflict of interest is Sirolimus solubility dmso personal (e.g., stock ownership) or non-personal (such as involvement in a study through an academic institution) and whether the conflict is specific or not to

the vaccine in question. In most cases, authors report that committee recommendations are advisory and not legally binding. However, in five countries the committee has some form of legal responsibility for determining some or all policy related to the topics under their mandate. In Iran, for example, the government is obliged to implement committee recommendations, although no law requires this. In Oman and Sri Lanka, the government is legally

obligated to implement recommendations. Recommendations from the United Kingdom also carry legal weight but a recommendation may be made only if economic data Neratinib supplier are convincing (as described above); otherwise, findings are considered advisory and are not legally binding. Lastly, the United States NITAG recommendations are advisory in most instances. The exception is the Vaccine for Children’s Act, which regulates financing of vaccines for low income children; in this case, committee

decisions determine which vaccines will be funded under this program. Some countries specifically state that not all recommendations are followed, such as South Africa, South Korea, and Thailand, where budget limitations are the most common reason for lack of implementation of recommendations. Other countries, such as Honduras and Switzerland, report that decisions do not carry legal force but to date all recommendations have been implemented. GBA3 Almost all committees identified areas for improvement. Of great interest is that this is the area with the greatest variation in results, with very little overlap between committees. The most commonly identified area for improvement (mentioned in eight reports) is in the realm of economic data including lack of policies regarding how to weigh economic data, lack of economic expertise on the committee, and insufficient weight given to economic data. The second most commonly identified area for improvement (mentioned in five reports) is lack of overall necessary expertise to reach optimal evidence-based decisions, followed by insufficient data availability, an increasing level of work, and insufficient committee independence from the pharmaceutical industry (three reports each) (Table 1).

Des modulations de ce profil de base peuvent être apportées par l’influence contraire de l’IMC sur le taux plasmatique de SHBG [71] ou par l’apparition d’une neuropathie qui peut participer à la constitution d’un déficit testiculaire primaire [72]. L’ensemble des données de la littérature suggère donc que, par des mécanismes complexes, le déficit en androgènes soit un des facteurs favorisant l’émergence

d’un diabète ou l’aggravation d’un diabète préexistant. Ainsi que cela est désormais recommandé par l’American Diabetes Association (ADA) MAPK inhibitor ceci incite à rechercher l’existence d’un hypogonadisme chez le patient dont le diabète est connu. Il convient également de détecter l’émergence d’un diabète ou l’aggravation d’un diabète préexistant chez le patient dont la pathologie relève d’un traitement par blocage androgénique. Les conséquences des modifications métaboliques associées à l’hypogonadisme ne sont pas négligeables. S’inscrit au premier rang le risque vasculaire. Une importante étude de cohorte chez des septuagénaires a montré, après ajustement pour les autres principaux facteurs confondants, que le risque de survenue d’un accident vasculaire cérébral ou

d’un accident ischémique transitoire était deux fois plus élevé lorsque le taux de testostérone plasmatique total ou libre était bas [73]. Le phénotype métabolique retrouvé chez l’homme hypogonadique pourrait ainsi constituer le lien physiopathologique entre hypotestostéronémie et complications vasculaires et possiblement risque vital. Laughlin et al. ont mis en évidence dans une selleck cohorte d’hommes âgés suivis sur une période de dix ans que le risque de décès était presque doublé chez ceux

dont le taux de testostérone plasmatique à l’entrée dans l’étude était le plus bas [74]. Il faut remarquer que dans cette étude les causes cardiovasculaires de décès sont le plus fréquemment observées sans pouvoir bien sûr conclure qu’il y ait un lien direct entre testostéronémie et risque vital. Les résultats des évaluations transversales et longitudinales issues de la Framingham Heart Study, et recueillies chez plusieurs milliers d’hommes suivis all au long terme, montrent que l’association entre testostéronémie libre et SMet disparaît après ajustement pour l’âge, l’IMC et la sensibilité à l’insuline. Les liens identifiés entre testostérone totale, d’une part, SMet et risque vasculaire, d’autre part, s’expliquent en fait par la corrélation linéaire qui existe entre testostéronémie totale et taux plasmatique de SHBG [75]. Les taux de testostérone plasmatique totale sont étroitement liés à ceux de la SHBG. Le taux plasmatique de la protéine de transport apparaît être un facteur indépendant associé au risque de survenue d’un SMet [76]. Ce dernier, modulé par l’âge, l’IMC et le degré d’insulino-résistance, apparaît donc comme un marqueur plus fiable de ce risque.

2, 95% CI 1.1 to 4.4), but not at 12 months. No significant intervention effect was demonstrated for mobility capacity (Table 4), attitude towards sports (Table 5) and the other secondary outcomes (Tables 6 and 7) at 4 months, 6 months or 12 months. (See eAddenda for Tables 6 and 7.) A positive trend was found for the GMFM-66 at 6 months (mean between-group difference 2.8, 95% CI 0.2 to 5.4), but not at 12 months, and for the 1-minute walk test at 4 months (mean between-group difference 5 m, 95% CI 0 to 9), but not at 6 months or 12 months. For attitude towards sports, when compared to the control group, EX 527 concentration there was also a trend for

reporting greater agreement with possible advantages of sports at 12 months (p = 0.04) but not at 6 months, and a borderline significant greater disagreement with possible disadvantages of sports at 6 months (p = 0.02) but not at 12 months. There was no significant effect of the intervention on AC220 price physical activity, so the hypothesis that counselling, home-based physiotherapy and fitness training would work synergistically to improve physical activity could not be confirmed. This was against our expectations, previous studies in cerebral palsy showed (non-significant) positive trends towards improving physical activity in children and adolescents with cerebral palsy

after either counselling,11 or fitness training only.9 Nevertheless, the present findings are in agreement with research involving typically developing children where evidence is equivocal. No evidence has been found for the effectiveness of family-based and community-based physical activity interventions that combine exercise programs with the provision of information.29 Another review has pointed out that physical activity among typically developing children can be increased by means of school-based interventions.30 The authors of that review indicated that the highest-quality studies with positive effects on physical second activity were characterised by a multicomponent intervention (education, focus on behavioural change and involvement of parents) and a minimum intervention

duration of one school year. Therefore, it is possible that our 6-month program was too short to elicit changes in such a complex behaviour as physical activity. Whether a longer counselling period, with periodical attention to physical activity, may be needed to improve physical activity in children with cerebral palsy should be examined in further research. Another explanation for the intervention’s lack of effect on physical activity might be insufficient contrast between groups, which could arise from three possible sources. First, the families who chose to participate in the study were likely to be more interested in (increasing) physical activity than those who refused to participate, as illustrated by the parents’ already very positive attitude towards sports in both groups.

The CAMPRAL® Compound C order enteric coated tablets containing 333 mg Acamprosate per tablet, were obtained from Forest pharmaceuticals, INC, USA. The 1200 Series HPLC system (Agilent Technologies, Waldbronn, Germany), Mass spectrometry API 4200 triple quadrupole instrument (ABI-SCIEX, Toronto, Canada) and data processing was performed on Analyst 1.5.1 software package (SCIEX). The mass spectrometer was operated in the multiple reaction monitoring (MRM) mode. Sample introduction and ionization were electrospray

ionization in the negative ion mode. Sources dependent parameters optimized were as follows: nebulizer gas flow: 20 psi; Heatergas flow 40 psi; curtain gas flow: 8 psi; ion spray voltage (ISV): 5500 V; temperature (TEM): 650 °C. The compound dependent parameters such as the declustering potential (DP), focusing potential (FP), entrance potential (EP), collision energy (CE), cell exit potential Selleck BLZ945 (CXP) were optimized during tuning as 55, 30, 10, 18, 12 eV for Acamprosate and Acamprosate D12, respectively. The collision activated dissociation (CAD) gas was set at 5 psi using nitrogen gas. Quadrupole 1 and quadrupole 3 were both maintained at a unit resolution and dwell time was set at 600 ms for Acamprosate and Acamprosate D12. The mass transitions were selected as m/z 180.0 → 79.9 for Acamprosate and m/z 186.1→ 79.9 for

Acamprosate D12. The parent and product ion spectra for Acamprosate and Acamprosate D12 are represented in Figs. 2a and b, 3a and b respectively. The data acquisition was ascertained by Analyst 1.5.4software. Waters Atlantis, HILIC, 50 × 2.1 mm, 3 μm, was selected as the analytical column connected with Guard column Waters Atlantis, HILIC, 10 × 2.1 mm, 3 μm. Column temperature was set at 40 °C. Mobile phase composition was 10 mM Ammonium formate pH 3.5: Acetonitrile (10:90 v/v). Source flow rate 250 μL/min without split. Injection volume of 10 μL. Acamprosate and Acamprosate D12 were eluted at 2.1 ± 0.2 min, with a total run time of 3.0 min for each sample. Acamprosate and Acamprosate D12 standard stock solutions 100 μg/mL each were prepared by dissolving the appropriate standard in methanol. From the Acamprosate

stock solution calibration and quality control standards were prepared by using screened human blank plasma as diluent. Calibration standards were prepared at concentration levels of 1.00, Tryptophan synthase 2.00, 5.00, 25.00, 50.00, 100.00, 150.00, 200.00 and 250.00 ng/mL. Quality control standards were prepared at concentration levels of 1.00, 3.00, 125.00 and 175.00 ng/mL for Acamprosate. Internal standard spiking solution at 50 ng/mL concentration was prepared by using 50% methanol solution from Acamprosate D12 standard stock solution. Calibration and quality control standards were prepared from two separate stock solutions of Acamprosate and stored at −30 °C. Internal standard spiking solution was stored in refrigerator conditions at 2–8 °C until analysis.

Additional versions: Nil. Expert working group: 16 individuals representing health care professional groups

(medical specialties, nursing, pharmacy), consumers, and guideline developers. Funded by: National Health and Medical Research Council of Australia. The guidelines were developed by the National Institute of Clinical Studies (NICS). Consultation with: External input was indicated in the guideline development process, but Proteases inhibitor details were not provided. Approved by: National Health and Medical Research Council of Australia. Location: http://www.nhmrc.gov.au/_files_nhmrc/file/nics/programs/vtp/guideline_prevention_venous_thromboembolism.pdf Description: This is a 157 page document that presents evidence-based recommendations related to the prevention of venous thromboembolis in patients admitted to Australian hospitals. The primary options for thrombophylaxis considered in this guideline were pharmacological and mechanical, which included knee or thigh

length graduated compression stockings, knee or thigh length intermittent pneumatic compression, or venous foot pumps. A 7-page summary of recommendations is provided from page 4. These recommendations are presented by clinical procedure (e.g. total hip replacement), or medical condition (e.g. stroke). Specific recommendations are provided for cancer patients (surgical and non-surgical) and pregnancy and childbirth. There is also a clear 1-page summary of the evidence GSK2118436 for the use of thromboprophylactic agents by clinical category (e.g. abdominal surgery) on page 25. The body of the guideline provides the detailed evidence that underpins the

recommendations, including the level and grade of evidence and the related references. A list of the 392 references included in the document is provided. “
“Latest update: August 2010. Next update: Within 3–5 years. Patient and group: Patients aged over 18 years presenting with a stroke or TIA. Intended audience: Health professionals, administrators, funders and policy makers who plan, organise and deliver health care for people with stroke in all phases of recovery. Additional versions: This document updates and amalgamates two previous Australian guidelines: Clinical Guidelines for Acute Stroke Management (2007) and Clinical Guidelines for Stroke Rehabilitation and Recovery (2005). Expert working group: 35 individuals representing 17 health care professional groups including medical specialties, nursing, physiotherapy, occupational therapy, speech pathology, and other professions. Funded by: National Stroke Foundation of Australia, Department of Health & Ageing. Consultation with: Public consultation about the draft document was undertaken over one month, with numerous stakeholder groups specifically targeted for feedback. Approved by: National Health and Medical Research Council of Australia, National Stroke Foundation. Location: http://www.strokefoundation.com.

To verify N. caninum immunostaining, IFAT was performed with mouse sera collected at 45 d.a.i. as previously described [29]. Slides Dasatinib containing formolized tachyzoites were incubated with serum samples diluted 1:50, and then with FITC-labeled goat anti-mouse IgG (1:50; Sigma). Slides were overlaid with buffered glycerol and examined in fluorescence microscope (EVOS, Advanced Microscopy Group, Inc., Mill Creek, WA). Two weeks after the last immunization (45 d.a.i.), three mice from each group were euthanized and

their spleens were aseptically removed for cell culture and cytokine production assay. Mouse spleens were dissociated in RPMI medium and cell suspensions were washed in medium, treated with lysis buffer (0.16 M NH4Cl and 0.17 M Tris–HCl, pH 7.5), washed again and resuspended in complete RPMI medium containing 10% CFS. Viable cells (2 × 105 cells/200 μl/well) were cultured in triplicate in

96-well plates in the presence of antigen (NLA, 10 μg/ml), mitogen (Concanavalin A – ConA, 2.5 μg/ml) or medium alone and incubated at 37 °C in 5% CO2. After 48 h, cell-free supernatants were collected and stored at −70 °C for cytokine quantification. IL-10 and IFN-γ measurements were carried out by sandwich ELISAs according to manufacturer’s selleck inhibitor instructions (R&D Systems, Minneapolis, MN). The limit of detection for each assay was 31 pg/ml and intra-assay variation coefficients were below 15%. After 30 days of the last immunization (60 d.a.i.), the remaining animals of each group (10 per group) were challenged intraperitoneally (200 μl/mouse) with 2 × 107 low-passage Nc-1 tachyzoites. Animals were observed daily for clinical signs through morbidity scores, body weight changes

and mortality during 30 days post-infection (d.p.i.). Morbidity scores were calculated as described elsewhere [32], with minor modifications as follows: sleek/glossy coat, bright and active (score 0); ruffled coat (score 1); hunched, tottering gait, starry stiff coat (score 2), reluctance to move (score 3). Results were expressed as the mean of the scores given daily to each animal for each group. After 30 days of challenge, surviving animals were euthanized and blood PAK6 samples and brain tissues were collected. Serum samples were tested for N. caninum serology and brain tissues were sliced longitudinally, being half of them stored at −70 °C for polymerase chain reaction (PCR) assay. The remaining tissue was fixed in 10% buffered formalin, embedded in paraffin and routinely processed for immunohistochemical and histological assays. Brain parasite load was determined by quantitative real-time PCR as previously described [29], using primer pairs (sense 3′ GCTGAACACCGTATGTCGTAAA-5′; antisense 3′-AGAGGAATGCCACATAGAAGC-5′) to detect the N. caninum Nc-5 sequence through SYBR green detection system (Invitrogen, San Francisco, CA). DNA extraction was performed from 20 mg of murine brain tissues (Genomic DNA kit, Promega Co.

The effluent was analysed by APHA, 1981.3 The fresh material of plant was collected from both sites non-polluted (ALTT Centre) and polluted (cycles manufacturing unit) area of Ghaziabad, UP, India. For colour reaction test Cromwell, 19554 & Trease and Evans, 19835 were followed. TLC was done According to the WHO, Geneva, 1998.6 Chlorophyll a, b and total chlorophyll (a + b) were determined according to Arnon, 1949.7 The effluent was analysed and the results are given in Table 1. The result shows the presence of alkaloids, saponin, tannin, lignin, protein, carbohydrate, suberin, glucoside, oil, sugars, steroids and absence of flavanoids in both the cases. Degree of change in colour reaction tests are

tabulated in Table 2. From the observation of TLC, it is found that the number of spots were higher in non-polluted plants than the polluted plants (Plate 1). The RF values are tabulated in Table 3. Chlorophyll a, chlorophyll b and Autophagy inhibitor total chlorophyll were observed 76.98%, 86.29% and 80.10% of control leaves samples (Plate 2). The results are tabulated in Table 4. The effluent samples collected from the industry selected for this study was

analysed for different physico-chemical parameters which showed higher values as compared to the standard values recommended by the Indian Standard Institute (I.S.I.; 1974, 1974 and 1977). Similar results were also obtained by Kumar, et al,1988.8 A critical observation on the data studied clearly indicate that plants growing at polluted sites were badly affected and there were a significant reduction R428 ic50 in number of parameters studied as compared to the plants growing at the control sites. Major qualitative changes, noticed under the impact of industrial effluent, are reduction in chlorophyll level, photosynthesis rate, accumulation of heavy metals, alternation in pH, BOD, COD, Colour, Temp, Odour, TS, TDS. Heavy metals resulted into reduced growth and yield in comparison to plant species growing at non-polluted sites. The impact of industrial effluent on the qualitative and quantitative

values of medicinal plants does not appear to have been undertaken much till now. Colour reaction tests showed the degree of changes in plants of polluted sites. From the observations some alteration in the bio-chemical parameters were also recorded in plants growing nearly near the industrial effluent. The amount of chemical constituents found to have decreased in those plants which were growing in polluted areas. From the observations of TLC, it was seen that the number of spots were decreased in the plant samples of polluted sites. From the findings of this investigation it may be ascertained that there had been qualitative and quantitative alternations in the chemical constituents in the plants growing in industrial areas. It can also be stated that industrial pollution may also have lowered the drug potency of the plants growing in the vicinity of industries.

As negative control, medium of uninfected MDCK cells was used (Mock).

As positive control, cells were stimulated with 2 μg/ml Con A (Sigma). All stimulations were performed in a total volume of 0.6 ml and were incubated for 20 h at 37 °C. After incubation, all (0.6 ml) supernatant of the PBMC was collected in cryotubes (Simport) and stored at −80 °C for determining cytokine concentrations. Remaining cells were lysed with 0.25 ml lysis buffer (150 mM NaCl, 15 mM Tris, 1% Triton X-100), transferred to Eppendorf tubes and stored at −80 °C for measuring granzyme B concentrations. Frozen cell lysates were subjected to three freeze/thaw cycles to enable release of granzyme B from the granules. www.selleckchem.com/products/Fludarabine(Fludara).html Granzyme B standards were prepared in duplicate

in a 96-well plate (Corning) and the cell lysates (20 μl/well) were added in duplicate to the same plate. Subsequently, 80 μl enzyme reaction buffer containing 400 μM acetyl-Ile-Glu-Pro-Asp-paranitroanilide (Ac-IEPD-pNA, Calbiochem), 100 mM HEPES pH 7.5 (Sigma), 10% (w/v) sucrose (Sigma), 0.1% (w/v) CHAPS (Roche), and 10 mM DTT (Sigma) was added per sample. The plate was covered with thermo-plastic seal and www.selleckchem.com/products/pexidartinib-plx3397.html incubated in a dark humidified chamber at 37 °C for 20 h. After incubation, the plate was read at 405 nm. Granzyme B units in the cell lysates were calculated using a fourth-order polynomial curve (y = ax4 + bx3 + cx2 + dx + e) with a log (concentration) − log (absorbance) plot. The granzyme B units were corrected for protein content in the lysates (granzyme B units/mg protein). The protein concentration of the lysates was determined by the BCA protein assay (ThermoScientific). To prevent batch-to-batch variation of test kits, all laboratories used the same lot-number of the Th1/Th2 cytokine multiplex kit (Bio-Rad, Cat#171A11081, Batch#5008594), IL-17 singleplex kit (Bio-Rad, Cat#171B14076, through Batch#5008678), and Reagent kit (Bio-Rad, Cat#171304000, Batch#310002936). The multiplex assay was performed according to the protocol of the manufacturer with some modifications. The

plate was measured on the Bio-Plex suspension array system (Bio-Plex 100 or 200) under low photo multiplier tube settings. Calculation of cytokine concentrations in unknown samples was determined by 5-parameter fit regression analysis of the standard curve. The validation plan was designed to assess a range of parameters (Fig. 1) [33]. To determine linearity, range, detection limit, specificity, accuracy, and precision, PBMC were stimulated with mock, influenza H3N2 or concanavalin A (Con A, Sigma) and used for generating a bulk amount of cell lysate and culture supernatant for testing in the granzyme B and cytokine assay, respectively. Generation of this bulk amount of cell lysate and culture supernatant was performed by NVI, The Netherlands.