The action cycle consists of the following phases: identifying a problem and the relevant knowledge; adapting the knowledge to the local context; assessing barriers (and enablers) to using the knowledge; implementing tailored interventions

to promote use of the knowledge; VX-770 nmr monitoring knowledge use; evaluating the outcomes of using the knowledge and sustaining ongoing knowledge use (Figure ​(Figure11). Figure 1 Model of Knowledge to Action (18). Dissemination Strategies A multi-faceted dissemination plan targeted at stakeholders will be executed Inhibitors,research,lifescience,medical at the completion of the Agenda project. The primary means of conveying the results of the study will be through publication of peer-reviewed manuscripts in journals that appeal to the target Inhibitors,research,lifescience,medical scientific and provider audience. Second, submissions will be made to prominent EMS trade journals targeting front-line paramedics, medical directors and EMS operators. Third, presentations at regional, provincial, and national EMS conferences will be used to communicate the results to attendees. Lastly, key organizations will be notified of the results and asked to convey them to their members through free electronic media and posting

of the results on their free-access websites. As well the research agenda will be circulated to potential granting agencies, foundations, industrial and academic partners and follow up meetings with these constituents will Inhibitors,research,lifescience,medical enable full discussion and identify opportunities to lobby for change, influence priority and new funding initiatives. Once the Agenda has been widely disseminated

in Canada, it is expected Inhibitors,research,lifescience,medical it will serve as a guide for EMS stakeholders to implement a local and national strategy to address barriers to research and to maximize Inhibitors,research,lifescience,medical on existing and future opportunities. It may be used to inform budget planning and grant allocation and human resources for research, and to lobby for change in the way funding is directed in existing funding agencies. The implementation strategy is likely to include efforts to develop and protect researchers to conduct studies on questions found to be important, and will ignite collaboration across the country, which will allow for studies with sample sizes sufficient to answer vital questions. Potential Challenges in Knowledge Translation Once this study is complete, and results are for disseminated through the mediums described above, we anticipate challenges in the following components of the knowledge translation action cycle: adaptation to the local context, monitoring knowledge use, and evaluating the outcomes of using the knowledge. EMS systems in Canada have had variable research experiences, ranging from no exposure to studies to being involved with the conduction of randomized controlled trials. Because of this variability, the implementation of the Canadian EMS Research Agenda will need to be adapted to individual services and settings.

In adults, doses ranged from 50 to 300 mg/day and fluvoxamine was overall well-tolerated. Fluoxetine Larger studies of fluoxetine have not found it to be effective in the www.selleckchem.com/screening/pi3k-signaling-inhibitor-library.html treatment of repetitive behaviors in children. The drug has proven to be more effective in adults and adolescents with autism. Adolescents appear to experience adverse effects more frequently than adults. The Study of Fluoxetine in Autism (SOFIA), the largest double-blind, placebo-controlled

trial of an SRI in children with autism to date, concluded that Inhibitors,research,lifescience,medical fluoxetine is not effective for the treatment of repetitive behaviors in children.25 Prior to this, a study found liquid fluoxetine superior to placebo in decreasing repetitive behaviors in children and adolescents, with minimal adverse effects.26 Inhibitors,research,lifescience,medical Two males with Asperger’s disorder, aged 9 and 10 years old, exhibited initial improvements in compulsive behaviors and reduced irritability, respectively, but later experienced episodes of hypomania with fluoxetine

20 mg/day.27 A retrospective review of 7 subjects with autism, aged 9 to 20 years (mean age, 16 years), revealed improvement in stereotypy, irritability, lethargy, and inappropriate speech during fluoxetine treatment.28 An open-label study of individuals with autism, Inhibitors,research,lifescience,medical aged 7 to 28 years (mean age, 15 years), showed favorable responses in the treatment of perseverative and compulsive behaviors, although the presence of comorbid Axis I diagnoses in many subjects Inhibitors,research,lifescience,medical makes it difficult to generalize these results.29 Twenty-three percent of subjects experienced significant adverse effects that interfered with drug continuation. Some case reports of adults describe reductions in repetitive behaviors, obsessive-compulsive symptoms, and temper outbursts with fluoxetine.30,31 However, a case series that included adults and adolescents together observed poor responses in treating repetitive symptoms but improvements in depressive symptoms.32 The adolescent subjects exhibited anxiety and agitation above 20 mg/day of fluoxetine, Inhibitors,research,lifescience,medical so therapeutic doses in these studies remained near

that level. Another case report of a 25-year-old male with Asperger’s disorder, OCD, major depression and 45,X/46,XY mosaicism described poor response to fluoxeine in the treatment of OCD.33 A recent double Terminal deoxynucleotidyl transferase -blind, placebo-controlled study in 37 adults with autism, aged 18 to 60 years (mean age, 34 years), showed moderate efficacy in the management of repetitive behaviors, with a 50% response rate compared to 8% in the placebo-treated group.34 Unless otherwise noted, fluoxetine was dosed 20 to 80 mg/day in the studies above and adverse effects were generally milder in adults than children. Sertraline Sertraline is moderately effective and relatively well-tolerated in the management of repetitive behaviors and aggression in adults with ASDs.

BF-2 cells (from bluegill fry, Lepomis macrochirus; ATCC CCL-91) were used for antibody neutralizing tests. Both cell lines were grown in MEM (Gibco) culture medium supplemented with penicillin (100 IU ml−1), streptomycin (100 μg ml−1) and 10% FCS at 20 °C. For the construction of the this website IPNV DNA vaccine (pIPNV-PP), the inhibitors polyprotein gene was amplified by a polymerase chain reaction (PCR)

from a cDNA sample obtained from the spleen of a trout infected with IPNV Sp strain using specific primers (Table 1), containing both the start and stop codons. The PCR product was cloned into the expression vector pcDNA3.1/V5-His-TOPO according to manufacturer’s instructions (Invitrogen) and used to transform One Shot TOP10 Escherichia coli cells (Invitrogen). A clone containing the pIPNV-PP was identified by PCR screening, and the proper orientation was verified by sequencing. A religated empty pcDNA3.1/V5-His-TOPO plasmid (pcDNA3.1) was used as a negative control. The pMCV1.4-G plasmid used as a VHSV DNA vaccine consisted of the gene encoding the glycoprotein

G of VHSV PF-01367338 in vitro under the control of the long cytomegalovirus (CMV) promoter, previously described [22]. The effectiveness of this VHSV vaccine has been previously demonstrated [23] and [24]. The empty vector (pMCV1.4) was used as a control. To ensure that cloned polyprotein gene could express protein in vitro, the pIPNV-PP plasmid was used as template in the transcend non-radioactive transcription/translation quick coupled system (Promega), which allows a biotinylated detection of proteins synthesized in vitro. The viral protein(s) expressed were separated on a SDS-polyacrylamide science gel electrophoresis, transferred to nitrocellulose membranes and the biotinylated proteins visualized by binding streptavidin–horsedish peroxidase, followed by colorimetric detection. Confluent cultures of actively growing EPC cells were trypsined and dispensed into 24-well plates

at a concentration of 6 × 105 cells ml−1. After 24 h of incubation at 28 °C, cells were transfected by the addition of 3 μl of Fugene 6 (Roche) complexed with either 0.5 μg of pIPNV-PP or the empty plasmid (control). After a further 72 h of incubation at 28 °C, cells were trypsined and processed for RNA isolation or electron microscopy. Expression of the plasmid by the EPC cells was confirmed by VP2 gene expression by semi-quantitative PCR whilst induction of the EPC-antiviral Mx gene was evaluated by real-time PCR (see below). For electron microscopy, cells were fixed in 1% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2) for 2 h at 4 °C, then postfixed in 1% osmium tetroxide in 0.1 M cacodylate buffer (pH 7.2) for 1 h at 4 °C and embedded in Epon. Ultrathin sections were obtained with a Reichert-Jung ultramicrotome, contrasted with uranyl acetate and lead citrate and examined with a Zeiss EM 10C electron microscope.

Although both groups showed improvements in depressive symptoms, the ziprasidone group did not significantly differ from the placebo group in the total score of the HAMD-17 or the MADRS. This is in contrast to an open-label trial of ziprasidone monotherapy in bipolar depression, in which at 8 weeks post treatment, significant improvement

was seen on both of these measures [Liebowitz et al. 2009]. Inhibitors,research,lifescience,medical Interestingly, significant improvement was observed on the HAMA and CGI-S. Moreover, correlation analysis demonstrated a significant correlation between increase in the SWS duration and improvement in CGI-S score. This finding is important as it shows a relation between change in sleep architecture and improvement in illness severity. However, this correlation did not withstand Bonferroni correction. Hence, a significant correlation between these two factors is not sufficient to say that ziprasidone’s sleep-consolidating properties Inhibitors,research,lifescience,medical are causative of the improvement in overall illness severity. The main limitations of this study are the small sample

size and the use of concomitant medications. Participants were click here taking a variety of concomitant medications, Inhibitors,research,lifescience,medical including antidepressants, mood stabilizers, and benzodiazepines, which may affect the key neurotransmitters involved in sleep–wake Inhibitors,research,lifescience,medical manipulation. Furthermore, sleep studies such as this acquire PSG data at distinct time points, which may not be representative of the entire time period. Conclusion A close association exists between sleep architectural abnormalities and affective disorders, and patients with bipolar depression who continue to experience Inhibitors,research,lifescience,medical sleep disturbances face a high risk of relapse. AAs such as olanzapine, quetiapine, and risperidone, which are often used in augmentation strategies in the treatment

of bipolar depression, have been shown to have sleep-consolidating properties. Ziprasidone augmentation in bipolar depression alters sleep architecture and improves overall global illness severity. As far as we are aware, this is the first study to date to have investigated the effects of ziprasidone treatment on both objective and subjective sleep in a clinical population. A clear correlation Chlormezanone was found between change in SWS and overall illness severity. Although this association is not causative, the suggestion that part of ziprasidone’s mechanism of action may be achieved through the restoration of sleep architecture merits further investigation with further randomized investigations with large sample sizes. Acknowledgments The assistance of Dr Meshal Khaled Alaqeel is gratefully acknowledged.

(B) Overlay lesion plot of the patients with left-sided lesions. The number of I-BET151 mw overlapping lesions is illustrated by different colors coding increasing frequencies from violet (n = 1) to red … Vestibular testing SVV as a measure of tonic vestibular otolith perception as well as HT, skew deviation, and OT were tested as previously described

(Dieterich and Brandt 1993; Brandt et al. 1994). A mean binocularly determined deviation of more Inhibitors,research,lifescience,medical than 2.5° of the static SVV was considered as abnormal (Dieterich and Brandt 1993). MRI scans In all patients MRI scans were performed with a mean time interval of 5 days between lesion onset and MRI (SD 1 day). We used diffusion-weighted imaging (DWI) within the first 48 h poststroke and fluid-attenuated Inhibitors,research,lifescience,medical inversion-recovery (FLAIR) sequences when imaging was conducted 48 h or later. Lesion mapping using the Brunner–Munzel test implemented in MRicron and MRI processing using the normalization algorithm of SPM8 (http://fil.ion.ucl.ac.uk/spm/) was conducted as described previously (Rorden et al. 2007; Baier et al. 2012). To prevent a rise in the probability of familywise error, we computed a false discovery rate (FDR) correction. The results Inhibitors,research,lifescience,medical of the lesion analysis were combined with the probabilistic

maps of the posterior IC with the aid of the SPM Anatomy Toolbox (Eickhoff et al. 2005; Kurth et al. 2010) whereas the anterior insular and peri-insular regions were defined by the anatomical

maps provided (Bense et al. 2001; Tzourio-Mazoyer et al. 2002). In the previously analyzed sensory data for our correlation analysis in the Inhibitors,research,lifescience,medical subgroup of 20 patients which were described above, the statistical voxelwise lesion behavior mapping (VLBM) for a binary comparison (CDT [cold detection threshold]; WDT [warm detection threshold]) was performed using the Liebermeister statistics (Rorden et al. 2007). Statistical analysis was conducted using SPSS 15.0 for Windows (SPSS Inc., Chicago, IL). Inhibitors,research,lifescience,medical For correlation analysis we used the Spearman rho analysis. In addition, we conducted a bivariate linear regression analysis to indicate prediction of one variable from another. Thermal perception adapted for our previous analysis In the previous study, which was submitted unless for publication elsewhere, we performed quantitative sensory testing (QST) according to a protocol of the German Research Network on Neuropathic Pain (DFNS) (B. Baier, P. zu Eulenburg, C. Geber, R. Rohde, R. Rolke, C. Maihöfner, F. Birklein, M. Dieterich, unpubl. ms; Rolke et al. 2006) in 20 patients for the actual subgroup analysis (Rolke et al. 2006). We found employing a statistical VLBM with Liebermeister statistics (Rorden et al. 2007) that warm and cold perception thresholds contralateral to the stroke were strongly associated to lesions in the posterior IC (B. Baier, P. zu Eulenburg, C. Geber, R. Rohde, R. Rolke, C. Maihöfner, F. Birklein, M. Dieterich, unpubl. ms.).

For all non-responders, two weeks following the reminder e-mail, a paper version of the survey will be mailed, with addressed stamped envelopes. The data from the paper surveys will be reconciled with the data from the electronic surveys. Study population There are currently 23 medical dispatch centres in Ontario that employ approximately 850 call takers [personal communication, Ontario Ministry of Health and Long-Term Care, [8]. We will approach click here managers of the selected medical dispatch centres and ask for their assistance in recruiting call takers to participate in Phase One (iterative

surveys) and Phase Three (survey). Inhibitors,research,lifescience,medical In Phase Three, the finalized survey will Inhibitors,research,lifescience,medical be circulated to our target population – all 9-1-1 call takers in the province of Ontario. Sample Size Power calculations for multiple regression analysis of Phase Three survey will depend on the number of cases per predictor variable. A minimum sample size of 50 + 8 m, where m is the number of predictor variables, is recommended for testing the multiple correlation, and 100

+ 8 m for testing Inhibitors,research,lifescience,medical individual predictors [37,38]. The survey will be organized using the theoretical constructs of the theory of planned behaviour which measure: behavioural intentions, attitudes, subjective norms, and perceived behavioural control. Assessments for each of the four theoretical constructs for both behaviours under study will include direct and indirect belief-based measures; each measure

Inhibitors,research,lifescience,medical will use a minimum of three items on a 7-point Likert scale. Our survey should measure approximately 10–12 items, requiring a minimum sample size of 146 to test the multiple correlation, or 196 to test individual predictors. Methods of measurement For Phase One, the audio-tapes of the interviews will be transcribed verbatim and verified by the interviewer prior Inhibitors,research,lifescience,medical to analysis. Data will be analyzed to identify themes and codes, including intentions to perform the behaviour, attitudes, subjective norms and perceived behavioural control of the behaviour. Two researchers will independently analyse the content of the responses, identify themes, and list them in order of frequency about [39-42]. In Phase Two and Three, the survey data will be entered into a secure database developed by the Data Methods Centre at the Ottawa Hospital Research Institute. Upper and lower limits will be set for each variable and logical and range errors will be detected immediately by the program and highlighted for correction. Ten percent of case records, randomly selected, will be re-entered to assess data entry accuracy.

La réalisation des PEM peut compléter ce premier bilan. La réalisation systématique d’une étude du LCS ne fait pas l’objet d’un consensus. La réalisation d’autres tests, notamment biologiques, est guidée par le contexte clinique : bilan phosphocalcique ;

dosage des folates, de la vitamine B12 ; sérologie de la maladie de Lyme, du VIH, de la syphilis ; dosage de TSH. Au cours d’un bilan immunologique, peut être réalisé le dosage des AC antigangliosides, des AC antinucléaires et dans certaines situations des AC antineuronaux (anti-HU, etc.), des AC antirécepteurs à l’acétylcholine. Enfin, une exploration Selleck GSK1120212 plus spécifique pourra être demandée devant des particularités cliniques. les auteurs déclarent ne pas avoir de conflits d’intérêts en relation avec cet article. “
“Seas and oceans cover about 70% of the Earth’s surface and they are now viewed by the scientific community as the last great frontier for natural source of bioactive compounds.1 One of the resources is coral reef ecosystem. Coral reef ecosystem is a part of marine ecosystems where a vast amount of marine biota lives. In the coral reef ecosystem live more than 300 species of reefs, more than 200 species of fish, tens of mollusks, crustaceans, sponges, alga,

sea grasses, and many other species of biota. Sponges play a role in constructing the coral reefs since they contain crotamiton active compounds. Moreover, active compounds in the sponges are higher that those

produced by land vegetations. Sponges are also marine invertebrates that are most actively investigated in the efforts Epacadostat nmr of finding marine natural products with anticancer properties.2 Relatively few works were carried out to investigate antioxidant and cytotoxic properties of sponges from inhibitors Pecaron Bay, Situbondo, Indonesia. This study describes a screening for cytotoxicity and antioxidant of hydro-ethanolic extracts derived from eight sponge species collected at the Tanjung Pecaron, East Java. Cytotoxic and antioxidant activities were evaluated in order to improve the knowledge on the pharmacological potential of the sponge fauna from the East Java, Indonesia. Sponge samples were taken from Pecaron Bay 2009 on the last July 2009 using SCUBA diving in 5–20 m depth from 500 of m length from coastline. They were photographed under water for helping the identification and finally samples were preserved by ethanol solution 70% for specimen and morphology identification. The specimen and morphology identification were conducted in the Laboratory of Zoology Institute of Technology Surabaya. The method for morphology identification used the determination key.3 The DPPH radical scavenging effects of the total extract and compounds were performed by using a modified version of the previously established methodology.

For human subjects, two tests were considered, the tube task and the bimanual Brinkman board task, as well as the questionnaire indicating their self-assessed hand preference. For the tube task, the preferred hand was defined as the hand used to grasp the reward into the tube, playing the manipulative role, whereas the other hand, holding the tube, played the Adriamycin datasheet postural role. The preferred hand (left Inhibitors,research,lifescience,medical hand or right hand) was determined for each tube task trial performed by the subject (humans and monkeys), in order to calculate the handedness index (HI) (see below). For the bimanual board task, the subjects (humans and monkeys) used two different strategies to retrieve the reward. In the first one, the

hand above the board pushed the reward while the other hand collected it below the board. In the second one, the hand positioned below the

board pushed up the reward using one Inhibitors,research,lifescience,medical finger (usually the index finger) and the other hand grasped it above the board, performing the precision grip. In the first strategy (adopted in more than 98% of trials in five out of eight monkeys), the preferred hand is the one pushing the reward. Indeed its role is manipulative, whereas the role of the other hand is postural. For the second strategy, the preferred hand is the one retrieving the reward, as its action is more manipulative and more challenging Inhibitors,research,lifescience,medical (precision grip), as compared to the role of the other hand (one finger used). Additionally, the board has an inclination, making this movement still more difficult. This second strategy was used in about half of the trials in one monkey (Mk-MI) and it was predominant Inhibitors,research,lifescience,medical in two other monkeys

(Mk-CA and Mk-AN; 68% and 98%, respectively). For the reach and grasp drawer task (in monkeys only), the preferred hand is the hand grasping the reward (manipulative role) while the other hand, the postural one, holds the drawer. For these three tasks (bimanual Brinkman board task, reach and grasp drawer task, tube task), we computed the HI (Westergaard et al. 1997; Spinozzi et al. 1998; Hopkins et al. 2004; Schmitt et al. 2008), defined as follows: the number of trials the right hand (R) Inhibitors,research,lifescience,medical was used as preferred hand minus the number of times the left hand (L) was used as preferred hand, divided by the total number of trials: Consequently, a negative HI reflects a left bias whereas a positive HI reflects a right bias. The HI (lateralization) ranges between +1 (strongly right-handed) and −1 (strongly left-handed). much For the modified Brinkman board task, we measured the score in 30 sec when the animal was free to use both hands, and counted the number of pellets grasped with each hand. The hand with the highest score is considered as the preferred hand. For the questionnaire, we calculated a handedness score by using the criteria of MacManus (2009): “Laterality scores (laterality indices): Score all the items as −1 = Always left, −0.5 = Usually left, 0 = Either, +0.

The Lin group found decreased choline in HCC and cirrhosis patient sera compared with normal sera, although they did not compare HCC and cirrhotic patients [48]. In HCC tissue, choline was found upregulated [27], which is consistent with previous in vivo MRS studies [49]. Generally, choline is an essential

metabolite in the Z-VAD-FMK nmr synthesis of phospholipids for cancer cell membranes [50]. This metabolism has been studied and monitored by NMR previously [51,52,53]. Choline is also associated with many cancer types. For example, Inhibitors,research,lifescience,medical it has shown to be associated with colorectal cancer [54], high grade gliomas [55], and breast cancer [56]. Thus, the metabolism of the membrane phospholipids caused by accelerated cell proliferation could be a reason for elevated choline in the sera of HCC patients [27]. 5. Conclusions 1H NMR metabolic profiling of serum samples has been shown to differentiate HCC from HCV patients. In addition to a good separation based on broad lipid signals in the NMR spectra, three metabolites, Inhibitors,research,lifescience,medical creatinine, valine and choline, were found to differentiate the two disease groups and each metabolite has some precedence as a potential HCC biomarker in human serum or urine. In addition, these metabolites are readily

detected in serum by a number Inhibitors,research,lifescience,medical of analytical methods, indicating that upon further validation they could be straightforwardly translated into clinical practice. A distinguishing feature of this study is that it focuses on a particularly challenging patient cohort, i.e., those with underlying HCV. It is extremely difficult to differentiate HCC patients with underlying Inhibitors,research,lifescience,medical HCV from HCV patients for several reasons: 1) mediators associated with inflammation often overlap with those associated with cancer and therefore teasing out cancer specific differences is difficult; 2) changes associated with fibrosis also overlap with cancer

and the majority of HCV patients do not develop Inhibitors,research,lifescience,medical cancer until the liver has become severely fibrotic; and 3) confirmation of cancer requires pathologic evidence that is not found in cases where resection or transplant has not been performed or where occult disease is present, but only detected from the most sophisticated tests. Patients with HCV were of particular interest for this study since they represent the largest cohort of HCC patients within the US and are at the highest Florfenicol risk for developing HCC during their lifetimes. The results of this study indicate the promise of developing metabolite profiles for the detection of HCC. Future studies will focus on adding MS detected biomarker candidates and expansion of the studies with additional sample cohorts. We anticipate that additional metabolite biomarkers will significantly improve the detection model and provide an alternative to current modalities. Acknowledgments This work was supported by funding from the National Cancer Institute, (1R21CA133770) and the Purdue Research Foundation.

MPI Research is accredited by the Association for Assessment and Accreditation of Laboratory Cytoskeletal Signaling inhibitor Animal Care International (AAALAC International), and was under guidance of IACUC. Modulators Vaccinations with the nanoparticle vaccine and saline control were administered by injection between the skin and underlying layers of tissue in the thigh region of each animal. The same injection site on each animal was used for each administration unless a reaction at the injection site indicated that another site must be used. All injection sites were marked and identified throughout the course of

the study. The dose was administered by bolus injection. Monkeys were immunized (N = 10 per group) on days −78 and −48 with a combined pediatric diphtheria/tetanus

toxoid vaccine, and then immunized on days 1, 29, and 57 with saline, or escalating doses of 1 mL of nanoparticle vaccine at 0.5, 2.0, 8.0 and 16.0 mg/mL. Blood was collected on days shown, prior to immunization (day 1) and then on days 29, 57, 85, 113, and 141 to test for anti-nicotine antibodies. Peripheral blood was collected on day 85 for T cell recall analysis (3 mL) and PBMC isolated by percoll centrifugation. Briefly, human peripheral blood mononuclear cells (PBMCs) were isolated from normal human donors (Research Blood Components, Cambridge, MA). Blood was DAPT diluted 1:1 in phosphate buffered saline and then 35 mL overlaid on top of 12 mLs Ficoll-Paque premium

(GE Healthcare, Pittsburgh, PA) in a 50 mL centrifuge tube. Tubes were spun at 1400 RPM for 30 min, and the transition phase PBMCs collected, diluted in PBS with 2% fetal calf serum and spun at 1200 rpm for 10 min. Cells were re-suspended in cell freezing media (Sigma–Aldrich, St. Louis, MO) and immediately frozen at −80 °C. For long term storage, cells were transferred to liquid nitrogen. For rhesus monkey PBMC isolation the protocol was the same except 5 mL of blood was collected and processed. from For cynomolgus monkey PBMC, 3 mL of blood was processed, buffy coat was collected and overlaid on 60% Percoll (GE Healthcare), centrifuged 30 min at 1755 rpm, washed and frozen as described above. Frozen PBMC were thawed (37 °C water bath), re-suspended in PBS 10% FCS, spun down and re-suspended to 5 × 106 cells/mL in tissue culture media (RPMI), supplemented with 5% heat inactivated human serum (Sigma–Aldrich), l-glutamine, penicillin and streptomycin, (Gibco, Grand Island, NY). For memory T cell recall response assays, cells (0.6–1.0 mL) were cultured in 24-well plates with 4 μM peptide (GenScript) at 37 °C 5% CO2 for 2 h. One μL of 1000× Brefeldin A (BD, San Jose, CA) per mL of culture media was then added and cells returned to a 37 °C incubator for 4–6 h. Cells were then incubated at 27 °C, 5% CO2 for 16 h.