In good agreement with the collapsed mitochondrial potentials, tr

In good agreement with the collapsed mitochondrial potentials, treated cells showed an increase in the oxidized CL (Fig. 6, lower panel). Moreover, we decided to evaluate a correlation between erythroid differentiation and mitochondrial impairment. In particular, the involvement of the mitochondrial pathway

via activation of caspase-3 and caspase-9 was evaluated; to this aim, K562 cells were irradiated in the presence of the pancaspase inhibitor z-VAD.fmk and then benzidine test was performed. As shown in Fig. 7, z-VAD.fmk suppressed erythroid differentiation induced by all furocumarins. We also Duvelisib solubility dmso studied the possible erythroid differentiation activity of irradiated mixtures of some tested furocoumarins. These compounds were 5′-MP, 4′,5′-DMP and 5,5′-DMP and were chosen on the basis of their higher

sensitivity to UV-A photodegradation LY2157299 purchase (followed by UV–vis spectroscopy- data not shown). After their irradiation in methanol solution with different UV-A doses (0, 8, 16 and 32 J/cm2), psoralens were concentrated by solvent evaporation and then resuspended in methanol. The erythroid differentiation of photoproducts was investigated by benzidine test incubating K562 with psoralen irradiated mixtures at two different concentrations (50 and 200 μM) for 5–7 days. Cell growth was also evaluated using the MTT assay after 6 days of treatment (Table 3). After 6 days of incubation, cells treated with 50 μM pre-irradiated mixtures

did not show a clear increase of benzidine positive cells (Fig. 8, upper panel) nor a decrease in cellular viability in comparison to control (Table 3); on the contrary, using the higher concentration, an induction of erythroid differentiation (26–36% benzidine positive cells) (Fig. 8, lower panel) together with a reduction of cellular viability was over observed only with 5,5′-DMP (Table 3); the other POP mixtures exhibited low activity or were inactive. The irreversibility of the erythroid differentiation induction by 5,5′-DMP photoproduct mixtures was also assessed. The first 6 days of treatment were sufficient for K562 cells to differentiate irreversibly since during additional 4 days of culturing in the absence of the inducer of washed cells, the population of benzidine-positive cells still increased (from 26.3 ± 3.1 to 44.3 ± 2.2 in the case of 8 J and from 35.1 ± 2.0 to 40.5 ± 1.1 in the case of 16 J). RT-qPCR was also employed to quantify the expression of globin mRNA following treatment of K562 cells with 5,5′-DMP photoproducts. There is a clear positive relationship between UV-A doses used to obtain the photoproducts and the extent of increased globin mRNAs in respect to control K562 cells (Fig. 9). As far as a possible differential activity of furocoumarin photoproducts on globin gene expression is concerned, the data clearly indicate that accumulation of both the α-like α-globin mRNA and ζ-globin mRNA are strongly induced.

Finally talc was added as an anti-sticking agent based on the sol

Finally talc was added as an anti-sticking agent based on the solid dry weight of the polymers with continuous stirring for approximately 10 min. In this

way all the coating dispersions were prepared and was sprayed onto the drug loaded pellets until the pellets achieved desired coating level. Compositions were given in Table 3. The above pellets were evaluated for various parameters like particle size analysis, size distribution, shape and surface roughness, flow properties, drug content and in vitro dissolution profile. this website Particle size analysis was done by optical microscopy method. Drug content was carried out by UV method. 4, 14 and 15 The particle size of drug loaded formulations were measured by an optical microscope fitted with an ocular see more and stage micrometer and particle size

distribution was calculated. The Weswox model having resolution of 45× was used for this purpose. The instrument was calibrated at 1 unit of eyepiece micrometer was equal to 30.07 μm. Angle of repose (θ) was assessed to know the flowability of pellets, by a fixed funnel method using the formula: Angleofrepose(θ)=tan−1(h/r) Tap density and bulk density of the pellets were determined using tap density tester. The percentage Carr’s index (I, %) was calculated using the formula: Carr’sindex(I,%)=Tappeddensity−Bulkdensity/Tappeddensity Hausner’s ratio was measured by the ratio of tapped density to bulk density. Hausner’sratio=Tappeddensity/Bulkdensity The whatever friability test was performed on the pellets to ensure their mechanical strength. Lower friability values indicate good mechanical strength. Pellets of known mass

were placed in a Roche Friability tester and subjected to impact testing at 25 RPM for 5 min. Prior to and following the test, the weights of the formulation were accurately recorded and friability ratios were calculated with the given equation. F=W1−W2/W1×100F=W1−W2/W1×100where, W1 = Initial weight of the formulation, W2 = Final weight of the formulation. Shape and morphological features of pellets were observed by scanning electron microscopy (SEM). Surface and shape of the formulated pellets were observed to be varying depending on composition of polymer and plasticizer. The shape of the pellets was investigated by JEOL, JSM-6610LL, Scanning electron microscope, Japan. Compatibility of aceclofenac with polymers EC N50 and HPMC E5 in 1:1 ratio of physical mixtures were analyzed by Fourier transform-infrared spectroscopic analysis (FT-IR) and the IR spectra were taken. The aceclofenac content of the pellet formulation was evaluated over accurately weighed 100 mg pellets which were dissolved in a little quantity of ethanol and then the volume was made upto the mark with pH 6.8 phosphate buffer. The resulted solution was analyzed spectrophotometrically at 274 nm (LAB INDIA, UV-3092) after suitable dilution with pH 6.8 phosphate buffer.

This article reviews the role of coronary computed tomography (CT

This article reviews the role of coronary computed tomography (CT) angiography in the assessment of coronary risk, and its usefulness in the emergency department in facilitating appropriate disposition decisions. Also discussed is coronary artery calcification incidentally found on CT scans when done for indications such as evaluation of pulmonary embolism or lung cancer. The evidence base and clinical applications for both techniques are described, together with cost-effectiveness and radiation exposure considerations. Ozlem Soran Medically refractory angina pectoris (RAP) is defined by presence of severe angina with objective evidence of ischemia and failure to relieve

symptoms with coronary revascularization. Medication and invasive revascularization are the most common approaches for buy CT99021 treating coronary artery disease (CAD). Although symptoms are eliminated or alleviated by these invasive approaches, the disease and its causes are present after treatment. New treatment approaches are needed to prevent the disease from progressing and symptoms from recurring. External enhanced counterpulsation therapy provides a treatment modality in the management of CAD and can complement invasive revascularization procedures. Data support that it should be considered as a first-line treatment of RAP. Doron Aronson and Elazer R. Edelman Diabetes mellitus (DM) is a major NLG919 ic50 risk factor for cardiovascular

disease. Near-normal glycemic control does not reduce cardiovascular events. For many patients with 1- or 2-vessel coronary artery disease, there is little benefit from any revascularization procedure over optimal medical therapy. For multivessel coronary disease, randomized trials demonstrated the superiority of coronary artery bypass grafting over multivessel percutaneous coronary intervention in patients with treated DM. However, selection of the optimal myocardial revascularization strategy requires a multidisciplinary team approach (‘heart team’). This review summarizes the current evidence regarding the effectiveness of various medical

therapies and revascularization strategies in patients with DM. A. Pieter Kappetein, Nicolas M. van Mieghem, and Stuart J. Head Coronary artery bypass grafting (CAGB) is superior to percutaneous coronary intervention (PCI) in reducing mortality in certain patients Liothyronine Sodium and improving the composite end points of angina, recurrent myocardial infarction, and repeat revascularization procedures. However, CABG is associated with a higher perioperative stroke risk. For patients with less complex disease or left main coronary disease, PCI is an acceptable alternative to CABG. Lesion complexity is an essential consideration for stenting, whereas patient comorbidity is an essential consideration for CABG. All patients with complex multivessel coronary artery disease should be reviewed by a heart team including a cardiac surgeon and interventional cardiologist. Shilpa Agrawal, Puja K.

From this subset of 118 responses, five themes were identified th

From this subset of 118 responses, five themes were identified that indicated implicit weight stigma: negative language when speaking about weight in overweight patients (n = 41,

35%); focus on weight management to the detriment of other important considerations (n = 12, 10%); weight assumed to be individually controllable (n = 69, 58%); directive or prescriptive responses rather than collaborative (n = 96, learn more 81%); and complexity of weight management not recognised (n = 98, 83%). The first theme was illustrated by negative terms used about body weight: a patient who was overweight had a ‘weight issue/weight problem’ that ‘needed to be/must be/should be’ ‘managed/addressed’. The second theme was most evident in the case study of the patient in an aged care setting. Weight management was often mentioned for this patient with a reduced focus (in comparison to

the normal weight presentation) on other important factors such as social support. The third theme (assumed controllability of weight) was evident in that diet and/or exercise were almost the only weight management strategies mentioned. The fourth theme of directive communication was demonstrated in the choice of language such as ‘speak to them about weight management’ or ‘he should lose weight’. Finally, the fifth theme identified a lack of recognition of the complexity of weight management. Specifically, only three (3%) responses questioned BMI selleck kinase inhibitor as a measurement of adiposity or health, three (3%) mentioned weight management strategies other than diet or exercise (referral to GP, referral to naturopath, mood), and six (5%) responses considered the psychological sensitivity next of weight. This paper explored whether physiotherapists demonstrate weight stigma and whether this might negatively influence patient treatment. The total Anti-Fat Attitudes questionnaire scores indicated that physiotherapists, in line with studies on many other health professionals,1 demonstrate explicit weight stigma. The scores on the subscales provided more insight

into the nature of this stigma and its likely implications for behaviour towards patients who are overweight. The Dislike subscale had a relatively low score, however responses were notably high in answer to the question ‘If I were an employer, I might avoid hiring an overweight person’, suggesting that physiotherapists’ negative attitudes may result in discriminatory behaviours. In contrast, the quantitative responses to the case studies showed little evidence of discriminatory behaviours. In fact, responses to one question (feeling similar to a patient) indicated a greater liking of patients who were overweight. A similar effect is noticeable elsewhere in physiotherapists’ attitudes.28 This apparent contradiction is possibly explained by the ‘jolly fat stereotype’,40 which fits with the stereotype content model.


the investigation of its use in children and a


the investigation of its use in children and adolescents with AIDS might provide information about the response to this vaccine, as well as its potential for preventing meningococcal disease. The main objective of this Palbociclib in vitro study was to evaluate the antibody response to the meningococcal serogroup C conjugate vaccine in HIV-infected children, adolescents, and young adults. Additional objectives included determining whether the immunity acquired correlated with clinical, viral, and immunological parameters of infection; analysing the response to a second dose of the vaccine, if necessary; and reporting any side effects of the vaccine. This was a prospective clinical trial involving a cumulative sample of HIV-infected children, adolescents, and young adults (HIV+ group) and age-matched non-HIV-individuals (HIV− group). The sample

size was calculated considering an expected rate of 60% of subjects with a post-vaccination serum bactericidal antibody (SBA) titer ≥8, assuming an actual proportion of 90%. To compensate for a potential loss of 20%, we selected 40 subjects for inclusion in each group. The method employed was hypothesis testing for comparing two proportions [20]. All subjects were recruited among patients treated at the Instituto da Criança do Hospital das Clínicas da Universidade de São Paulo, Department of Pediatric Infectious Diseases – Brazil or at the Centro de Referência e Treinamento em DST/Aids – Programa Estadual São Paulo – Brazil, both located in the city of São Paulo, Brazil. Patients SCR7 supplier were considered eligible if they were between 10 and 20 years of age, had never been vaccinated with meningococcal serogroup C conjugate vaccine, had no prior history of meningococcal disease or meningitis of undetermined etiology, had not used corticosteroids at immunosuppressive doses, had not

below been treated with immunosuppressive therapy or chemotherapy, and presented with no evidence of significant dyslipidemia [21]. The inclusion criteria for the HIV+ group were being HIV-infected, having a CD4 count ≥100 cells/mm3, and not having received immunoglobulin therapy within the last six months. The inclusion criterion for the HIV− group was having no underlying disease that would result in immunosuppression or would require immunosuppressive therapy. The HIV− group patients with unknown HIV serologic status were submitted to a rapid HIV test to confirm that status. We collected demographic, clinical, viral and immunological data at inclusion. All patients underwent an initial blood test to determine pre-vaccination SBA titers, after which they were vaccinated with the meningococcal serogroup C conjugate vaccine in isolation (i.e., no other vaccine was administered).


the majority of individuals achieve an i


the majority of individuals achieve an independent gait after stroke, many do not reach a walking level that enables them to perform all their daily activities (Flansbjer et al 2005). Typically, the mean walking speed for the majority of community-dwelling people after stroke ranges from 0.4 m/s to 0.8 m/s (Duncan et al 1998, Eng et al 2002, Green et al 2002, Pohl et al 2002, Ada et al 2003). This slow speed frequently prevents their full participation in community activities. Additionally, people report a lack of ability BMN 673 purchase to cover long distances after stroke, restricting their participation in work and social activities (Combs et al 2012). Moreover, walking ability has been found selleck kinase inhibitor to be related to community

participation (Robinson 2011). While the goal of inpatient rehabilitation is independent and safe ambulation, once individuals return home, rehabilitation aims to enhance community ambulation skills by increasing walking speed and endurance. Lord et al (2004) found that the ability to confidently negotiate uneven terrain, private venues, malls and other public venues is the most relevant predictor of community ambulation. Therefore, in order to enhance community participation, rehabilitation has focused on identifying the best approach to optimise walking speed and walking distance. One approach to improving gait is the use of mechanically assisted walking devices, such as treadmills or gait trainers. Two Cochrane systematic reviews have examined

these devices separately: Moseley et al (2005) reported on treadmill training and Mehrholz (2010) examined electromechanically-assisted training. We wanted to examine all devices that will help improve walking in the one review. In ambulatory stroke, mechanically assisted walking, whether by treadmills or gait trainers, allows an intensive amount of stepping practice by working as a ‘forced use’. Mechanically assisted walking also facilitates the practice of a more normal walking pattern because it forces appropriate timing between lower limbs, promotes hip extension during the stance phase of walking and discourages common compensatory behaviours PAK6 such as circumduction (Harris-Love et al 2001, Ada et al 2003, Moore et al 2010). We have already taken this approach in What is already known on this topic: Mechanically assisted walking training, which can involve interventions such as treadmill training or electromechanical gait trainers, increases independent walking among people who have been unable to walk after stroke. However, previous systematic reviews have not drawn clear conclusions about the effect of treadmill training or gait trainers among ambulatory stroke survivors specifically. What this study adds: Compared with no intervention or with an intervention with no walking training component, treadmill training improved walking speed and distance among ambulatory people after stroke.

Strong negative associations with intention were found for having

Strong negative associations with intention were found for having an omission bias, holding naturalistic views, for the disbelief in scientific

evidence that influenza vaccination is effective, BMS907351 and the disbelief in the relevance of the flu shot. Results of the multinominal logistic regression are shown in Table 4. HCP were more likely to have no intention to get vaccinated vs. not having made a clear decision when they reported a negative attitude towards influenza vaccination and high feelings of autonomy, when they showed a stronger omission bias, a lesser sense of personal responsibility to protect patients by getting vaccinated, when they reported high self-protection motives, and lower frequency of influenza GW786034 vaccinations in the past. When comparing having a high intention vs. not having made a clear decision, we found that HCP with a positive attitude towards influenza vaccination and a higher frequency of influenza vaccinations in the past were more likely to have a high intention

vs. not having made a clear decision. No other significant unique contributions to the prediction of having a high intention were found. The variables in the regression model explained 80% of the variance in intention (pseudo R2 = .80), with a classification accuracy of 82%. In an exploratory manner we excluded the most influential variable, attitude, from the multinominal analysis, because we hypothesized that it might overrule the (indirect) influence of other variables on intention. Only one additional significant predictor appeared Oxymatrine in this analysis: higher sense of personal responsibility significantly predicts a high intention to get vaccinated as opposed to an unclear decision when attitude is excluded. We next tested whether attitude mediates the relationship between personal responsibility and high intention vs. an unclear decision. To test for mediation, we used the SPSS macros that Preacher and Hayes [28] provide for a binary logistic regression with bootstrapping technique. The bias corrected and accelerated

(BCa) confidence intervals were set at .95 with 5000 resamples. The mediation analysis revealed that there is a meaningful indirect effect of attitude on the relationship between personal responsibility and intention (b = 1.29, BCa 95% CI [.874; 1.856]), only for participants in the categories high intention vs. no clear decision (N = 274). The fact that zero falls outside this interval indicates a significant mediation effect. For the regression coefficients for the relationship between personal responsibility and intention (high/unsure) as mediated by attitude, see Fig. 1. Table 5 shows that amongst the HCP that got vaccinated against influenza, the majority had reported to have a high intention to get vaccinated at baseline (N = 68, 73.9%). The percentage of participants that were vaccinated differed by intention, χ2 (2, N = 458) = 224.42, p < .001. Of the HCP who participated in the follow-up survey (N = 458), 90 (19.

3) of ACEL(0 15) and ACEL(0 30) also suggested that both the prop

3) of ACEL(0.15) and ACEL(0.30) also suggested that both the proportions exhibited a singe step weight loss at about 200 °C. The X-ray powder diffraction patterns of ACT, ACEU and ACEL are shown in Fig. 4. Intense and sharp diffraction peaks at 9.9°, 21.8°, 24.9° and 29.5° 2θ and weak and diffused peaks at 16.5°, 17.3°, 18° and 23.6° 2θ; in addition to peaks at 20.3° and 20.9° 2θ in the diffraction pattern of ACT confirmed its crystalline polymorphic form A.12 ACT also showed additional diffused peak at 12.1° and an intense peak at 31.6° 2θ. Characteristic hump shaped diffraction pattern in the range of 10–20° 2θ for EPO confirmed

its amorphous nature, whereas a sharp and intense peak at 19.1° 2θ as well as a diffused and weak peak at 23.3° 2θ for POL confirmed its semi-crystalline nature. 1:2 proportion of ACEU could be differentiated from 1:1 proportion on the grounds that the principal peaks Entinostat mouse were observed with much lower intensity and significant broadening in 1:2 proportion and it

was assessed to provide relatively more extent of amorphisation. Amorphous character of ACT in ACEL was significantly improved by the addition of find more POL as evident by XRPD profiles. XRPD profile of ACEL(0.30) distinctly showed a halo diffraction pattern and absence of all the principal peaks corresponding to crystalline ACT, unlike that of ACEL(0.15), which confirmed that the drug was molecularly dispersed in the polymer–plasticiser matrix and the extrudates heptaminol so formed were homogeneous, amorphous solid solution.

Percent content of ACT in ACEU and ACEL was found to be in the range of 98.3 ± 0.16%–99.1 ± 0.23% (n = 3) of theoretical proportion of the drug in the respective solid dispersions. The intrinsic solubility and in vitro dissolution rate of ACT, ACEU and ACEL in 0.1 N HCl is shown in Table 1 and Fig. 5, respectively. As compared to pure drug, both the proportions of ACEU exhibited considerable enhancement in intrinsic solubility; with more than 90% drug release in about 60 min. This could be attributed to high mass transfer associated with increased surface area of the drug by the high shear during extrusion process. Furthermore, both the proportions of ACEL reported about 7–10 folds enhancement in intrinsic solubility and more than 90% drug release within ∼20 min. Such enhancement in solubility characteristics could be attributed to decreased recrystallisation of the drug within plasticised polymer and lack of strong intramoleular bonds within ACT and existence of week intermolecular hydrogen bonds between the drug and plasticised polymer molecules. These randomly arranged molecules required less energy to separate and dissolve as compared to crystalline ACT. In addition, poloxamer being non-ionic surfactant further improved wettability of the dispersed drug particles. The hydrophilic polyoxyethylene segment of the copolymer also prevented aggregation or agglomeration of individual drug particles, thus improving solid–liquid surface tension.

, 2008),

, 2008), find protocol providing one potential mechanism for stress-induced deficits in memory recall (Chen et al., 2010). Similarly, using transcranial two-photon microscopy to image the dynamic remodeling of postsynaptic dendritic spines in the living, developing cortex (Liston and Gan, 2011), we found that glucocorticoids have rapid effects on both spine formation and elimination within hours of exposure. Surprisingly, low-dose dexamethasone (0.1 mg/kg), a synthetic glucocorticoid that inhibits endogenous corticosteroid synthesis without penetrating the blood/brain barrier

(Karssen et al., 2005), effectively prevented developmental spine formation and pruning. It is important to note that studies in neuronal cultures and in the developing cortex are investigating spine remodeling under conditions of heightened plasticity, so additional work will be needed to understand how the results apply to the adult brain. However, these experiments indicate that glucocorticoids play an unexpected, necessary role in facilitating physiological spine maturation in the developing adolescent brain, acting on timescale of selleck screening library minutes to hours to facilitate spine remodeling. These unexpectedly

rapid effects also suggest that circadian glucocorticoid oscillations may contribute to synaptic plasticity during learning and development. To test this hypothesis, we conducted a series of two-photon imaging studies in mice before and after training on a RotaRod motor skill-learning paradigm, and found that

circadian glucocorticoid peaks and troughs play critical, complementary roles in facilitating experience-dependent spine remodeling (Fig. 2c–g) (Liston et al., 2013). Specifically, circadian glucocorticoid peaks enhanced spine formation rapidly in the hours after learning, acting through a glucocorticoid receptor-dependent, non-transcriptional mechanism. In accord with prior reports (Yang et al., 2009), training increased formation rates but only if it occurred during the circadian peak. In mice that were trained during the circadian trough, spine formation rates were equivalent to those of others untrained mice, and memory retention was reduced one week later. Furthermore, circadian troughs were necessary for stabilizing a subset of learning-related spines and pruning a corresponding set of pre-existing synapses. Memory retention and the long-term survival of learning-related spines required intact circadian troughs in the days after learning, which enhanced learning-related spine pruning through a distinct, mineralocorticoid receptor-dependent, transcriptional mechanism. In this way, circadian glucocorticoid oscillations were critical for maintaining homeostasis in synaptic density, by balancing formation and pruning after learning to maintain relatively stable synaptic densities despite repeated bouts of learning-related remodeling.

After 30 min incubation in the dark, cells were washed and analyz

After 30 min incubation in the dark, cells were washed and analyzed by flow cytometry. Antigen presenting cells (SmyleDCs, SmartDCs or PBMCs) were irradiated with 30Gy, and CD3+ T cells isolated with immunobeads (Miltenyi Biotech) were used as responders. Different APC or PBMC ratios were co-cultured with 1 × 105 allogeneic CD3+ T cells (2, 5 and 20) in rounded-bottom 96-well plates in a total volume of

200 μL Cellgro medium. Triplicate wells were set up for each reaction and ratio. The reactions were incubated for 6 days at 37 °C. For the last 18 h of the culture, the supernatants from each reaction were collected for find more multiplex luminex bead kit. 1 μCi/well of [3H] Thymidine was added and [3H] Thymidine incorporation in the cells was measured on a β-scintillation counter. The stimulatory

capacity was determined with stimulation index (SI) = counts per minute (cpm) of stimulated T cells and stimulators/cpm of unstimulated T cells. To determine the production of cytokines by NK cells stimulated with iDCs, autologous NK cells were freshly isolated from PBMCs and co-incubated with 7 day SmyleDCs or SmartDCs at 1–5 ratio for 15–17 h. Staining of surface antigens on stimulated CD3−CD56+ NK cells was performed at 4 °C for 30 min. For SB431542 ic50 analysis of IFN-γ and TNF-α intracellular staining, cells were washed and fixed with 4% paraformaldehyde others for 10 min. After fixation, cells were permeabilized with saponin buffer (PBS supplemented with 0.1% saponin and 10 mM HEPES) and stained with IFN-γ and TNF-α mAb. After 30 min incubation, cells were washed three times and the percentage of IFN-γ and TNF-α positive NK cells was determined by flow cytometry. SmyleDCs and SmartDCs kept in culture for 7 and 14 days were analyzed for their DC immunophenotype. Cell were harvested and washed once with PBS and blocked with mouse IgG (50 μg/mL) on ice for 15 min followed by staining with a combination of monoclonal antibodies; FITC-conjugated anti-human

CD209, APC-conjugated anti-human CD86, PE-conjugated anti-human CD80, PerCP-conjugated anti-human HLA-DR, PE-conjugated anti-human CD14 and PerCP-conjugated anti-human CD123 (Becton Dickinson) for 30 min in the dark. After washing off the unbound antibodies, cells were then resuspended in 1% paraformaldehyde for fixation and further analyzed with a FACS Calibur apparatus (Becton Dickinson), using CellQuest software. Total viable cells were gated and 20,000 cells in gate were acquired. 7-day conventional IL-4-DCs or IFN-α-DCs or iDCs (non-matured) or 5-day iDCs further incubated for 2 days with 200 IU/ml rhTNF-α, 5 ng/ml rhIL-1B, 10 ng/ml rhIL-6 and 1 mg/ml PGE2 (matured) were harvested and loaded with 10 μg/ml PepTivator CMV-pp65 overlapping peptide pool (Miltenyi Biotec). After 2 h, excess unloaded peptides were washed off.