Therefore a major limitation of the BrdU assay is that only cells

Therefore a major limitation of the BrdU assay is that only cells that have progressed through the S-phase during this short incubation period may be detected. In contrast, cells express Ki67 in all active phases of the cell cycle. Therefore, Ki67 appears to be a more sensitive marker for the detection of rare T cell responses, and may reflect the extent of in vitro antigen-specific proliferation more accurately than BrdU incorporation. Cellular proliferation in PBMC samples is routinely evaluated by dye dilution methods, using CFSE or derivatives such as OG (Robinson and Amara, 2005). A recent non-human primate study has proposed measurement of in vitro proliferation

by the combined analysis of Ki67 and side scatter properties of cells ( Shedlock et al., 2010). The authors demonstrate a correlation between this assay and the CFSE dilution assay. In this study, we show that the proliferation events detected by loss of OG dye are virtually identical to Carfilzomib the Ki67+ events. From this we reasoned that Ki67 expression is an accurate measure of T cell proliferation as only

cells that have completed cycling display a decrease in OG fluorescence intensity. Limitations of many protein reactive dye compounds include cellular toxicity ( Last’ovicka et al., 2009 and Shedlock et al., 2010) and sensitivity to pH and light ( Wallace et al., 2008). The Ki67 ITF2357 molecular weight proliferation assay requires no incubation or washing steps prior to or during the culture, and exposure of cells to toxic compounds is eliminated. Additionally, since labelling of cells is not required before antigen stimulation, detection of Ki67 by flow cytometry can be performed on antigen-stimulated cells after cryopreservation. A limitation of Ki67 as a proliferation marker is its inability to resolve the number of proliferation cycles that cells have undergone, Ketotifen as can be done with dye dilution assays ( Parish, 1999 and Lyons and Doherty, 2004). Enumeration of cell cycles enables calculation of

the original precursor frequency of specific cells, since the number of cells and their respective number of divisions are known ( Givan et al., 1999). Monitoring vaccine-induced T cell proliferative potential is important for determining vaccine take, memory function and long-term persistence of vaccine-specific responses. Previous studies have quantified Ki67 expression directly ex vivo as a measure of the vaccine-induced proliferative response ( Miller et al., 2008), or in combination with activation markers to identify antigen-specific T cells ( Stubbe et al., 2006). To detect increases in the expression of Ki67, these studies relied on low-level Ki67 expression before vaccination in healthy adults. Direct ex vivo detection of antigen-specific Ki67 expression may thus be challenging in individuals with high levels of in vivo T cell proliferation — such as those resulting from recent vaccinations or infections.

The following day, the coverslips with the labeled cells were was

The following day, the coverslips with the labeled cells were washed four times in PBT for 5 min and mounted with Mowiol (anti-fading medium). Images were obtained by using see more either fluorescence microscopy and a digital camera or multiple confocal sections by Zeiss LCM 5100. Two-month old cultures were incubated with 0.001% acridine orange diluted in IPL41 for 1 h. After washing cells three times in 1 mL PBS, cells were observed using an epifluorescence microscope to check for viability. A total of 300 cultured cells from three wells were analyzed for fluorescent nuclei. For comparative morphological analysis, cultured cells were also stained with 0.01% Giemsa

solution and observed under a light microscope. Through the use of a simple series of dissecting methods we were able to establish primary oenocyte cultures isolated from Ae. aegypti pupa. Oenocytes were free of other

cells as demonstrated by our microscopy analyses, and a number of cellular characters were assessed. Oenocytes were analyzed both in vivo and in vitro via light microscopy, SEM, TEM and LCM. Serial sections obtained from the abdomen of Ae. aegypti pupa revealed that oenocytes were detected as clusters of large cells within the fat body or in close proximity to the integument ( Fig. 1a). In fresh preparations the oenocytes were completely detached from other tissues and could be easily distinguished and sorted from trophocytes ( Fig. 1b). Under TEM, pupa oenocytes were clustered and enclosed by a basal lamina (Fig. 2a). These cells had a central nucleus with a well-developed nucleolus and the condensed chromatin appeared in irregular Inhibitor Library mw granular clumps, especially around the edge of the nucleus (Fig. 2b). The cytoplasm is replete with mitochondria and translucent rounded shape vesicle-like structures with different sizes (referred simply as vesicles) (Fig. 2a and b). The mitochondria were strongly electron-dense

with distinct profiles (Fig. 2c), while these vesicles were closely associated in bundles and not dispersed through the cytoplasm Pyruvate dehydrogenase (Fig. 2b). In addition, the cytoplasm was almost filled with numerous narrow, coiled and tubular structures of the smooth endoplasmic reticulum (SER) (Fig. 2c, inset). Plasma membrane protrusions touching the delicate basal lamina also were detected (Fig. 2d), and labeling with ruthenium red indicated that such protrusions surround the cell cortex, forming the lymph space, except in the intercellular space (Fig. 2d–f). Once in culture, oenocytes could be kept viable for at least two months. The two-old month cultured oenocytes observed under phase contrast microscope (Fig. 3a) and stained by Giemsa (Fig. 3b) confirmed the presence of a single type of adhered cells, isolated or in clusters. Cell clusters were consistently greater in number than isolated cells. The SEM confirmed the presence of clusters and of isolated oenocytes (Fig. 4a–d).

3 However, no information has been provided on the long-term effe

3 However, no information has been provided on the long-term effect of DSD on institutionalization in older patients admitted to a rehabilitation PD0325901 concentration settings and on the importance of DSD on long-term mortality in a large sample population in these settings. To address the paucity of data in this area, the purposes of this study were to evaluate (1) the association between DSD and functional outcomes, specifically walking recovery at discharge and at 1-year follow-up;

and (2) the association among DSD, institutionalization, and mortality at 1-year follow-up in a cohort of older inpatients in a rehabilitation unit. This was a prospective cohort study of inpatients aged 65 and older consecutively admitted to a rehabilitation unit between January 2002 and December 2006 either after acute hospitalization or directly from home. The

study was conducted in the Department of Rehabilitation and Aged Care (DRAC) at the “Ancelle della Carità” Hospital (Cremona, Italy), an 80-bed unit staffed by geriatricians; psychiatrists; neuropsychologists; nurses; and physical, speech, and occupational therapists. The characteristics of this clinical setting have been previously described.26 The Ethics Committee of Gerontological Sciences of the Geriatric Research Group approved the study. Informed consent was obtained from each patient at admission or an available proxy. Demographics included age and sex. Comorbidity was defined according to the Charlson Comorbidity Index (CCI).27 Admission diagnoses to the Belinostat in vitro Non-specific serine/threonine protein kinase DRAC were recorded. Overall functional status was assessed with the Barthel Index (BI)28 and 29 through patient and surrogate interview referring to 3 time points: (1) 1 month before the rehabilitation admission; (2) admission to the rehabilitation facility; and (3) at discharge. Presence of delirium at the time of admission was screened for with the Confusion Assessment Method (CAM) algorithm and it was confirmed by a gold standard clinical assessment using the Diagnostic and Statistical Manual of Mental Disorders (4th edition, text revision [DSM-IV-TR]) by 3 geriatricians (G.B., F.G., R.T.) trained in delirium and dementia assessment.

The presence of dementia was ascertained during inpatient rehabilitation by a consensus of 2 out of 3 geriatricians (G.B., F.G., R.T.) and 1 out of 2 neuropsychologists (E.L, S.M.) in accordance with the Diagnostic and Statistical Manual of Mental Disorders (3rd edition, revised [DSM-III-R, 1987]) criteria using a standardized approach, including assessment of cognitive and functional capacity, reviews of previous clinical and neuropsychological charts, and scores on Mini Mental State Examination (MMSE) and/or other neuropsychological tests. The DSM-III-R criteria were used instead of the DSM-IV-TR because they do not require a differentiation between subtypes of dementia and so defines the presence or absence of dementia per se.

10 Owing to the similarity in the ambient conditions and compara

10. Owing to the similarity in the ambient conditions and comparable

parameters at the simulated overflow, the shape of the θ-S curve and the magnitude of the temperature maximum are in good agreement with this generalisation. The results in this section expand on the Rudels and Quadfasel, 1991 schematic and describe the response in the mixing to variations in volume transport at the sill (see Fig. 8(b)). The maximum bottom temperature along the plume path is mainly a function of the flow rate (see Fig. 9(a)). The depth at which the temperature maximum occurs, on the other hand, is mainly a function of the inflow salinity. To explain these results we consider the processes and factors affecting the temperature maximum on the slope: (i) downslope advection of AW by the plume, (ii) Selleckchem DAPT the plume’s momentum arising from its density gradient, (iii) mixing of the plume with Atlantic Water, (iv) the smallness of the thermal expansion coefficient at low temperatures, and (v) the total thermal capacity of the plume water. In the following, we investigate how the salinity S   and flow rate Q   of the dense water inflow affect the plume’s final depth level. We quantify the downslope penetration of SFOW by calculating how much passive tracer (PTRC) is resident within a given www.selleckchem.com/products/i-bet-762.html depth range by the end of the model run. The concentration of tracer is integrated over a given volume to give the mass of PTRC, MPTRCMPTRC.

Astemizole The penetration of the cascade into a given depth range is calculated as a percentage of MPTRCMPTRC within the given range compared to the total MPTRCMPTRC over the entire domain. A model run and its dense water supply can then be characterised according to the depth range containing more than 50% of PTRC that has been injected over 90 days. In Fig. 11 we plot the results against S and Q for each of the 45 model runs. The final tracer percentage

present within the given depth range is shaded in a contour plot where the S-Q combination of each experiment is marked by a black dot. In those model runs where the majority of PTRC is present between 500 and 1000 m at the end of the experiment the plume has intruded into the Atlantic Layer and into the AW-NSDW interface, but not retained a strong enough density contrast to flow deeper. The combinations of S and Q producing this result are emphasised in Fig. 11(a) as the dots within the red shading indicating a tracer penetration greater than 50%. In the S-Q parameter space these runs are arranged in a curved band from low-S/high-Q via medium-S/medium-Q towards high-S/low-Q. In runs with lower S/lower Q (towards the lower left corner of the graph) the majority of the plume waters is trapped at shallower depths. In experiments with higher S/higher Q (towards the upper right corner of the graph) the plume reaches deeper as shown in Fig. 11(b) which is plotted for the presence of PTRC below 1000 m. Fig.

McPhaden and Hayes (1991) find that the one-day

lag in th

McPhaden and Hayes (1991) find that the one-day

lag in the correlation between SST and wind (pseudostress, zonal speed, and work) in the Western Pacific Warm Pool is due almost entirely to the surface heat fluxes and not from entrainment by wind-driven this website turbulent eddies. During intense but infrequent westerly wind burst events in the Western Pacific, wind-deepening of the boundary layer to the thermocline is hypothesized but latent heat fluxes at the surface are still thought to predominate ( Lukas and Lindstrom, 1991). Meridional advection may also contribute to SST during these exceptional wind events ( Feng et al., 1998). Insensitivity to Ri0 in the Western Pacific relative to the Central Selleck Metformin and Eastern Pacific ( Fig. 12) supports the hypothesis that interior diffusivity due to shear, and therefore entrainment, is not playing a role in the τ-SST correlation in that region. The sensitivity tests indicates that, given the uncertainty in the Tropical Pacific wind forcing

from Reanalysis products, calibration by comparison to data using the correlation cost alone would not be advisable. From the perspective of the unbiased “perfect model” the signal of the large perturbations to individual KPP parameters cannot be distinguished from the effect of changing between wind forcing products. Attempts to calibrate the KPP parameterizations using the cost function would yield wide probability distributions for the parameters. There are several potential sources of bias in our comparison between model and data. Because the atmosphere is not coupled to the ocean in the model, prescribed surface air temperature and specific humidity must, to some extent, control the heat flux across the ocean surface and therefore influence SST. All variables except wind speed and direction are held fixed at their NCAR/NCEP values across the alternative wind forcing experiments, so that the effect of this control over SST does not change from one wind experiment second to another. However, given that wind speed and direction are likely correlated with other prescribed variables (e.g. short wave

radiation), the default NCAR/NCEP forcing for variables other than wind may still affect the τ-SST correlation in the perturbed wind experiments. Missing processes or feedbacks may also contribute to the bias. On time scales on the order of a month, the τ-SST correlation is actually positive in the Tropical Pacific because of the atmospheric response to SST ( Bryan et al., 2010). Any feedbacks that may exist on the 40–160 h time scale used in this paper will not be represented because of the lack of a coupled atmosphere. Another possible source of bias in R could be related to the difference in spatial scales between model and data. The model has much less variability in SST than the data, even after band pass filtering ( Fig. 2).

[18], 15 patients with single episode of MDD (MDDs), 22 with recu

[18], 15 patients with single episode of MDD (MDDs), 22 with recurrent MDD (MDDr) and 15 with ADDM. Reduced BR echogenicity was found in 54% of the patients with Z-VAD-FMK research buy MDD and ADDM, but only in four

(8%) of the healthy subjects. BR echogenicity scores did not differ among patients with MDDs, MDDr, or ADDM, and pair-wise group comparisons failed to show differences between diagnostic groups with respect to frequency of reduced BR echogenicity. TCS findings of this study showed that reduced echogenicity of pontomesencephalic BR is frequent in depressive states, irrespective of diagnostic category. As a result of the present study, the hypothesis that BR echogenicity might distinguish patients with MDD and patients with ADDM had to be rejected. Reduced BR echogenicity is found with

similar frequency in MDDs, MDDr, and ADDM. This is in agreement with results of clinical and neurophysiological studies suggesting common pathophysiological mechanisms in MDD and ADDM [28]. Bipolar affective disorders are characterized by recurrent episodes of depression Lapatinib order as well as mania or hypomania [26]. In histological studies, subtle structural deficits in the dorsal raphe with a regional reduction in the synthesis of noradrenalin have been described in patients with bipolar disorder. The first TCS study evaluated BR alterations in patients with bipolar affective disorders, revealed normal or even increased echogenicity of BR in bipolar disorder, irrespective of the existing disease conditions. This observation led to the assumption that reduced echogenicity of BR may be specific to unipolar depression [3]. Recently, Krogias et al. found

the BR hypoechogenicity in 36.1% of the 36 patients with bipolar I disorder (14 depressed, 8 manic, 14 euthymic) and in 20% of the 35 healthy controls. Compared to the SB-3CT control group, frequency of altered BR echogenicities did not reach statistical significance. Hypoechogenicity of BR was depicted in six (42.9%) of the depressed, in three (37.5%) of the manic and in four (28.6%) of the euthymic bipolar patients, with no significant difference between the three subgroups [29]. The width of third ventricle was significantly larger in the patient group (3.8 ± 2.1 mm vs. 2.7 ± 1.2 mm). Depressed bipolar patients with reduced BR echogenicity showed significantly higher scores on the Hamilton Depression Rating Scale as well as the Montgomery-Åsberg Depression Rating Scale [29]. Relating to echogenicity of SN, a strong trend of more frequent SN hyperechogenicities in the depressed subgroup was identified. Hyperechogenic SN was seen in six patients (16.7%): five (35.7%) of the depressed, in none (0%) of the manic and in one (7.1%) of the euthymic patients, indicating cyclical dysregulation in quantitative dopaminergic transmission as one of the underlying pathologies in the pathogenesis of bipolar disorder.

This framework could account for the strong concreteness effects

This framework could account for the strong concreteness effects observed at the “edges” of the temporal lobe (i.e., STG and PHG) in terms of their relative specialisations for verbal versus visual inputs, while predicting equi-modal activations in the centre (ITG and the vATL). Importantly, this framework assumes graded specialisation within a single functional system in the ATL, rather than an absolute selleck products division into separate processing modules. Finally, we consider C > A activations observed in other areas of the brain. As in previous studies and meta-analyses, we found C > A effects in angular gyrus, posterior cingulate and mid-PHG. As discussed

in the previous section, the activation of PHG most likely reflects retrieval of stored visual characteristics

of concepts, which is only possible for highly imageable, concrete words (D’Esposito et al., 1997 and Sabsevitz et al., 2005). C > A effects in the angular gyrus and posterior cingulate are harder to interpret. The role of posterior cingulate in semantic cognition is unclear, though it has been suggested that it may be involved in the interface between semantic knowledge and episodic memory (Binder et al., 2009). Stronger claims have been made about the function of BI 6727 price angular gyrus. Binder et al. (2009) proposed that the angular gyrus is critically involved in semantic representation Adenylyl cyclase and that concrete regions activate this region strongly because they have more detailed semantic representations. It is therefore interesting that the activation profile of angular gyrus diverged strongly from that of the vATL, for which a similar representational function has been proposed. There were three findings that suggest the function of the angular gyrus is very different to that of vATL. First, the angular gyrus was not activated in the main contrast of semantics over numbers; in fact, the contrast in this region slightly favoured the numbers (see Fig. 3). This suggests that, in addition to any

putative role in semantic processing, the angular gyrus is at least equally involved in the processing of numerical magnitudes. This was not the case for vATL. Second, angular gyrus showed a clear C > A activation pattern, while the activation in the vATL slightly favoured abstract words (though this effect varied elsewhere in the ATL region). Finally, angular gyrus and vATL showed very different activation patterns with respect to rest, with angular gyrus significantly deactivated by the semantic task while vATL, along with other elements of the semantic network, were positively activated. This result is consistent with the status of angular gyrus as a key element of the default mode network. Binder et al., 2009 and Binder et al.

These results also fit well with recent local field potential (LF

These results also fit well with recent local field potential (LFP) recordings in human STN that show activity changes on successful stop trials preceding SSRT [30]. However, for a number of reasons, these single unit recordings in the basal ganglia need to be replicated in non-human primates. First, lesion studies in primates have not in all Selleck ERK inhibitor cases conclusively supported the gating hypothesis [31•]. Second, the STN activity pattern found in the rat is different than activity pattern in primate STN in a

related impulse control task, in which an automatic response had to be overridden to generate a voluntary response 32 and 33]. In particular, the primate STN neurons responded selectively when the automatic response was successfully suppressed and this activity difference predicted behavior. This is hard to reconcile with the unspecific response in the rodent STN neurons. Third, the head movement that has to be controlled by the rat is a very complex movement, involving multiple body parts and a number of different muscles. In contrast, most other stop signal experiments in monkeys have used simpler movements involving just one effector. Because of this, it is not entirely clear at what level of

motor representation the basal ganglia neurons operate that were recorded in the rat. They could either represent the entire movement on a more abstract level or they could represent the control of specific muscles. Lastly, given the mixed results in motor cortex, it is very important Meloxicam to understand the mechanistic relationship and timing between the gating INCB018424 in vitro in the basal ganglia and changes in motor cortex. A final potential

locus of inhibitory control in the skeletomotor system is the spinal cord [34]. Recording in the spinal interneurons of behaving monkeys showed evidence for inhibitory suppression of motor activity during an instructed delay preceding a wrist response [35]. These neurons might be the closest functional equivalent to omnipause cells that has been found so far in the skeletomotor system. It will be very important to understand the input controlling these inhibitory interneurons and to better understand their role in the suppression of motor responses. Cognitive control operates via two distinct operating modes: proactive control and reactive control 36 and 37]. Proactive control relies upon the anticipation and prevention of interference before it occurs, whereas reactive control relies upon the detection and resolution of interference after its onset. Reactive control is recruited as a late correction mechanism and has been the focus of most of the stop signal research. In contrast, proactive control adjusts the response selection and preparation process in anticipation of known task demands. In the context of the stop signal task, proactive control is mostly related to a regulation of the level of excitability of the motor system.

Strains of SP with penicillin MIC’s of <0 06 μg/ml were considere

Strains of SP with penicillin MIC’s of <0.06 μg/ml were considered susceptible, MIC's of >0.1 μg/ml are regarded as non-sensitive. Results were analyzed using Statistica programme. The following indexes were calculated for NP culture in relation to MEF cultures: sensitivity, specificity, positive predictive values (PPV) and negative predictability value (NPV). For three main AOM pathogen S. pneumoniae, H. influenzae and M. catarrhalis plus Str. pyogenes positive NP cultures were obtained in 68 from 123 episodes of AOM (60.1%). MEF were simultaneously positively cultured in 48/123 (39.0%) cases (only one of the AOM pathogens was found in each specimen). Among bacterial pathogens cultured

in NP there were following isolations: 33/69 (47.9%) of S. pneumoniae, 20/69 (28.9%) of H. influenzae, 14/69 (11.4%) of M. catarrhalis and 2/69 of Streptococcus pyogenes. In MEF the buy Sirolimus following bacterial pathogens were found: S. pneumonia in 28/48 (58.3%), H. influenzae in 14/48 (29.1%), M. catarrhalis www.selleckchem.com/products/XL184.html in 4/48 (0.08%). The sterility of nasopharynx and MEF was found in 7/123

and 9/123 respectively. Remaining NP and MEF cultures contained bacterial species considered to be normal nasopharyngeal flora (Str. viridans, Neisseria spec., Klebsiella, Staph. aureus) or as a contamination (Staphylococcus epidermidis, Enterococcus faecalis, Enterobacter cloacae). In 5/123 NP and in 5/123 MEF AOM pathogens were accompanied with bacterial species which can be considered as a contamination. An agreement between NP and MEF cultures was found in 36/123 cases (29.8%), [S. pneumonia in 20/123 (16%), H. influenzae in 12/123 Amisulpride (10%), M. catarrhalis in 4/123 (0.8%), Str. pyogenes 1/123 (0.8%)]. The sensitivity, specificity and both positive and negative predictive values of NP culture for recovering in MEF culture the same AOM pathogen is presented in table I. Total analysis of all positive and negative NP and MEF cultures revealed moderate sensitivity, low specificity, poor PPV and high NPV of NP cultures in relation to MEF culture which is considered as the AOM etiology. The separate analysis for SP, HI and MC showed relatively

high specificity for each species, moderate sensitivity for SP and moderately good sensitivity for HI and MC, PPV was low for SP and HI and very low for MC. Our data demonstrated relatively high rate (56%) of nasopharyngeal colonization in the course of AOM and this rate was higher than the rate of positive MEF cultures (39.9%). In remaining the MEF was sterile or contaminated with skin saprophytic bacteria (Staph. epidermidis and Staph. aureus). In our study all children initially presented signs of viral nasopharyngeal infections and clinical signs suggesting bacterial superinfection and relative indications for tympanocentesis. In nearly 60% bacteria pathogens were not revealed and in fact these cases did not require antibiotic therapy. The potential pathogens of AOM were absent in nasopharynx in 44% cases.

An Annexin V FITC Apoptosis Kit was purchased from Calbiochem Al

An Annexin V FITC Apoptosis Kit was purchased from Calbiochem. All the solvents and other chemicals used were of analytical grade from Gibco™, Invitrogen™, Sigma–Aldrich and Merck. All solutions were prepared with Fluorouracil price water purified by the Milli-Q® system (Millipore). BlL was purified according to the protocol previously described by Nunes et al. (2011). The cell lines used in the cytotoxicity assays were K562 (chronic myelocytic

leukemia), NCI-H292 (human lung mucoepidermoid carcinoma cells) and Hep-2 (human larynx epidermoid carcinoma cells) obtained from the Instituto Adolfo Lutz (São Paulo, Brazil). The non-tumorigenic cell line (HaCaT), derived from human keratinocytes was purchased from Cell Line Service (CLS, Heidelberg, Germany). The cells were maintained in DMEM supplemented with 10% fetal bovine serum, 2 mM glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin and maintained at 37 °C with 5% CO2. Cytotoxicity of BlL was tested in tumor cell lines (K562, NCI-H292 and Hep-2) and in non-tumorigenic cell line (HaCaT). PFT�� mouse The cells (105 cells/mL for adherent cells or 0.3 × 106 cells/mL for suspended cells) were plated in 96-well microtiter plates and after 24 h, BlL (0.07–50 μg/mL) dissolved in DMSO was added to each well and incubated for 72 h at 37 °C. Then, MTT (5.0 mg/mL) was

added to the plate and growth of tumor cells was estimated by the ability of living cells to reduce the yellow tetrazolium to a blue formazan

product (Mosmann, HSP90 1983; Alley et al., 1988). Negative control groups received only DMSO; etoposide (1.25–20 μg/mL) was used as a positive control. After 3 h (for suspend cells) or 2 h (for adherent cells), the formazan product was dissolved in DMSO and absorbance was measured using a multi-plate reader (Multiplate Reader Thermoplate). The BlL effect was quantified as the percentage of control absorbance of reduced dye at 450 nm. The K562 suspension (0.3 × 106 cells/mL) was seeded in 96-well microtiter plates and incubated at 37 °C at 5% CO2 for 24 h; after this period, BlL at IC50 was added. After 48 h the cells were stained with annexin V and propidium iodide using Annexin V–FITC Kit (Calbiochem®) following the protocol provided by the manufacturer and analyzed by an epifluorescence microscope (Carl Zeiss, Gottingen, Germany) at 1000× magnification under oil immersion with filters for LP 515 nm emission and BP 450–490 nm for excitement. A minimum of 200 cells was counted in every sample. Mitochondrial depolarization was evaluated by incorporation of JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide), a fluorescent lipophilic cationic probe (Kang et al., 2002; Guthrie and Welch, 2006). The probe JC-1 is freely permeable to cells and undergoes reversible transformation from a monomer to an aggregate form (Jagg). K562 suspension (0.