The adjuvant effect of including CaP in PCMCs was confirmed for both antigens ( Table 1). This was particularly marked for the anti-CyaA* response as only one mouse in the 0% CaP group produced a detectable anti-CyaA* IgG titre at each time point investigated. Increasing the CaP content did not significantly further inhibitors increase the antigen-specific IgG titres or alter the duration of antibody response. The attempted prime-boost check details formulation failed to enhance immunogenicity compared to other CaP PCMC formulations. J774.2 cells were incubated with equal amounts of either soluble BSA-FITC or BSA-FITC formulated

as 0% or 8% CaP PCMCs. Uptake of fluorescent antigen was visualised by confocal laser-scanning microscopy (Fig. 5, panels A–C) and quantified by flow cytometry (panels D–F). Confocal microscopy showed that soluble BSA-FITC was poorly phagocytosed, with J774.2 cells containing low levels of fluorescence (Fig. 5A). In contrast, loading BSA-FITC onto PCMCs increased phagocytosis, with cells displaying punctate regions of green fluorescence (Fig. 5B) and this was further enhanced with CaP PCMCs (Fig. 5C). These observations were confirmed by flow cytometry. The P2 daughter population was derived

from the parent population P1. The increase in MFI of the P2-gated population of the cells upon exposure buy LY2109761 to BSA-FITC PCMCs (Fig. 5E) and the further increase in the presence of CaP-modified PCMCs (Fig. 5F) indicates a greater phagocytosis of these particles compared to soluble BSA-FITC (Fig. 5D). These results, in combination with published data, demonstrate that PCMC formulations are suitable for vaccine applications and may address problems associated with current vaccines. Moreover, CaP PCMCs were shown to be immunogenic and to promote a more

until mixed Th1/Th2 response in comparison to traditional formulations and to soluble PCMCs [5] and [7]. Modification of the surface of PCMC with an outer layer of CaP altered the particle morphology from planar discs to rod-like structures and significantly decreased the rate of antigen release in vitro. PCMCs without CaP released antigen almost immediately in aqueous buffers whereas increasing the CaP loading progressively decreased the rate of antigen release. This is consistent with release being controlled by dissolution of an outer layer of CaP, the thickness of which is expected to increase with CaP loading. This suggests that CaP PCMCs would potentially show enhanced immunogenicity due to a depot effect in vivo as has been proposed for other adjuvants [2] and [15]. Surprisingly, mice immunised with DT formulated into soluble PCMCs showed enhanced immunogenicity compared to soluble DT antigen. The in vitro solubility data indicated that this enhanced immunogenicity was not due to a depot effect.

Platelet count was determined using an automated cell counter, and prothrombin time and partial thromboplastin kaolin time (PTTK) were measured by a coagulometer. A bleeding time of 2–7 min, a clotting time of 4–9 min, a platelet count of 1.5–4 lacs/mm3,

a prothrombin time of 11–16 s (as per the control) and a PTTK of 30–40 s were considered normal. The data obtained were analysed using descriptive statistics and paired Student’s t-test to compare the results from baseline. Statistical analysis This was a preliminary, exploratory study to assess the effects of escitalopram and fluoxetine on coagulation profile Inhibitors,research,lifescience,medical in patients with major depression. Patient data were analysed on an intent-to-treat basis in Excel. Nominal data (e.g. sex) were expressed as number and percentage, and continuous data (e.g. age and coagulation parameters) as mean and standard deviation (SD). Paired Student’s t-test was used for a within-group Inhibitors,research,lifescience,medical (pre versus post) comparison. A p value of 0.05 or less was considered statistically significant. Results In both study groups, eight (40%) patients were men and 12 (60%) were women. The average age was 32 ± 10.89 years in the escitalopram group and 31.95 ± 9.45 years in the fluoxetine group. The coagulation Inhibitors,research,lifescience,medical profile for patients receiving escitalopram is given in Table 1. In the escitalopram

group, no significant differences in coagulation parameters were observed when compared with baseline. Table 1. Coagulation profile for patients receiving escitalopram. The coagulation profile for patients receiving fluoxetine is given in Table 2. In this group, there was a significant increase in the bleeding time after 3 months of treatment compared with baseline, but this was not beyond the normal range as seen in the table. For the other parameters Inhibitors,research,lifescience,medical – clotting time, platelet count, prothrombin

time and PTTK – no significant differences were observed. Table 2. Coagulation profile for patients receiving fluoxetine. Discussion Life-time risk of major Selleckchem IPI 145 depression is 5–10% and is twice as common in women compared with men [Baldessani et al. 2006]. SSRIs are one of the most widely used drugs for the treatment of Inhibitors,research,lifescience,medical depression. They are well tolerated and have fewer side effects than older tricyclic antidepressants and are thus preferred [Rang et al. 2007]. A recent increased incidence of epistaxis and ecchymosis with SSRI use has been reported, probably because of impairment of platelet function. Serotonin is one of the Megestrol Acetate mediators released during platelet release reaction, causing platelet aggregation. In one study, five children aged between 8 and 15 years developed bruising or epistaxis 1 week to 3 months after starting SSRI treatment. It is possible that the impact of SSRIs on platelet function are causing these effects or a separate coagulopathy exists in these patients [Lake et al., 2000]. Gastric blood loss due to NSAIDs can be increased by SSRIs [Dalton et al. 2003, 2006; Weinreib et al.

Astrocytes play a crucial role in the CNS, supporting normal neuronal activity by maintaining CNS homeostasis and controlling the concentrations of neurotransmitters and ions in the extracellular space (Vernadakis 1988; Wang and Bordey 2008; Belanger and Magistretti 2009). Ethanol regulation of the heat shock cascade and gene expression in astrocytes, therefore, Inhibitors,research,lifescience,medical may have profound implications for neuronal physiology. While there has been no work directly addressing this issue, several studies have shown that HSPs are involved in protecting the brain

from a variety of insults, including ischemia and neurodegeneration (Yenari 2002). In particular, it was found that overexpression of HSP72 in astrocytes prior to ischemia prevented astrocytic glutamate transporter dysfunction

and subsequent neuronal death in the CA1 region of the hippocampus (Xu et al. 2010). These findings suggest that ethanol activation of the heat shock cascade and induction of the Hsp Inhibitors,research,lifescience,medical genes in astrocytes may actually protect nearby neurons from any deleterious effects of alcohol exposure, as well as from future insults. Future studies will Inhibitors,research,lifescience,medical investigate these secondary effects of alcohol on neurons in order to identify changes in astrocytic gene expression and pathways that may be associated with the neuroprotective effects of alcohol. Acknowledgments We thank Heather Durham (McGill Inhibitors,research,lifescience,medical University, Montreal, Quebec, Canada) for providing the Hsf1 constructs and Richard Voellmy (University of Miami, Miami, FL) for permission to use them. This work was supported by National Institutes of Health (NIH)/NIAAA grants (R21 AA018783) to N. L. H. Conflict of Interest None declared. Supporting Information Additional Supporting Information

may be found in the online version of this article: Figure S1. Primary cell cultures Inhibitors,research,lifescience,medical from embryonic mouse PD98059 research buy cortex result in almost pure astrocyte populations. Figure S2. Normalization with two different housekeeping genes does not affect ethanol-induced expression of Hsp70 in primary astrocyte culture. Figure S3. Efficiency of transfection of DNA constructs in primary culture of mouse astrocytes. Click here to view.(1.6M, others docx) Table S1. Genes significantly activated by acute ethanol (E, 60 mmol/L, 1 h) and heat shock (HS, 42°C, 1 h) in primary cultures of astrocytes. Click here to view.(109K, docx)
Pain is defined as an unpleasant sensory and emotional experience, associated with real or potential tissue damage (Merskey and Bogduk 1994) and including both physical (i.e., nociception which means the detection of pain-producing stimuli by primary sensory neurons) and affective aspects (i.e. suffering) (Kupers et al. 2005).

Footnotes Conflict of interest: No potential conflict of interest relevant to this article was reported.
The medical field mostly relies on chemicals to treat illness, but since neurons use electrical signaling, electrical currents can alter their activity—a phenomenon increasingly exploited to treat neurological disorders. The first evidence for the use of electrical stimulation to treat chronic pain comes from antiquity, in the Compositiones Medicam entorum, the early guide to drugs and recipes written in 47 CE by Scribonius Largus, the court

physician of the Roman emperor check details Claudius.2 Inhibitors,research,lifescience,medical He described using electrical currents to treat headaches and gout by applying electric torpedo fish to the painful regions. Inhibitors,research,lifescience,medical This treatment was popular for seizures, depression, and pain until the eighteenth century. Electricity-based therapies later multiplied, based on the work of Luigi Galvani, Charles Le Roy, Duchenne de Boulogne, Beard and Rockwell, and others.3 Obviously, not all such treatments were well-grounded. Electrical stimulation was also applied to treat refractory chronic pain, with deep brain stimulation (DBS) as the first modern method. In DBS, small electrodes are surgically implanted in precise brain locations to deliver tiny electrical currents to neurons immediately adjacent to the electrode. Inhibitors,research,lifescience,medical Thus, unlike

with medications, there are no distant adverse effects (e.g. rashes, gastrointestinal upset, allergies). Since only nearby neurons are affected, most brain functions continue unperturbed. A battery is implanted subcutaneously to power the electrode using technology based on cardiac pacemakers. Inhibitors,research,lifescience,medical A 1960 Inhibitors,research,lifescience,medical article by Heath and Mickle reported that DBS applied to the septum between the lateral ventricles of the brain produced immediate pain relief in a series

of six patients with intractable pain, results duplicated by other early studies.4–6 In 1977, Richardson and Akil reported analgesic efficacy of DBS of the periaqueductal and periventricular gray matter.7,8 Stimulation of another deep target involved in pain sensation, the periventricular gray matter of the posterior Non-specific serine/threonine protein kinase thalamus, brought good pain relief to patients with cancer pain.9 Despite these encouraging results, high costs and rates of complication have limited DBS use; 3.9% of patients developed permanent neurological deficits, thalamic hemorrhage, or death, while 19.1% of patients had temporary complications, including neurological deficits, infection, and hardware malfunction.10 Epidural brain stimulation then emerged as a less invasive alternative. Here the electrodes are implanted under the skull, but outside the dura, so the brain itself is not disturbed and the risk is lower, although only superficial areas of the brain can be reached.

Another study showed that anesthesiology rotation

is an essential part of EM training and that the optimal duration is approximately 6 months, preferably in the first 2 years of advanced training [6]. The duration of the rotation on acquisition of airway management skills is probably not long enough and should be extended. In the future it is also important to study the find more effects of increased length and quality of training programs for EPs on the success and failure rates for airway management carefully in the ED. It should also be remembered that the acquisition Inhibitors,research,lifescience,medical of anesthesiology-related knowledge and skills is not confined to an anesthesiology rotation. In fact, many of these may be learned more effectively in other settings. For example, knowledge of the pharmacology related to this area is Inhibitors,research,lifescience,medical an important part of the primary examination curriculum. Intubation of trauma and unstable patients may be best learned in EDs under appropriate supervision, because these patients differ from those encountered in elective anesthetic practice

[6]. It is widely accepted that orotracheal intubation in the emergency room is significantly more hazardous and is more frequently associated with an increased rate of Inhibitors,research,lifescience,medical difficult intubation and failed intubation than in the operating room [13]. Maintenance of skills in emergency airway management is also currently a subject of considerable debate. There is no objective scientific data to support a minimum requirement of Inhibitors,research,lifescience,medical numbers of emergency or rapid sequence intubations (RSI) to be performed by EPs (or indeed anesthetists) to maintain competency in this area. The figures from the Trauma Audit Research Network suggest a maximum of approximately 6 RSIs per month and per department (based Inhibitors,research,lifescience,medical on an average of 68,000 patients per year) which if staffed by 4 consultants in EM equates to 1-2 RSIs per consultant per month. The Scottish Trauma Audit Group data also suggest that the individual consultant in EM is likely to be involved in

unless 2-3 RSIs per month [7]. Due to the presence of a large number of patients at our ED (nearly 6000 patients per month); the individual skills maintenance will not be difficult for the practicing EMRs. In the US, several studies have reported on the requirements and experience of their residents in Emergency Medicine. Although accepting that there are always variations between people and it is difficult to be clear on exact numbers, it would seem that novice anesthesiology residents require 80 or more intubations to achieve reasonably consistent skills in orotracheal intubation [7]. Hayden and Panacek reported that the mean number of intubations per trainee was 75 (95% confidence intervals 62 to 87); this is over a three-year residency in emergency medicine in a US setting [14].

In SULT1E1 knock-out mice, loss of SULT1E1 causing an excess of estrogens leads to the formation of smaller patches from white fat and insulin resistance [97]. In type 2 diabetes, induction of hepatic SULT1E1 is most

frequently observed. Loss of SULT1E1 improves the metabolic function in a female mouse model of type 2 diabetes, restores insulin sensitivity, and blocks hepatic gluconeogenesis and lipogenesis [98]. Since in diabetes, upregulation of SULT1E1 decreases E2 levels, inactivation of the enzyme will prevent loss of estrogens and normalize estrogenic activity in the liver. Inhibitors,research,lifescience,medical This beneficial effects of SULT1E1 inactivation were absent in ovariectomized mice. These effects were also sex specific, as SULT1E1 loss in males worsened the diabetic phenotype and led to a decreased islet beta-cell mass, failure Inhibitors,research,lifescience,medical of glucose-stimulated insulin secretion, increased macrophage infiltration, and inflammation in white adipose tissue. The authors suggest that inhibition of SULT1E1 at least in females may represent a novel approach in the therapy of type 2 diabetes [98, 99]. However, it has to be considered that type 2 diabetes mostly occurs in women after the menopause when local formation of steroid hormones from adrenal precursors becomes important. Since extragonadal Inhibitors,research,lifescience,medical estrogen production is typical for primates [2], the benefit of increasing levels of active estrogens

by reducing SULT1E1 may have to be studied in a proper model for type 2 diabetes in this group. In any case, higher estrogen levels are thought to have beneficial effects on type 2 diabetes, but the risk of the induction of hormone-sensitive cancers may be considered as well. 5. Steroid Sulfatase Inhibitors as Agents for a Therapy of Hormone-Sensitive Tumors Inhibitors,research,lifescience,medical Hormone therapy is used to treat both early and advanced breast Inhibitors,research,lifescience,medical cancer and to prevent breast cancer in women who are at high-risk of developing the

disease. Lonafarnib order Currently, the most widely used therapies for the treatment of hormone-dependent cancer is to block the action of steroid hormones. Adjuvant endocrine therapy with the selective estrogen receptor modulator (SERM) tamoxifen is recommended for premenopausal women with a history of atypical hyperplasia to reduce breast cancer risk. Raloxifene, another selective PAK6 estrogen receptor modulator, was found to be equivalent to tamoxifen in reducing the risk of developing invasive breast cancer in postmenopausal women. However, it did not provide the same level of risk reduction for developing noninvasive breast cancer. Aromatase inhibitors, which block the conversion of androstenedione to estrone, are applied in postmenopausal women. Currently, third-generation aromatase inhibitors, which comprise the nonsteroidal compounds anastrozole and letrozole, and the steroidal exemestane are finding widespread application in the clinic (for reviews see [100, 101]).

Structural MRI Structural MRI is increasingly accepted as a surrogate for anatomic phenotype in neuroscience research. In many areas, anatomic MRI has replaced the need for analysis of the postmortem brain in order to elucidate relationships between structure and function. It is not hard to find examples in which anatomic MRI has transformed the entire research landscape of a field: cerebrovascular disease, epilepsy, multiple sclerosis, and other inflammatory conditions, cerebral developmental disorders, to some extent psychiatric disorders, and neurodegenerative disorders. Animal model studies at high magnetic fields have made unique contributions to

this development. Morphological images of brain tissue rely largely on proton Inhibitors,research,lifescience,medical density, T1 and T2 differences between tissue types (eg, white matter vs. gray matter, cortex vs. subcortical nuclei etc.). Proton density is clearly a magnetic field independent parameter. However, relaxation times T1 and T2 are field dependent, Inhibitors,research,lifescience,medical generally increasing63,64 and decreasing1,9,65,66 respectively, with higher magnetic fields Inhibitors,research,lifescience,medical (see review in ref 67). It was recently shown that, contrary to expectations, the Kinase Inhibitor Library screening dispersion in T1 increases

with increasing magnetic fields in the brain, leading to superior T1-weighted structural Images at the higher magnetic fields.68 Lengthening of T1 with increasing magnetic field also holds true Inhibitors,research,lifescience,medical for blood. Blood T1 is virtually insensitive to its oxygenation state. Ex vivo measurements have shown that blood T1 varies linearly with field strength going from 1.5 T to 9.4 T according to T1= 1.226+ 0.134B0.69 This imparts a clear benefit in time-of-flight type vascular imaging, as well as perfusion imaging using spin labeling techniques. Mapping signals in all hemodynamic-based functional imaging

methods, such as fMRI and optical Inhibitors,research,lifescience,medical imaging with intrinsic signals, are mediated through the vasculature. Consequently, vascular components in these methods are of utmost significance in determining the ultimate spatial and temporal accuracy of the neural activity maps produced by these methods. Therefore, it is important to be able to image vasculature in great detail, and ideally together with functional data in order Parvulin to understand more precisely the source of the fMRI signals and their spatial correlation with the volume of altered neuronal activity. Vascular imaging with high resolution is also of paramount importance in other fields of biomedical research such as tumor biology, where angiogenesis is a necessary component of tumor growth. Taking advantage of the gains in SNR and longer T1 values, the feasibility of obtaining high-resolution MR images of intracortlcal vessels was demonstrated in the cat brain.70 This accomplishment relied on a combination of time-of -flight MR angiography and T2*-weighted contrast based on both endogenous BOLD effect and exogenous iron-oxide particles.

e., “red” printed in red ink) and EGFR inhibitor incongruent stimuli (i.e., “green” in red ink) and measures interference between cognitive processes by requiring the participant to name the color (“red”) regardless of the word (“red” or “green”; Stroop 1935). It is hypothesized that the slower the speed of color-naming during incongruent stimuli, the more important the cognitive interference component. Consequently, in the Drug Stroop, the slower the speed of color-naming during stimuli associated with drug cues, the stronger the attentional bias toward

the drug-related stimuli (Cox et al. 2006). Inhibitors,research,lifescience,medical For example, 24-h abstinent smokers showed higher attentional bias for smoking cues than current smokers (Waters and Feyerabend 2000). The Dot Probe task also measures attentional bias toward drug-related stimuli. Here, two stimuli (one drug-related and one neutral) are presented side by side, after which Inhibitors,research,lifescience,medical the images disappear and a dot appears for a short time. Fast responding toward the dot where a drug-related stimulus was previously shown is a measure for increased attentional bias. Smokers showed Inhibitors,research,lifescience,medical greater attentional bias toward smoking-cues than nonsmokers during a Dot Probe task (Ehrman et al. 2002). In addition, compared with current smokers, 12-h abstinent smokers showed increased attentional bias for smoking cues (Gross et al. 1993),

and ex-smokers showed an intermediate level of attentional bias compared with current smokers and nonsmokers measured with the Dot Probe task (Ehrman et al. 2002). Using a related measure, abstinent crack-cocaine dependent patients had faster eye-movements toward cocaine-related pictures Inhibitors,research,lifescience,medical as compared to neutral pictures, Inhibitors,research,lifescience,medical and this correlated with self-reported intensity of cocaine craving (Rosse et al. 1997). It should

be noted that the drug Stroop and the Dot Probe task both measure selective attention (i.e., to drug stimuli), but the Stroop task requires more cognitive effort and flexibility, which might be responsible for different findings when using these different Farnesyltransferase paradigms. Cue-reactivity is also an import aspect of drug addiction and refers to the physiological and related subjective reactions (craving) that occur in the presence of drug-related stimuli, and can ultimately lead to relapse. Cue-reactivity is generally investigated using a cue-exposure or cue-reactivity task. Unlike other neurocognitive tasks, cue-reactivity paradigms employed during functional imaging only require the participant to watch drug-related pictures or videos (without any cognitive effort), although some cue-reactivity tasks include easy binary tasks to control for attention differences, in which baseline trials are usually incorporated requiring similar motor responses.

Sprague-Dawley rats were given a single intravenous administration of crosslinked daunorubicin

micelle, uncrosslinked daunorubicin micelle, or free daunorubicin at a 10mg/kg dose. Plasma … Figure 6 Pharmacokinetics of crosslinked BB4007431 micelles in rats. Sprague-Dawley rats were given a single intravenous administration of crosslinked BB4007431 micelle, or free BB4007431 at a 25mg/kg dose. Plasma was analyzed for BB4007431 concentration … 4. Discussion Improving stability of therapeutic molecules is a well-established aim in the field Inhibitors,research,lifescience,medical of drug delivery. An ideal drug-loaded nanoparticle would be stable to dilution in biological media, possess stealth-like properties to avoid uptake by the RES, and release the drug only in the area of diseased tissue. The data presented in this paper describe a versatile polymer micelle drug delivery system that has been engineered to efficiently Inhibitors,research,lifescience,medical encapsulate a wide variety of hydrophobic drugs. In addition, the stabilization technology

built-in to the micelle is dependent on pH, such that the micelle is stable at physiological pH, and unstable at low pH, thus providing a mechanism to release the drug in the tumor microenvironment or in endosomes, which are both slightly acidic environments. A vast number of drugs exist today that possess potent Inhibitors,research,lifescience,medical anticancer activity; however, many of them are unable to be utilized in the clinic due to their inability to be dissolved in aqueous solutions [27]. Some hydrophobic drugs can be solubilized with excipients; however, such vehicles have been shown to cause toxicity to the patient [28]. Inhibitors,research,lifescience,medical The core block of the triblock copolymer (poly(D-leucine-co-tyrosine)) was rationally designed and chosen to encapsulate

hydrophobic molecules. A key factor leading to the versatility arises from the use of both D and L stereoisomers of amino acids in the core block, which disrupts the secondary structure of the polypeptide. Replacing the rod-like helical nature of the polypeptide Inhibitors,research,lifescience,medical with the flexibility of a random coil allows for significant increases in drug loading efficiency. The ability of drugs to be encapsulated within the triblock copolymer was the related to its LogP value, such that only hydrophobic drugs could be encapsulated. This result is logical as hydrophilic molecules would prefer to associate with the hydrophilic part of the polymer versus the hydrophobic core, leading to inefficient drug encapsulation. Crosslinking was performed using metal acetate chemistry, specifically, iron (II) chloride. The crosslinking dialysis assay determined that 40–90% of the drug remained in the crosslinked micelle after six hours. Typically, 10% of the drug or less was VE-821 mw retained in uncrosslinked micelles examined using the same crosslinking dialysis assay. Although there was a correlation between LogP and encapsulation ability, there was no clear correlation between LogP and the crosslinking retention or the particle size.

31 Several drugs are known to delay repolarization and to be associated with a prolongation of QT interval and possibly torsade de pointes: tricyclic antidepressants, antipsychotics (thioridazine, chlorpromazinc), antihistamines (terfcnadine, astemizolc), antiinfectives (erythromycin, chloroquine, halofantrine), and miscellaneous drugs (cisapride, terodiline, furosemide,

prednisolone, and beta-agonists).32-34 Prolongation of cardiac repolarization is easily identified using ECG. Increased QT intervals in a patient, are indicative of prolonged cardiac repolarization. However, because the QT interval is dependent on heart, rate, it has to be corrected into a new variable independent of heart Inhibitors,research,lifescience,medical rate, called the corrected QT interval (QTc). Various equations have been Inhibitors,research,lifescience,medical proposed for this. The most widely used is Bazett’s formula (QTc = QT/√RR). This formula gives an excellent correction for a heart rate value of 60 bpm. However, it overestimates (undercorrects) QTc at

low heart rate and underestimates (overcorrects) QTc at high heart rate values.35 Fridericia’s formula (QTc = QT/3√RR) seems to have better predictive properties than Bazett’s formula.36-38 In 1997, the European Agency for the Evaluation of Medicinal products (EMEA) proposed a “points to consider” document for the assessment of the potential for QT interval prolongation by a Inhibitors,research,lifescience,medical A-1210477 purchase noncardiovascular medicinal product.39 Several papers in the literature also emphasize the need to assess cardiac repolarization.40,41 During phase 1, ECGs are collected from healthy, normal subjects, Inhibitors,research,lifescience,medical usually males, several times before, during, and after drug administration. The potential for QT interval prolongation of a noncardiovascular NCE should be assessed in a randomized, double-blind, Inhibitors,research,lifescience,medical placebo-controlled study, and with a sufficient number of doses to be able to characterize

the dose-response relationship, including doses sufficiently higher than the proposed therapeutic dose to demonstrate a no-effect outcome. The time course of ECG effects should be evaluated according to the pharmacokinetic profile of the parent compound, as well as its active Calpain (toxic) metabolites if appropriate, after a single dose as well as at steady-state plasma concentrations. This timing should coincide with the expected Cmax of the NCE or when the maximum concentration in the target cardiac cell is expected. The EMEA document also emphasized that, at present, automatic readings from 12-lead ECGs are generally not considered sufficiently accurate and reliable. Holter may be useful to assess the occurrence of arrhythmia, but this is also inaccurate and not reliable enough for QTc readings, as it. does not. correlate sufficiently well with 12-lead ECG recorded at, the same time. Therefore, manual reading of QT intervals by trained personnel is recommended.