In some cases, such as in Rwanda, no expansion was deemed necessary. In other countries national-level interviewees reported that there had been an expansion or modernisation of the cold chain in preparation for the introduction, although this was generally at the national and sub-national levels, rather selleck than in facilities. There was a discrepancy between some national- and facility-level

responses, with the former reporting cold chain expansion whilst the latter reported none. It is not clear whether this discrepancy was Libraries because expected expansions had not occurred, or whether facility staff had not realised that new equipment received (sometimes up to a year earlier) was for a particular vaccine introduction. In four countries, the presentation of other vaccines had changed (pentavalent in Cameroon, Kenya and Mali, and PCV in Rwanda), which reduced their cold chain requirement, making capacity available for the new vaccine. Finally, some districts and a minority of facilities reported using adaptive strategies, such as more frequent vaccine deliveries, in order to manage their cold chain space. “There is a problem with the cold chain because the volume [of vaccines] is bigger and districts

are struggling with the cold chain… there is no space. They selleck inhibitor [the health centres] have to take small quantities; we send them the remainder when there is an opportunity. This creates a risk of stock outs Guatemala was an exception in that no assessment was conducted before the introduction and there was no nationally-organised cold chain expansion. Some equipment was reported to have been procured at sub-national levels after the introduction. Interviewees in most countries reported no effect on regulatory policies, with some exceptions. In Kenya, WHO worked to strengthen the country’s Pharmacy and Poisons Board in order to register the new vaccine. It was felt that this would be beneficial for future vaccines. In Mali, the national regulatory process was bypassed for both Men A and PCV vaccines. In Idoxuridine doing so, some interviewees argued that this weakened national ownership and

domestic regulatory processes. In most countries the new vaccines were not thought to have affected the functioning of their ICCs. However, in Mali (for Men A) and in Rwanda, membership of the committees was extended to additional stakeholders. In Ethiopia some interviewees felt that the ICC had been strengthened by the introduction, particularly because of highly active thematic sub-committees. Vaccination is, in general, well accepted and this was the case for the new vaccines too, with high acceptance and demand reported. Only a minority of facilities reported that they had experienced any resistance from the community regarding the new vaccine – this was most common in Rwanda for the HPV vaccine, or because of a fear of the effect of receiving two vaccinations at once (e.g. in Ethiopia, where PCV and pentavalent were given at the same time).

The reasons for this are possibly the large inter-individual blood concentration levels of antidepressants in general [Hiemke et al. 2011]. During pregnancy, there are changes in pharmacokinetic parameters such as increased plasma

volume, decreased concentration of plasma albumin, changes in hepatic metabolism of some drugs (induction of CYP2D6 and CYP3A4) and gastrointestinal changes in absorbance [Ververs et al. Inhibitors,research,lifescience,medical 2009; Freeman et al. 2008; Altshuler and Hendrick, 1996; Sit et al. 2008]. Another problem is the interpretation of blood levels against clinical efficacy, because clear relationships are lacking, especially for selective serotonin reuptake inhibitors (SSRIs) [Rasmussen and Brosen, 2000]. Therefore, in this population, blood level concentration as a method to measure adherence does not seem to be a good alternative for MEMS. Despite the possibility of predictors for poor adherence in our data with logistic regression analyses, our results are in line with Inhibitors,research,lifescience,medical ten Doesschate and colleagues [ten Doesschate et al. 2009]. The strength of our study is that we compared different methods in a specific population. We found that pill count was a good alternative to measure the adherence in our population and that this method

can be easily implemented into daily practice by the community pharmacist or specialized pharmacists in our situation. Our study has some limitations. First, women were aware of the MEMS cap Inhibitors,research,lifescience,medical function and needed to be instructed, which may have influenced the adherence rate in a positive way; however, from other studies we know that this method has no impact on adherence Inhibitors,research,lifescience,medical over a longer period [Hugen et al. 2002; Kastrissios et al. 1998]. Second, our study population was small. Despite the high number of patients visiting our clinic, we could not include most women because the time of inclusion was set at less than 3 months of pregnancy. They had their first visit at the PCS professional within the second or third trimester of pregnancy. Third, in our setting the PCS team delivers high standards of care, which may not be representative of other clinics. It is possible that high standard of care and the frequent from Inhibitors,research,lifescience,medical visits (see Table

2) leads to higher adherence in general. In a pilot experiment, using Medication Possession Ratio we found a lower number of adherent women, where adherence decreased from 62% to 46% during selleckchem pregnancy [home-PW, 2012]. These results are in contrast to the findings presented here. In our earlier retrospective study, women were not aware of participation at time of inclusion. It could be that participating in a trial leads to higher adherence [van Onzenoort et al. 2011]. This has to be further evaluated in larger studies. Conclusion Adherence to antidepressant use during pregnancy using MEMS is 86%. Compared with MEMS, pill counts show good agreement. Therefore, this method may serve as a good alternative that can be easily implemented into daily practice.

152 IL-6 may be involved in the modulation of the HPA axis.153 Activation of the HPA axis is one of the best-documented changes in MD.154 Furthermore, the relationship between psychological or physical stress and an enhanced IL-6 secretion in the peripheral immune system seems to be well established.155-158 Impaired stress coping Inhibitors,research,lifescience,medical is often observed in depressed patients. Thus, the high number of reports of elevated peripheral IL-6 levels in MD patients may be related to psychological stress. On the other hand, there is evidence for a relationship between high peripheral IL-6 levels and elevated CNS 5-HT availability.

IV or IP administration of IL-6 induced not only an activation of the HPA axis, but also an increase in brain tryptophan and 5-HT metabolism, whereas the norepinephrine metabolism was unaffected.113

Accordingly, IL-6 seems to mediate the activation of the HPA axis and the 5-HT CNS after administration of the endotoxin LPS.112 Thus, elevated plasma levels of IL-6 do not fit with the hypothesis of a Inhibitors,research,lifescience,medical 5-HT deficiency in MD. Rather, it should be recognized that an inherent heterogeneity exists in the etiology of depression and different neurotransmitter systems may be disturbed. On the basis of the commonly accepted Inhibitors,research,lifescience,medical idea that MD may be a heterogeneous group of disease entities, the group of Arolt and Rothermundt www.selleckchem.com/products/chir-99021-ct99021-hcl.html investigated the difference between melancholic and nonmelancholic MD regarding their cytokine expression patterns.159 They detected profound differences between these diagnostic subgroups: nonmelancholic patients showed increased counts of leukocytes, lymphocytes, and natural killer (NK) cells in the acute stage of

disease Inhibitors,research,lifescience,medical and after 2 and 4 weeks of treatment. However, their in vitro production of the cytokines IL-2, IL-10, and IFN-γ was unchanged compared with that of healthy controls. Melancholic patients on the other hand demonstrated normal Inhibitors,research,lifescience,medical cell counts, but a decreased in vitro production of IL-2, IFN-γ, and IL-10 during the acute stage of disease. Following clinical improvement, cytokine production patterns normalized in these patients. Schizophrenia A pathophysiological role of cytokines is also discussed in the other major psychiatric disorder, schizophrenia. The reports of the psychotic symptoms inducing effects of IL-2 in cancer patients attracted attention Mephenoxalone of this Th1-like cytokine to the immunopsychiatric schizophrenia research. Early studies reported elevated IL-2 levels in cerebrospinal fluid of schizophrenia patients,160,161 but others failed to replicate these intriguing findings.162-165 IL-2 levels in serum were reported to be either increased166 or decreased.167 A significant decrease in the production of IL-2 by peripheral lymphocytes is one of the best-replicated immunological findings in schizophrenia.

In another

study in cocaine abusers, Wexler et al. (2001) found higher ACC activity both preceding and following the onset of craving while watching a cocaine video, but not when watching happy and sad video tapes, compared to HCs. In addition, cocaine selleck compound abusers showed lower activation in various prefrontal and temporal areas compared with HCs during the cocaine-cue video. In contrast to Childress et al. (1999), the authors concluded that there was a fundamental neurobiological difference between craving and normal emotional states, most probably due to an imbalance between Inhibitors,research,lifescience,medical limbic and prefrontal cortical activity. During craving, cocaine-dependent subjects showed greater activity than HCs in regions that were found to be active in HCs when viewing sad video tapes compared to happy tapes, suggesting a physiological link between

cocaine cue-responses Inhibitors,research,lifescience,medical and normal dysphoric states rather than normal euphoric states (Wexler et al. 2001). In smokers, greater activation was found after exposure to smoking-related images compared with neutral images in several limbic brain regions (part of the mesocorticolimbic dopamine (DA) reward pathway), as well as in regions part of the visuospatial attention circuitry, compared to HCs (Due et al. 2002). The authors suggest that the reward Inhibitors,research,lifescience,medical and visuospatial attention circuitry act in concert to increase and direct attention to potentially important stimuli, such as smoking stimuli Inhibitors,research,lifescience,medical in deprived smokers (Due et al. 2002). This study thus replicated findings of increased limbic activation during processing of cocaine cues. However,

in comparison to the previous studies performed during craving in abstinent cocaine-dependent individuals, the findings from this study may additionally reflect the effects of craving during acute (nicotine) withdrawal, which might be Inhibitors,research,lifescience,medical different from the effects of craving during long-term abstinence. David et al. (2005) failed to observe significant differences in overall brain activation in a small study with smokers, suggesting Thiamine-diphosphate kinase that the absence of whole-brain group differences was due to wide inter-individual variability in magnitude and location of activation, indicating the need for larger sample sizes. In a secondary ROI-analysis, greater ventral striatum/nucleus accumbens (VS/NcA) activation was in smokers, but, however, no correlation was found between NcA activation and self-reported craving, which might be due to a ceiling effect due to nicotine withdrawal during the study (David et al. 2007). Also, Okuyemi et al. (2006) found significant group (smokers vs. HCs) by condition (smoking vs. neutral) interaction effects in medial PFC, right lateral OFC, and bilateral VLPFC activation.

Nonpharinacological approaches to Alzheimer’s disease As emphasized by Reichman,147 pharmacological approaches can be combined with behavioral and environmental interventions that assist patients

in maintaining the highest, possible level of function. Patients in the early stages of dementia may benefit from support groups and other constructive environments that provide information and feedback on the cognitive and behavioral symptoms. Attempts to improve cognitive function Inhibitors,research,lifescience,medical in AD patients through reality orientation, reminiscence, and memory retraining have had some limited success.148 Reality orientation was developed primarily to reduce confusion and disorientation in dementia patients in institutionalized settings. A key feature of reality orientation is Inhibitors,research,lifescience,medical to remind patients of who and where they are, provide feedback on time of day, day of week, etc, comment, on and describe what is happening at a given moment in time, and generally reinforce the patient’s awareness of their environment. Recent studies have observed improvements on the MMSE following sustained Inhibitors,research,lifescience,medical treatment, with reality orientation.149,150 However, such changes are often observed on the

orientation components of the MMSE-, and reality orientation does not appear to significantly impact behavioral functioning and, despite improvement, in cognition, improvements in IADL were not observed in several studies.150,151 There are a variety of memory training techniques that have been employed with some success in nondemented older adults, and we discuss these in detail Inhibitors,research,lifescience,medical below. These techniques are typically not effective in patients with dementia since their success relies upon utilization of many of the information-processing systems, which are no longer intact, in dementia. However, prosthetic memory aids such as diaries, memory wallets, and well-placed lists around the house and garden have been found to be helpful, particularly for early-stage patients who

can benefit from the Inhibitors,research,lifescience,medical type of mnemonic check details cueing such aids provide.152,153 Reminiscence therapy has also been postulated to be a potentially effective therapy for patients with dementia since studies suggest, that memories for remote events remain intact, longer than other forms of memory. Reminiscence through therapy aims to facilitate recall of past experiences with the overall goal of enhancing well-being. Few systematic studies of the effectiveness of reminiscence therapy in dementia patients exist, but the limited data available suggest that this technique may be more beneficial to interpersonal communication than cognitive processing.154-156 Indeed, many of the aforementioned techniques can also frustrate the dementia patient, by underscoring the limitations of their cognitive functioning. Behavioral therapy approaches aimed at, decreasing agitation, negative thoughts, and depression, and improving self-care have been quite successful.

Indeed, results of this study confirm that utilization of PLGA polymers to encapsulate Risperidone allows both the researcher and the clinician to customize therapy for schizophrenic patients. Additionally, these dosage forms can eliminate patient compliance issues, minimize costs associated with therapy, and improve the quality of life for patients and caregivers. Hence, a proper choice of polymer properties to manufacture long acting injections Inhibitors,research,lifescience,medical of atypical antipsychotics shows great promise to efficiently and effectively treat patients suffering from schizophrenia. 4. Conclusions The study demonstrated that sustained release microspheres of Risperidone utilizing two PLGA copolymers with varying lactide:glycolide

ratios (50:50 and 75:25) as well as molecular weights had a strong potential to be excellent Inhibitors,research,lifescience,medical for providing initial and maintenance levels of Risperidone and its active metabolite. Results from the simulation study indicate that, when utilizing the superposition principle, simulations of weekly http://www.selleckchem.com/products/MK-1775.html continual dosing of Formulations A and B and 15-day administration of Formulations C and D could be an effective approach for

sustained delivery of this molecule and a possible alternative to the currently available combination therapy. Thus, proper selection of polymer properties to prepare long acting dosage forms with atypical antipsychotics will ensure patient compliance, reduce side effects, Inhibitors,research,lifescience,medical and improve the quality of life for patients who suffer

from schizophrenia. Acknowledgments The research described in this paper Inhibitors,research,lifescience,medical was performed while the authors were affiliated with the University of Kentucky, Lexington, KY. The authors wish to thank Oakwood Labs, Oakwood, OH, and the Graduate School, University of Kentucky, Lexington, KY, for their financial support. Conflict of Interests The authors declare that there is no conflict of interests regarding Inhibitors,research,lifescience,medical the publication of this paper.
Drug delivery systems (DDS) are designed to increase the therapeutic properties of a drug and reduce its side effects. Poly lactic-co-glycolic acids (PLGA), which have been approved by the US FDA, are frequently used as biomaterials for drug delivery Isotretinoin due to their excellent biocompatibility and biodegradability [1]. PLGA particles are prepared by single- or double emulsion-solvent evaporation. In particular, a water-in-oil-in-water (w/o/w) method is widely used to encapsulate water soluble drugs [2]. The mechanism of degradation of PLGA particles generally involves a hydrolytic process. The maximum effect of a drug can only be achieved by strictly controlling target cell specificity. Moreover, reduced exposure of nontargeted cells to the drug may prevent undesirable side effects. In the context of in vivo distribution of PLGA “particles,” visualization of the particles themselves is feasible when markers such as fluorescent dyes are used [3–5].

More specifically, three smaller trials have

investigated the long-term effect on cognitive function of intervention of escitalopram compared with placebo in healthy individuals. Two of these studies found no significant effect of escitalopram. Wingen and coworkers [Knorr and Kessing, 2010; Wingen et al. 2005, 2006] investigated doses of escitalopram 10–20 mg/day versus placebo for 15 days in a crossover design in 18 participants with an unknown Etoposide price family history of depression and found no effect on actual driving performance, psychomotor performance or Inhibitors,research,lifescience,medical visual memory performance. Paul and colleagues [Paul et al. 2007] investigated escitalopram 20 mg/day versus placebo for 14 days in a crossover design in 24 participants with an unknown family history of depression and found no effect on psychomotor performance evaluated by multiple tests. In the third and most recent trial, Drueke and colleagues Inhibitors,research,lifescience,medical [Drueke et al. 2009] administered 10 mg of escitalopram for a period of 7 days in a crossover design to 20 healthy male participants with no family history of major mental disorder Inhibitors,research,lifescience,medical suggesting a differential effects of serotonergic manipulation depending on whether the test was applied

for the first or the second time The diversity of symptoms in MDD suggests that many areas of the brain are involved in the pathophysiology of the disorder. The serotonin transporter is expressed abundantly in the raphe nucleus and in the limbic system which may be the main site of action for SSRIs [Sierksma et al. 2009]. It is, however, not clear whether treatment with SSRIs results in a direct improvement of cognition or whether Inhibitors,research,lifescience,medical the effect of SSRIs on cognitive function is secondary to the effect of Inhibitors,research,lifescience,medical SSRIs on depressive symptoms. A neuropsychological hypothesis of antidepressant drug action suggests that, at the neuropsychological level, antidepressants work

by remediating negative affective biases in depression and anxiety and that these actions occur relatively quickly following drug administration [Harmer et al. 2009a, 2009b; Miskowiak et Bay 11-7085 al. 2007]. To disentangle the effect of antidepressant treatment from the effect of recovery from the depressive disorder per se, we investigated the effect of a SSRI on cognitive function in healthy first-degree relatives of patients with MDD. As revealed previously, such individuals may present with cognitive disturbances intermediate to those found in patients with depression and those in healthy individuals without a family history of affective disorders [Mannie et al. 2009; Christensen et al. 2006]. We hypothesized that 4 weeks of treatment with escitalopram would improve cognitive function compared with placebo.

1 The clear message was that, although there is, as yet, no cure for urologic CPPS (UCPPS), urologists can help to ameliorate the pain and improve the quality of life for patients using the treatments we currently have available. As the chairperson of the session, Dr. J. Curtis Nickel (Queen’s University, Kingston, Ontario, Canada) stated that these conditions are very prevalent (2% to 4% of men and women), represent a significant proportion Inhibitors,research,lifescience,medical of urological outpatient practice (> 5%), and yet remain the most enigmatic and frustrating conditions that urologists have to deal with in daily clinical practice. The patients’ quality

of life is dismal, mirroring that of other major chronic medical conditions such as active Crohn’s disease, insulin-dependent Inhibitors,research,lifescience,medical diabetes, and congestive heart failure. Because it affects patients of all ages, the condition results in an enormous expense in terms of direct and indirect costs to both society and individual patients. The diagnosis is one of exclusion (which surgeons do not

like) and the treatment regimens and strategies, to date, have been rather dismal. Inhibitors,research,lifescience,medical There are only two US Food and Drug Administration (FDA)-indicated interventions for BPS (oral pentosanpolysulfate sodium and intravesical Osimertinib order dimethyl sulfoxide [DMSO]). At best, they provide only modest benefit in a small percentage of patients. And for men with CPPS, there are no FDA-indicated medical or other interventions. So, not only does this condition Inhibitors,research,lifescience,medical represent the greatest unmet need in urology, it also represents the greatest opportunity for advances. During the panel discussion, the speakers outlined how these conditions should be evaluated. Their recommendations are described in Table 1 and Table 2. Table 1 Evaluation of a Man With Chronic Prostatitis/Chronic Pelvic Pain Syndrome Table 2 Evaluation of a Patient (Male or Female) With Interstitial

Cystitis/Bladder Pain Syndrome Dr. Nickel presented the Inhibitors,research,lifescience,medical evidence from available randomized, placebo-controlled clinical trials for CP/CPPS therapy using a unique network meta-analytical approach and indicated that, although our standard medical therapies provide statistically Parvulin significant treatment effects, they are, at most, barely clinically significant and, furthermore, there is a disconnect between overall benefit in the entire population and individual responses (Table 3). Therefore, traditional therapies can remain as part of our CP/CPPS treatment strategy, but monotherapy is not really effective. Table 3 Traditional Medical Therapies for Chronic Prostatis/Chronic Pelvic Pain Syndrome The UPOINT phenotype system was introduced as a clinical tool, using our standard urologic evaluation, to differentiate patients into one or more of six distinct phenotypic domains (Table 4). The traditional therapies are then directed, in a multimodal fashion, toward the different phenotypes identified in each individual patient. Dr.

Pulmonary vascular responses to sustained alveolar hypoxia have not been addressed in the isolated perfused rat lung. In this study, we investigated the effect of sustained hypoxic ventilation on pulmonary artery pressure in the present of phenylephrine, an α1-receptor agonist, under the above condition. Methods: We performed this study in the isolated perfused rat lung. After preparation, the lungs were divided randomly into five groups of normoxic-normocapnia, hypoxic-normocapnia, phenylephrine pre- or post-treated hypoxic-normocapnia

and phenylephrine pre-treated normoxic-normocapnia. Pulmonary hemodynamic, airway pressure Inhibitors,research,lifescience,medical and lung weight were measured during 60 min of the experiment for each group. Results: In the phenylephrine-pre-treated hypoxic-normocapnia group we observed a gradual increase in pulmonary artery pressure which approximated the results seen in the phenylephrine-pre-treated normoxic-normocapnia group. In contrast, in the Inhibitors,research,lifescience,medical phenylephrine-post-treated hypoxic-normcapnic group, pulmonary artery pressure did not change during the first 3 min of hypoxic-normocapnia. However at 1.5 min after administration of phenylephrine, this Inhibitors,research,lifescience,medical pressure began to increase sharply and continued

until the end of the experiment. This response was biphasic (0-10 min: acute phase, 10-60 min: sustained phase) with significantly higher pulmonary artery pressure ALK assay compared to the other groups. Conclusion: This study, for the first time, showed Inhibitors,research,lifescience,medical biphasic hypoxic pulmonary vasoconstriction in the isolated perfused rat lung with the sole administration of phenylephrine after but not before hypoxic gas ventilation. This finding suggested a facilitative role of alveolar hypoxia on pulmonary vasoconstriction induced by an α1-receptor agonist. Keywords: Hypoxia, Rat lung, Phenylephrine Introduction Investigations over

several decades have shown that numerous lung diseases and respiratory system disorders may disrupt alveolar ventilation and induce alveolar hypoxia, which may increase pulmonary resistance. Inhibitors,research,lifescience,medical This many response is known as hypoxic pulmonary vasoconstriction (HPV) which can regulate pulmonary blood flow distribution when it occurs in the local region of the lung, and, pulmonary hypertension during global and persistent alveolar hypoxia. Although HPV has been described since 1946,1 its underlying mechanism(s) remain unclear. Many scientists have established in vivo as well as in vitro models to study the mechanism of this physiological response.2 The isolated perfused lung is one of the basic methods for determining pulmonary hemodynamic and biochemical events associated with endothelial/epithelial interactions and physiological conditions compared with an in vivo study.3-5 It has been shown that HPV in the rabbit isolated perfused lung and isolated rat artery rings is biphasic with acute and sustained phases.

4) [3]. Propidium iodide (PI), α-mannnosidase, β-mannnosidase, endoglycosidase H, and rhodamine 6G were selleck compound obtained from Sigma-Aldrich (St. Louis, MO, USA). The “Annexin V-PE Apoptosis Detection Kit I” which contains Annexin V-PE and 7-amino-actinomycin D (7-ADD) was obtained from Becton Dickinson Biosciences (Franklin Lakes, NJ, USA). The caspase assay system was purchased from Promega

(Madison, WI, USA). Fluorescein isothiocyanate, isomer I (FITC), Span 80, cholesterol, and lecithin from soybeans were obtained from Wako Pure Chemical Industries (Osaka, Japan). The lecithin was purified by acetone precipitation [23]. The phospholipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl) (SuPE) Inhibitors,research,lifescience,medical was obtained from Avanti Polar Lipids Inhibitors,research,lifescience,medical (Alabaster, AL, USA). DSPE-PEG2000

was from NOF Corporation (Tokyo, Japan). PBS (phosphate buffered saline) was composed of 137mM NaCl, 2.7mM KCl, 10mM Na2HPO4 and 2mM KH2PO4, (pH = 7.4). 2.2. Cells and Cell Cultures Human osteosarcoma Takase (OST) cells were offered by Dr. Katsuro Tomita (Department of Orthopaedic Surgery, Kanazawa University School of Medicine, Japan), cultured in either ERDF medium (Kyokuto Pharmaceutical Industrial, Tokyo, Japan) or Dulbecco’s Modified Eagle Medium (D-MEM) (Wako Pure Chemical Industries, Osaka, Japan) supplemented with 10% of fetal bovine serum (FBS) at 37°C in a humidified Inhibitors,research,lifescience,medical atmosphere consisting of 5% CO2. Murine osteosarcoma cell line (LM8 cells) was obtained from RIKEN (RIKEN BRC Cell Bank). These LM8 cells were grown in D-MEM supplemented with 10% of FBS at 37°C in a humidified atmosphere consisting of 5% CO2. 2.3. Cell Viability Assay OST cells and LM8 cells were inoculated in 6-well Inhibitors,research,lifescience,medical culture plates at a cell density of 2.0 × 105cells/mL Inhibitors,research,lifescience,medical suspended in D-MEM with 10% FBS. After 16 hours, the medium in each plate was exchanged with 10% FBS D-MEM containing various concentration of ESA. After

incubation during one day, the cell number and the viabilities of both types of cells were evaluated by means of the “Propidium Iodide Nucleic Acid Stain” using flow cytometry [24]. The viability assay of OST others cells for EPV was also performed by the same way as above. In a similar way, time-courses of the viability of both OST cells and LM8 cells were experimentally measured in medium with ESA at a concentration of 50μg/mL. 2.4. Apoptosis Assay Apoptosis was analyzed by using the “Annexin V-PE Apoptosis Detection Kit I” according to a previously published protocol [25–27]. OST cells or LM8 cells, at a concentration of 2 × 105cells/mL, were suspended in D-MEM containing 10% FBS, and then inoculated in 6-well culture plates. After 16 hours inoculation, the medium in each plate was exchanged with 10% FBS, D-MEM containing 50μg/mL ESA. The cell lines in each plate were incubated for different time periods, followed by twice washing with cold PBS.