Let me take a look at the Professor’s work from another angle, i.e., from the viewpoint of child neurology and the JSCN. He started his career at the Department of Pediatrics, University of Tokyo in April 1960, and was soon active, along with myself, as a part of the child neurology team. However, our time together was limited, as four years later, he completed a graduate course and then moved to Unites

States in July 1964. During this 4 years period, he gained Selleckchem DAPT his PhD with a thesis on a neuropathologic study of an autopsied MLD case [4]. This case became the first example of MLD in Japan. The most impressive article for me in early days is a report on neuropathology of a FCMD case published in 1976 [5]. This is the first orthodox, English-written paper on FCMD in the world. FCMD is a new entity discovered by myself in 1960, and numerous supportive investigations had been published inside Japan already; however, nearly all papers were written only in Japanese, so that

the disease entity of FCMD had been seldom recognized outside Japan. Kamoshita’s paper opened a window to the world for the first time. During the period in United States (1964–1968) he engaged in the study of developmental neuropathology at the Department of Pathology, Children’s Hospital of Los Angeles and the University of Southern California School of Medicine (chief: Dr. Benjamin H Landing) for 3 years, and at the Departments of Neurology and Pathology, Albert Einstein College of Medicine (chief: Dr. Kinuko Suzuki IWR-1 cell line and Dr. Kunihiko Suzuki). He contributed multiple original reports on neuropathology of several neurometabolic-degenerative disorders such as infantile neuroaxonal dystrophy with neonatal onset [6], infantile Niemann–Pick disease [7], lipidoses, ataxia telangiectasia, etc. His articles http://www.selleck.co.jp/products/azd9291.html are characterized by keen observations and precise descriptions, but always they included some novel viewpoints and hypotheses. On the other hand, as you see from Table 3, his relationship with the JSCN was both long and deep, through 43 years of membership. In particular, he served as

the president of the 25th Annual Meeting of JSCN in 1983, and, for another six years (1993–1999) he executed heavy responsibilities of the chief director with distinction. His resolute posture as he provided concise and appropriate comments from the moderator’s seat at the meetings each year remains vivid in our brain. He was a productive and proficient author, and published innumerable original articles and reviews in the field of child neurology, in addition to some in general pediatrics. He was an educator and mentor at a top ranked position, and, as a consequence, numerous excellent pupils grew up under his guidance to become leaders of the next generation in various field of pediatrics throughout Japan [8].

klinitox.de/index.php?id=3). Chemical incidents warrant a rapid decision whether HBM shall be applied and clear strategies for collection of biological samples, HBM analysis and communication on the outcomes of a HBM study to an individual or group in the aftermath. From a European perspective two alternative approaches are offered: the German “public interest–legal liability approach for the application of chemical incident HBM” (Empfehlungen des Umweltbundesamtes, 2006; this article) and the Dutch “pre-defined

transparent procedure for early decision-making concerning application of HBM following chemical incidents” (Scheepers et al., 2011; selleck chemicals Scheepers et al., 2014, this issue). Both procedures share important features, nevertheless there are also obvious differences. With respect to the selection of agents the first selleck approach covers a list of 50 chemical substances and substance groups (Burbiel et al., 2009). In creating this compilation special emphasis was laid on a civil protection point of view through considering the abuse of chemicals for terrorist attacks. In addition to the toxicity data Burbiel et al. designed a scoring system to evaluate the key parameters “availability”, “application” and “socio–economic

impact” to establish a ranking of importance. The second approach comprises of 15 chemical substances and substance groups from a public health point of view. The selection is partially based on practical toxicological experiences and considerations,

e.g., substances being important see more constituents of process emissions and fires or identification as acute exposure threshold level case study substances. Moreover, the key parameter “availability” plays an important role as the relevance of the chemical substances and substance groups was assessed based on the Dutch “Register Risk Situations Hazardous Substances”. The registry highlights nationwide the frequency of occurrence of chemical substances using the format of risk maps (http://www.risicokaart.nl). The use of the identical criterion “availability” in both procedures results in 47% match (7/15 of the Dutch list) of identified hazardous substances, namely acrylonitrile, arsine, benzene, dioxine, ethylene oxide, hydrogen cyanide and hydrogen fluoride. This may form a nucleus for a future European consensus list. The two approaches supply for each chemical substance or substance group CAS-number(s), basic toxicity data, IVERs (especially US EPA AEGL-2 values), occupational air and biological threshold values and HBM procedure data.

Os corticoides e os anti-histamínicos podem ser usados no tratamento das formas ligeiras e moderadas. A necessidade de os inibidores da protease serem ingeridos com alimentos faz com que a disgueusia seja um sintoma que deve ser monitorizado com cuidado. O boceprevir e, sobretudo, o telaprevir são metabolizados pelo citocromo p450 3A4 e 3A5 (CYP3A4/5). Existe, portanto, o risco de interação com medicamentos metabolizados pelas mesmas vias. Assim, chamamos a atenção para fármacos que podem induzir o CYP3A e, desse modo, baixar a concentração plasmática dos inibidores da protease como, por exemplo, a rifampicina, fenitoína e a buy Cetuximab carbamazepina; outros medicamentos são inibidores, competitivos ou não,

do CYP3A, diminuindo o metabolismo do boceprevir e do telaprevir, originando a sua sobredosagem como, por

exemplo, os antifúngicos. Os inibidores da protease podem, por outro lado, ter um efeito inibidor sobre diversos medicamentos, o que pode originar sobredosagem desses fármacos, como find more é o caso dos antiarrítmicos amiodarona, os derivados da ergotamina, as benzodiazepinas e as estatinas; outros, ainda, podem ter a sua eliminação aumentada com a consequente redução da sua eficácia, como é o caso dos anovulatórios, escitalopram, desipramina e zolpidem. Consoante as situações, durante a terapêutica com os inibidores da protease pode ser necessário descontinuar alguns fármacos ou proceder a modificações da dosagem. “
“Diarreia crónica define-se como uma alteração no trânsito intestinal caracterizada pela alteração da consistência das fezes, aumento do número de frequência das dejeções (mais de dejeções diárias) e peso fecal superior a 200 g/24 h, prologando-se por mais de

4 semanas. O diagnóstico diferencial pode ser muito complexo e abrangente, pois pode ter inúmeras etiologias: causas infecciosas, GBA3 endócrino-metabólicas, neoplásicas, funcionais e medicamentosas. Doente do sexo feminino, de 46 anos, caucasiana, residente em Leiria. Referenciada à consulta de Medicina Interna por diarreia crónica com estudo inconclusivo, episódios de lipotimia e incontinência esfincteriana. Referia o início do quadro há 8 anos (1997) com cerca de 6 dejeções diarreicas/dia, líquidas, por vezes com resíduos alimentares, diurnas e noturnas, sem sangue, sem muco e sem tenesmo ou falsas vontades. Negava fatores desencadeantes ou outros sintomas acompanhantes, nomeadamente náuseas, vómitos, febre, artralgias, alterações cutâneas, dor abdominal, esteatorreia. Os antecedentes pessoais eram irrelevantes. Do historial familiar, a doente era filha única de pai incógnito e negava patologia relevante da linha materna. Não existia igualmente registo de viagens recentes ou hábitos medicamentosos previamente ao início da diarreia. Dos exames complementares, realizados na altura, fez hemograma, bioquímica completa, estudo hormonal incluindo FSH, LH, PTH, cortisol, TSH, T3 e T4 totais e livres, que se encontravam dentro dos valores normais.

Only 2 patients (0.9%) reported a rectal toxicity of grade 2 (moderate diarrhea in both cases), which resolved in 1 patient and improved significantly in the second patient, shortly after treatment. No patients reported acute GI Grade 3 or 4 GI toxicity. The 7-year incidence of Grade 2 and 3 late rectal toxicities were 1% and 0.4%, respectively. One patient (0.4%) reported Grade 3 GI toxicity (daily rectal bleeding requiring transfusion, which resolved after cauterization). Approximately 1 year after Selleckchem Y-27632 completing radiation therapy, 1 patient was found to have

a midsigmoid stricture with fibrosis and angulation of the sigmoid distally on regular screening colonoscopy. The patient did not complain of abdominal pain and had regular bowel movements. The NU7441 area of the stricture was laparoscopically resected and final pathology was consistent with diverticulitis and abscess formation. The location of the stricture was inside the treatment field of the EBRT, but outside of the high dose region of the brachytherapy treatment volume. In the management of patients with intermediate- and high-risk prostate adenocarcinoma, dose-escalation studies have demonstrated an improvement in tumor control, disease-free survival, and freedom from DMs [1], [2], [3], [4], [5] and [19]. Yet, the benefits of dose escalation must be weighed against the risks of toxicity to the surrounding normal tissue structures. For patients with

disease localized to the prostate, HDR brachytherapy has been shown to be a favorable method of increasing the intraprostatic dose while minimizing the dose to peripheral sensitive structures. Our results indicate that a treatment regimen combining EBRT with a HDR brachytherapy boost is associated with a low likelihood of developing Grade 3 or higher GU or GI toxicities. An interesting finding in our report was the observation of improved outcomes in the high-risk patient cohort when higher BED doses were delivered with the HDR. Among patients with BED doses >190 Gy (α/β ratio of 2), the 7-year PSA relapse-free survival outcome for high-risk patients was 81% compared with 60% for patients who received

lower dose levels (p = 0.02). In addition, dose escalation for this high-risk selleck chemicals llc cohort was also associated with a reduction in improvement in the 7-year DMs-free survival outcomes from 60% to 89% for those who received lower and higher BED dose levels. These improved biochemical control outcomes for high-risk patients using higher doses appear to be consistent with what has been reported in the literature (See Table 5). Martinez et al. (20) had reported the outcomes of a cohort of 472 patients with intermediate- and high-risk disease treated with HDR brachytherapy and supplemental EBRT who were followed for a median of 8 years. The authors noted improved biochemical control and DMs-free survival outcomes with higher BED values. In that report, an α/β ratio of 1.

Fica por esclarecer, neste doente, a data exata de início do quadro de EE. O diagnóstico é por vezes tardio, principalmente pela semelhança ou coexistência de RGE. A nível histológico ficam ainda por esclarecer

os dados que permitem ao patologista afirmar o grau de eosinofilia20. Importante refletir sobre o objetivo do tratamento. Se pretendemos uma melhoria clínica ou uma melhoria histológica (prevenção de impacto alimentar, prevenção de fibroestenose, risco de infeções HSV)15 and 21. Quais serão os marcadores de maior risco ou maior gravidade da doença. Será que a estenose se traduz numa doença de mais difícil controlo? As complicações major da EE são a remodelação e estreitamento esofágico, que devemos evitar. Para isso são necessárias estratégias para monitorizar a doença. Para já, acompanhamento destes doentes é curto. Não há ainda evidência de associação a malignidade. De salientar a realização de biópsia para o diagnóstico Nutlin-3a clinical trial desta patologia, o tratamento de 8 semanas com IBP, que não só inibe a secreção ácida como também diminui citoquinas (IL‐5 this website e eotaxina 3), e posterior repetição de EDA com biópsia. Só aí se chega a conclusões definitivas e se considera a necessidade de estudos adicionais3, 7 and 8. É uma doença com uma boa

correlação clínico‐histológica, questiona‐se assim sobre a necessidade de repetições seriadas de EDA e biópsia no doente assintomático. Ainda por definir nos consensos mundiais os timings para a sua realização 21. Este artigo pretende salientar a importância de pensar neste diagnóstico, para a melhoria da qualidade de vida do doente, redução dos riscos/impactação e prevenção de danos irreversíveis most (remodelação do tecido). Devemos ponderar esta etiologia em casos de má evolução ponderal e dificuldades alimentares5. A terapêutica de ser adequada ao doente e em articulação com o alergologista7, 11, 12 and 18. Os autores declaram que para esta investigação não se realizaram experiências em seres humanos e/ou animais. Os autores declaram ter seguido os protocolos

do seu centro de trabalho acerca da publicação dos dados de pacientes. Os autores declaram ter recebido consentimento escrito dos pacientes e/ou sujeitos mencionados no artigo. O autor para correspondência deve estar na posse deste documento. Os autores declaram não haver conflito de interesses. “
“Renal cell carcinoma (RCC) has the potential to metastasize to almost any site. Metastatic disease may be present in up to 25% of patients at the time of diagnosis while another 50% develop metastasis during follow-up.1 and 2 Its tendency to vascular spreading as well as expansible ostetolytic skeletal metastization is well known. However, it is less expected that RCC presents as an unpredictable tumour which can manifest with very late metastases at unexpected sites, even a long period after successful resection of early stage lesion.

Na série de Sugiyama et al. nenhum dos doentes assintomáticos com NMPI-DS revelou presença de tecido maligno invasivo. De facto, a revisão dos trabalhos publicados mostra que doentes com NMPI-DS assintomáticos apresentam baixo risco de malignidade (0-5% dos casos) comparativamente aos doentes sintomáticos (30%)16.

Paralelamente, vários trabalhos identificaram aspetos morfológicos associados a maior risco de malignidade, nomeadamente, lesões superiores a 3 cm, presença de nódulos murais e paredes ou septos espessados3 and 16. Assim, o consenso da Associação Internacional Buparlisib cell line de Pancreatologia publicado em 2006 considerou razoável o controlo evolutivo imagiológico destas lesões em doentes assintomáticos e sem estigmas de elevado risco de malignidade (> 3 cm, presença de nódulos murais

ou citologia positiva para malignidade)16. Este controlo deverá ser feito através de TC ou CPRMN periódicas ou alternativamente com USE, esta última eventualmente com maior importância num futuro AZD2281 próximo, caso o doseamento do CEA no líquido das lesões se revele um fator discriminativo da sua natureza benigna ou maligna, como alguns trabalhos recentes sugerem, não existindo, contudo, consenso até à data21, 22 and 23. Todavia, a decisão de vigilância deverá ser individualizada, considerando a idade do doente, eventuais comorbilidades e a vontade do mesmo em cumprir o programa de PRKACG vigilância16 and 24. A necessidade de vigilância destas lesões acresce pelo relato de casos de aumento do risco de adenocarcinoma ductal pancreático, como lesões síncronas ou metácronas, aparentemente independentes das NMPI9, 16 and 25. No segundo caso apresentado,

dada a presença de uma NMPI-DS sem aparente envolvimento do ducto principal e sem estigmas de malignidade, optou-se por uma estratégia conservadora, mantendo a doente em vigilância clínica e imagiológica regulares. Outra constatação importante é a associação destas lesões a um elevado número de neoplasias extra-pancreáticas, nomeadamente gástricas e colorretais, identificadas em cerca de 30% dos casos16 and 24. Embora não se saiba se há um verdadeiro risco acrescido ou se se trata somente de uma associação fortuita com patologia mais frequente neste grupo etário, os clínicos deverão estar alerta para esta possibilidade, de forma a estimular a adesão aos programas de rastreio neoplásico existentes e a proceder à adequada investigação de sintomas extra-pancreáticos concomitantes. Ainda com várias questões em aberto, o conhecimento crescente sobre as NMPI observado nos últimos anos tem-se revelado fundamental para uma melhor abordagem clínica destas lesões e, desta forma, garantir o melhor prognóstico para estes doentes. Os autores declaram não haver conflito de interesses.

Evidence based on sputum culture results suggests that bacterial infection may be responsible for around half of AE-COPD,15 with a clear relationship being demonstrated between sputum purulence and the presence of bacteria.16 and 17 For this reason, current guidelines recommend acute antibiotic therapy for patients with more severe symptoms

of AE-COPD, with treatment typically lasting for 5–7 days.18, 19, 20 and 21 In particular, guidelines issued by the Global Initiative for Chronic Obstructive Lung Disease (GOLD) and the Joint Task Force of the European Respiratory Society and the European Society for Clinical Microbiology and Infectious Diseases advocate antibiotic use for check details those with Anthonisen type I (worsening dyspnoea with increased sputum volume and purulence) or type II (change in any two of these symptoms, particularly if one of these symptoms is increase in sputum purulence) episodes,18, 20 and 21 while the Canadian Thoracic Society suggests that antibiotics are beneficial for severe purulent AE-COPD (i.e. new increased expectoration of mucopurulent sputum and dyspnoea).19 Nevertheless, while such treatment has been shown to reduce the risk of subsequent exacerbations, relapse is common.22 Failure may be

related to this website inadequate antibiotic efficacy, which through incomplete resolution of the initial exacerbation and persistent bacterial infection is likely to influence risk of relapse.23, 24, 25, 26 and 27 Indeed, confirmed bacterial eradication following antibiotic therapy has been shown to be associated with higher clinical cure rates in patients with AE-COPD.28 Effective treatment of the acute exacerbation and reducing the risk of a subsequent bacterial exacerbation are thus important therapeutic goals for

antimicrobial treatment in COPD that may improve, in addition to other conventional treatments (e.g. long-acting bronchodilators and inhaled corticosteroids), the patients’ quality of life. The rate Meloxicam at which exacerbations occur appears to reflect an independent susceptibility phenotype.5 and 29 Furthermore, exacerbations appear to cluster together, with some patients remaining at high risk for recurrent exacerbation for some weeks after the initial exacerbation,5, 9, 30 and 31 possibly due to ongoing lung and systemic inflammation.32 While acquisition of new strains of respiratory pathogens is an important mechanism underlying acute COPD exacerbations,33 chronic microbial colonisation of the lower respiratory tract is also relevant.34, 35 and 36 This colonisation is likely to contribute to chronic inflammation and progressive loss of lung function in COPD due to increased rate of exacerbations.33, 35, 36, 37, 38 and 39 Treatments aimed at reducing bacterial colonisation, which may be regarded as chronic infection in the presence of an inflammatory response,40 may, therefore, help reduce the progression of the disease.

Table 1a and Table 1b lists the SQGs that were applied and those used to develop them. The DaS program contains four LAL values, for Cd, Hg, tPAH and tPCB (CEPA, 1999). For other contaminants of interest, the DaS program may look to the CCME Interim Sediment Quality Guideline (ISQG) list (CCME, Epacadostat molecular weight 2002), and then to SQGs from other jurisdictions. The metal values in the ISQG list are based upon the threshold effects levels (TELs) and probable effects levels (PELs) from MacDonald et al. (1996), but

without the inclusion of Ni for which no ISQG was available. As many other dredging programs include Ni in their lists, the TEL and PEL values, including Ni, are also applied in test protocols. However, the DaS and ISQG lists do not address all of the

other organics (e.g. pesticides, TBT) that were evaluated in this study and some of the ‘other’ organic SQG values used come from sources other than the GSK J4 molecular weight CCME. To compare sediment data to a full list of SQGs in this study, a range of dredging program LAL and UAL values (IMO, 2009), as well as non-dredging sediment threshold and probable effects values (Buchman, 2008), were collected (Table 1a and Table 1b). A “Consensus” set of LAL and UAL values was generated by calculating the geometric mean of all dredging LAL values for a given parameter. If no dredging values were available, or, if the only dredging-value was the CCME value (which is largely based on the non-dredging TEL and PEL values), then the geometric mean of the relevant non-dredging threshold or probable effects SQGs was used. It is not suggested that these values should be taken up as regulatory values. SQGs from different countries are developed based upon different sediment size fractions, and different analytical methods. As most (but not all) sediment contaminants tend to associate with the fine-grained Dichloromethane dehalogenase sediment fraction, these differences could result in different analytical results and pass/fail interpretations in various countries. However, it has been noted that overall sediment pass/fail outcomes using different SQG sets with the same narrative intent (e.g., LAL, UAL) do not differ

nearly as much as outcomes using different analyte sets and decision rules (Apitz et al., 2007, Apitz, 2008, Apitz, 2011 and Wenning et al., 2005). The “Consensus” LAL and UAL values developed for this paper provide a consistent set of hypothetical SQGs for the full suite of contaminants in this study. There are countless potential analyte and SQG lists that could be tested; in this paper we present a subset of plausible values to provide insight into how a range of choices affects potential regulatory outcomes. As various analyte and action level lists are selected by Environment Canada in future, the implications of these specific choices could be tested using the database. As noted above, the DaS PCB LAL is based upon aroclor, rather than congener values.

For example, one recent fMRI study [38••] suggests that the hippocampus supports the transfer of monetary value across related experiences through additional recruitment of reward regions. The researchers PI3K inhibitor cancer showed greater reactivation of prior related knowledge during encoding

of new reward information for stimuli that showed more evidence of subsequent preference shifts, a behavioral index of value transfer. Hippocampal–striatal functional coupling was also associated with value-related preference changes [38••], suggesting that hippocampus may interact with domain-specific regions (e.g., striatum in value learning tasks) in service of integration. Consistent with a domain-general role for hippocampus in memory integration, rodent work [39] has found that the hippocampus was necessary for updating a known goal location with new value information. These updated memories may then be transferred to neocortex, as mPFC was necessary for retaining the updated this website knowledge to support performance on the next day [39]. Thus, integrated memories incorporating value information may be maintained as memory

models in mPFC that will later bias behavior. We note that this role for mPFC is likely also domain-general given its documented involvement in a number of tasks lacking an explicit value component. Recent attention has focused on the behavioral benefits conferred by memory schema. For instance, research in rodents has shown that prior knowledge of a spatial layout (i.e., a spatial schema) can both facilitate acquisition of new related memories and speed their consolidation 40 and 41. Echoing these results, a number of human studies have reported behavioral benefits in learning and memory when new information can be incorporated into an existing schema 42•, 43 and 44. Application of a schema to a new Tenoxicam scenario has also been shown to recruit hippocampus 45 and 46. For example, one fMRI study [46] found that while engagement and connectivity of hippocampus and ventral mPFC was enhanced during generation of a task schema, the application of schema to guide behavior

in a novel but similarly structured task selectively recruited hippocampus. Rodent [41] and human 26, 42• and 43 work further suggests that mPFC may be activated along with hippocampus during learning of schema-related information. Recent empirical data indicate that one factor that may influence the relative engagement of MTL and mPFC is the degree of consistency between new information and existing schema. Specifically, one study [42•] demonstrated that mPFC engagement was more predictive of subsequent memory for information congruent with existing schema, perhaps reflecting direct encoding1 of new content into prior knowledge. By contrast, MTL engagement was more predictive of successful encoding of incongruent information.

The characteristics of these cell lines are listed in Table W1. BO-1509 (3-(4-methoxyphenyl)-9H-pyrrolo[1,2-a]indole-1,2-diyl)bis(methylene) bis(ethylcarbamate) was synthesized as previously described www.selleckchem.com/products/ABT-888.html [28]. The PI3K inhibitor LY294002 was purchased from Cayman Chemical Company (Ann

Arbor, MI). For the cytotoxicity assays, 3000 cells were seeded into each well of a 96-well plate, incubated overnight at 37°C, and then treated for 72 hours with various concentrations of BO-1509, LY294002, or a combination of both compounds. At the end of the treatment, 20 μl of Alamar Blue solution (AbD Serotec, Kidlington, United Kingdom) was added to each well and then incubated for 6 hours. Cell viability was assessed by measuring the absorbance at 570

and 600 nm according to the manufacturer’s instructions. The concentration of drug that resulted in a 50% inhibition of cell growth (IC50) was determined for each drug, and the combination index (CI) was determined using the CompuSyn software (version 1.0.1; CompuSyn, Inc, Paramus, NJ) and the median effect principle and plot [43]. The IC50 values were presented as means ± SD of three independent experiments. Western blot analysis MK-1775 solubility dmso was performed as previously described [29] and was adopted to determine the intracellular protein levels in response to drug treatment. Briefly, cells were harvested after drug treatment and lysed in electrophoresis sample

buffer. Proteins were then electrophoretically separated on a sodium dodecyl sulfate–polyacrylamide gel and transferred onto polyvinylidene difluoride membranes (Amersham Biosciences, GE Healthcare Bio-Sciences Corp, Piscataway, NJ). Protein-conjugated membranes were incubated with primary antibodies overnight at 4°C and then incubated with HRP-conjugated anti-rabbit or anti-mouse secondary antibody for 1 hour at room temperature. Western blot signals were visualized by chemiluminescence using SuperSignal West Pico chemiluminescence reagent (Pierce, Rockford, IL). Antibodies against AKT, phospho-AKT, Mre11, and FANCD2 were obtained Org 27569 from Santa Cruz Biotechnology (Dallas, TX), whereas antibodies against Nbs1, phospho-Nbs1 (pNbs1), Rad50, Rad51, and β-actin were from Genetex (San Antonio, TX). Antibodies against caspase-3, caspase-7, and poly(ADP-ribose) polymerase (PARP) and secondary antibodies against rabbit and mouse Ig were purchased from Cell Signaling Technology (Danver, MA). The anti-γH2AX antibody was obtained from Millipore (Billerica, MA). The induction of apoptosis by the treatment of cells with BO-1509, LY294002, or a combination of both agents was detected by flow cytometry using 4′,6-diamidino-2-phenylindole (DAPI) staining (1 μg/ml; Merck Millipore, Darmstadt, Germany) and the Annexin V–FITC Apoptosis Detection Kit (Calbiochem, La Jolla, CA) according to the manufacturer’s instructions.