Late cardiotoxicity after low dose of anthracycline therapy for acute lymphoblastic leukemia in childhood

Introduction Late cardiotoxicity is a known complication of anthracycline therapy but the long-term effects of low cumulative doses are not well documented. We studied late cardiotoxicity in survivors of childhood acute lymphoblastic leukemia (ALL) treated with low anthracycline doses 10 to 20 years earlier. Methods Seventy-seven ALL survivors who received a cumulative anthracycline dose <250 mg/m(2) and were at least 10 years after treatment were evaluated for signs of clinical heart failure. Cardiac function was assessed by echocardiography including tissue Doppler measurements of the septal mitral annulus in 37 ALL survivors 10.6-18.3 years (median 13.3 years) after anthracycline treatment with cumulative doses of 180 (n=19) or 240 mg/m(2) (n=18). The control group consisted of 30 healthy volunteers matched for age, sex, BSA, and BMI. Results No clinical relevant cardiotoxicity was found. Left ventricular shortening fraction (SF) was significantly reduced in male ALL survivors. Three of the 19 male ALL survivors had an SF below 30%. Male ALL survivors showed a significantly lower early filling velocity to atrial contraction velocity ratio but myocardial velocity during early filling was comparable between patients and controls. ALL survivors had a significantly longer isovolumetric relaxation time (IVRT). Thirty percent of the ALL survivors have an abnormal IVRT compared to the normal range of the controls. Conclusion and implications for cancer survivors At a median of 13.3 years after exposure to cumulative doses of anthracyclines of 180 or 240 mg/m(2), no clinical relevant cardiotoxicity was found but subclinical cardiac abnormalities were present in 30% of the patients

Anticancer prodrugs of butyric acid and formaldehyde protect against doxorubicin-induced cardiotoxicity

Formaldehyde has been previously shown to play a dominant role in promoting synergy between doxorubicin (Dox) and formaldehyde-releasing butyric acid (BA) prodrugs in killing cancer cells. In this work, we report that these prodrugs also protect neonatal rat cardiomyocytes and adult mice against toxicity elicited by Dox. In cardiomyocytes treated with Dox, the formaldehyde releasing prodrugs butyroyloxymethyl diethylphosphate (AN-7) and butyroyloxymethyl butyrate (AN-1), but not the corresponding acetaldehyde-releasing butyroyloxydiethyl phosphate (AN-88) or butyroyloxyethyl butyrate (AN-11), reduced lactate dehydrogenase leakage, prevented loss of mitochondrial membrane potential (ΔΨm) and attenuated upregulation of the proapoptotic gene Bax. In Dox-treated mice, AN-7 but not AN-88 attenuated weight-loss and mortality, and increase in serum lactate dehydrogenase. These findings show that BA prodrugs that release formaldehyde and augment Dox anticancer activity also protect against Dox cardiotoxicity. Based on these observations, clinical applications of these prodrugs for patients treated with Dox warrant further investigation

Anti-inflammatory agents and monoHER protect against DOX-induced cardiotoxicity and accumulation of CML in mice

Cardiac damage is the major limiting factor for the clinical use of doxorubicin (DOX). Preclinical studies indicate that inflammatory effects may be involved in DOX-induced cardiotoxicity. Nɛ-(carboxymethyl) lysine (CML) is suggested to be generated subsequent to oxidative stress, including inflammation. Therefore, the aim of this study was to investigate whether CML increased in the heart after DOX and whether anti-inflammatory agents reduced this effect in addition to their possible protection on DOX-induced cardiotoxicity. These effects were compared with those of the potential cardioprotector 7-monohydroxyethylrutoside (monoHER)

Candidate reference genes and their annotated functions.

<p>^a<i>Araucaria angustifolia</i> transcriptome database (Elbl et al. 2015).</p><p>^b Encoded-protein function according to TAIR database (<a href="http://www.arabidopsis.org/" target="_blank">http://www.arabidopsis.org/</a>).</p><p>Candidate reference genes and their annotated functions.</p

Ranking of <i>Araucaria angustifolia</i> candidate reference genes based on GeNorm analysis.

<p>^a Stability coefficient is the mean of the variation of two internal control genes between an individual and all other tested genes. The most stable gene has the lowest <i>M</i> value (cut-off < 1.5).</p><p>^b Pairwise variation values <i>Vn/n+1</i> < 0.15 mean that use of the two most stable genes is sufficient to normalize the expression of a test gene in the corresponding set of samples. <i>n</i>: number of genes.</p><p>Ranking of <i>Araucaria angustifolia</i> candidate reference genes based on GeNorm analysis.</p

Identification and Evaluation of Reference Genes for Quantitative Analysis of Brazilian Pine (<i>Araucaria angustifolia</i> Bertol. Kuntze) Gene Expression

<div><p>Quantitative analysis of gene expression is a fundamental experimental approach in many fields of plant biology, but it requires the use of internal controls representing constitutively expressed genes for reliable transcript quantification. In this study, we identified fifteen putative reference genes from an <i>A</i>. <i>angustifolia</i> transcriptome database. Variation in transcript levels was first evaluated <i>in silico</i> by comparing read counts and then by quantitative real-time PCR (qRT-PCR), resulting in the identification of six candidate genes. The consistency of transcript abundance was also calculated applying geNorm and NormFinder software packages followed by a validation approach using four target genes. The results presented here indicate that a diverse set of samples should ideally be used in order to identify constitutively expressed genes, and that the use of any two reference genes in combination, of the six tested genes, is sufficient for effective expression normalization. Finally, in agreement with the <i>in silico</i> prediction, a comprehensive analysis of the qRT-PCR data combined with validation analysis revealed that <i>AaEIF4B-L</i> and <i>AaPP2A</i> are the most suitable reference genes for comparative studies of <i>A</i>. <i>angustifolia</i> gene expression.</p></div

cDNA quality (a) and primer (b) test.

<p>Amplification products of PCR analyses using genomic DNA (gDNA) or a pool of all cDNA samples (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136714#pone.0136714.g001" target="_blank">Fig 1</a>) and <i>UBI</i> intron-flanking specific primers (A). Amplicons obtained by PCR using a pool of all cDNA samples and specific primers for the reference genes (B). bp = base pairs.</p

Primers used for gene amplification.

<p>^a Genes used for reference gene validation.</p><p>Primers used for gene amplification.</p

Validation of the most stable reference genes.

<p>(A) Relative expression of <i>AaADC</i>, <i>AaCAT</i>, <i>AaTPS3</i> and <i>AaUGP</i> in the samples used in this study, normalized with different combinations of reference genes. (B) Co-variation patterns by neural network analysis, performed by applying the *omeSOM software [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136714#pone.0136714.ref043" target="_blank">43</a>].</p