Small patients, significant findings:Electrophysiological properties of Bachmann's bundle in pediatric patients

Background: Bachmann's bundle (BB) may potentially play a role in the earlier onset and faster progression of atrial fibrillation in adult patients with congenital heart disease (CHD). It is unknown whether electrophysiological alterations already exist at BB in pediatric patients with CHD and whether they are related to aging. Objective: This study aimed to investigate BB electrophysiology in pediatric patients with CHD and assess the impact of age on BB electrophysiology. Methods: BB mapping was conducted in 55 patients (0.2–17.5 years). Activation patterns, potential voltages, low-voltage areas (LVAs), potential morphology, and conduction disorders of BB were analyzed and correlated with age. Results: Right-to-left activation across BB occurred in 96.4% of patients. Potential voltage was on average 7.2 ± 3.0 mV, and LVAs occurred in 85.4% of patients. Median local conduction velocity was 96.6 (72.9–121.0) cm/s, and conduction block occurred in 56.4% of patients. Most potentials were single and short-double potentials; long-double and fractionated potentials were recorded in 49.1% and 72.7% of patients, respectively. Age was weakly correlated with potential voltages (r = 0.312, P = .020) and moderately with local conduction velocity (r = 0.439, P &lt; .001), but not with potential morphology or conduction block. Conclusion: In pediatric patients with CHD, BB already contains a considerable amount of conduction disorders, LVAs, and potentials with complex morphology. The prevalence of these early electrophysiological alterations is not age related and does not differ among the right, left, and middle parts of BB.</p

Regulation of MYCN expression in human neuroblastoma cells

Contains fulltext : 81722.pdf (publisher's version ) (Open Access)BACKGROUND: Amplification of the MYCN gene in neuroblastoma (NB) is associated with a poor prognosis. However, MYCN-amplification does not automatically result in higher expression of MYCN in children with NB. We hypothesized that the discrepancy between MYCN gene expression and prognosis in these children might be explained by the expression of either MYCN-opposite strand (MYCNOS) or the shortened MYCN-isoform (DeltaMYCN) that was recently identified in fetal tissues. Both MYCNOS and DeltaMYCN are potential inhibitors of MYCN either at the mRNA or at the protein level. METHODS: Expression of MYCN, MYCNOS and DeltaMYCN was measured in human NB tissues of different stages. Transcript levels were quantified using a real-time reverse transcriptase polymerase chain reaction assay (QPCR). In addition, relative expression of these three transcripts was compared to the number of MYCN copies, which was determined by genomic real-time PCR (gQPCR). RESULTS: Both DeltaMYCN and MYCNOS are expressed in all NBs examined. In NBs with MYCN-amplification, these transcripts are significantly higher expressed. The ratio of MYCN:DeltaMYCN expression was identical in all tested NBs. This indicates that DeltaMYCN and MYCN are co-regulated, which suggests that DeltaMYCN is not a regulator of MYCN in NB. However, the ratio of MYCNOS:MYCN expression is directly correlated with NB disease stage (p = 0.007). In the more advanced NB stages and NBs with MYCN-amplification, relatively more MYCNOS is present as compared to MYCN. Expression of the antisense gene MYCNOS might be relevant to the progression of NB, potentially by directly inhibiting MYCN transcription by transcriptional interference at the DNA level. CONCLUSION: The MYCNOS:MYCN-ratio in NBs is significantly correlated with both MYCN-amplification and NB-stage. Our data indicate that in NB, MYCN expression levels might be influenced by MYCNOS but not by DeltaMYCN

Involvement of the Pleiotropic Drug Resistance Response, Protein Kinase C Signaling, and Altered Zinc Homeostasis in Resistance of Saccharomyces cerevisiae to Diclofenac ▿

Diclofenac is a widely used analgesic drug that can cause serious adverse drug reactions. We used Saccharomyces cerevisiae as a model eukaryote with which to elucidate the molecular mechanisms of diclofenac toxicity and resistance. Although most yeast cells died during the initial diclofenac treatment, some survived and started growing again. Microarray analysis of the adapted cells identified three major processes involved in diclofenac detoxification and tolerance. In particular, pleiotropic drug resistance (PDR) genes and genes under the control of Rlm1p, a transcription factor in the protein kinase C (PKC) pathway, were upregulated in diclofenac-adapted cells. We tested if these processes or pathways were directly involved in diclofenac toxicity or resistance. Of the pleiotropic drug resistance gene products, the multidrug transporter Pdr5p was crucially important for diclofenac tolerance. Furthermore, deletion of components of the cell wall stress-responsive PKC pathway increased diclofenac toxicity, whereas incubation of cells with the cell wall stressor calcofluor white before the addition of diclofenac decreased its toxicity. Also, diclofenac induced flocculation, which might trigger the cell wall alterations. Genes involved in ribosome biogenesis and rRNA processing were downregulated, as were zinc-responsive genes. Paradoxically, deletion of the zinc-responsive transcription factor Zap1p or addition of the zinc chelator 1,10-phenanthroline significantly increased diclofenac toxicity, establishing a regulatory role for zinc in diclofenac resistance. In conclusion, we have identified three new pathways involved in diclofenac tolerance in yeast, namely, Pdr5p as the main contributor to the PDR response, cell wall signaling via the PKC pathway, and zinc homeostasis, regulated by Zap1p

Deletion of DDR genes <i>MMS2</i>, <i>UBC13</i>, and <i>RFX1</i> confers resistance to APAP.

Fivefold serial dilutions of a cell suspension of the mutant strains with optical density OD600 = 0.05 were plated on YPD plates containing 0, 50 and 75 mM APAP and incubated for one (control) and three days at 37°C. After three days, the 75 mM plate was transferred to room temperature (RT) for 6 days. The strains used were WT (BY4741), Δmms2, Δubc13, Δrad5 and Δrfx1.</p

Predictors for trismus in patients receiving radiotherapy

Background: Trismus, a restricted mouth opening in head and neck cancer patients may be caused by tumor infiltration in masticatory muscles, radiation-induced fibrosis or scarring after surgery. It may impede oral functioning severely. The aims of our study were to determine: (1) the incidence of trismus at various time points; and (2) the patient, tumor, and treatment characteristics that predict the development of trismus after radiotherapy in head and neck cancer patients using a large database (n=641).Methods: Maximal mouth opening was measured prior to and 6, 12, 18, 24, 36, and 48 months after radiotherapy. Patient, tumor, and treatment characteristics were analyzed as potential predictors for trismus using a multivariable logistic regression analysis.Results: At six months after radiotherapy, 28.1% of the patients without trismus prior to radiotherapy developed trismus for the first time. At subsequent time points the incidence declined. Over a total period of 48 months after radiotherapy, the incidence of trismus was 3.6 per 10 person years at risk. Patients who had tumors located in the oral cavity, oropharynx or nasopharynx, and the salivary glands or ear, and who had a longer overall treatment time of radiotherapy, were more likely to develop trismus in the first six months after radiotherapy. Maximal mouth opening was a predictor for developing trismus at all time points.Conclusion: Incidence of trismus is 3.6 per 10 person years at risk. Tumor localization and overall treatment time of radiotherapy are predictors for developing trismus the first six months after radiotherapy. Maximal mouth opening is a significant predictor for developing trismus at all time points. Regular measurements of maximal mouth opening are needed to predict trismus

APAP affects levels of free ubiquitin in yeast.

<p>The WT, Δ<i>ubi4</i>, Δ<i>doa1</i> and Δ<i>ubp6</i> cells were grown exponentially and treated with 0, 25 or 50 mM APAP for 2 hrs at 37°C before harvesting. 4A) Western blot with antibodies against ubiquitin. 4B) Western blot with anti-actin antibodies was used as an internal control for loading.</p