bluuebunny/crossref_metadata_2025 huggingface dataset:
Inactivation of the putative suppressor gene DOK1 by promoter hypermethylation in primary human cancers
Authors: Saulnier Amandine , Vaissière Thomas , Yue Jiping , Siouda Maha , Malfroy Marine , Accardi Rosita , Creveaux Marion , Sebastian Sinto , Shahzad Naveed , Gheit Tarik , Hussain Ishraq , Torrente Mariela , Maffini Fausto Antonio , Calabrese Luca , Chiesa Fausto , Cuenin Cyrille , Shukla Ruchi , Fathallah Ikbal , Matos Elena , Daudt Alexander , Koifman Sergio , Wünsch‐Filho Victor , Menezes Ana M.B. , Curado Maria‐Paula , Zaridze David , Boffetta Paolo , Brennan Paul , Tommasino Massimo , Herceg Zdenko , Sylla Bakary S.
Published: July 2011

The DOK1 gene is a putative tumour suppressor gene located on the human chromosome 2p13 which is frequently rearranged in leukaemia and other human tumours. We previously reported that the DOK1 gene can be mutated and its expression down‐regulated in human malignancies. However, the mechanism underlying DOK1 silencing remains largely unknown. We show here that unscheduled silencing of DOK1 expression through aberrant hypermethylation is a frequent event in a variety of human malignancies. DOK1 was found to be silenced in nine head and neck cancer (HNC) cell lines studied and DOK1 CpG hypermethylation correlated with loss of gene expression in these cells. DOK1 expression could be restored via demethylating treatment using 5‐aza‐2′deoxycytidine. In addition, transduction of cancer cell lines with DOK1 impaired their proliferation, consistent with the critical role of epigenetic silencing of DOK1 in the development and maintenance of malignant cells. We further observed that DOK1 hypermethylation occurs frequently in a variety of primary human neoplasm including solid tumours (93% in HNC, 81% in lung cancer) and haematopoietic malignancy (64% in Burkitt's lymphoma). Control blood samples and exfoliated mouth epithelial cells from healthy individuals showed a low level of DOK1 methylation, suggesting that DOK1 hypermethylation is a tumour specific event. Finally, an inverse correlation was observed between the level of DOK1 gene methylation and its expression in tumour and adjacent non tumour tissues. Thus, hypermethylation of DOK1 is a potentially critical event in human carcinogenesis, and may be a potential cancer biomarker and an attractive target for epigenetic‐based therapy.
Identification of a novel recurrent 1q42.2‐1qter deletion in high risk MYCN single copy 11q deleted neuroblastomas
Authors: Fieuw Annelies , Kumps Candy , Schramm Alexander , Pattyn Filip , Menten Björn , Antonacci Francesca , Sudmant Peter , Schulte Johannes H. , Van Roy N , Vergult Sarah , Buckley Patrick G. , De Paepe A , Noguera Rosa , Versteeg Rogier , Stallings Raymond , Eggert Angelika , Vandesompele Jo , De Preter K , Speleman Frank
Published: July 2011

Neuroblastoma is an aggressive embryonal tumor that accounts for ∼15% of childhood cancer deaths. Hitherto, despite the availability of comprehensive genomic data on DNA copy number changes in neuroblastoma, relatively little is known about the genes driving neuroblastoma tumorigenesis. In this study, high resolution array comparative genome hybridization (CGH) was performed on 188 primary neuroblastoma tumors and 33 neuroblastoma cell lines to search for previously undetected recurrent DNA copy number gains and losses. A new recurrent distal chromosome 1q deletion (del(1)(q42.2qter)) was detected in seven cases. Further analysis of available array CGH datasets revealed 13 additional similar distal 1q deletions. The majority of all detected 1q deletions was found in high risk 11q deleted tumors without MYCN amplification (Fisher exact test p = 5.61 × 10−5). Using ultra‐high resolution (∼115 bp resolution) custom arrays covering the breakpoints on 1q for 11 samples, clustering of nine breakpoints was observed within a 12.5‐kb region, of which eight were found in a 7‐kb copy number variable region, whereas the remaining two breakpoints were colocated 1.4‐Mb proximal. The commonly deleted region contains one miRNA (hsa‐mir‐1537), four transcribed ultra conserved region elements (uc.43‐uc.46) and 130 protein coding genes including at least two bona fide tumor suppressor genes, EGLN1 (or PHD2) and FH. This finding further contributes to the delineation of the genomic profile of aggressive neuroblastoma, offers perspectives for the identification of genes contributing to the disease phenotype and may be relevant in the light of assessment of response to new molecular treatments.
PI3K class IB controls the cell cycle checkpoint promoting cell proliferation in hepatocellular carcinoma
Authors: Dituri Francesco , Mazzocca Antonio , Lupo Luigi , Edling Charlotte E. , Azzariti Amalia , Antonaci Salvatore , Falasca Marco , Giannelli Gianluigi
Published: July 2011

Alterations of the cell cycle checkpoint frequently occur during hepatocarcinogenesis. Dysregulation of the phosphatidylinositol‐3‐kinases (PI3K) signaling pathway is believed to exert a potential oncogenic effect in hepatocellular carcinoma (HCC), ultimately promoting tumor cell proliferation. However, the impact of PI3K on cell cycle regulation remains unclear. We used a combined loss‐ and gain‐of‐function approach to address the involvement of p110γ in HCC cell proliferation, apoptosis and the cell cycle. We also investigated the correlation between p110γ and Ki‐67 in 24 HCC patients. Finally, we analyzed the expression levels of p110γ and cell cycle regulators in HCC tissues. We found that PI3K class IB, but not class IA, is required for HCC cell proliferation. In particular, we found that knock‐down of p110γ inhibits cell proliferation because of an arrest of the cell cycle in the G0‐G1 phase. This effect is associated with an altered expression of proteins regulating the cell cycle progression, including p21, and with an increased apoptosis. By contrast, we found that ectopic expression of p110γ promotes HCC cell proliferation. Tissues analysis performed in HCC patients showed a positive correlation between the expression of p110γ and Ki‐67, a marker of proliferation, and, even more importantly, that p21 expression is up‐regulated in HCC patients with a lower p110γ expression. Our results emphasize the role of p110γ as a promoter of HCC proliferation and unveil an important cell cycle regulation function of this molecule.
Epigenetic regulation by decitabine of melanoma differentiation in vitro and in vivo
Authors: Alcazar Oscar , Achberger Susan , Aldrich Wayne , Hu Zhenbo , Negrotto Soledad , Saunthararajah Yogen , Triozzi Pierre
Published: July 2011

Apoptosis genes, such as TP53 and p16/CDKN2A, that mediate responses to cytotoxic chemotherapy, are frequently nonfunctional in melanoma. Differentiation may be an alternative to apoptosis for inducing melanoma cell cycle exit. Epigenetic mechanisms regulate differentiation, and DNA methylation alterations are associated with the abnormal differentiation of melanoma cells. The effects of the deoxycytidine analogue decitabine (5‐aza‐2′‐deoxycytidine), which depletes DNA methyl transferase 1 (DNMT1), on melanoma differentiation were examined. Treatment of human and murine melanoma cells in vitro with concentrations of decitabine that did not cause apoptosis inhibited proliferation accompanied by cellular differentiation. A decrease in promoter methylation, and increase in expression of the melanocyte late‐differentiation driver SOX9, was followed by increases in cyclin‐dependent kinase inhibitors (CDKN) p27/CDKN1B and p21/CDKN1A that mediate cell cycle exit with differentiation. Effects were independent of the TP53, p16/CDKN2A and also the BRAF status of the melanoma cells. Resistance, when observed, was pharmacologic, characterized by diminished ability of decitabine to deplete DNMT1. Treatment of murine melanoma models in vivo with intermittent, low‐dose decitabine, administered sub‐cutaneously to limit high peak drug levels that cause cytotoxicity and increase exposure time for DNMT1 depletion, and with tetrahydrouridine to decrease decitabine metabolism and further increase exposure time, inhibited tumor growth and increased molecular and tumor stromal factors implicated in melanocyte differentiation. Modification of decitabine dose, schedule and formulation for differentiation rather than cytotoxic objectives inhibits the growth of melanoma cells in vitro and in vivo.
Molecular history of Richter syndrome: origin from a cell already present at the time of chronic lymphocytic leukemia diagnosis
Authors: Rossi Davide , Spina Valeria , Forconi Francesco , Capello Daniela , Fangazio Marco , Rasi Silvia , Martini Maurizio , Gattei Valter , Ramponi Antonio , Larocca Luigi M. , Bertoni Francesco , Gaidano Gianluca
Published: July 2011

Richter syndrome (RS) represents the transformation of chronic lymphocytic leukemia to aggressive lymphoma. We explored intraclonal diversification (ID) of immunoglobulin genes in order to (i) follow the evolutionary history of the RS clone (ii) compare the role of ID in clonally related RS vs. clonally unrelated cases. Most (10/11, 90.9%) clonally related RS stem from the predominant clone observed at CLL diagnosis. One single RS had a transformation pattern compatible with sequential evolution from a secondary CLL subclone. Once RS transformation had occurred, all secondary CLL subclones disappeared and were substituted by the dominant RS clone with its own descendants. These observations suggest that genetic lesions associated with RS transformation are acquired by a cell belonging to the original CLL clone, rather than being progressively accumulated by later CLL subclones. Accordingly, most (9/11, 81.1%) clonally related RS harbored a genetic lesion disrupting TP53 that was already present, though at subclonal levels, in 5/11 (45.5%) samples of the paired CLL phase. A fraction of clonally related RS switched off ID (4/11, 36.4%) or reduced the levels of ID (5/11, 45.4%) at transformation. Conversely, all clonally unrelated RS harbored ID and were characterized by a significantly higher mutation frequency compared to clonally related RS (median: 1.18 × 10−3
vs. 0.13 × 10‐3; p =0.002). These data indicate that (i) clonally related RS stems from a cell that is already present within the initial CLL clone and (ii) clonally unrelated and clonally related RS are biologically distinct disorders also in terms of antigen affinity maturation.
The lymphangiogenic factor SOX 18: A key indicator to stage gastric tumor progression
Authors: Eom Bang Wool , Jo Min Jung , Kook Myeong‐Cherl , Ryu Keun Won , Choi Il Ju , Nam Byung‐Ho , Kim Young‐Woo , Lee Jun Ho
Published: July 2011

SOX group F genes are important regulators of angiogenesis and lymphangiogenesis. The aim of the present study was to examine the relationships between Sox group F expression and clinicopathological factors in gastric cancer. Three hundred and fifteen gastric cancer tissues and the corresponding normal gastric tissue were obtained from the tumor bank at the National Cancer Center, Korea. SOX group F mRNA levels in these tissues were evaluated by reverse transcriptase polymerase chain reaction (RT‐PCR). The serum levels of SOX 18 proteins in 219 gastric cancer patients and in 30 healthy volunteers were also measured by enzyme‐linked immunosorbent assay. Furthermore, immunohistochemistry (IHC) was performed on 679 gastric cancer tissues and the clinicopathological characteristics, as well as the survival rates of SOX 18 positive and negative gastric cancers were compared. RT‐PCR showed that SOX group F mRNA was increased in the gastric cancer tissues compared to the normal gastric tissues (p < 0.001, respectively). The serum levels of SOX 18 protein were also increased in gastric cancer patients compared to healthy volunteers. IHC showed that of the 679 gastric cancer cases, 177 (26.1%) were positive for SOX 18 expression in their tumor stroma, and the frequencies of both lymphovascular invasion and lymph node metastases were higher in the SOX 18 positive than in the negative group. Both the 5‐year survival and the recurrence‐free survival were shorter for SOX 18 positive tumors (p = 0.023 and 0.012, respectively). SOX 18 expression might be a prognostic tumor marker and a potential therapeutic target in gastric cancer.
A four‐gene methylation marker panel as triage test in high‐risk human papillomavirus positive patients
Authors: Eijsink J.J.H. , Lendvai Á. , Deregowski V. , Klip H.G. , Verpooten G. , Dehaspe L. , de Bock G.H. , Hollema H. , van Criekinge W. , Schuuring E. , van der Zee A.G.J. , Wisman G.B.A.
Published: July 2011

Cervical neoplasia‐specific biomarkers, e.g. DNA methylation markers, with high sensitivity and specificity are urgently needed to improve current population‐based screening on (pre)malignant cervical neoplasia. We aimed to identify new cervical neoplasia‐specific DNA methylation markers and to design and validate a methylation marker panel for triage of high‐risk human papillomavirus (hr‐HPV) positive patients. First, high‐throughput quantitative methylation‐specific PCRs (QMSP) on a novel OpenArray™ platform, representing 424 primers of 213 cancer specific methylated genes, were performed on frozen tissue samples from 84 cervical cancer patients and 106 normal cervices. Second, the top 20 discriminating methylation markers were validated by LightCycler® MSP on frozen tissue from 27 cervical cancer patients and 20 normal cervices and ROCs and test characteristics were assessed. Three new methylation markers were identified (JAM3, EPB41L3 and TERT), which were subsequently combined with C13ORF18 in our four‐gene methylation panel. In a third step, our methylation panel detected in cervical scrapings 94% (70/74) of cervical cancers, while in a fourth step 82% (32/39) cervical intraepithelial neoplasia grade 3 or higher (CIN3+) and 65% (44/68) CIN2+ were detected, with 21% positive cases for ≤CIN1 (16/75). Finally, hypothetical scenario analysis showed that primary hr‐HPV testing combined with our four‐gene methylation panel as a triage test resulted in a higher identification of CIN3 and cervical cancers and a higher percentage of correct referrals compared to hr‐HPV testing in combination with conventional cytology. In conclusion, our four‐gene methylation panel might provide an alternative triage test after primary hr‐HPV testing.
Co‐expression of hepatocyte growth factor and c‐Met predicts peritoneal dissemination established by autocrine hepatocyte growth factor/c‐Met signaling in gastric cancer
Authors: Toiyama Yuji , Yasuda Hiromi , Saigusa Susumu , Matushita Kouhei , Fujikawa Hiroyuki , Tanaka Koji , Mohri Yasuhiko , Inoue Yasuhiro , Goel Ajay , Kusunoki Masato
Published: July 2011

Epithelial–mesenchymal transition (EMT) promotes and facilitates migration and invasion of epithelial tumor cells. EMT is induced by factors such as hepatocyte growth factor (HGF). This study aimed to establish whether the HGF/c‐Met pathway is associated with gastric cancer metastasis; especially peritoneal dissemination. HGF and c‐Met expression and EMT‐related molecules were evaluated using real‐time PCR and immunohistochemistry. The role of the HGF/c‐Met pathway in EMT and anoikis was determined, and kinase inhibitor SU11274 was tested for its ability to block HGF‐induced biological effects. In HGF−/c‐Met+ gastric cancer cells, recombinant HGF promoted an EMT phenotype that was characterized by morphology, impaired E‐cadherin and induction of vimentin. HGF promoted cell growth, invasiveness and migration and inhibition of anoikis. SU11274 blocked HGF‐induced EMT and biological effects in vitro. In HGF+/c‐Met+ gastric cancer cells, HGF did not affect the biological outcome of EMT and anoikis, but SU11274 exerted the same inhibitory effects as in HGF−/c‐Met+ cells. In vivo, HGF+/c‐Met+ gastric cancer cells only established peritoneal dissemination and SU11274 inhibited tumor growth. Clinically, HGF expression was significantly correlated with c‐Met expression in gastric cancer. Increased HGF and c‐Met had a significant association with poor prognosis and predicted peritoneal dissemination. We demonstrated that the HGF/c‐Met pathway induces EMT and inhibition of anoikis in gastric cancer cells. Co‐expression of HGF and c‐Met has the potential to promote peritoneal dissemination in gastric cancer. Blockade of the autocrine HGF/c‐Met pathway could be clinically useful for the treatment of peritoneal dissemination in gastric cancer.
Knockdown of metallopanstimulin‐1 inhibits NF‐κB signaling at different levels: The role of apoptosis induction of gastric cancer cells
Authors: Yang Zhong‐Yin , Qu Ying , Zhang Qing , Wei Min , Liu Chuan‐Xu , Chen Xue‐Hua , Yan Min , Zhu Zheng‐Gang , Liu Bing‐Ya , Chen Guo‐Qiang , Wu Ying‐Li , Gu Qin‐Long
Published: July 2011

The ribosomal protein S27 (metallopanstimulin‐1, MPS‐1) has been reported to be a multifunctional protein, with increased expression in a number of cancers. We reported previously that MPS‐1 was highly expressed in human gastric cancer. Knockdown of MPS‐1 led to spontaneous apoptosis and repressed proliferation of human gastric cancer cells in vitro and in vivo. However, how does MPS‐1 regulate these processes is unclear. Here we performed microarray and pathway analyses to investigate possible pathways involved in MPS‐1 knockdown‐induced apoptosis in gastric cancer cells. Our results showed that knockdown of MPS‐1 inhibited NF‐κB activity by reducing phosphorylation of p65 at Ser536 and IκBα at Ser32, inhibiting NF‐κB nuclear translocation, and down‐regulating its DNA binding activity. Furthermore, data‐mining the Gene‐Regulatory‐Network revealed that growth arrest DNA damage inducible gene 45β (Gadd45β), a direct NF‐κB target gene, played a critical role in MPS‐1 knockdown‐induced apoptosis. Over‐expression of Gadd45β inhibited MPS‐1 knockdown‐induced apoptosis via inhibition of JNK phosphorylation. Taken together, these data revealed a novel pathway, the MPS‐1/NF‐κB/Gadd45β signal pathway, played an important role in MPS‐1 knockdown‐induced apoptosis of gastric cancer cells. This study sheds new light on the role of MPS‐1/NF‐κB in apoptosis and the possible use of MPS‐1 targeting strategy in the treatment of gastric cancer.
Developmental evolution of the insect retina: insights from standardized numbering of homologous photoreceptors
Authors: Friedrich Markus , Wood Emily J. , Wu Meng
Published: July 2011

The canonical number of eight photoreceptors and their arrangement in the ommatidia of insect compound eyes is very conserved. However significant variations exist in selective groups, such as the Lepidoptera and Hymenoptera, which independently evolved additional photoreceptors. For this and historical reasons, heterogeneous labeling conventions have been in use for photoreceptor subtypes, despite developmentally and structurally well‐defined homologies. Extending earlier efforts, we introduce a universal photoreceptor subtype classification key that relates to the Drosophila numbering system. Its application is demonstrated in major insect orders, with detailed information on the relationship to previous conventions. We then discuss new insights that result from the improved understanding of photoreceptor subtype homologies. This includes evidence of functionally imposed ground rules of differential opsin expression, the underappreciated role of R8 as ancestral color receptor, the causes and consequences of parallel R7 photoreceptor addition in Hymenoptera and Lepidoptera, and the ancestral subfunctionalization of outer photoreceptors cells, which may be only developmentally recapitulated in Drosophila. We conclude with pointing out the need for opsin expression data from a wider range of insect orders. J. Exp. Zool. (Mol. Dev. Evol.) 316:484–499, 2011. © 2011 Wiley Periodicals, Inc.
Evaluation of a 188Re‐radiolabeled Arg‐Gly‐Asp peptide for tumor overexpressed αvβ3 receptors
Authors: Ma Yufei , Yu Junfeng , Han Yanjiang , Wang Cheng , Li Jianbo , Shen Hua , Wang Ni , Yin Duanzhi
Published: July 2011

To develop a peptide‐based radiopharmaceutical for the therapy of αvβ3 receptors overexpressed tumors, we have prepared a novel Arg‐Gly‐Asp (RGD) peptide (HCRGDCF(D)CRGDC, P12) radiolabeled with 188Re. With His acid at the end of the peptide containing RGD, the label efficiency was more than 95% within 30 min. The peptide binds to human glioblastoma U87MG cells with high affinity [IC50 = 86.3 nm]. The stability of 188Re‐P12 in vitro was also investigated. More than 80% of radioactivity was kept in the peptide after 4 h incubation in phosphate buffer solution (pH = 7.4) or calf serum under physiological conditions. Biodistribution of this radiocompound was carried out in mice bearing S180 tumor. Fast clearance of 188Re‐peptide from blood and specific uptakes by tumors realized higher tumor‐to‐blood ratio (1.80) 4 h post‐injection. Obvious difference was observed between the blocking and unblocking experiments in whole body autoradiography imaging. These results have demonstrated the potential of 188Re‐labeling RGD as a radiotherapeutic agent.
Spring temperature variability in northern Fennoscandia AD 1693–2011
Authors: Loader Neil J. , Jalkanen Risto , Mccarroll Danny , Moberg Anders
Published: July 2011

A reconstruction of spring (April–May) temperature for northern Fennoscandia developed from the Tornionjoki (Tornio river) long cryophenological record of ice break‐up dates, back to AD 1693, is presented. The record is strongly climatically sensitive and explains 67% of the variance in the instrumental data over the last 150 years. The record exhibits a stepped decrease in the duration of the river's ice cover by 14 days, equivalent to an increase in April–May mean temperature of approximately 2.5°C over the last three centuries. The relationship between the date of ice break‐up, and accumulated daily mean temperatures (>0°C) is investigated. Uncertainty in the observation of ice break‐up is also considered in addition to the potential of this time series for regional climate model validation. Copyright © 2011 John Wiley & Sons, Ltd.
Physico‐chemical and toxicological assessment of liquid wastes from olive processing‐related industries
Authors: Pierantozzi Pierluigi , Zampini Catiana , Torres Mariela , Isla María I , Verdenelli Romina A , Meriles José M , Maestri Damián
Published: July 2011

BACKGROUND: In the last few years, agricultural uses of waste waters from olive processing‐related industries have been gaining interest mainly with a view to composting or bio‐fertilizers. The present work examines physico‐chemical, toxicological and geno‐toxicological properties of three liquid wastes, namely olive mill wastewater (OMWW), olive wet husk and olive brine. The effect of OMWW spreading on soil microbial activity and biomass was also evaluated.

RESULTS: Data from Artemia salina and Lactuca sativa toxicity tests indicated high levels of lethality, and inhibitory effects on seed germination and seedling growth of all olive wastes. The genotoxicity assays using Allium cepa tests showed contrasting results. At high concentrations, olive wastes caused inhibition or suppression of mitosis. However, they did not produce induced anaphase aberrations. Data on reversion of Salmonella thyphimurium strains using the Ames test indicated that the olive wastes did not present mutagenic activity. Results from the field experiment showed that OMWW at a 500 m3 ha−1 had the highest values of both soil microbial activity and biomass after 3 months of the amendment application.

CONCLUSION: This work adds new data for environmental risk assessment of olive industrial wastes. Direct use of olive wastes for agricultural purposes should be limited owing to their possible chemotoxic, phytotoxic and antimicrobial effects. Copyright © 2011 Society of Chemical Industry
Effects of freeze‐dried Mitsuokella jalaludinii culture and Natuphos® phytase supplementation on the performance and nutrient utilisation of broiler chickens
Authors: Lan Ganqiu , Abdullah Norhani , Jalaludin Syed , Ho Yin Wan
Published: July 2011

BACKGROUND: Phytate‐bound phosphorus (P) in poultry diets is poorly available to chickens. Hence exogenous phytase is often added to their diets. Mitsuokella jalaludinii is a rumen bacterial species that produces high phytase activity. In this study the effects of freeze‐dried active M. jalaludinii culture (FD‐AMJC) and Natuphos® phytase (phytase N) supplementations on the growth performance and nutrient utilisation of broiler chickens fed a low‐available P (aP) diet were evaluated.

RESULTS: Supplementation of FD‐AMJC or phytase N to the low‐aP diet improved the feed intake, feed conversion rate, body weight gain, dry matter (DM) digestibility and P, Ca and Mn retention, increased the tibia bone ash content, Ca and P concentrations in tibia DM and P and Zn concentrations in plasma and reduced the P excretion of broiler chickens. However, the feed conversion rate, P and Ca retention, DM digestibility and reduction of P excretion were better with FD‐AMJC than phytase N supplementation. Supplementation of FD‐AMJC to the low‐aP diet also improved the apparent metabolisable energy value of the diet, Cu and Zn retention and crude protein digestibility, but phytase N supplementation did not.

CONCLUSION: FD‐AMJC supplementation was more efficient in improving nutrient utilisation and reducing P excretion in chickens than phytase N supplementation. Copyright © 2011 Society of Chemical Industry
Method validation for the determination of propellant components by Soxhlet extraction and gas chromatography/mass spectrometry
Authors: Fryš Ondřej , Bajerová Petra , Eisner Aleš , Mudruňková Martina , Ventura Karel
Published: July 2011

A fast, sensitive, and accurate GC/MS method for the quantification of aliphatic nitroesters (ethylene glycol dinitrate, nitroglycerin, and triethylene glycol dinitrate) and aromatic amines (diphenylamine, 2‐nitrodiphenylamine, and triphenylamine) in propellants was developed and validated. This method comprises a Soxhlet extraction step with dichloromethane, followed by separation on a capillary column MDN‐5. Ionization of the analytes is carried out using electron ionization. The limit of quantification of the method was 1% w/w for aliphatic nitroesters and 0.1% w/w for aromatic amines (diphenylamine and triphenylamine). Values of repeatability and reproducibility for analyzed compounds were smaller than values of the maximum allowed tolerances of the Horwitz‐equation RSDmax and 2/3 RSDmax. Values of accuracy for selected compounds were below the acceptable threshold of 15% for all tested levels in the range of calibration curve excepting the lowest concentration of calibration curve for nitroglycerin and aromatic amines. During the validation of method, temperature instability in injection port of gas chromatograph and column was observed for 2‐nitrodiphenylamine. Hence, it follows worse results of accuracy and linearity and 2‐nitrodiphenylamine was not validated successfully.
Hot-topic debate on kidney function: Renal-sparing approaches are beneficial
Authors: Durand François
Published: July 2011

Key Points

1. Renal function is frequently compromised in candidates for transplantation with advanced cirrhosis. These patients frequently have chronic and irreversible kidney changes at the time of transplantation.

2. The accumulated incidence of chronic renal failure is high in liver transplant recipients. Chronic renal failure has a deleterious impact on the outcome.

3. Calcineurin inhibitor (CNI)–based immunosuppression is highly effective at preventing rejection. However, CNI nephrotoxicity has a central role in the occurrence of chronic renal failure.

4. Renal function impairment frequently occurs within the first year after transplantation. Once renal function is significantly impaired [glomerular filtration rate (GFR) < 60 mL/minute/1.73 m2], any intervention is unlikely to result in a return to normal renal function. Early interventions are needed to prevent chronic and irreversible kidney injury.

5. De novo CNI minimization has been proven to be effective at reducing the rate of impaired renal function after transplantation. The reduction in the CNI doses should be offset by the addition of mycophenolate mofetil or enteric-coated mycophenolate sodium.

6. Delayed CNI minimization in patients with established renal insufficiency may result in a significant improvement in the GFR, even though the increase in the GFR after minimization is generally modest.

7. Mammalian target of rapamycin (mTOR) inhibitors are considered nonnephrotoxic immunosuppressive agents. They may be an option for improving renal function in liver transplant recipients. However, not all patients with renal dysfunction benefit from a switch to mTOR inhibitors. In addition, the benefits in terms of renal function should be balanced against specific side effects.

8. New immunosuppressive agents without intrinsic nephrotoxicity are currently under development for solid organ transplantation. These agents could help to reduce the burden of impaired renal function in transplantation in the near future.
Pain in patients with myotonic dystrophy type 2: A postal survey in finland
Authors: Suokas Kimmo I. , Haanpää Maija , Kautiainen Hannu , Udd Bjarne , Hietaharju Aki J.
Published: July 2011

Introduction:

Widespread musculoskeletal pain is a well‐known symptom of myotonic dystrophy type 2 (DM2), but so far it has been addressed in only a few studies.

Methods:

A postal survey for all traceable DM2 patients (n = 132) was conducted. A specific questionnaire, and severity and interference subscales of the Brief Pain Inventory, quality of life (RAND‐36), and modified Beck Depression Inventory were completed.

Results:

The response rate was 70%. The mean age of respondents was 53 years, 59% of whom were women. Current pain was reported by 54%. Lifetime prevalence of pain was 76%. The mean intensity of pain at its highest in the last week was 5.9, and 2.3 at its lowest (on a numerical rating scale of 0–10). Quality of life was lower in DM2 patients who reported pain. In 18%, the depression score was noticeably different.

Conclusions:

Pain of moderate severity and unpleasant muscular symptoms are common in DM2. DM2 should be taken into consideration in the differential diagnosis of musculoskeletal pain. Muscle Nerve 45: 70–74, 2012
NMDA receptors at the endplate of rat skeletal muscles: Precise postsynaptic localization
Authors: Malomouzh Artem I. , Nurullin Leniz F. , Arkhipova Svetlana S. , Nikolsky Evgeny E.
Published: July 2011

In this study we demonstrate expression of the N‐methyl‐D‐aspartate receptor NR1 subunit in the rat neuromuscular junction of skeletal muscles of different functional types (extensor digitorum longus, soleus, and diaphragm muscles) using fluorescence immunocytochemistry. Electron microscopic immunocytochemistry has shown that the NR1 subunit is localized solely on the sarcolemma in the depths of the postsynaptic folds. These findings suggest participation of the glutamatergic signaling system in functioning of the adult mammalian neuromuscular junction. Muscle Nerve, 2011
Classification of single‐voxel 1H spectra of brain tumours using LCModel
Authors: Raschke F. , Fuster‐Garcia E. , Opstad K. S. , Howe F. A.
Published: July 2011

This study presents a novel method for the direct classification of 1H single‐voxel MR brain tumour spectra using the widespread analysis tool LCModel. LCModel is designed to estimate individual metabolite proportions by fitting a linear combination of in vitro metabolite spectra to an in vivo MR spectrum. In this study, it is used to fit representations of complete tumour spectra and to perform a classification according to the highest estimated tissue proportion. Each tumour type is represented by two spectra, a mean component and a variability term, as calculated using a principal component analysis of a training dataset. In the same manner, a mean component and a variability term for normal white matter are also added into the analysis to allow a mixed tissue approach. An unbiased evaluation of the method is carried out through the automatic selection of training and test sets using the Kennard and Stone algorithm, and a comparison of LCModel classification results with those of the INTERPRET Decision Support System (IDSS) which incorporates an advanced pattern recognition method. In a test set of 46 spectra comprising glioblastoma multiforme, low‐grade gliomas and meningiomas, LCModel gives a classification accuracy of 90% compared with an accuracy of 95% by IDSS. Copyright © 2011 John Wiley & Sons, Ltd.
Frequency, Risk‐Factors and Survival of Children With Atypical Teratoid Rhabdoid Tumors (AT/RT) of the CNS Diagnosed between 1988 and 2004, and Registered to the German HIT Database
Authors: von Hoff Katja , Hinkes Bernward , Dannenmann‐Stern Elke , von Bueren André O. , Warmuth‐Metz Monika , Soerensen Niels , Emser Angela , Zwiener Isabella , Schlegel Paul G. , Kuehl Joachim , Frühwald Michael C. , Kortmann Rolf D. , Pietsch Torsten , Rutkowski Stefan
Published: July 2011

Purpose

To analyze the frequency, prognostic factors, and outcome of children with atypical teratoid/rhabdoid tumors (AT/RT), a rare and highly malignant embryonal brain tumor.

Materials and Methods

Clinical data of patients diagnosed between 1988 and 2004 with AT/RT who were registered to the German HIT trial center, were correlated with outcome. Patient numbers for AT/RT were compared to numbers for primitive neuroectodermal tumors and medulloblastomas (PNET/MB) registered to the population‐based HIT trials.

Results

We identified 56 patients with the centrally confirmed histopathological diagnosis of AT/RT with a median age of 1.2 years (range, 0.1–14.0 years). The AT/RT:PNET/MB ratio was 1:12.2 for all children, and 1:1.5 for children younger than 1 year at diagnosis. Three‐year overall survival (OS) and event‐free survival (EFS) for all patients were 22% and 13%, respectively. Eight patients (14%) are considered long‐term event‐free survivors (follow‐up 1.4–10.6 years). By univariable analyses, younger age, metastatic disease, infratentorial location, and less than complete remission at the end of chemotherapy were identified as negative influencing factors for OS. By multivariable analyses, younger age (OS, EFS) and metastatic disease (OS) were identified as independent risk factors.

Conclusion

The incidence of AT/RT in children below 1 year is higher than previously reported. A subset of patients with favorable clinical risk factors profits from intensive multimodal treatment. Prospective clinical and biological studies are needed to further define prognostic factors and optimize therapy. Pediatr Blood Cancer 2011; 57: 978–985. © 2011 Wiley‐Liss, Inc.
Young adults with SCD in US children's hospitals: Are they different from adolescents?
Authors: Dickerson Amanda K. , Klima Jennifer , Rhodes Melissa M. , O'Brien Sarah H.
Published: July 2011

Background

The period of transition from pediatric to adult care is a vulnerable time for patients with sickle cell disease (SCD). The optimal time for transition is unknown and there is no standard of care regarding this timing in the United States.

Procedures

We collected administrative data from the Pediatric Health Information System for all SCD admissions from 2000 to 2009. We compared reasons for hospitalization and resulting charges in adolescents (13–17 years) and young adults (18–21 years).

Results

We identified 25,371 admissions of adolescents (n = 18,299) and young adults (n = 7,072) with SCD. Median admissions per patient per year was higher in young adults (0.6) compared to adolescents (0.2, P < 0.001), but reasons for hospitalization were similar between the two age groups. Complications of adult SCD such as nephropathy and pulmonary hypertension were rare (<2.5% of discharges) but more frequent in older patients (P = 0.001). Although length of stay was similar between the two groups (median = 4 days), young adults tended to incur higher charges (median +$1,314, P < 0.001) and were less likely to utilize private insurance (P < 0.001). Deaths (0.2% of admissions) were rare and similar across age groups (P = 0.7).

Conclusion

In a national sample of US children's hospitals, adolescents (13–17 years) and young adults (18–21 years) with SCD had similar reasons for hospitalization and low mortality. Further studies are needed to investigate whether extending the age of transition to ≥21 years as a national standard may decrease morbidity and mortality, improve health‐related quality of life, or increase readiness for transition in patients with SCD. Pediatr Blood Cancer 2012; 58: 741–745. © 2011 Wiley Periodicals, Inc.
Treatment of relapsed acute lymphoblastic leukemia: Approaches used by pediatric oncologists and bone marrow transplant physicians
Authors: Burke Michael J. , Lindgen Bruce , Verneris Michael R.
Published: July 2011

Background

Management of relapsed B‐precursor acute lymphoblastic leukemia (ALL) is challenging and varied. We hypothesized that treatment approaches differ between pediatric oncologists and bone marrow transplant (BMT) physicians.

Procedure

A survey addressing management of relapsed ALL was sent to pediatric oncologists (n = 883) and BMT (n = 86) physicians across North America.

Results

A number of similarities in treatment approaches were identified including: choice of chemotherapy for re‐induction/consolidation, preference for unrelated donors (URDs) in very early marrow relapse and the choice to not use URD donors in late marrow relapse. However, differences between the two disciplines were noted. For patients who relapsed 18–36 months from diagnosis, the majority of oncologists (53.7%) would retreat with chemotherapy while a majority BMT physicians (70.3%) recommended URD transplant (P < 0.001). Oncologists were also less likely to use minimal residual disease (MRD) in relapse assessment compared to BMT physicians (52% vs. 67.2%; P = 0.028) and more oncologists believed MRD testing was experimental and/or not proven in relapsed ALL (27.1% vs. 12.3%; P = 0.011).

Conclusions

This study highlights management differences in children with ALL between pediatric oncologists and BMT physicians, identifying opportunities for collaborative clinical trials. Pediatr Blood Cancer 2012; 58: 840–845. © 2011 Wiley Periodicals, Inc.
Anti‐tumor activity of the HSP90 inhibitor SNX‐2112 in pediatric cancer cell lines
Authors: Chinn Danielle C. , Holland William S. , Yoon Janet M. , Zwerdling Theodore , Mack Philip C.
Published: July 2011

Background

HSP90 plays a central role in stabilizing client proteins involved in malignant processes. SNX‐2112 is an orally administered potent HSP90 inhibitor that has demonstrated pre‐clinical anti‐tumor activity in adult malignancies. As many childhood tumors depend upon HSP90 client proteins, we sought to test the pre‐clinical efficacy of SNX‐2112 in a panel of pediatric cancer cell lines both as a single‐agent and in combination with cisplatin (CP).

Procedure

Eight cell lines (from osteosarcoma, neuroblastoma, hepatoblastoma, and lymphoma) were studied. Short‐ and long‐term effects of SNX‐2112 were assessed by MTT and clonogenic assays. Cell cycling was measured using flow cytometry. Status of HSC70, HSP72, AKT1, C‐Raf, and PARP was assessed by immunoblotting. Efficacy of SNX‐2112 in combination with CP was assessed using median‐effect analysis.

Results

Cell lines studied demonstrated sensitivity to SNX‐2112 with IC50 values ranging from 10–100 nM. Low dose treatments (12 nM) resulted in a cytostatic response with a minimal increase in sub‐G1 content. A higher dose (70 nM) exhibited a more prolonged inhibition and larger sub‐G1 accumulation. Observed levels of AKT1 and C‐Raf were markedly reduced over time along with an increase in PARP cleavage. In concurrently administered combination treatments, SNX‐2112 and CP synergistically inhibited cell growth.

Conclusions

SNX‐2112 showed marked single‐agent activity in pediatric cancer cell lines with downstream effects on HSP90 client proteins. The combination of SNX‐2112 and CP showed synergistic activity in two cell lines tested. Further studies of HSP90 inhibitors such as SNX‐2112 as a single agent or in combination with chemotherapy are warranted in pediatric cancer. Pediatr Blood Cancer 2012; 58: 885–890. © 2011 Wiley Periodicals, Inc.
Hearing loss among survivors of childhood brain tumors treated with an irradiation‐sparing approach
Authors: Orgel Etan , Jain Shveta , Ji Lingyun , Pollick Liora , Si Stephanie , Finlay Jonathan , Freyer David R.
Published: July 2011

Background

Intensive, irradiation‐sparing chemotherapy regimens for malignant brain tumors have improved survival and neurocognitive outcomes in very young children. Platinum compounds are pivotal to this approach's success but are associated with hearing loss that markedly reduces quality of life for survivors. The purpose of this study was to determine the prevalence and severity of ototoxicity associated with Head Start and similar irradiation‐sparing regimens.

Procedure

A retrospective cohort study was conducted of children treated for malignant brain tumors at Children's Hospital Los Angeles using irradiation‐sparing regimens. Patient and treatment characteristics were ascertained. Primary outcomes were post‐treatment hearing status, need for hearing aids, and hearing threshold change.

Results

Twenty‐nine patients were evaluable. The most common diagnosis was medulloblastoma (n = 14). The median age at diagnosis was 2.0 years (range, 0.2–9.2). Median time from diagnosis to most recent hearing assessment was 1.1 years (mean 2.4; range, 0.2–17.5). Cumulative cisplatin and carboplatin dose was 281 ± 88 mg/m2 and 1,205 ± 277 mg/m2, respectively. All patients had aminoglycoside exposure. Following treatment, 18 patients (62.1%) had abnormal hearing and 11 (37.9%) required hearing aids. At 4,000 Hz, statistically significant hearing loss was documented in the range of 30–40 dB.

Conclusions

While eliminating cranial irradiation has dramatically improved survival and neurocognitive and neuroendocrine outcomes in this population, clinically significant hearing loss is now the leading late effect due to the necessity of platinum‐based chemotherapy. Our results document the need for audiometric monitoring and developing otoprotective strategies in this vulnerable population. Pediatr Blood Cancer 2012; 58: 953–958. © 2011 Wiley Periodicals, Inc.
Using MS‐MLPA as an efficient screening tool for detecting 9p21 abnormalities in pediatric acute lymphoblastic leukemia
Authors: Gardiner R. Brett , Morash Barbara A. , Riddell Christie , Wang Hao , Fernandez Conrad V. , Yhap Margaret , Berman Jason N.
Published: July 2011

Background

Characterization of recurrent genetic lesions in childhood acute lymphoblastic leukemia (ALL) has enabled therapeutic stratification with improved outcomes. The tumor suppressor genes, CDKN2A and CDKN2B, encoding p16INK4a, p14ARF, and p15INK4b have been localized to 9p21. Abnormalities of 9p21 have been reported in 10–30% of childhood ALL using conventional cytogenetics and fluorescence in situ hybridization (FISH). The incidence of 9p21 using more sensitive techniques, such as methylation specific multiplex ligation‐dependent probe amplification (MS‐MLPA), remains uncertain, and thus also the prognostic significance.

Procedure

We investigated the incidence and prognostic importance of 9p21 abnormalities in pediatric ALL patients using MS‐MLPA and compared these results to FISH.

Results

In total, MS‐MLPA or FISH detected aberrations (both dosage and methylation abnormalities) at 9p21 in a remarkable 32/48 (67%) patients in contrast to a much lower rate of only 8% of patients identified to have deletions by standard G banding cytogenetics. MS‐MLPA identified five deletions not found by FISH. Aberrant methylation at CDKN2B was found in 19 (46%) patients. 9p21 abnormalities were associated with National Cancer Institute (NCI) high‐risk criteria (P = 0.04) and were present in all five patients with T‐cell disease. Four pre‐B‐cell ALL patients relapsed, three of whom had prior 9p21 abnormalities.

Conclusions

MS‐MLPA had a higher detection rate for 9p21 abnormalities than previously reported for other techniques. Given the ease of processing, minimal equipment and low cost of MS‐MLPA, our results suggest that previous reports may have underestimated the true frequency of 9p21 abnormalities and their potential impact upon ALL outcome. Pediatr Blood Cancer 2012; 58: 852–859. © 2011 Wiley Periodicals, Inc.
Understanding Heritability: What it is and What it is Not
Authors: Johnson Wendy , Penke Lars , Spinath Frank M.
Published: July 2011

Commentators generally found our exposition of the concept of heritability helpful for psychologists, suggesting that we largely accomplished our primary goal. Many provided supplemental and helpful perspectives on concepts we addressed. A few of the comments indicated that we may not have been completely successful in making clear our secondary goal, which was to outline how heritability estimates confound a plethora of influences. In this response, we thus emphasize that we do not claim that specific kinds of complexity, or, even worse, intractable complexity, pervade the genetics of behavioural traits. Rather, our claim is that genetics is riddled with complexity of many degrees and kinds, and heritability is a poor indicator of either degree or kind of underlying genetic complexity. Copyright © 2011 John Wiley & Sons, Ltd.
Creating a human brain proteome atlas – 14th HUPO BPP workshop September 20–21, 2010, Sydney, Australia
Authors: Gröttrup Bernd , Marcus Katrin , Grinberg Lea T. , Lee Sang K. , Meyer Helmut E. , Park Young M.
Published: July 2011

The HUPO Brain Proteome Project (HUPO BPP) held its 14th workshop during the HUPO 9th Annual World Congress in Sydney, Australia. The principal aim of this project is to discover prognostic and diagnostic biomarkers associated with neurodegenerative diseases and brain aging, with the ultimate objective of obtaining a better understanding of these conditions and creating roads for the development of novel diagnostic techniques and effective treatments. The attendees came together to discuss progress in the human clinical neuroproteomics and to define the needs and guidelines required for more advanced proteomics approaches.
Structure of a novel thermostable GH51 α‐L‐arabinofuranosidase from Thermotoga petrophila RKU‐1
Authors: Souza Tatiana A.C.B. , Santos Camila R. , Souza Angelica R. , Oldiges Daiane P. , Ruller Roberto , Prade Rolf A. , Squina Fabio M. , Murakami Mario T.
Published: July 2011

α‐L‐arabinofuranosidases (EC 3.2.1.55) participate in the degradation of a variety of L‐arabinose‐containing polysaccharides and interact synergistically with other hemicellulases in the production of oligosaccharides and bioconversion of lignocellulosic biomass into biofuels. In this work, the structure of a novel thermostable family 51 (GH51) α‐L‐arabinofuranosidase from Thermotoga petrophila RKU‐1 (TpAraF) was determined at 3.1 Å resolution. The TpAraF tertiary structure consists of an (α/β)‐barrel catalytic core associated with a C‐terminal β‐sandwich domain, which is stabilized by hydrophobic contacts. In contrast to other structurally characterized GH51 AraFs, the accessory domain of TpAraF is intimately linked to the active site by a long β‐hairpin motif, which modifies the catalytic cavity in shape and volume. Sequence and structural analyses indicate that this motif is unique to Thermotoga AraFs. Small angle X‐ray scattering investigation showed that TpAraF assembles as a hexamer in solution and is preserved at the optimum catalytic temperature, 65°C, suggesting functional significance. Crystal packing analysis shows that the biological hexamer encompasses a dimer of trimers and the multiple oligomeric interfaces are predominantly fashioned by polar and electrostatic contacts.