bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of telaprevir on the pharmacokinetics of cyclosporine and tacrolimus
Authors: Garg Varun , van Heeswijk Rolf , Eun Lee Jee , Alves Katia , Nadkarni Priya , Luo Xia
Published: May 2011

Abstract:

The hepatitis C virus protease inhibitor telaprevir is an inhibitor of the enzyme cytochrome P450 3A, responsible for the metabolism of both cyclosporine and tacrolimus. This Phase I, open-label, nonrandomized, single-sequence study assessed the effect of telaprevir coadministration on the pharmacokinetics of a single dose of either cyclosporine or tacrolimus in two separate panels of 10 healthy volunteers each. In Part A, cyclosporine was administered alone as a single 100-mg oral dose, followed by a minimum 8-day washout period, and subsequent coadministration of a single 10-mg oral dose of cyclosporine with either a single dose of telaprevir (750 mg) or with steady-state telaprevir (750 mg every 8 hours [q8h]). In Part B, tacrolimus was administered alone as a single 2-mg oral dose, followed by a minimum 14-day washout period, and subsequent coadministration of a single 0.5-mg dose of tacrolimus with steady-state telaprevir (750 mg q8h). Coadministration with steady-state telaprevir increased cyclosporine dose-normalized (DN) exposure (DN_AUC0-∞) by approximately 4.6-fold and increased tacrolimus DN_AUC0-∞ by approximately 70-fold. Coadministration with telaprevir increased the terminal elimination half-life (t½) of cyclosporine from a mean (standard deviation [SD]) of 12 (1.67) hours to 42.1 (11.3) hours and t½ of tacrolimus from a mean (SD) of 40.7 (5.85) hours to 196 (159) hours. Conclusion: In this study, telaprevir increased the blood concentrations of both cyclosporine and tacrolimus significantly, which could lead to serious or life-threatening adverse events. Telaprevir has not been studied in organ transplant patients; its use in these patients is not recommended because the required studies have not been completed to understand appropriate dose adjustments needed for safe coadministration of telaprevir with cyclosporine or tacrolimus, and regulatory approval has not been obtained. (HEPATOLOGY 2011;)
c-Met represents a potential therapeutic target for personalized treatment in hepatocellular carcinoma
Authors: You Hanning , Ding Wei , Dang Hien , Jiang Yixing , Rountree Bart C.
Published: May 2011

c-Met, a high-affinity receptor for hepatocyte growth factor (HGF), plays a critical role in cancer growth, invasion, and metastasis. Hepatocellular carcinoma (HCC) patients with an active HGF/c-Met signaling pathway have a significantly worse prognosis. Although targeting the HGF/c-Met pathway has been proposed for the treatment of multiple cancers, the effect of c-Met inhibition in HCC remains unclear. The human HCC cell lines Huh7, Hep3B, MHCC97-L, and MHCC97-H were used in this study to investigate the effect of c-Met inhibition using the small molecule selective c-Met tyrosine kinase inhibitor PHA665752. MHCC97-L and MHCC97-H cells demonstrate a mesenchymal phenotype with decreased expression of E-cadherin and increased expression of c-Met, fibronectin, and

Zeb2

compared with Huh7 and Hep3B cells, which have an epithelial phenotype. PHA665752 treatment blocked phosphorylation of c-Met and downstream phosphoinositide 3-kinase/Akt and mitogen-activated protein kinase/Erk pathways, inhibited cell proliferation, and induced apoptosis in c-Met–positive MHCC97-L and MHCC97-H cells. In xenograft models, administration of PHA665752 significantly inhibited c-Met–positive MHCC97-L and MHCC97-H tumor growth, and PHA665752-treated tumors demonstrated marked reduction of both c-Met phosphorylation and cell proliferation. c-Met–negative Huh7 and Hep3B cells were not affected by c-Met inhibitor treatment

in vitro

or

in vivo.

In addition, c-Met–positive MHCC97-L and MHCC97-H cells demonstrated cancer stem cell–like characteristics, such as resistance to chemotherapy, tumor sphere formation, and increased expression of

CD44

and

ABCG2,

and PHA665752 treatment suppressed tumor sphere formation and inhibited CD44 expression.

Conclusion:

c-Met represents a potential target of personalized treatment for HCC with an active HGF/c-Met pathway. (HEPATOLOGY 2011;)
Diagnostic accuracy and reliability of ultrasonography for the detection of fatty liver: A meta-analysis
Authors: Hernaez Ruben , Lazo Mariana , Bonekamp Susanne , Kamel Ihab , Brancati Frederick L. , Guallar Eliseo , Clark Jeanne M.
Published: May 2011

Ultrasonography is a widely accessible imaging technique for the detection of fatty liver, but the reported accuracy and reliability have been inconsistent across studies. We aimed to perform a systematic review and meta-analysis of the diagnostic accuracy and reliability of ultrasonography for the detection of fatty liver. We used MEDLINE and Embase from October 1967 to March 2010. Studies that provided cross-tabulations of ultrasonography versus histology or standard imaging techniques, or that provided reliability data for ultrasonography, were included. Study variables were independently abstracted by three reviewers and double checked by one reviewer. Forty-nine (4720 participants) studies were included for the meta-analysis of diagnostic accuracy. The overall sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio of ultrasound for the detection of moderate-severe fatty liver, compared to histology (gold standard), were 84.8% (95% confidence interval: 79.5-88.9), 93.6% (87.2-97.0), 13.3 (6.4-27.6), and 0.16 (0.12-0.22), respectively. The area under the summary receiving operating characteristics curve was 0.93 (0.91-0.95). Reliability of ultrasound for the detection of fatty liver showed kappa statistics ranging from 0.54 to 0.92 for intrarater reliability and from 0.44 to 1.00 for interrater reliability. Sensitivity and specificity of ultrasound was similar to that of other imaging techniques (i.e., computed tomography or magnetic resonance imaging). Statistical heterogeneity was present even after stratification for multiple clinically relevant characteristics.

Conclusion:

Ultrasonography allows for reliable and accurate detection of moderate-severe fatty liver, compared to histology. Because of its low cost, safety, and accessibility, ultrasound is likely the imaging technique of choice for screening for fatty liver in clinical and population settings. (HEPATOLOGY 2011; 54:1082–1090)
Nuclear expression of S100A4 calcium-binding protein increases cholangiocarcinoma invasiveness and metastasization
Authors: Fabris Luca , Cadamuro Massimiliano , Moserle Lidia , Dziura James , Cong Xiangyu , Sambado Luisa , Nardo Giorgia , Sonzogni Aurelio , Colledan Michele , Furlanetto Alberto , Bassi Nicolò , Massani Marco , Cillo Umberto , Mescoli Claudia , Indraccolo Stefano , Rugge Massimo , Okolicsanyi Lajos , Strazzabosco Mario
Published: May 2011

Cholangiocarcinoma (CCA) carries a severe prognosis because of its strong invasiveness and early metastasization. In several patients, otherwise eligible for surgical resection, micrometastasis are already present at the time of surgery. The mechanisms responsible for CCA invasiveness are unclear. S100A4, a member of the S100 family of small Ca2+-binding proteins, is expressed in mesenchymal cells, regulates cell motility in several cell types, and is expressed in some epithelial cancers. Thus, we aimed to study the role of S100A4 in CCA invasiveness and metastasization. The expression of S100A4 was studied by immunohistochemistry in 93 human liver samples of CCA patients undergoing surgical resection and correlated with metastases development (67 cases) and patient survival following surgery using log rank tests and multivariate analysis. S100A4 expression was studied in EGI-1 and TFK-1, human CCA cell lines with and without nuclear S100A4 expression, respectively. Metastatic properties of CCA cells were assessed by xenotransplantation in severe combined immunodeficiency (SCID) mice after transduction with lentiviral vectors encoding firefly luciferase gene. Proliferation, motility (wound healing), invasiveness (Boyden chamber), and metalloproteinases (MMPs) secretion were studied in CCA cells, with or without lentiviral silencing of S100A4. Nuclear expression of S100A4 by neoplastic ducts was a strong predictor of metastasization and reduced survival after resection

(P

< 0.01). EGI-1 CCA cells showed stronger metastatic properties than TFK-1 when xenotransplanted in SCID mice. S100A4-silenced EGI-1 cells showed significantly reduced motility, invasiveness, and MMP-9 secretion

in vitro,

without changes in cell proliferation.

Conclusion:

Nuclear S100A4 identifies a subset of CCA patients with a poor prognosis after surgical resection. Nuclear expression of S100A4 increases CCA cells invasiveness and metastasization, indicating that S100A4 may also represent a potential therapeutic target. (HEPATOLOGY 2011; 54:890–899)
Direct numerical simulation of multiphase flows involving dispersed components with deformable interfaces
Authors: Kajishima Takeo , Takeuchi Shintaro
Published: May 2011

Recent developments in direct numerical simulation of dispersed multiphase flows are reviewed. A current trend is to use a fixed Cartesian grid, as it is suitable for dealing with multiple solids, bubbles, and/or droplets (dispersed components). The features of several typical methods for gas–liquid interfaces and fluid–solid boundaries are discussed. For gas–liquid interfaces, we categorize the existing techniques into fitting, tracking, capturing, and indicator‐transportation methods in order to clarify the characteristics of the methods. For solid objects moving in a fluid, on the other hand, methods on a fixed Cartesian grid are highlighted. Our primary interest is in the conservation properties of the algorithms. From this point of view, we show a combination of our conservative momentum exchange (CMX) algorithm for immersed solid objects and volume‐of‐fluid (VOF) method for gas–liquid interface. This combination enables simulation of interaction problems between particles and bubbles of comparable sizes (three‐phase flows). As for interaction problems of a deformable solid object with a fluid, we demonstrate the successful coupling of CMX and a finite element method. © 2011 Wiley Periodicals, Inc. Heat Trans Asian Res; Published online in Wiley Online Library (wileyonlinelibrary.com/journal/htj). DOI 10.1002/htj.20345
Dual inhibition of phosphatidylinositol 3‐kinase/Akt and mammalian target of rapamycin signaling in human nonsmall cell lung cancer cells by a dietary flavonoid fisetin
Authors: Khan Naghma , Afaq Farrukh , Khusro Fatima H. , Mustafa Adhami Vaqar , Suh Yewseok , Mukhtar Hasan
Published: May 2011

Lung cancer is one of the most commonly occurring malignancies. It has been reported that mammalian target of rapamycin (mTOR) is phosphorylated in lung cancer and its activation was more frequent in tumors with overexpression of phosphatidylinositol 3‐kinase (PI3K)/Akt. Therefore, dual inhibitors of PI3K/Akt and mTOR signaling could be valuable agents for treating lung cancer. In the present study, we show that fisetin, a dietary tetrahydroxyflavone inhibits cell growth with the concomitant suppression of PI3K/Akt and mTOR signaling in human nonsmall cell lung cancer (NSCLC) cells. Using autodock 4, we found that fisetin physically interacts with the mTOR complex at two sites. Fisetin treatment was also found to reduce the formation of A549 cell colonies in a dose‐dependent manner. Treatment of cells with fisetin caused decrease in the protein expression of PI3K (p85 and p110), inhibition of phosphorylation of Akt, mTOR, p70S6K1, eIF‐4E and 4E‐BP1. Fisetin‐treated cells also exhibited dose‐dependent inhibition of the constituents of mTOR signaling complex such as Rictor, Raptor, GβL and PRAS40. There was an increase in the phosphorylation of AMPKα and a decrease in the phosphorylation of TSC2 on treatment of cells with fisetin. We also found that treatment of cells with mTOR inhibitor rapamycin and mTOR‐siRNA caused decrease in phosphorylation of mTOR and its target proteins which were further downregulated on treatment with fisetin, suggesting that these effects are mediated in part, through mTOR signaling. Our results show that fisetin suppressed PI3K/Akt and mTOR signaling in NSCLC cells and thus, could be developed as a chemotherapeutic agent against human lung cancer.
Pathological complete remission in patients with oesophagogastric cancer receiving preoperative 5‐fluorouracil, oxaliplatin and docetaxel
Authors: Homann Nils , Pauligk Claudia , Luley Kim , Werner Kraus Thomas , Bruch Hans‐Peter , Atmaca Akin , Noack Frank , Altmannsberger Hans‐Michael , Jäger Elke , Al‐Batran Salah‐Eddin
Published: May 2011

The aim of this study was to determine the pathological complete remission (pCR) rate, and its relationship to clinical outcome, in patients with adenocarcinoma of the stomach or oesophagogastric junction receiving preoperative 5‐fluorouracil, leucovorin, oxaliplatin and docetaxel (FLOT) every 2 weeks. Data from these patients who received at least one cycle of preoperative FLOT followed by surgery were prospectively collected in three German centres. Outcome analyses were conducted and tumour samples were evaluated for pathological remission by a central pathologist. A total of 46 patients were included in this analysis. All patients had clinical T3‐ and/or N+‐stages and 11 (23.9%) had distant metastases (M1). After a median of 4 (range 2–8) preoperative cycles, 8 of 46 patients (17.4%) achieved a pCR. The pCR rate was highest in tumours of intestinal type histology (30.8%) and in those located in the oesophagogastric junction (30.4%) and lowest in patients with diffuse/mixed type tumours (0%) or tumours located in the stomach (4.3%; p < 0.05 for both comparisons). Patients with pCR had 100% probability of overall and disease‐free survival (DFS) during the observation period, which was significantly higher (p = 0.037 and p = 0.009, respectively) than the survival probability in patients without pCR. In conclusion, treatment intensification using FLOT was associated with significant pCR rates in patients with oesophagogastric cancer. The distribution of pCR appeared to be significantly different according to histological type and location of the tumours.
Lymph node CEA and MUC2 mRNA as useful predictors of outcome in colorectal cancer
Authors: Ohlsson Lina , Israelsson Anne , Öberg Åke , Palmqvist Richard , Stenlund Hans , Hammarström Marie‐Louise , Hammarström Sten , Lindmark Gudrun
Published: May 2011

The aim was to explore the utility for staging and prognostic impact of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20), guanylyl cyclase C (GCC), CUB (complement protein subcomponents C1r/C1s, urchin embryonic growth factor, and bone morphogenic protein 1) containing domain protein 1 (CDCP1) and mucin 2 (MUC2) mRNA levels in mesenteric lymph nodes of colorectal cancer (CRC) patients. Lymph nodes were collected at surgery and bisected; one half was subjected to biomarker mRNA analysis using real‐time quantitative RT‐PCR and the other half to routine histopathology. Lymph nodes from 174 CRC patients and 24 controls were analyzed. The median follow‐up time was 59 (range 17–131) months. Cut‐off levels were defined by analyzing quintiles by Cox regression model. CEA mRNA showed the best discriminating power between patients with recurrence in CRC after surgery and patients who were apparently disease‐free (p = 0.015). The risk of recurrence for the CEA(+) patients was 4.6 times greater than for the CEA(−) patients (p < 0.0001). The other biomarkers gave lower hazard ratios. Cumulative survival analysis demonstrated that the average survival time was 99 months for CEA(−) patients compared to 39 months for CEA(+) patients, a difference of 60 months (p < 0.0001). Six to nine percent of the Stage I and Stage II patients [H&E(−)] had CEA(+), CK20(+), GCC(+) and/or MUC2(+) lymph nodes. Two of these patients died from recurrent CRC. Low lymph node MUC2/CEA mRNA ratio identified patients with high risk for recurrence (p = 0.011). Thus, quantitative reverse transcriptase‐polymerase chain reaction of CEA mRNA is a sensitive method to identify tumor cells in lymph nodes of CRC patients and, in combination with MUC2 mRNA, allows improved prediction of clinical outcome.
Cisplatin increases B‐cell‐lymphoma‐2 expression via activation of protein kinase C and Akt2 in endometrial cancer cells
Authors: Rouette Alexandre , Parent Sophie , Girouard Julie , Leblanc Valérie , Asselin Eric
Published: May 2011

Human carcinomas often show resistance to cisplatin and Bcl‐2 is associated with resistance to cisplatin. However, Bcl‐2 regulation on cisplatin treatment in human cancers is unknown. Here, we show a novel mechanism by which cisplatin treatment promotes resistance by increasing the expression of Bcl‐2 mRNA. Bcl‐2 mRNA and protein expression was increased in cisplatin‐resistant endometrial cancer cell lines (KLE and HEC‐1‐A), but not in cisplatin‐sensitive cell line (Ishikawa). Cisplatin‐mediated increase in Bcl‐2 expression was blocked by combination with either actinomycin D or cycloheximide. In addition, Bcl‐2 inhibition by HA14‐1 led to increased cisplatin‐induced apoptosis in KLE and HEC‐1‐A, whereas Bcl‐2 overexpression in Ishikawa led to decreased cisplatin‐induced apoptosis. Inhibition of protein kinase C (PKC) activity prevented cisplatin‐dependant increase in Bcl‐2 mRNA, and induced apoptosis in KLE cells. Furthermore, PKC inhibition was associated with decreased Akt and NF‐κB activity. Cells stably expressing shRNA for Akt isoforms revealed that Akt2 was involved in cisplatin‐dependant increase in Bcl‐2 and apoptosis. Overexpression of Akt2 in Akt2‐deficient cells led to increased Bcl‐2 expression on cisplatin treatment. Our data suggest a novel regulation pathway of Bcl‐2 by cisplatin, via the activation of PKC and Akt2, which has a profound impact on resistance to cisplatin‐induced apoptosis in endometrial cancer cells.
Dietary lignan and proanthocyanidin consumption and colorectal adenoma recurrence in the Polyp Prevention Trial
Authors: Bobe Gerd , Murphy Gwen , Albert Paul S. , Sansbury Leah B. , Lanza Elaine , Schatzkin Arthur , Cross Amanda J.
Published: May 2011

Lignans and proanthocyanidins are plant polyphenols that have shown protective properties against colorectal neoplasms in some human studies. Using logistic regression, we estimated odds ratios (ORs) and 95% confidence intervals (CIs) to prospectively evaluate the association between lignan and proanthocyanidin intake, estimated from databases linked to a food frequency questionnaire, and adenoma recurrence in 1,859 participants of the Polyp Prevention Trial. Overall, individual or total lignans or proanthocyanidins were not associated with colorectal adenoma recurrence. However, in sex‐specific analyses, total lignan intake was positively associated with any adenoma recurrence in women (highest vs. lowest lignan intake quartile OR = 2.07, 95% CI: 1.22–3.52, p trend = 0.004) but not in men (p interaction = 0.04). To conclude, dietary lignan and proanthocyanidin consumption were not generally related to colorectal adenoma recurrence; however, high lignan intake may increase the risk of adenoma recurrence in women.
Is mortality after childhood cancer dependent on social or economic resources of parents? A population‐based study
Authors: Syse Astri , Lyngstad Torkild Hovde , Kravdal Oystein
Published: May 2011

Diagnostic and treatment protocols for childhood cancer are generally standardized, and therefore, survival ought to be fairly equal across social strata in societies with free public health care readily available. Nevertheless, our study explores whether there are disparities in mortality after childhood cancer in Norway depending on socioeconomic status of parents. Limited knowledge on differentials exists from earlier analyses. Discrete‐time hazard regression models for all‐cause mortality for the first 10 years after diagnosis were estimated for all Norwegian children (younger than 20 years), who were diagnosed with cancer during 1974–2007 (N = 6,280), using data from five national registers. Mortality was reduced by about 15% for children with highly educated mothers and children without siblings. These effects were most pronounced for cancers predicted to encompass intense, long‐lasting treatments resulting in chronic health problems. Neither earnings nor the marital status of parents affected children's survival. This large, registry‐based study suggests that time constraints and various noneconomic rewards of parents from their education appears to have an impact on childhood cancer survival. It may be that children with resourceful parents are healthier at the outset and/or are more likely to avoid later health problems. It may also be that children of well‐informed and strongly involved parents are offered better treatment or are able to make better use of what is offered, for instance, by adhering more closely to recommendations for follow‐up treatment. The possibility of such differentials in offered and actual treatment should be addressed in future research.
Kinesin family member 14: An independent prognostic marker and potential therapeutic target for ovarian cancer
Authors: Thériault Brigitte L. , Pajovic Sanja , Bernardini Marcus Q. , Shaw Patricia A. , Gallie Brenda L.
Published: May 2011

The novel oncogene KIF14 (kinesin family member 14) shows genomic gain and overexpression in many cancers including OvCa (ovarian cancer). We discovered that expression of the mitotic kinesin KIF14 is predictive of poor outcome in breast and lung cancers. We now determine the prognostic significance of KIF14 expression in primary OvCa tumors, and evaluate KIF14 action on OvCa cell tumorigenicity in vitro. Gene‐specific multiplex PCR and real‐time QPCR were used to measure KIF14 genomic (109 samples) and mRNA levels (122 samples) in OvCa tumors. Association of KIF14 with clinical variables was studied using Kaplan–Meier survival and Cox regression analyses. Cellular effects of KIF14 overexpression (stable transfection) and inhibition (stable shRNA knockdown) were studied by proliferation (cell counts), survival (Annexin V immunocytochemistry) and colony formation (soft‐agar growth). KIF14 genomic gain (>2.6 copies) was present in 30% of serous OvCas, and KIF14 mRNA was elevated in 91% of tumors versus normal epithelium. High KIF14 in tumors independently predicted for worse outcome (p = 0.03) with loss of correlation with proliferation marker expression and increased rates of recurrence. Overexpression of KIF14 in OvCa cell lines increased proliferation and colony formation (p < 0.01), whereas KIF14 knockdown induced apoptosis and dramatically reduced colony formation (p < 0.05). Our findings indicate that KIF14 mRNA is an independent prognostic marker in serous OvCa. Dependence of OvCa cells on KIF14 for maintenance of in vitro colony formation suggests a role of KIF14 in promoting a tumorigenic phenotype, beyond its known role in proliferation.
Let‐7 modulates acquired resistance of ovarian cancer to Taxanes via IMP‐1‐mediated stabilization of multidrug resistance 1
Authors: Boyerinas Benjamin , Park Sun‐Mi , Murmann Andrea E. , Gwin Katja , Montag Anton G. , Zillhardt Marion , Hua You‐Jia , Lengyel Ernst , Peter Marcus E.
Published: May 2011

Ovarian cancer patients frequently develop resistance to chemotherapy regiments using Taxol and carboplatin. One of the resistance factors that protects cancer cells from Taxol‐based therapy is multidrug resistance 1 (MDR1). micro(mi)RNAs are small noncoding RNAs that negatively regulate protein expression. Members of the let‐7 family of miRNAs are downregulated in many human cancers, and low let‐7 expression has been correlated with resistance to microtubule targeting drugs (Taxanes), although little is known that would explain this activity. We now provide evidence that, although let‐7 is not a universal sensitizer of cancer cells to Taxanes, it affects acquired resistance of cells to this class of drugs by targeting IMP‐1, resulting in destabilization of the mRNA of MDR1. Introducing let‐7g into ADR‐RES cells expressing both IMP‐1 and MDR1 reduced expression of both proteins rendering the cells more sensitive to treatment with either Taxol or vinblastine without affecting the sensitivity of the cells to carboplatin, a non‐MDR1 substrate. This effect could be reversed by reintroducing IMP‐1 into let‐7g high/MDR1 low cells causing MDR1 to again become stabilized. Consistently, many relapsed ovarian cancer patients tested before and after chemotherapy were found to downregulate let‐7 and to co‐upregulate IMP‐1 and MDR1, and the increase in the expression levels of both proteins after chemotherapy negatively correlated with disease‐free time before recurrence. Our data point at IMP‐1 and MDR1 as indicators for response to therapy, and at IMP‐1 as a novel therapeutic target for overcoming multidrug resistance of ovarian cancer.
Male circumcision is associated with a lower prevalence of human papillomavirus‐associated penile lesions among Kenyan men
Authors: Backes Danielle M. , Bleeker Maaike C.G. , Meijer Chris J.L.M. , Hudgens Michael G. , Agot Kawango , Bailey Robert C. , Ndinya‐Achola J.O. , Hayombe Juma , Hogewoning Cornelis J.A. , Moses Stephen , Snijders Peter J.F. , Smith Jennifer S.
Published: May 2011

Human papillomavirus (HPV)‐associated penile lesions in men may increase the risk of HPV transmission to their female partners. Risk factor data on HPV‐associated penile lesions are needed from regions with a high burden of cervical cancer. Visual inspection of the penis was conducted using a colposcope at the 24‐month visit among participants in a randomized controlled trial of male circumcision in Kenya, from May 2006 to October 2007. All photos were read independently by two observers for quality control. Penile exfoliated cells sampled from the glans/coronal sulcus and the shaft were tested for HPV DNA using GP5+/6+ PCR and for HPV16, 18 and 31 viral loads using a real time PCR assay. Of 275 men, 151 were circumcised and 124 uncircumcised. The median age was 22 years. Circumcised men had a lower prevalence of flat penile lesions (0.7%) versus uncircumcised (26.0%); adjusted odds ratio (OR) = 0.02; 95% confidence interval (CI) = 0.003–0.1. Compared to men who were HPV negative, men who were HPV DNA positive (OR = 6.5; 95% CI = 2.4–17.5) or who had high HPV16/18/31 viral load (OR = 5.2; 95% CI = 1.1–24.4) had higher odds of flat penile lesions. Among men with flat penile lesions, HPV56 (29.0%) and 16 (25.8%) were the most common types within single or multiple infections. Flat penile lesions are much more frequent in uncircumcised men and associated with higher prevalence of HPV and higher viral loads. This study suggests that circumcision reduces the prevalence of HPV‐associated flat lesions and may ultimately reduce male‐to‐female HPV transmission.
Demethylation of the FOXP3 gene in human melanoma cells precludes the use of this epigenetic mark for quantification of Tregs in unseparated melanoma samples
Authors: Lucas Sophie , van Baren Nicolas , de Smet Charles , Coulie Pierre G.
Published: May 2011

The human suppressive T cells that stably express transcription factor FOXP3, or regulatory T cells (Tregs), are thought to suppress antitumor immune responses. The most specific marker for human Tregs is the demethylation of CpG dinucleotides located in the first intron of FOXP3 (FOXP3i1). FOXP3i1 is completely methylated in other hematopoietic cells, including nonsuppressive T cells that transiently express FOXP3 after activation. Previously, we and others reported estimations of the frequency of Tregs in the blood of melanoma patients using a FOXP3i1 methylation‐specific qPCR assay. Here, we attempted to quantify Tregs inside tumor samples using this assay. However, we found demethylated FOXP3i1 sequences in the melanoma cells themselves. This demethylation was not associated with substantial FOXP3 mRNA or protein expression, even though the demethylation extended to the promoter and terminal regions of the gene in some melanoma cells. Our results imply that analyzing Treg frequencies by quantification of demethylated FOXP3i1 will require that tumor‐infiltrating T cells be separated from melanoma cells.
IGF binding protein‐6 expression in vascular endothelial cells is induced by hypoxia and plays a negative role in tumor angiogenesis
Authors: Zhang Chunyang , Lu Ling , Li Yun , Wang Xianlei , Zhou Jianfeng , Liu Yunzhang , Fu Ping , Gallicchio Marisa A. , Bach Leon A. , Duan Cunming
Published: May 2011

Hypoxia stimulates tumor angiogenesis by inducing the expression of angiogenic molecules. The negative regulators of this process, however, are not well understood. Here, we report that hypoxia induced the expression of insulin‐like growth factor binding protein‐6 (IGFBP‐6), a tumor repressor, in human and rodent vascular endothelial cells (VECs) via a hypoxia‐inducible factor (HIF)‐mediated mechanism. Addition of human IGFBP‐6 to cultured human VECs inhibited angiogenesis in vitro. An IGFBP‐6 mutant with at least 10,000‐fold lower binding affinity for IGFs was an equally potent inhibitor of angiogenesis, suggesting that this action of IGFBP‐6 is IGF‐independent. The functional relationship between IGFBP‐6 and vascular endothelial growth factor (VEGF), a major hypoxia‐inducible angiogenic molecule, was examined. While VEGF alone increased angiogenesis in vitro, co‐incubation with IGFBP‐6 abolished VEGF‐stimulated angiogenesis. The in vivo role of IGFBP‐6 in angiogenesis was tested in flk1:GFP zebrafish embryos, which exhibit green fluorescence protein in developing vascular endothelium, permitting visualization of developing blood vessels. Injection of human IGFBP‐6 mRNA reduced the number of embryonic inter‐segmental blood vessels by ∼40%. This anti‐angiogenic activity is conserved in zebrafish because expression of zebrafish IGFBP‐6b had similar effects. To determine the anti‐angiogenic effect of IGFBP‐6 in a tumor model, human Rh30 rhabdomyosarcoma cells stably transfected with IGFBP‐6 were inoculated into athymic BALB/c nude mice. Vessel density was 52% lower in IGFBP‐6‐transfected xenografts than in vector control xenografts. These results suggest that the expression of IGFBP‐6 in VECs is up‐regulated by hypoxia and IGFBP‐6 inhibits angiogenesis in vitro and in vivo.
Three‐residue loop closure in proteins: A new kinematic method reveals a locus of connected loop conformations
Authors: Nekouzadeh Ali , Rudy Yoram
Published: May 2011

The closure of a three‐residue loop was studied using a developed kinematic method. It was shown that there are infinite number of three‐residue loops (a locus of conformations), which can connect two segments of a polypeptide. This adds to the current understanding of a finite number of conformations for three‐residue loop‐closure. In the developed method, some of the equations can be solved analytically to reduce the computation cost. Benefiting from the reduced computation time, we determined all the relative positions of two polypeptide segments that can be connected by a three‐residue loop. © 2011 Wiley Periodicals, Inc. J Comput Chem 2011
Attenuation of MicroRNA‐22 derepressed PTEN to effectively protect rat cardiomyocytes from hypertrophy
Authors: Xu Xu‐Dong , Song Xiao‐Wei , Li Qing , Wang Guo‐Kun , Jing Qing , Qin Yong‐Wen
Published: May 2011

Cardiac hypertrophy, which is characterized by the enlargement of cell size, reactivation of fetal genes, remains one of the most important triggers to heart failure. Increasing evidence shows that microRNA (miRNA) is extensively involved in the pathogenesis of cardiac hypertrophy. But the effects of miRNAs on cardiomyocyte hypertrophy have not been completely solved yet. Here, we showed that a collection of miRNAs was aberrantly expressed in hypertrophic cardiomyocytes induced by phenylephrine (PE) or angiotensin II (Ang II). Among them, miR‐22 was the most strikingly up‐regulated miRNA. To investigate the role of miR‐22 in hypertrophy, both over‐expression and knock‐down assays were performed on cardiomyocytes. The results showed that up‐regulation of miR‐22 significantly increased the cell size and markedly influenced the expression of hypertrophic markers, including induction of nppa and reduction of myh6. In contrast, reduction of miR‐22 level attenuated either PE‐ or Ang II‐induced hypertrophic reaction. Furthermore, several genes, including PTEN, were identified as potential targets of miR‐22 by bioinformatic algorithms. Using luciferase analysis, miR‐22 could significantly suppress the luciferase activity of reporter fused with 3′ untranslated region of PTEN mRNA. Furthermore, up‐regulation of miR‐22 could suppress the protein level of PTEN and reduction of miR‐22 level markedly increased the protein level of PTEN in cardiomyocytes by Western blot analysis, suggesting that the contribution of miR‐22 to cardiomyocyte hypertrophy may be partially through targeting PTEN. Taken together, miRNAs were dynamically regulated in cardiomyocyte hypertrophy and attenuation of miR‐22 in rat cardiomyocytes efficiently protected from hypertrophic effects through derepressing PTEN. J. Cell. Physiol. 227: 1391–1398, 2012. © 2011 Wiley Periodicals, Inc.
Cdc42 negatively regulates intrinsic migration of highly aggressive breast cancer cells
Authors: Zuo Yufeng , Wu Yuexiu , Chakraborty Chandan
Published: May 2011

The small GTPase Cdc42 has been implicated as an important regulator of cell migration. However, whether Cdc42 plays similar role in all cancer cells irrespective of metastatic potential remains poorly defined. Here, we show by using three different breast cancer cell lines with different metastatic potential, the role of Cdc42 in cell migration/invasion and its relationship with a number of downstream signaling pathways controlling cell migration. Small interfering RNA (siRNA)‐mediated knockdown of Cdc42 in two highly metastatic breast cancer cell lines (MDA‐MB‐231 and C3L5) resulted in enhancement, whereas the same in moderately metastatic (Hs578T) cell line resulted in inhibition of intrinsic cellular migration/invasion. Furthermore, Cdc42 silencing in MDA‐MB‐231 and C3L5 but not Hs578T cells was shown to be accompanied by increased RhoA activity and phosphorylation of protein kinase C (PKC)‐δ, extracellular signal regulated kinase1/2 (Erk1/2), and protein kinase A (PKA). Pharmacological inhibition of PKCδ, MEK‐Erk1/2, or PKA was shown to inhibit migration of both control and Cdc42‐silenced MDA‐MB‐231 cells. Furthermore, introduction of constitutively active Cdc42 was shown to decrease migration/invasion of MDA‐MB‐231 and C3L5 but increase migration/invasion of Hs578T cells. This decreased migration/invasion of MDA‐MB‐231 and C3L5 cells was also shown to be accompanied by the decrease in the phosphorylations of PKCδ, Erk1/2, and PKA. These results suggested that endogenous Cdc42 could exert a negative regulatory influence on intrinsic migration/invasion and some potentially relevant changes in phosphorylation of PKCδ, Erk1/2, and PKA of some aggressive breast cancer cells. J. Cell. Physiol. 227: 1399–1407, 2012. © 2011 Wiley Periodicals, Inc.
Hyposmotic stress causes ATP release and stimulates Na,K‐ATPase activity in porcine lens
Authors: Shahidullah M. , Mandal A. , Beimgraben C. , Delamere N.A.
Published: May 2011

Purinergic receptors in lens epithelium suggest lens function can be altered by chemical signals from aqueous humor or the lens itself. Here we show release of ATP by intact porcine lenses exposed to hyposmotic solution (200 mOsm). 18α‐glycyrrhetinic acid (AGA) added together with probenecid eliminated the ATP increase. N‐ethylmaleimide (200 µM), an exocytotic inhibitor, had no significant effect on ATP increase. Lenses exposed to hyposmotic solution displayed a ∼400% increase of propidium iodide (PI) entry into the epithelium. The increased ability of PI (MW 668) to enter the epithelium suggests possible opening of connexin and/or pannexin hemichannels. This is consistent with detection of connexin 43, connexin 50, and pannexin 1 in the epithelium and the ability of AGA + probenecid to prevent ATP release. Na,K‐ATPase activity doubled in the epithelium of lenses exposed to hyposmotic solution. The increase of Na,K‐ATPase activity did not occur when apyrase was used to prevent extracellular ATP accumulation or when AGA + probenecid prevented ATP release. The increase of Na,K‐ATPase activity was inhibited by the purinergic P2 antagonist reactive blue‐2 and pertussis toxin, a G‐protein inhibitor, but not by the P2X antagonist PPADS. Hyposmotic solution activated Src family kinase (SFK) in the epithelium, judged by Western blot. The SFK inhibitor PP2 abolished both SFK activation and the Na,K‐ATPase activity increase. In summary, hyposmotic shock‐induced ATP release is sufficient to activate a purinergic receptor‐ and SFK‐dependent mechanism that stimulates Na,K‐ATPase activity. The responses might signify an autoregulatory loop initiated by mechanical stress or osmotic swelling. J. Cell. Physiol. 227: 1428–1437, 2012. © 2011 Wiley Periodicals, Inc.
Hepatitis B virus X protein induces IKKα nuclear translocation via Akt‐Dependent phosphorylation to promote the motility of hepatocarcinoma cells
Authors: Huang Wei‐Chien , Chen Wen‐Shu , Chen Yun‐Ju , Wang Li‐Yun , Hsu Sheng‐Chieh , Chen Ching‐Chow , Hung Mien‐Chie
Published: May 2011

Hepatitis B virus (HBV) X protein (HBx) has been implicated in HBV‐associated carcinogenesis through activation of IκB kinase (IKK)/nuclear factor kappa B (NF‐κB) signaling pathway. Besides activating NF‐κB in the cytoplasm, IKKα was found in the nucleus to regulate gene expression epigenetically in response to various stimuli. However, it is unknown whether nuclear IKKα plays a role in HBx‐associated tumor progression. Moreover, the molecular mechanism underlying IKKα nuclear transport also remains to be elucidated. Here, we disclosed HBx as a new inducer of IKKα nuclear transport in hepatoma cells. HBx induced IKKα nuclear transport in an Akt‐dependent manner. HBx‐activated Akt promoted IKKα nuclear translocation via phosphorylating its threonine‐23 (Thr23). In addition, IKKα ubiquitination enhanced by HBx and Akt also contributed to the IKKα accumulation in the nucleus, indicating the involvement of ubiquitination in Akt‐increased IKKα nuclear transport in response to HBx. Furthermore, inhibition of IKKα nuclear translocation by mutation of its nuclear localization signal and Thr23 diminished IKKα‐dependent cell migration. Taken together, our findings shed light on the molecular mechanism of IKKα nuclear translocation and provide a potential role of nuclear IKKα in HBx‐mediated hepatocellular carcinoma (HCC) progression. J. Cell. Physiol. 227: 1446–1454, 2012. © 2011 Wiley Periodicals, Inc.
Cooperation between Shh and IGF‐I in promoting myogenic proliferation and differentiation via the MAPK/ERK and PI3K/Akt pathways requires smo activity
Authors: Madhala‐Levy D. , Williams V.C. , Hughes S.M. , Reshef R. , Halevy O.
Published: May 2011

Sonic Hedgehog (Shh) has been shown to promote adult myoblast proliferation and differentiation and affect Akt phosphorylation via its effector Smoothened (Smo). Here, the relationship between Shh and insulin‐like growth factor I (IGF‐I) was examined with regard to myogenic differentiation via signaling pathways which regulate this process. Each factor enhanced Akt and MAPK/ERK (p42/44) phosphorylation and myogenic factor expression levels in a dose‐responsive manner, while combinations of Shh and IGF‐I showed additive effects. Blockage of the IGF‐I effects by neutralizing antibody partially reduced Shh's effects on signaling pathways, suggesting that IGF‐I enhances, but is not essential for Shh effects. Addition of cyclopamine, a Smo inhibitor, reduced Shh‐ and IGF‐I‐induced Akt phosphorylation in a similar manner, implying that Shh affects gain of the IGF‐I signaling pathway. This implication was also examined via a genetic approach. In cultures derived from Smomut (MCre;Smoflox/flox) mice lacking Smo expression specifically in hindlimb muscles, IGF‐I‐induced Akt and p42/44 phosphorylation was significantly reduced compared to IGF‐I's effect on Smocont cells. Moreover, remarkable inhibition of the stimulatory effect of IGF‐I on myogenic differentiation was observed in Smomut cultures, implying that intact Smo is required for IGF‐I effects in myoblasts. Immunoprecipitation assays revealed that tyrosine‐phosphorylated proteins, including the regulatory unit of PI3K (p85), are recruited to Smo in response to Shh. Moreover, IGF‐IR was found to associate with Smo in response to Shh and to IGF‐I, suggesting that Shh and IGF‐I are already integrated at the receptor level, a mechanism by which their signaling pathways interact in augmenting their effects on adult myoblasts. J. Cell. Physiol. 227: 1455–1464, 2012. © 2011 Wiley Periodicals, Inc.
Clozapine impairs insulin action by up‐regulating AKT phosphorylation and Ped/Pea‐15 protein abundance
Authors: Panariello Fabio , Perruolo Giuseppe , Cassese Angela , Giacco Ferdinando , Botta Ginevra , Barbagallo Alessia P.M. , Muscettola Giovanni , Beguinot Francesco , Formisano Pietro , de Bartolomeis Andrea
Published: May 2011

Clinical and experimental evidence indicates that atypical antipsychotics impair glucose metabolism. We investigated whether clozapine may directly affect insulin action by analyzing insulin signaling in vitro and in vivo. Clozapine reduced insulin‐stimulated glucose uptake in PC12 and in L6 cells, representative models of neuron and skeletal muscle, respectively. Consistently, clozapine reduced insulin effect on insulin receptor (IR) by 40% and on IR substrate‐1 (IRS1) tyrosine phosphorylation by 60%. Insulin‐stimulated Akt phosphorylation was also reduced by about 40%. Moreover, insulin‐dependent phosphorylation of protein kinase C‐ζ (PKC‐ζ) was completely blunted in clozapine‐treated cells. Interestingly, clozapine treatment was accompanied by an insulin‐independent increase of Akt phosphorylation, with no change of IR, IRS1, and PKC‐ζ basal phosphorylation. The cellular abundance of Ped/Pea‐15, an Akt substrate and inducer of insulin resistance, was also increased following clozapine exposure, both in the absence and in the presence of cyclohexymide, a protein synthesis inhibitor. Similar as in cellular models, in the caudate–putamen and in the tibialis muscle of clozapine‐treated C57/BL/KsJ mice, Akt phosphorylation and Ped/Pea‐15 protein levels were increased and PKC‐ζ phosphorylation was decreased. Thus, in these experimental models, clozapine deranged Akt function and up‐regulated Ped/Pea‐15, thereby inhibiting insulin stimulation of PKC‐ζ and of glucose uptake. J. Cell. Physiol. 227: 1485–1492, 2012. © 2011 Wiley Periodicals, Inc.
Antennal sensilla of Apanteles cypris Nixon (Hymenoptera: Braconidae), a larval endoparasitoid of Cnaphalocrocis medinalis Guenée (Lepidoptera: Pyralidae)
Authors: Zhou Hui , Wu Wei‐Jian , Zhnag Zhen‐Fei , Zhnag Yang
Published: May 2011

Apanteles cypris is an endoparasitoid of rice pest Cnaphalocrocis medinalis. To better understand its host location mechanism, we examined the external morphology of antennal sensilla of A. cypris via scanning electron microscopy. The antennae of male and female A. cypris were filiform in shape, which consist of scape, pedicel, and flagellum with 18 antennomeres. Eleven types of sensilla were recorded in both sexes, including two types of chaetica sensilla, trichodea sensilla without pore, basiconica sensilla I with a tip pore, basiconica sensilla II with wall pores, basiconica sensilla III with a mastoid at the tip, coeloconica sensilla with apparent fingerlike projections, campaniform sensilla with a sprout tip pore, clavatea sensilla, smell pore, and placodea sensilla (PLA) with the multiporous cuticular structure. We also compared the morphology, number, and distribution of the sensory receptors between the two sexes. The length and width of the antennae on the males were significantly greater than that of females. In addition, males of A. cypris showed a larger number and a higher density of PLA compared to females. The morphology and structure of these sensilla were compared to that found in other braconid wasps, and their possible functions in host location were discussed. These observations could facilitate future studies on the biology of olfaction in A. cypris. Microsc. Res. Tech. 2011. © 2011 Wiley Periodicals, Inc.
Cytotoxicity of fipronil on mice liver cells
Authors: De Oliveira Patrícia Rosa , Bechara Gervásio Henrique , Denardi Sandra Eloisi , Oliveira Rodrigo Juliano , Mathias Maria Izabel Camargo
Published: May 2011

In the present study, mice livers were examined following exposure to different doses of fipronil (15, 25, and 50 mg/kg). Histological and histochemical techniques were used to determine the cytotoxic potential of this compound and to assess the damage it caused to livers. Mice were divided into four groups: control group and groups I, II, and III were exposed to 15, 25, and 50 mg/kg fipronil, respectively. Our findings revealed cytological, morphohistological, and histochemical alterations in liver cells of animals from groups I, II, and III compared to group control animals. These changes included Kupffer‐cell proliferation, hepatocyte hypertrophy, accumulation and distribution of proteins, polysaccharides, lipids, and vacuoles in the cytoplasm of hepatocytes, and congestion of blood vessels. These phenotypes mainly characterize the following: (a) autophagic processes, (b) steatosis, and (c) cell death by necrosis, which demonstrate the damage caused by fipronil on nontarget organisms in artificial conditions. Microsc. Res. Tech., 2012. © 2011 Wiley Periodicals, Inc.
A prospective longitudinal study of BK virus infection in 120 Czech renal transplant recipients
Authors: Girmanova Eva , Brabcova Irena , Bandur Stepan , Hribova Petra , Skibova Jelena , Viklicky Ondrej
Published: May 2011

Polyomavirus BK (BKV) is a common human polyomavirus that rarely causes clinical symptoms in immunocompetent individuals. However, BK virus reactivation occurs in 20–40% of kidney transplant patients and 1–10% of cases present with BK virus‐associated nephropathy (BKVN) and reduced kidney allograft survival. In this study, 120 consecutive renal allograft recipients were monitored for BK virus replication by real–time PCR (qPCR) in the blood and urine during the first year post‐transplantation and risk factors for BK viremia, viruria, and polyoma BKV‐associated nephropathy were evaluated. Receiver operating characteristic curve analysis was used to determine the cutoff points for assessing the risk of developing BKVN. In total, 1,243 samples were tested. BK‐DNAuria >107 copies/ml and BK‐DNAemia >104 copies/ml were found in 25.8% and 5% of the samples screened, respectively, during the 12 month follow‐up period. BKVN was confirmed histologically in 3/120 patients and viremic patients were treated with dialysis for longer time periods and had higher levels of anti‐BKV‐reactive antibodies. Patients with viruria were also treated longer with dialysis and had impaired graft function 12 months post‐transplantation. Patients with sustained viruria exhibited more acute rejection episodes than patients with transient viruria. Using receiver operating characteristic curve analysis, the cutoff point for viremia and viruria was redefined to 103 copies/ml serum for BK viremia and a cutoff point of 6.7 × 107 copies/ml in urine. In conclusion, polyoma BK viremia and viruria are frequent findings in kidney transplant recipients that warrant intensive monitoring as a means of preventing graft rejection. J. Med. Virol. 83:1395–1400, 2011. © 2011 Wiley‐Liss, Inc.