bluuebunny/crossref_metadata_2025 huggingface dataset:
Akt-Mediated Phosphorylation and Activation of Estrogen Receptor α Is Required for Endometrial Neoplastic Transformation in Pten+/− Mice
Authors: Vilgelm Anna , Lian Zenglin , Wang Hong , Beauparlant Stephen L. , Klein-Szanto Andres , Ellenson Lora Hedrick , Di Cristofano Antonio
Published: April 2006

PTEN is a tumor suppressor gene frequently mutated in human cancers. In vitro and in vivo studies have shown that PTEN can exert its tumor suppressive function through a variety of mechanisms, including regulation of cell death and cell proliferation. However, it is still unclear which of the many downstream pathways are critical in each different tissue, in vivo. Loss of PTEN is the earliest detectable genetic lesion in the estrogen-related type I (endometrioid) endometrial cancer. Pten+/− mice develop endometrial neoplastic lesions with full penetrance, thus providing a model system to dissect the genetic and biochemical events leading to the transition from normal to hyperplastic and neoplastic endometrial epithelium. Here, we show that loss of Pten in the mouse endometrium activates Akt and results in increased phosphorylation of estrogen receptor α (ERα) on Ser167. ERα phosphorylation results, in turn, in the activation of this nuclear receptor both in vivo and in vitro, even in the absence of ligand, and in its increased ability to activate the transcription of several of its target genes. Strikingly, reduction of endometrial ERα levels and activity dramatically reduces the neoplastic effect of Pten loss in the endometrium, in contrast to complete estrogen depletion. Thus, we provide for the first time in vivo evidence supporting the hypothesis that loss of Pten and subsequent Akt activation result in the activation of ERα-dependent pathways that play a pivotal role in the neoplastic process. (Cancer Res 2006; 66(7): 3375-80)
Targeting the RNA Splicing Machinery as a Novel Treatment Strategy for Pancreatic Carcinoma
Authors: Hayes Gregory M. , Carrigan Patricia E. , Beck Alison M. , Miller Laurence J.
Published: April 2006

Aberrant patterns of pre-mRNA splicing have been established for many human malignancies, yet the mechanisms responsible for these tumor-specific changes remain undefined and represent a promising area for therapeutic intervention. Using immunohistochemistry, we have localized the expression of a central splicing regulator, serine-arginine protein kinase 1 (SRPK1), to the ductular epithelial cells within human pancreas and have further shown its increased expression in tumors of the pancreas, breast, and colon. Small interfering RNA–mediated down-regulation of SRPK1 in pancreatic tumor cell lines resulted in a dose-dependent decrease in proliferative capacity and increase in apoptotic potential. Coordinately, the disruption of SRPK1 expression resulted in enhanced sensitivity of tumor cells to killing by gemcitabine and/or cisplatin. A dose-dependent reduction in the phosphorylation status of specific SR proteins was detected following the down-regulation of SRPK1 and is likely responsible for the observed alterations in expression of proteins associated with apoptosis and multidrug resistance. These data support SRPK1 as a new, potential target for the treatment of pancreatic ductular cancer that at present remains largely unresponsive to conventional therapies. Furthermore, these results support the development of innovative therapies that target not only specific splice variants arising during tumorigenesis but also the splice regulatory machinery that itself may be abnormal in malignant cells. (Cancer Res 2006; 66(7): 3819-27)
The p53 Pathway Promotes Efficient Mitochondrial DNA Base Excision Repair in Colorectal Cancer Cells
Authors: Chen Dexi , Yu Zhiyong , Zhu Zhiyi , Lopez Charles D.
Published: April 2006

The tumor suppressor p53 plays a central role in the DNA damage response. p53 enhances base excision repair (BER), in part, through direct interaction with the repair complex. Mitochondrial DNA (mtDNA) is repaired by a mtBER pathway. Many colorectal cancers harbor mtDNA mutations that are associated with poor prognosis. In addition to modulating the apoptotic response, mitochondria-localized p53 also stimulates mtBER. However, the mechanisms by which p53 enhances colorectal cancer mtBER after stress remain unclear. To explore this, we used colorectal cancer cells isogenic for p53 (HCT116p53+/+ and HCT116p53−/−). p53+/+ cells more efficiently repaired H2O2 damaged DNA in vivo as measured by semiquantitative mtDNA displacement loop PCR. Mitochondrial extracts from p53+/+ cells more efficiently stimulated 32P-dCTP incorporation into a uracil-oligonucleotide. Recombinant p53 complemented p53−/− mitochondrial extract repair of uracil or 8-oxo-G–containing oligonucleotides. As a measure of DNA glycosylase activity, p53+/+ mitochondrial extracts more efficiently incised uracil or 8-oxo-G oligonucleotides, although recombinant p53 could not stimulate oligonucleotide incision. p53 did not influence mitochondrial apurinic/apyrimidinic endonuclease activity measured by incision of a tetrahydrofuran-oligonucleotide. p53+/+ mitochondrial extracts had higher DNA polymerase-γ activity measured by 32P-dCTP incorporation into a single-nucleotide gap oligonucleotide, and recombinant p53 complemented p53−/− mitochondrial extract DNA polymerase-γ activity. mtDNA ligase activity was not affected by p53 status. p53 protein was detected in an inner mitochondrial membrane subfraction containing components of the mtBER complex. Our data suggest that an intact p53 pathway stimulates specific mtBER steps and provides mechanistic insight into the development of mtDNA mutations in colorectal cancer. (Cancer Res 2006; 66(7): 3485-94)
High-Resolution Analysis of Chromosomal Breakpoints and Genomic Instability Identifies PTPRD as a Candidate Tumor Suppressor Gene in Neuroblastoma
Authors: Stallings Raymond L. , Nair Prakash , Maris John M. , Catchpoole Daniel , McDermott Michael , O'Meara Anne , Breatnach Fin
Published: April 2006

Although neuroblastoma is characterized by numerous recurrent, large-scale chromosomal imbalances, the genes targeted by such imbalances have remained elusive. We have applied whole-genome oligonucleotide array comparative genomic hybridization (median probe spacing 6 kb) to 56 neuroblastoma tumors and cell lines to identify genes involved with disease pathogenesis. This set of tumors was selected for having either 11q loss or MYCN amplification, abnormalities that define the two most common genetic subtypes of metastatic neuroblastoma. Our analyses have permitted us to map large-scale chromosomal imbalances and high-level amplifications at exon-level resolution and to identify novel microdeletions and duplications. Chromosomal breakpoints (n = 467) generating imbalances >2 Mb were mapped to intervals ranging between 6 and 50 kb in size, providing substantial information on each abnormality. For example, breakpoints leading to large-scale hemizygous loss of chromosome 11q were highly clustered and preferentially associated with segmental duplications. High-level amplifications of MYCN were extremely complex, often resulting in a series of discontinuous regions of amplification. Imbalances (n = 540) <2 Mb long were also detected. Although the majority (78%) of these imbalances mapped to segmentally duplicated regions and primarily reflect constitutional copy number polymorphisms, many subtle imbalances were detected that are likely somatically acquired alterations and include genes involved with tumorigenesis, apoptosis, or neural cell differentiation. The most frequent microdeletion involved the PTPRD locus, indicating a possible tumor suppressor function for this gene. (Cancer Res 2006; 66(7): 3673-80)
Frequent Met Oncogene Amplification in a Brca1/Trp53 Mouse Model of Mammary Tumorigenesis
Authors: Smolen Gromoslaw A. , Muir Beth , Mohapatra Gayatry , Barmettler Anne , Kim Woo J. , Rivera Miguel N. , Haserlat Sara M. , Okimoto Ross A. , Kwak Eunice , Dahiya Sonika , Garber Judy E. , Bell Daphne W. , Sgroi Dennis C. , Chin Lynda , Deng Chu-Xia , Haber Daniel A.
Published: April 2006

In a screen for gene copy number alterations in mouse mammary tumors initiated by loss of the Brca1 and Trp53 genes, we observed that the majority (11 of 15; 73%) had high-level amplification of wild-type Met, encoding a growth factor receptor implicated in tumor progression. Met amplification was localized to unstable double minute chromosomes and was uniquely found in mouse breast tumors driven by loss of Brca1 and Trp53. Whereas analogous MET amplification was not found in human breast cancers, the identification of a dominant somatic genetic lesion in the Brca1/Trp53 mouse model suggests that recurrent secondary hits may also exist in BRCA1-initiated human breast cancer. (Cancer Res 2006; 66(7): 3452-5)
Transcription Analysis of Human Equilibrative Nucleoside Transporter-1 Predicts Survival in Pancreas Cancer Patients Treated with Gemcitabine
Authors: Giovannetti Elisa , Del Tacca Mario , Mey Valentina , Funel Niccola , Nannizzi Sara , Ricci Sergio , Orlandini Cinzia , Boggi Ugo , Campani Daniela , Del Chiaro Marco , Iannopollo Mauro , Bevilacqua Generoso , Mosca Franco , Danesi Romano
Published: April 2006

Gene expression analysis may help the management of cancer patients, allowing the selection of subjects responding to treatment. The aim of this study was the characterization of expression pattern of genes involved in gemcitabine activity in pancreas tumor specimens and its correlation with treatment outcome. The role of drug transport and metabolism on gemcitabine cytotoxicity was examined with specific inhibitors, whereas transcription analysis of human equilibrative nucleoside transporter-1 (hENT1), deoxycytidine kinase (dCK), 5′-nucleotidase (5′-NT), cytidine deaminase (CDA), and ribonucleotide reductase subunits M1 and M2 (RRM1 and RRM2) was done by quantitative reverse transcription-PCR in tumor tissue isolated by laser microdissection from surgical or biopsy samples of 102 patients. Association between clinical outcome and gene expression levels was estimated using Kaplan-Meier method and Cox's proportional hazards model. Transport and metabolism had a key role on gemcitabine sensitivity in vitro; moreover, hENT1, dCK, 5′-NT, CDA, RRM1, and RRM2 were detectable in most tumor specimens. hENT1 expression was significantly correlated with clinical outcome. Patients with high levels of hENT1 had a significantly longer overall survival [median, 25.7; 95% confidence interval (95% CI), 17.6-33.7 months in the higher expression tertile versus median, 8.5; 95% CI, 7.0-9.9 months in the lower expression tertile]. Similar results were obtained with disease-free survival and time to disease progression, and the multivariate analysis confirmed the prognostic significance of hENT1. This study suggests that the expression levels of hENT1 may allow the stratification of patients based on their likelihood of survival, thus offering a potential new tool for treatment optimization. (Cancer Res 2006; 66(7): 3928-35)
Tumor B7-H1 Is Associated with Poor Prognosis in Renal Cell Carcinoma Patients with Long-term Follow-up
Authors: Thompson R. Houston , Kuntz Susan M. , Leibovich Bradley C. , Dong Haidong , Lohse Christine M. , Webster W. Scott , Sengupta Shomik , Frank Igor , Parker Alexander S. , Zincke Horst , Blute Michael L. , Sebo Thomas J. , Cheville John C. , Kwon Eugene D.
Published: April 2006

B7-H1 participates in T-cell costimulation functioning as a negative regulator of immunity. Recent observations suggest that B7-H1 is expressed by renal cell carcinoma (RCC) tumor cells and is associated with poor prognosis. However, outcome analyses have been restricted to patients with fresh-frozen tissue and limited follow-up. We report the clinical effect of B7-H1 in RCC patients with a median of 10 years of follow-up. Between 1990 and 1994, 306 patients underwent nephrectomy for clear cell RCC and had paraffin tissue available for review. We did immunohistochemistry with anti-B7-H1 and conducted outcome analyses. Among the 306 patients, 73 (23.9%) harbored tumors with B7-H1 expression. Patients with tumor B7-H1 were at a significantly increased risk of both death from RCC [risk ratio (RR), 3.92; P < 0.001] and overall mortality (RR, 2.37; P < 0.001). The 5-year cancer-specific survival rates were 41.9% and 82.9% for patients with and without tumor B7-H1, respectively. In a multivariate model, tumor B7-H1 remained associated with cancer-specific death even after adjusting for tumor-node-metastasis stage, grade, and performance status (RR, 2.00; P = 0.003). In the subset of 268 patients with localized RCC, tumor B7-H1 was significantly associated with metastatic cancer progression (RR, 3.46; P < 0.001) and death from RCC (RR, 4.13; P < 0.001) even after adjusting for stage, grade, and performance status (RR, 1.78, P = 0.036). RCC patients with tumor B7-H1 are at significant risk of rapid cancer progression and accelerated rates of mortality. B7-H1 may function as a key determinant in RCC, abrogating immune responses directed against this immunogenic tumor. (Cancer Res 2006; 66(7): 3381-5)
Activation of Mitogen-Activated Protein Kinase in Estrogen Receptor α–Positive Breast Cancer Cells In vitro Induces an In vivo Molecular Phenotype of Estrogen Receptor α–Negative Human Breast Tumors
Authors: Creighton Chad J. , Hilger Amy M. , Murthy Shalini , Rae James M. , Chinnaiyan Arul M. , El-Ashry Dorraya
Published: April 2006

Breast cancer presents as either estrogen receptor α (ERα) positive or negative, with ERα+ tumors responding to antiestrogen therapy and having a better prognosis. By themselves, mRNA expression signatures of estrogen regulation in ERα+ breast cancer cells do not account for the vast molecular differences observed between ERα+ and ERα− cancers. In ERα− tumors, overexpression of epidermal growth factor receptor (EGFR) or c-erbB-2, leading to increased growth factor signaling, is observed such that mitogen-activated protein (MAP) kinase (MAPK) is significantly hyperactivated compared with ERα+ breast cancer. In ERα+/progesterone receptor–positive, estrogen-dependent MCF-7 breast cancer cells, we stably overexpressed EGFR or constitutively active erbB-2, Raf, or MAP/extracellular signal-regulated kinase kinase, resulting in cell lines exhibiting hyperactivation of MAPK, estrogen-independent growth, and the reversible down-regulation of ERα expression. By global mRNA profiling, we found a “MAPK signature” of ∼400 genes consistently up-regulated or down-regulated in each of the MAPK+ cell lines. In several independent profile data sets of human breast tumors, the in vitro MAPK signature was able to accurately distinguish ER+ from ER− tumors. In addition, our in vitro mRNA profile data revealed distinct mRNA signatures specific to either erbB-2 or EGFR activation. A subset of breast tumor profiles was found to share extensive similarities with either the erbB-2-specific or the EGFR-specific signatures. Our results confirm that increased MAPK activation causes loss of ERα expression and suggest that hyperactivation of MAPK plays a role in the generation of the ERα− phenotype in breast cancer. These MAPK+ cell lines are excellent models for investigating the underlying mechanisms behind the ERα− phenotype. (Cancer Res 2006; 66(7): 3903-11)
Highly Efficacious Nontoxic Preclinical Treatment for Advanced Metastatic Breast Cancer Using Combination Oral UFT-Cyclophosphamide Metronomic Chemotherapy
Authors: Munoz Raquel , Man Shan , Shaked Yuval , Lee Christina R. , Wong John , Francia Giulio , Kerbel Robert S.
Published: April 2006

Metronomic antiangiogenic chemotherapy, the prolonged administration of relatively low drug doses, at close regular intervals with no significant breaks, has been mainly studied at the preclinical level using single chemotherapeutic drugs, frequently in combination with a targeted antiangiogenic drug, and almost always evaluated on primary localized tumors. We tested a “doublet” combination metronomic chemotherapy treatment using two oral drugs, UFT, a 5-fluorouracil (5-FU) prodrug administered by gavage, and cyclophosphamide, for efficacy and toxicity in a new mouse model of advanced, terminal, metastatic human breast cancer. The optimal biological dose of each drug was first determined by effects on levels of circulating endothelial progenitor cells as a surrogate marker for angiogenesis, which was assessed to be 15 mg/kg for UFT and 20 mg/kg for cyclophosphamide. A combination treatment was then evaluated in mice with advanced metastatic disease using a serially selected metastatic variant of the MDA-MB-231 breast cancer-cell line, 231/LM2-4. UFT or cyclophosphamide treatment showed only very modest survival advantages whereas a combination of the two resulted in a remarkable prolongation of survival, with no evidence of overt toxicity despite 140 days of continuous therapy, such that a significant proportion of mice survived for over a year. In contrast, this striking therapeutic effect of the combination treatment was not observed when tested on primary orthotopic tumors. We conclude that combination oral low-dose daily metronomic chemotherapy, using cyclophosphamide and UFT, is superior to monotherapy and seems to be a safe and highly effective experimental antimetastatic therapy, in this case, for advanced metastatic breast cancer. (Cancer Res 2006; 66(7): 3386-91)
Estrogen Induces Lung Metastasis through a Host Compartment–Specific Response
Authors: Banka Carole L. , Lund Caren V. , Nguyen Mai T.N. , Pakchoian Andrew J. , Mueller Barbara M. , Eliceiri Brian P.
Published: April 2006

Direct proliferative effects of estrogen (E2) on estrogen receptor–positive tumors are well documented; however, the potential for E2 to mediate effects selective for the host (i.e., angiogenesis, vascular permeability, or stromal effects), which influence tumor growth and/or metastasis, has received less attention. In this study, we examine the capacity for E2 to promote tumor growth and/or metastasis independent of direct effects on tumor cells. In these studies, we distinguish host versus tumor compartment components of E2 action in tumor growth and metastasis by analysis of E2-nonresponsive tumor cells implanted in ovariectomized (OVX) mice that contain s.c. implants of placebo (OVX) or E2-containing slow-release pellets (OVX + E2). We show that the D121 lung carcinoma cell line is E2-nonresponsive, and following s.c. implantation in OVX versus OVX + E2 mice, E2 action on the host compartment leads to an increase in spontaneous metastasis but not primary tumor growth or neovascularization. Similarly, experimental lung metastasis of E2-nonresponsive 4T1 mammary carcinoma cells also leads to increased tumor burden in the lungs of OVX + E2 mice. These results suggest that the E2 status of the host compartment influences late steps in tumor cell metastasis that can provide important insights into the role of E2 in the tumor versus host compartments. (Cancer Res 2006; 66(7): 3667-72)
Mitosis-Independent Survivin Gene Expression In vivo and Regulation by p53
Authors: Xia Fang , Altieri Dario C.
Published: April 2006

Survivin is an essential mitotic gene, and this has been speculated to reflect its primary function in development and cancer. Here, we generated a knock-in transgenic mouse (SVVp-GFP) in which a green fluorescent protein (GFP) reporter gene was placed under the control of the survivin promoter that regulates transcription at mitosis. The expression of endogenous survivin was widespread in mouse tissues during development and shortly after birth. In contrast, GFP reactivity was undetectable in transgenic mouse embryos, and was largely limited postnatally to mitotic cells in the testes. Double transgenic mice generated in the tumor-prone Min/+ background exhibited intestinal adenomas that strongly expressed endogenous survivin, but only isolated GFP-positive cells. Conversely, dysplastic adenomas (16%) stained intensely for GFP, and revealed focal reactivity for mutant, but not wild-type, p53. The expression of GFP was increased by ∼10-fold in p53−/− as opposed to p53+/+ HCT116 colorectal cancer cells, and reintroduction of p53 in p53−/− cells abolished GFP expression. Therefore, the mitotic transcription of the survivin gene is highly restricted in vivo, and unexpectedly negatively regulated by p53. Contrary to a commonly held view, the dominant function(s) of survivin in development and tumor ontogeny are largely cell cycle–independent. (Cancer Res 2006; (66)7: 3392-5)
The Oxygen Sensor Factor-Inhibiting Hypoxia-Inducible Factor-1 Controls Expression of Distinct Genes through the Bifunctional Transcriptional Character of Hypoxia-Inducible Factor-1α
Authors: Dayan Frédéric , Roux Danièle , Brahimi-Horn M. Christiane , Pouyssegur Jacques , Mazure Nathalie M.
Published: April 2006

The function of the hypoxia-inducible factor-1 (HIF-1), the key transcription factor involved in cellular adaptation to hypoxia, is restricted to low oxygen tension (pO2). As such, this transcription factor is central in modulating the tumor microenvironment, sensing nutrient availability, and controlling anaerobic glycolysis, intracellular pH, and cell survival. Degradation and inhibition of the limiting HIF-1α subunit are intimately connected in normoxia. Hydroxylation of two proline residues by prolyl hydroxylase domain (PHD) 2 protein earmarks the protein for degradation, whereas hydroxylation of an asparagine residue by factor-inhibiting HIF-1 (FIH-1 or FIH) reduces its transcriptional activity. Indeed, silencing of either PHD2 or FIH in normoxia partially induced hypoxic genes, whereas combined PHD2/FIH silencing generated a full hypoxic gene response. Given the fact that HIF-1α possesses two transcriptional activation domains [TAD; NH2-terminal (N-TAD) and COOH-terminal (C-TAD)], we hypothesized on a possible bifunctional activity of HIF-1α that could be discriminated by FIH, an inhibitor of the C-TAD. In human cell lines engineered to overexpress or silence FIH in response to tetracycline, we show by quantitative reverse transcription-PCR that a set of hypoxic genes (ca9, phd3, pgk1, and bnip3) respond differently toward FIH expression. This finding, extended to 26 hypoxia-induced genes, indicates differential gene expression by the N-TAD and C-TAD in response to the hypoxic gradient. We propose that the oxygen-sensitive attenuator FIH, together with two distinct TADs, is central in setting the gene expression repertoire dictated by the cell pO2. (Cancer Res 2006; 66(7): 3688-98)
TMPRSS2:ETV4 Gene Fusions Define a Third Molecular Subtype of Prostate Cancer
Authors: Tomlins Scott A. , Mehra Rohit , Rhodes Daniel R. , Smith Lisa R. , Roulston Diane , Helgeson Beth E. , Cao Xuhong , Wei John T. , Rubin Mark A. , Shah Rajal B. , Chinnaiyan Arul M.
Published: April 2006

Although common in hematologic and mesenchymal malignancies, recurrent gene fusions have not been well characterized in epithelial carcinomas. Recently, using a novel bioinformatic approach, we identified recurrent gene fusions between TMPRSS2 and the ETS family members ERG or ETV1 in the majority of prostate cancers. Here, we interrogated the expression of all ETS family members in prostate cancer profiling studies and identified marked overexpression of ETV4 in 2 of 98 cases. In one such case, we confirmed the overexpression of ETV4 using quantitative PCR, and by rapid amplification of cDNA ends, quantitative PCR, and fluorescence in situ hybridization, we show that the TMPRSS2 (21q22) and ETV4 (17q21) loci are fused in this case. This result defines a third molecular subtype of prostate cancer and supports the hypothesis that dysregulation of ETS family members through fusions with TMRPSS2 may be an initiating event in prostate cancer development. (Cancer Res 2006; 66(7): 3396-400)
Sildenafil Citrate and Fetal Outcome in Pregnant Rats
Authors: Refuerzo Jerrie S. , Sokol Robert J. , Aranda Jacob V. , Hallak Mordechai , Hotra John W. , Kruger Michael , Sorokin Yoram
Published: April 2006

Objective: To determine the effects of sildenafil citrate on fetal growth in maternal rats exposed to hypoxia. Methods: Timed pregnant rats were randomized to either hypoxia or control on gestational days (GD) 18–20, and received either sildenafil (45 mg/kg) orally every 12 h on GD 18–21 or an equal volume of sterile water. Fetal pups were retrieved by laparotomy on GD 21. Pup weight and length were evaluated and cGMP measured in maternal and fetal blood. Results: In the non-hypoxic rats, sildenafil exposure was associated with a decrease in size(4.75 ± 0.43 vs. 5.11 ± 0.34 g, p = 0.00). In contrast, in the hypoxic rat model, sildenafil exposure was associated with increased size of the offspring (5.48 ± 0.45 vs. 5.16 ± 0.36 g, p = 0.016). Maternal cGMP levels were increased in the presence of both sildenafil and hypoxia (23.0 ± 10.5 vs. 15.6 ± 2.7 pmol/ml, p = 0.001). Conclusion: Exposure to sildenafil in a non-hypoxic setting results in a decrease in fetal size. Sildenafil in the presence of a stimulus, hypoxia, will lead to increased fetal size. These results suggest that sildenafil may have some influence on fetal growth. How these effects occur and by what mechanism remain to be determined.
Successful Treatment of Extremely Severe Fetal Anemia due to Anti-Jra Alloimmunization
Authors: Ishihara Yuki , Miyata Shigeki , Chiba Yoshihide , Kawai Takeshi
Published: April 2006

Background: Although Jra hemolytic disease of the newborn (HDN) is generally considered to be mild, we encountered a case of anti-Jra alloimmunization hydrops fetalis due to extremely severe fetal anemia that required aggressive intrauterine treatment. Case Report: A Japanese woman developed hydrops fetalis at the 29th week of gestation. Blood examination was highly positive for anti-Jra antibodies. Cordocentesis demonstrated that the fetus had anti-Jra antibodies and the lowest fetal hemoglobin concentration was 2.3 g/dl. After a total of four intravascular transfusions, fetal hydrops improved and a healthy girl of 2,120 g was delivered at the 35th week of gestation. The initial neonatal hemoglobin concentration at birth was 7.2 g/dl. After two straight transfusions, the infant’s anemia did not worsen. Anti-Jra antibodies became negative without developing hyperbilirubinemia. Conclusion: The incidence of Jra negativity can lead to underestimation of the risk of Jra alloimmunization. This case shows that anti-Jra alloimmunization may cause HDN and hydrops fetalis.
Inherited Thrombophilia: Treatment during Pregnancy
Authors: De Carolis Sara , Ferrazzani Sergio , De Stefano Valerio , Garofalo Serafina , Fatigante Gabriella , Rossi Elena , Leone Giuseppe , Caruso Alessandro
Published: April 2006

Objective: Inherited thrombophilia is associated with thromboembolic events and/or poor obstetric outcome. We evaluated the pregnancy outcome in women with inherited thrombophilia treated with low-molecular-weight heparin (LMWH). Methods: 38 thrombophilic women with a history of thromboembolic events and/or poor obstetric outcome were treated during their 39 consecutive pregnancies with LMWH from pregnancy verification until 4–6 weeks in puerperium. A fixed dose of enoxaparin 4,000 IU/day (except 1 case who required nadroparin 0.3 ml/day) was administered in most cases, adopting a higher dose (6,000 IU/day to 6,000 IU twice a day) in those with previous thromboembolic events. Results:In the treated women, all had a good obstetric outcome, whereas in the previous untreated pregnancies (n = 78), the rate of fetal loss (early and late) was 76.9%, only 12 live infants survived (66.6%). Moreover, birth weight resulted significantly higher in live infants born to treated pregnancies in comparison to that of previous untreated pregnancies (p = 0.009). No maternal thrombosis or major bleeding complications were recorded. Conclusions: The treatment with LMWH improved pregnancy outcome resulting effective and safe in thrombophilic women with a history of thromboembolic events and/or poor obstetric outcome.
Encouraging Early Clinical Experience with Deliberately Delayed Temporary Fetoscopic Tracheal Occlusion for the Prenatal Treatment of Life-Threatening Right and Left Congenital Diaphragmatic Hernias
Authors: Kohl Thomas , Gembruch Ulrich , Filsinger Barbara , Hering Rudolf , Bruhn Jörgen , Tchatcheva Kristina , Aryee Sebastian , Franz Axel , Heep Axel , Müller Andreas , Bartmann Peter , Loff Stefan , Hosie Stuart , Neff Wolfgang , Schaible Thomas
Published: April 2006

Objective: In order to assess the effect of deliberately delayed percutaneous fetoscopic tracheal occlusion on survival of fetuses with life-threatening congenital diaphragmatic hernia. Methods: Eight fetuses with life-threatening congenital diaphragmatic hernia underwent fetoscopic tracheal balloon occlusion between 29 + 0 and 32 + 4 weeks of gestation. Delayed occlusion was chosen in order to minimize potentially negative pulmonary effects from premature delivery as a result of fetal surgery. In addition, we wanted to become able to provide all available postnatal intensive care treatment means in these patients. Results: Six of the 8 fetuses survived to discharge from hospital. Conclusion: Delayed fetoscopic tracheal balloon occlusion may be rewarded with lung growth sufficient to allow survival of fetuses with life-threatening congenital diaphragmatic hernia.
Distal Hypospadias Repair in Adults: The Results of 97 Cases
Authors: Adayener Cuneyt , Akyol İlker
Published: April 2006

Introduction: We reviewed our 9-year experience on 97 men with distal hypospadias who had undergone hypospadias surgeries. Subjects and Methods: Patients were divided into two groups: group 1 included 17 men who had one or more unsuccessful surgeries in their childhood, and group 2 included 80 men with no previous hypospadias repairs. Meatal positions were glanular in 6, coronal in 35 and subcoronal in 56 patients. Our 97 hypospadias surgeries involved 42 meatal advancement and glanuloplasty, 41 Mathieu and 14 tubularized incised plate urethroplasty procedures. Results: The overall success rate was 88% (86 of the 97 patients). The success rate was 76.5% for group 1 (13 of the 17 patients) and 91.3% for group 2 (73 of the 80 patients). Better voiding function and good cosmetic appearance was achieved in all patients. Urinary flow rates were significantly higher in all groups postoperatively. The success rates were 91.4 and 85% for the patients with preoperative coronal and subcoronal meatal positions, respectively. The success rate was significantly lower in crippled cases and in cases with longer neourethra. Conclusion: Although the same techniques are used for children and adults in hypospadias surgery, the results in adults differ from those in children. The success rate for primary adult cases is quite acceptable, but it is decreased in patients having longer neourethra and having multiple previous interventions.
Cannabis-Induced Koro-Like Syndrome
Authors: Kalaitzi C.K. , Kalantzis A.
Published: April 2006

The genital-shrinking syndrome is a transient state of acute anxiety characterized by the triad of a deep-seated fear of penile shrinkage, its disappearance into the abdomen, and apprehension regarding inevitable impotence or even death. It is also known by the Malayo-Indonesian term ‘koro’. In the West, koro syndrome has been reported only in relation with various somatic, psychiatric and drug-induced disorders. The vast majority of non-psychiatric cases was related to neurological disorders or intoxication with cannabis or amphetamine. This is the first case of cannabis-induced koro-like syndrome ever reported in Greece.
Mouse Strains Differ in Their Susceptibility to Poststroke Infections
Authors: Schulte-Herbrüggen Olaf , Klehmet Juliane , Quarcoo David , Meisel Christian , Meisel Andreas
Published: April 2006

Objective: Severe infections, in particular pneumonia, have a major impact on the clinical management and outcome of stroke patients. In a mouse model we have recently demonstrated that stroke induces immunodepression which can result in life-threatening infections. Here, we investigated whether the susceptibility to infections after stroke is strain dependent. Methods and Results: Mice from 129SV, C57/B6, and Balb/C strains were subjected to experimental stroke by filament occlusion of the middle cerebral artery (MCAO) for 60 min. Infarct volumes were measured 3 days after MCAO. Microbiological assessment was based on cultures of bronchoalveolar lavage (BAL), lung tissue and blood of animals obtained 3 days after stroke. Three days after stroke 129SV mice did not only develop bacterial chest infection, but also had a strongly increased susceptibility to bacteremia. In contrast, C57BL/6 and Balb/C mice acquired bacterial lung infections only. In addition, bacterial load in BAL was significantly higher in 129SV mice than in the other mice strains. These differences in susceptibility to infection did not correlate with infarct volumes. Conclusions: Stroke-associated pneumonia developed in three commonly used mouse strains while severity of infections differed between strains. Since infections affect outcome, monitoring of infections is highly relevant for the interpretation of results in experimental stroke research.
Sensitization Potential of Citronellol
Authors: Hostýnek Jurij J. , Maibach Howard I.
Published: April 2006

The fragrance material citronellol has been cited as a moderately frequent cause of allergic contact dermatitis. A review of the literature shows that when the underlying clinical and experimental data are analyzed, a clear cause-effect relationship has infrequently or rarely been established. On the basis of its generally weak sensitizing potential in animals and human volunteers, coupled with its generally low exposure conditions, the prevalence of clinical cases would not be expected to be particularly high. This is not to say that citronellol is a frequent inducer of type IV allergy in members of the public. It remains to be seen, however, how often such allergy, once established, is responsible for any of the cases of allergic contact dermatitis commonly described in the literature. Indeed, in some cases, patch test conditions may not be optimal for differentiating between clinically relevant and irrelevant allergy to citronellol. Clarification of patch-test-related clinical relevance may be obtained utilizing retesting, dilution series and use tests.
Elevated Troponin Levels after Prolonged Supraventricular Tachycardia in Patient with Normal Coronary Angiography
Authors: Miranda Rafael C. , Machado Maurício N. , Takakura Isabela T. , da Mata Paula F. , da Fonseca Carlos Guilherme B. , Mouco Osana M.C.C. , Hernandes Mauro E. , Lemos Maria Angélica B.T. , Maia Lília N.
Published: April 2006

The European Society of Cardiology and the American College of Cardiology redefined the concept of myocardial infarction in the presence of highly positive markers of myocardial injury associated with at least one of the following: ischemic symptoms; development of pathologic Q waves on the ECG or ECG changes indicative of ischemia (positive or negative deviation of the ST segment), making troponins one of the most important aspects in the evaluation and stratification of patients with chest pain in the emergency room. However, although troponin gives excellent accuracy in the identification of myocardial necrosis, it is known that it can also be elevated in a series of nonatherosclerotic heart diseases. We present the case of a 49-year-old female patient admitted to the Chest Pain Unit with a history of supraventricular tachycardia associated with chest discomfort, nausea and diaphoresis. During risk stratification, the patient presented with a high serum troponin T level (0.143 ng/ml) but with a normal coronary angiography.
Recapitulation of Embryological Programmes in Renal Fibrosis – The Importance of Epithelial Cell Plasticity and Developmental Genes
Authors: Roxburgh Sarah A. , Murphy Madeline , Pollock Carol A. , Brazil Derek P.
Published: April 2006

Chronic fibrosis represents the final common pathway in progressive renal disease. Myofibroblasts deposit the constituents of renal scar, thus crippling renal function. It has recently emerged that an important source of these pivotal effector cells is the injured renal epithelium. This review concentrates on the process of epithelial-mesenchymal transition (EMT) and its regulation. The role of the developmental gene, gremlin, which is reactivated in adult renal disease, is the subject of particular focus. This member of the cysteine knot protein superfamily is critical to the process of nephrogenesis but quiescent in normal adult kidney. There is increasing evidence that gremlin expression reactivates in diabetic nephropathy, and in the diseased fibrotic kidney per se. Known to antagonize members of the bone morphogenic protein (BMP) family, gremlin may also act downstream of TGF-β in induction of EMT. An increased understanding of the extracellular modulation of EMT and, in particular, of the gremlin-BMP axis may result in strategies that can halt or reverse the devastating progression of chronic renal fibrosis.
Metabolic Syndrome as a Risk Factor for Contrast-Induced Nephropathy in Non-Diabetic Elderly Patients with Renal Impairment
Authors: Toprak Omer , Cirit Mustafa , Yesil Murat , Byrne Daniel W. , Postaci Nursen , Bayata Serdar , Majchrzak Karen M. , Esi Ertap
Published: April 2006

Background/Aims: Metabolic syndrome (MS) as a risk factor for contrast-induced nephropathy (CIN) has not been studied. The aim of the present study was to assess the influence of MS on the development of CIN in patients undergoing coronary angiography. Methods: This was a prospective cohort study. A total of 219 non-diabetic patients with reduced kidney function and age ≧60 years were divided into two groups (MS, n = 107 and non-MS, n = 112). CIN was defined as an increase of ≧25% in creatinine over the baseline value within 48 h of angiography. Results: CIN occurred in 14% of the MS group and 3.6% of the non-MS group (p = 0.006). Serum creatinine increased from 1.06 ± 0.17 to 1.12 ± 0.27 mg/dl in the MS group and from 1.03 ± 0.17 to 1.09 ± 0.23 mg/dl in the non-MS group (p < 0.001). MS was a risk indicator of CIN [odds ratio (OR) 4.26; 95% confidence interval (95% CI) 1.19–15.25; p = 0.026). Impaired fasting glucose (OR 4.72; 95% CI 1.53–14.56; p = 0.007), high triglyceride (OR 4.06; 95% CI 1.22–13.44; p = 0.022), and multivessel involvement (OR 3.14; 95% CI 1.07–9.82; p = 0.038) in the MS group were predictors of CIN. Conclusion: Our data support the hypothesis that patients with MS are at risk of developing CIN.
Relation between Different Treatment Modalities and Genomic Damage of End-Stage Renal Failure Patients
Authors: Kobras K. , Schupp N. , Nehrlich K. , Adelhardt M. , Bahner U. , Vienken J. , Heidland A. , Sebekova K. , Stopper H.
Published: April 2006

Background: Patients with end-stage renal disease display enhanced genomic damage. We investigated the relation between genomic damage and different treatment modalities. Methods: In a longitudinal study two groups of patients were analyzed in monthly intervals. We assessed the initiation of hemodialysis in 5 conservatively treated patients, and a switch from hemodialysis to hemodiafiltration in 7 patients. DNA damage was investigated in peripheral blood lymphocytes by micronucleus frequency and by comet assay analysis. With regard to potential genotoxicity of advanced glycation end products (AGEs), levels of imidazolone A and AGE-associated fluorescence (AGE-FL) were determined. Results: The initiation of hemodialysis did not alter the genomic damage. In patients who switched from hemodialysis to hemodiafiltration, a small but significant reduction in the comet assay but not in the micronucleus frequency was observed. Elevated plasmalevels of imidazolone A and AGE-FL were not influenced by the treatment modalities. Conclusion: In our small patient group no major reduction of the elevated genomic damage could be reached. Disease factors not influenced by altered dialysis modalities may have contributed considerably in our patient group. The persisting high levels of DNA damage suggest a need for further improvement. Inhibiting AGE formation may be one promising way for the future.
Increased Expression of Renal Aquaporin Water Channels in Spontaneously Hypertensive Rats
Authors: Lee JongUn , Kim SungWha , Kim Jin , Jeong Myung Ho , Oh YoonWha , Choi Ki Chul
Published: April 2006

Aims: The present study was aimed to determine whether there exists an altered regulation of aquaporin (AQP) water channels in hypertension. Methods: Male spontaneously hypertensive rats (SHR) aged 10–12 weeks were used. Age-matched Wistar-Kyoto (WKY) rats served as control. The abundance of AQP1–4 proteins in the kidney was determined by Western blot analysis. The protein expression and activity of adenylyl cyclase were also determined. Results: The medullary expression of AQP2 and AQP3 proteins was increased in SHR compared with that in WKY rats. The expression of AQP1 protein was also significantly increased in the inner medulla, while that of AQP4 was not. Immunohistochemistry of AQP2 revealed that principal cells of the collecting duct have strong immunoreactivity, the degree of which was augmented with prominent apical labeling in SHR. The plasma level of arginine vasopressin (AVP) was higher in SHR; the adenylyl cyclase activity stimulated by AVP was augmented, along with increased expression of type VI adenylyl cyclase. The urine was more concentrated with its volume decreased in SHR. Conclusion: The expression of AQP1–3 channels is increased in the kidney, in association with enhanced activity of the AVP/cAMP pathway, in SHR.
Endothelial Progenitor Cells in Chronic Renal Insufficiency
Authors: Herbrig Kay , Pistrosch Frank , Foerster Sarah , Gross Peter
Published: April 2006

There is growing evidence for a role of endothelial progenitor cells (EPCs) in the repair of damaged endothelium. It remains unclear which cell populations are most useful for clinical trials. Administration of drugs increasing EPC numbers and/or improving functional properties seems attractive. Further basic research is necessary to understand the mechanisms of mobilization, differentiation and homing of EPC in general and in particular under uremic conditions. Nephrologists should search for strategies to ameliorate EPC dysfunction of uremia. In this way it might be possible to test whether improved EPC biology is associated with decreased cardiovascular mortality in uremic humans. In any such studies the difficulties are going to be related to the complex procedures for EPC isolation, the testing of their identity and differentiation and their propagation before use.
Cystatin C – A Marker for Assessment of the Glomerular Filtration Rate in Patients with Cisplatin Chemotherapy
Authors: Benöhr P. , Grenz A. , Hartmann J.T. , Müller G.A. , Blaschke S.
Published: April 2006

Background: Cystatin C has recently been proposed as an ideal marker for glomerular filtration rate (GFR). In this study, cystatin C serum levels were evaluated in comparison to serum creatinine concentrations and inulin clearances in patients with normal kidney function receiving cisplatin-based chemotherapy to assess the validity of cystatin C as an alternative endogenous marker of GFR. Methods: Blood samples for the assessment of cystatin C, creatinine and inulin clearances were collected in patients before and after application of cisplatin in a clinical trial. Overall, 41 patients were included in the study, 35 of them were eligible receiving cisplatin in two different cisplatin-based chemotherapy schedules. Results: A 21% increase of cystatin C serum levels was demonstrated in the placebo group after application of cisplatin. Analysis of inulin clearances revealed a 23% loss of inulin clearance in patients of the placebo arm. In contrast, significant changes could not be detected by analysis of serum creatinine levels. Conclusions: Cystatin C represents a more sensitive clinical marker than serum creatinine for the early assessment of GFR damage caused by cisplatin. Changes in cystatin C serum concentrations correlate well to GFR decrease as measured by inulin clearance.
Mice Lacking Protein Kinase C Beta Present Modest Increases in Systolic Blood Pressure and NH4Cl-Induced Metabolic Acidosis
Authors: Yao Li-Jun , Leitges Michael , Vallon Volker
Published: April 2006

The conventional protein kinase C isoenzyme β (PKC-β) is expressed in various structures of mouse kidney. To get insights into the function, PKC-β knockout (–/–) and wild-type (+/+) mice were studied. Under basal conditions, PKC-β–/– mice exhibited a higher systolic blood pressure (in awake mice), normal plasma concentrations of Na+ and K+, and normal plasma pH. Urine osmolality and 24-hour excretion of fluid, Na+, K+ and albumin were not different between genotypes, but urine pH was more alkaline in PKC-β–/– mice. Inulin clearance experiments under anesthesia confirmed a higher systolic blood pressure and revealed normal glomerular filtration rate and fractional excretion of fluid, Na+ and K+ in PKC-β–/– mice. The ability to restrict renal Na+ excretion in response to a low Na+ diet was unaltered in PKC-β–/– mice. Chronic acid loading (NH4Cl) did not affect blood pH in PKC-β+/+ mice, but induced a modest metabolic acidosis in PKC-β–/– mice. In conclusion, first evidence is presented that (i) PKC-β contributes to the regulation of arterial blood pressure, and (ii) PKC-β is required for normal acid-base balance, which may relate to its expression and function in intercalated cells of the collecting duct.
Co-Expression of Interleukin-2 by a Bicistronic Plasmid Increases the Efficacy of DNA Immunization to Prevent Influenza Virus Infections
Authors: Henke Andreas , Rohland Nadine , Zell Roland , Wutzler Peter
Published: April 2006

A promising approach to protect susceptible individuals against severe diseases is the inoculation of plasmids. Such DNA vaccines against influenza virus infections were quite efficient in different animal models; but still this procedure is not in clinical use until today. The present study reports the generation and characterization of bicistronic plasmids which enables the expression of influenza A virus gene sequences together with immunostimulatory cytokines demonstrating that among these cytokines especially interleukin-2 (IL-2) was efficient to prevent a lethal influenza virus infection in mice.
Gonadotropin Secretion in Girls with Turner Syndrome Measured by an Ultrasensitive Immunochemiluminometric Assay
Authors: Chrysis Dionisios , Spiliotis Bessie E. , Stene Mark , Cacciari Emanuele , Davenport Marsha L.
Published: April 2006

Background/Aim: Gonadotropin levels measured by radioimmunoassays are high in girls with Turner syndrome (TS), but overlap significantly with those of normal girls. We hypothesized that gonadotropin levels would be above the normal range in TS when measured by ultrasensitive assays. Methods: Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were measured in 68 TS, and 133 control girls using ultrasensitive immunochemiluminometric assays (ICMA). Results: FSH levels in TS and normal girls were highest in early childhood (56.0 ± 39.7 and 2.3 ± 1.8 IU/l, respectively), declined at 6–10 years of age (11.3 ± 13.1 and 1.8 ± 0.9 IU/l, respectively), and then increased again (104.4 ± 68.9 and 4.9 ± 2.4 IU/l, respectively). FSH was in the normal range on 11 of 27 occasions in TS girls with ages 5–10 years, and on 3 of 44 occasions in >10 years. Although average LH values were higher than those of controls, they often overlapped the normal range. Conclusion: A significant number of TS girls have normal gonadotropins by ICMA. Spontaneous gonadotropin levels are not an adequate screening test for the diagnosis of TS but may prove useful for predicting the gonadal function and determining the appropriate timing of estrogen replacement therapy.