bluuebunny/crossref_metadata_2025 huggingface dataset:
Ventral Cochlear Nucleus Responses to Contralateral Sound Are Mediated by Commissural and Olivocochlear Pathways
Authors: Bledsoe Sanford C. , Koehler Seth , Tucci Debara L. , Zhou Jianxun , Le Prell Colleen , Shore Susan E.
Published: May 2009

In the normal guinea pig, contralateral sound inhibits more than a third of ventral cochlear nucleus (VCN) neurons but excites <4% of these neurons. However, unilateral conductive hearing loss (CHL) and cochlear ablation (CA) result in a major enhancement of contralateral excitation. The response properties of the contralateral excitation produced by CHL and CA are similar, suggesting similar pathways are involved for both types of hearing loss. Here we used the neurotoxin melittin to test the hypothesis that this “compensatory” contralateral excitation is mediated either by direct glutamatergic CN-commissural projections or by cholinergic neurons of the olivocochlear bundle (OCB) that send collaterals to the VCN. Unit responses were recorded from the left VCN of anesthetized, unilaterally deafened guinea pigs (CHL via ossicular disruption, or CA via mechanical destruction). Neural responses were obtained with 16-channel electrodes to enable simultaneous data collection from a large number of single- and multiunits in response to ipsi- and contralateral tone burst and noise stimuli. Lesions of each pathway had differential effects on the contralateral excitation. We conclude that contralateral excitation has a fast and a slow component. The fast excitation is likely mediated by glutamatergic neurons located in medial regions of VCN that send their commissural axons to the other CN via the dorsal/intermediate acoustic striae. The slow component is likely mediated by the OCB collateral projections to the CN. Commissural neurons that leave the CN via the trapezoid body are an additional source of fast, contralateral excitation.
Beta-Range EEG-EMG Coherence With Isometric Compensation for Increasing Modulated Low-Level Forces
Authors: Chakarov Vihren , Naranjo José Raúl , Schulte-Mönting Jürgen , Omlor Wolfgang , Huethe Frank , Kristeva Rumyana
Published: May 2009

Corticomuscular synchronization has been shown to occur in beta (15–30 Hz) and gamma range (30–45 Hz) during isometric compensation of static and dynamic (periodically modulated) low-level forces, respectively. However, it is still unknown to what extent these synchronization processes in beta and gamma range are modified with increasing modulated force. We addressed this question by investigating the corticomuscular coherence (CMC) between the electroencephalogram (EEG) and electromyogram (EMG) from the first dorsal interosseus muscle (FDI) as well as the cortical and muscular spectral power during a visuomotor task where different levels of a dynamic (modulated) force were used. Seven healthy right-handed female subjects compensated dynamic forces at 8, 16, and 24% of the maximal voluntary contraction (MVC) isometrically with their right index finger. Under the three conditions investigated, we found a broad-band CMC comprising both beta and gamma range and peaking at ∼22 Hz within the beta band. This broad-band coherence increased linearly with higher force level. A separate analysis of the gamma range CMC did not show significant modulation of the CMC by the force levels. EEG and EMG spectral power did not show any significant difference among the three force conditions. Our results favor the view that the function of beta range CMC is not specific for low-level static forces only. The sensorimotor system may resort to stronger and also broader beta-range CMC to generate stable corticospinal interaction during increased force level, as well as when compensating for dynamic modulated forces. This finding re-enforces the importance of the beta-range EEG-EMG coherence in sensorimotor integration.
Combination Therapies against Chronic Myeloid Leukemia: Short-term versus Long-term Strategies
Authors: Komarova Natalia L. , Wodarz Dominik
Published: May 2009

During therapy for chronic myeloid leukemia (CML), decline of the number of BCR-ABL transcripts has been shown to follow a biphasic pattern, with a fast phase followed by a slower phase. Hence, sustained remission requires a long phase of therapy. Data indicate that a combination of different available targeted drugs might prevent treatment failure due to drug resistance, especially at advanced stages of the disease. However, for long-term multiple-drug treatments, complications can arise from side effects. We investigate whether and how the number of drugs could be reduced during long-term therapy. Using computational models, we show that one or more drugs can be removed once the number of tumor cells is reduced significantly, without compromising the chances of sustained tumor suppression. Which drug to remove first depends on the number of mutations in the BCR-ABL gene that confer resistance to the drugs, as well as on how effectively the drugs inhibit Bcr-Abl protein tyrosine kinase activity and inhibit tumor growth. We further show that the number of CML cells at which the number of drugs can be reduced does not correlate with the two phases of decline of the BCR-ABL transcript numbers. Neither does it depend much on kinetic parameters of CML growth, except for the mutation rates at which resistance is generated. This is a significant finding because even without any information on most parameters, and using only the data on the number of cancer cells and the rate at which resistant mutants are generated, it is possible to predict at which stage of treatment the number of drugs can be reduced. [Cancer Res 2009;69(11):4904–10]
Differentiation of Malignant B-Lymphoma Cells from Normal and Activated T-Cell Populations by Their Intrinsic Autofluorescence
Authors: Pantanelli Seth M. , Li Zhuqing , Fariss Robert , Mahesh Sankaranarayana P. , Liu Baoying , Nussenblatt Robert B.
Published: May 2009

Patients with active posterior and intermediate uveitis have inflammatory cells in their vitreous; those with primary intraocular lymphoma have malignant B-lymphoma cells concomitantly. These cell types cannot be distinguished clinically. The goal of this study was to investigate intrinsic autofluorescence as a noninvasive way of differentiating immune and lymphomatous cell populations. Human primary T cells were stimulated with or without anti-CD3 plus anti-CD28 stimulation. B-lymphoma cells (CA46) were cultured separately. Five experimental groups were prepared: unstimulated T cells, stimulated T cells, CA46 cells, and stimulated T cells mixed with CA46 cells at a ratio of 1:3 or mixed at a ratio of 3:1. Samples were excited with three wavelengths and imaged with a confocal microscope. For each condition, the autofluorescent emissions from the sample were measured. In separate experiments, T cells or CA46 cells were injected into the anterior chamber of a BALB/c mouse eye and autofluorescence was measured. Pure T-cell and lymphoma populations were clearly distinguishable based on autofluorescence intensity spectra. CA46 cells were the least fluorescent when excited with 351-nm light, but most fluorescent when excited with longer wavelengths like 488 nm. Mixed populations of T cells and CA46 cells had emission intensities that fell predictably in between those of the pure populations. An ex vivo study showed that CA46 cells could be detected based on their intrinsic autofluorescence. Our studies showed that normal activated and malignant lymphocyte populations can be distinguished based on their intrinsic autofluorescent properties. Future work with in vivo models may prove useful in facilitating the diagnosis of uveitis and other ocular diseases. [Cancer Res 2009;69(11):4911–7]
Combined Inhibition of MEK and Mammalian Target of Rapamycin Abolishes Phosphorylation of Cyclin-Dependent Kinase 4 in Glioblastoma Cell Lines and Prevents Their Proliferation
Authors: Paternot Sabine , Roger Pierre P.
Published: May 2009

The Ras/Raf/MEK/extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (mTOR) signaling pathways are aberrantly activated in many tumors, including highly proliferative glioblastomas, but how they are wired with the cell cycle remains imperfectly understood. Inhibitors of MEK/ERK and mTOR pathways are tested as anticancer agents. They are generally considered to induce a G1 cell cycle arrest through down-regulation of D-type cyclins and up-regulation of p27kip1. Here, we examined the effect of targeting mTOR by rapamycin and/or MEK by PD184352 in human glioblastoma cell lines. In combination, these drugs cooperatively and potently inhibited the G1-S transition and retinoblastoma protein phosphorylation. Their cooperation could not be explained by their partial and differential inhibitory effects on cyclin D1 or D3 but instead by their synergistic inhibition of the activating T172 phosphorylation of cyclin-dependent kinase (CDK) 4. This appeared independent of p27 and unrelated to weak modulations of the CDK-activating kinase activity. The T172 phosphorylation of CDK4 thus appears as a crucial node integrating the activity of both MEK/ERK and mTOR pathways. Combined inhibition of both pathways should be considered as a promising strategy for treatment of tumors harboring a deregulated CDK4 activity. [Cancer Res 2009;69(11):4577–81]
Rifampicin as an Oral Angiogenesis Inhibitor Targeting Hepatic Cancers
Authors: Shichiri Masayoshi , Fukai Nozomi , Kono Yutaka , Tanaka Yujiro
Published: May 2009

Angiogenesis is an important therapeutic target in cancer, and to fully exploit its therapeutic potential, combination chemotherapeutic/antiangiogenic regimens should be optimized and delivered earlier to more patients. Ideally, this could be done by a single potent oral agent with established safety. Rifampicin, a semisynthetic antibiotic derived from the rifamycins, is one of the most commonly used pharmaceutical compounds worldwide in the treatment of tuberculosis. Here, we present the effects of oral rifampicin on human cancer progression and its antiangiogenic properties, which were comparable to the angiogenesis inhibitor endostatin. Clinically, low-dose p.o. administration of rifampicin to six high-risk patients with hepatitis C virus–related liver cirrhosis resulted in a single occurrence of hepatocellular carcinoma during the follow-up period of 97.3 ± 29.1 (mean ± SD) months. Experimentally, rifampicin rapidly and markedly down-regulated the expression of a wide spectrum of angiogenesis-associated genes in growing human microvascular endothelial cells, thereby suppressing endothelial cell proliferation and migration. Rifampicin, at higher concentrations, also directly inhibited the growth of a variety of human cancer cells. P.o. administration of rifampicin significantly inhibited in vivo growth and metastases of subcutaneous human cancer xenografts. Thus, the potent antiangiogenic properties of oral rifampicin therapy were effective in suppressing cancer progression. It provides a promising new addition to antiangiogenic strategies for designing human cancer therapies. Considering the clinical pharmacokinetics of rifampicin, which enters the enterohepatic circulation and undergoes subsequent hepatic accumulation, it may be especially beneficial as an antitumor agent targeting hepatobiliary tumors. [Cancer Res 2009;69(11):4760–8]
Identification of a Subpopulation of Macrophages in Mammary Tumor–Bearing Mice That Are Neither M1 nor M2 and Are Less Differentiated
Authors: Torroella-Kouri Marta , Silvera Risset , Rodriguez Dayron , Caso Raul , Shatry Alwi , Opiela Shannon , Ilkovitch Dan , Schwendener Reto A. , Iragavarapu-Charyulu Vijaya , Cardentey Yoslayma , Strbo Natasa , Lopez Diana M.
Published: May 2009

Systemic and local immune deficiency is associated with cancer, and the role of M2 tumor-associated macrophages in this phenomenon is well recognized. However, the immune status of macrophages from peripheral compartments in tumor hosts is unclear. Peritoneal macrophages (PEM) are derived from circulating monocytes and recruited to the peritoneal cavity where they differentiate into macrophages. We have previously shown that PEMs from mice bearing D1-DMBA-3 mammary tumors (T-PEM) are deficient in inflammatory functions and that this impairment is associated with diminished expression of transcription factors nuclear factor κB and CAAT/enhancer-binding protein. We now provide evidence that T-PEMs display neither M1 nor M2 phenotypes, yet exhibit deficiencies in the expression of several inflammatory cytokines and various proinflammatory signaling pathways. Moreover, due to nuclear factor κB down-regulation, increased apoptosis was observed in T-PEMs. We report for the first time that macrophage depletion is associated with increased macrophage progenitors in bone marrow. Furthermore, T-PEMs have a lower expression of macrophage differentiation markers F4/80, CD68, CD115, and CD11b, whereas Gr-1 is up-regulated. Our results suggest that T-PEMs are less differentiated and represent a newly derived population from blood monocytes. Lastly, we show that transforming growth factor-β and prostaglandin E2, two immunosuppressive tumor-derived factors, may be involved in this phenomenon. [Cancer Res 2009;69(11):4800–9]
Quantitative Metabolome Profiling of Colon and Stomach Cancer Microenvironment by Capillary Electrophoresis Time-of-Flight Mass Spectrometry
Authors: Hirayama Akiyoshi , Kami Kenjiro , Sugimoto Masahiro , Sugawara Maki , Toki Naoko , Onozuka Hiroko , Kinoshita Taira , Saito Norio , Ochiai Atsushi , Tomita Masaru , Esumi Hiroyasu , Soga Tomoyoshi
Published: May 2009

Most cancer cells predominantly produce energy by glycolysis rather than oxidative phosphorylation via the tricarboxylic acid (TCA) cycle, even in the presence of an adequate oxygen supply (Warburg effect). However, little has been reported regarding the direct measurements of global metabolites in clinical tumor tissues. Here, we applied capillary electrophoresis time-of-flight mass spectrometry, which enables comprehensive and quantitative analysis of charged metabolites, to simultaneously measure their levels in tumor and grossly normal tissues obtained from 16 colon and 12 stomach cancer patients. Quantification of 94 metabolites in colon and 95 metabolites in stomach involved in glycolysis, the pentose phosphate pathway, the TCA and urea cycles, and amino acid and nucleotide metabolisms resulted in the identification of several cancer-specific metabolic traits. Extremely low glucose and high lactate and glycolytic intermediate concentrations were found in both colon and stomach tumor tissues, which indicated enhanced glycolysis and thus confirmed the Warburg effect. Significant accumulation of all amino acids except glutamine in the tumors implied autophagic degradation of proteins and active glutamine breakdown for energy production, i.e., glutaminolysis. In addition, significant organ-specific differences were found in the levels of TCA cycle intermediates, which reflected the dependency of each tissue on aerobic respiration according to oxygen availability. The results uncovered unexpectedly poor nutritional conditions in the actual tumor microenvironment and showed that capillary electrophoresis coupled to mass spectrometry–based metabolomics, which is capable of quantifying the levels of energy metabolites in tissues, could be a powerful tool for the development of novel anticancer agents that target cancer-specific metabolism. [Cancer Res 2009;69(11):4918–25]
Targeting Focal Adhesion Kinase with Dominant-Negative FRNK or Hsp90 Inhibitor 17-DMAG Suppresses Tumor Growth and Metastasis of SiHa Cervical Xenografts
Authors: Schwock Joerg , Dhani Neesha , Cao Mary Ping-Jiang , Zheng Jinzi , Clarkson Richard , Radulovich Nikolina , Navab Roya , Horn Lars-Christian , Hedley David W.
Published: May 2009

Focal adhesion kinase (FAK), a nonreceptor protein tyrosine kinase and key modulator of integrin signaling, is widely expressed in different tissues and cell types. Recent evidence indicates a central function of FAK in neoplasia where the kinase contributes to cell proliferation, resistance to apoptosis and anoikis, invasiveness, and metastasis. FAK, like other signaling kinases, is dependent on the chaperone heat shock protein 90 (Hsp90) for its stability and proper function. Thus, inhibition of Hsp90 might be a way of disrupting FAK signaling and, consequently, tumor progression. FAK is expressed in high-grade squamous intraepithelial lesions and metastatic cervical carcinomas but not in nonneoplastic cervical mucosa. In SiHa, a cervical cancer cell line with characteristics of epithelial-to-mesenchymal transition, the stable expression of dominant-negative FAK-related nonkinase decreases anchorage independence and delays xenograft growth. FAK-related nonkinase as well as the Hsp90 inhibitor 17-dimethylaminoethylamino-17-demethoxygeldanamycin both negatively interfere with FAK signaling and focal adhesion turnover. Short-term 17-dimethylaminoethylamino-17-demethoxygeldanamycin treatment prolongs survival in a SiHa lung metastasis model and chronic administration suppresses tumor growth as well as metastatic spread in orthotopic xenografts. Taken together, our data suggest that FAK is of importance for tumor progression in cervical cancer and that disruption of FAK signaling by Hsp90 inhibition might be an avenue to restrain tumor growth as well as metastatic spread. [Cancer Res 2009;69(11):4750–9]
Quality of Reporting of Serious Adverse Drug Events to an Institutional Review Board: A Case Study with the Novel Cancer Agent, Imatinib Mesylate
Authors: Dorr David A. , Burdon Rachel , West Dennis P. , Lagman Jennifer , Georgopoulos Christina , Belknap Steven M. , McKoy June M. , Djulbegovic Benjamin , Edwards Beatrice J. , Weitzman Sigmund A. , Boyle Simone , Tallman Martin S. , Talpaz Moshe , Sartor Oliver , Bennett Charles L.
Published: May 2009

Purpose: Serious adverse drug event (sADE) reporting to Institutional Review Boards (IRB) is essential to ensure pharmaceutical safety. However, the quality of these reports has not been studied. Safety reports are especially important for cancer drugs that receive accelerated Food and Drug Administration approval, like imatinib, as preapproval experience with these drugs is limited. We evaluated the quality, accuracy, and completeness of sADE reports submitted to an IRB.

Experimental Design: sADE reports submitted to an IRB from 14 clinical trials with imatinib were reviewed. Structured case report forms, containing detailed clinical data fields and a validated causality assessment instrument, were developed. Two forms were generated for each ADE, the first populated with data abstracted from the IRB reports, and the second populated with data from the corresponding clinical record. Completeness and causality assessments were evaluated for each of the two sources, and then compared. Accuracy (concordance between sources) was also assessed.

Results: Of 115 sADEs reported for 177 cancer patients to the IRB, overall completeness of adverse event descriptions was 2.4-fold greater for structured case report forms populated with information from the clinical record versus the corresponding forms from IRB reports (95.0% versus 40.3%, P < 0.05). Information supporting causality assessments was recorded 3.5-fold more often in primary data sources versus IRB adverse event descriptions (93% versus 26%, P < 0.05). Some key clinical information was discrepant between the two sources.

Conclusions: The use of structured syndrome-specific case report forms could enhance the quality of reporting to IRBs, thereby improving the safety of pharmaceuticals administered to cancer patients.
ERBB1 and ERBB2 Have Distinct Functions in Tumor Cell Invasion and Intravasation
Authors: Kedrin Dmitriy , Wyckoff Jeffrey , Boimel Pamela J. , Coniglio Salvatore J. , Hynes Nancy E. , Arteaga Carlos L. , Segall Jeffrey E.
Published: May 2009

Purpose: The epidermal growth factor receptor (ERBB1) and related family member HER-2/neu (ERBB2) are often overexpressed in aggressive breast cancers and their overexpression is correlated with poor prognosis. Clinical studies using ERBB inhibitors have focused on tumor growth effects, but ERBBs can contribute to malignancy independent of their effects on tumor growth. Our studies were designed to evaluate the effect of ERBB inhibition on tumor cell motility and intravasation in vivo using clinically relevant small-molecule inhibitors.

Experimental Design: Using in vivo mouse models of breast cancer, we test the effects of ERBB1 and ERBB2 inhibitors AC480 and lapatinib, ERBB1 inhibitor gefitinib, and ERBB2 inhibitor AG825 on in vivo tumor cell invasive properties in mammary fat pad tumors.

Results: ERBB1 and ERBB2 inhibition rapidly (within 3 h) inhibits both tumor cell motility and intravasation. Using gefitinib, ERBB1 inhibition rapidly inhibits tumor cell motility and invasion but not intravasation, whereas ERBB2 inhibition by AG825 rapidly blocks intravasation.

Conclusions: ERBB1 and ERBB2 inhibition can rapidly block tumor cell invasive properties. In addition, we differentiate for the first time the contributions of ERBB1 and ERBB2 to the key metastatic properties of in vivo tumor cell invasion and intravasation. These experiments temporally and molecularly separate two key stages in tumor cell entry into blood vessels: invasion and intravasation. These results indicate that ERBB inhibition should be considered for blocking other tumor cell malignant properties besides growth.
Interaction of TFAP2C with the Estrogen Receptor-α Promoter Is Controlled by Chromatin Structure
Authors: Woodfield George W. , Hitchler Michael J. , Chen Yizhen , Domann Frederick E. , Weigel Ronald J.
Published: May 2009

Purpose: Transcriptional regulation of estrogen receptor-α (ERα) involves both epigenetic mechanisms and trans-active factors, such as TFAP2C, which induces ERα transcription through an AP-2 regulatory region in the ERα promoter. Attempts to induce endogenous ERα expression in ERα-negative breast carcinomas by forced overexpression of TFAP2C have not been successful. We hypothesize that epigenetic chromatin structure alters the activity of TFAP2C at the ERα promoter.

Experimental Design: DNA methylation, histone acetylation, and chromatin accessibility were examined at the ERα promoter in a panel of breast carcinoma cell lines. TFAP2C and polymerase II binding were analyzed by chromatin immunoprecipitation. Epigenetic chromatin structure was altered using drug treatment with 5-aza-2′-deoxycytidine (AZA) and trichostatin A (TSA).

Results: The ERα promoter in the ERα-negative lines MDA-MB-231, MCF10A, and MCF7-5C show CpG island methylation, histone 3 lysine 9 deacetylation, and decreased chromatin accessibility compared with ERα-positive cell lines MCF7 and T47-D. Treatment with AZA/TSA increased chromatin accessibility at the ERα promoter and allowed TFAP2C to induce ERα expression in ERα-negative cells. Chromatin immunoprecipitation analysis showed that binding of TFAP2C to the ERα promoter is blocked in ERα-negative cells but that treatment with AZA/TSA enabled TFAP2C and polymerase II binding.

Conclusion: We conclude that the activity of TFAP2C at specific target genes depends upon epigenetic chromatin structure. Furthermore, the combination of increasing chromatin accessibility and inducing TFAP2C provides a more robust activation of the ERα gene in ERα-negative breast cancer cells.
Satraplatin, an Oral Platinum, Administered on a Five-day Every-Five-Week Schedule: a Pharmacokinetic and Food Effect Study
Authors: Ricart Alejandro D. , Sarantopoulos John , Calvo Emiliano , Chu Quincy S. , Greene Douglas , Nathan Faith E. , Petrone Michael E. , Tolcher Anthony W. , Papadopoulos Kyriakos P.
Published: May 2009

Purpose: The study aimed to assess the pharmacokinetic behavior of satraplatin under fasted and fed conditions, and its safety and preliminary antitumor activity in adults with advanced solid tumors.

Experimental Design: Satraplatin was administered orally at 80 mg/m2 once daily with prophylactic antiemetics for 5 consecutive days every 5 weeks. Patients were randomized to receive day 1 and day 5 doses of satraplatin in either the fed or fasted state, the order being reversed for cycle 2. Pharmacokinetic sampling was done during the first two cycles. For all subsequent cycles, patients received satraplatin in the fasted state.

Results: Seventeen patients were treated with 60 total cycles of satraplatin. There was no dose-limiting toxicity during cycle 1. Severe hematologic toxicity was rare and the hematologic nadir occurred during week 4. Nausea, vomiting, and diarrhea were grade 1/2. No significant cardiac, renal, hepatic, or neurologic toxicity was observed. The hypothesis that food decreased ultrafiltrate platinum bioavailability could not be rejected, as the lower limit of the 90% confidence intervals for peak plasma concentration and area under the concentration-time curve from time 0 to 24 hours were 56.14% and 73.53%, respectively, both below the 80% bioequivalence acceptance criterion. One partial response (hormone refractory prostate cancer) and four durable stable diseases (breast, ovarian, parotid, and hormone refractory prostate cancer) were confirmed.

Conclusions: There is an effect of food on the pharmacokinetics of satraplatin, the clinical significance of which is unclear. It is recommended that satraplatin be administered in the fasting state. This 5-week interval schedule of satraplatin was well tolerated in heavily pretreated patients.
Proteasome Enzymatic Activities in Plasma as Risk Stratification of Patients with Acute Myeloid Leukemia and Advanced-Stage Myelodysplastic Syndrome
Authors: Ma Wanlong , Kantarjian Hagop , Bekele Benjamin , Donahue Amber C. , Zhang Xi , Zhang Zhong J. , O'Brien Susan , Estey Elihu , Estrov Zeev , Cortes Jorge , Keating Michael , Giles Francis , Albitar Maher
Published: May 2009

Purpose: Cytogenetic abnormalities are currently the most important predictors of response and clinical outcome for patients with acute myeloid leukemia (AML) or advanced-stage myelodysplastic syndrome (MDS). Because clinical outcomes vary markedly within cytogenetic subgroups, additional biological markers are needed for risk stratification.

Experimental Design: We assessed the utility of measuring pretreatment proteasome chymotrypsin-like, caspase-like, and trypsin-like activities in plasma to predict response and survival of patients with AML (n = 174) or advanced-stage MDS (n = 52).

Results: All three enzymatic activities were significantly (P < 0.001) increased in the plasma of patients with AML and MDS compared with normal controls. Both chymotrypsin-like and caspase-like activities, but not trypsin-like activity, correlated with outcome. Chymotrypsin-like and caspase-like activities, but not trypsin-like activity, predicted response in univariate analysis (P = 0.002). However, only chymotrypsin-like activity was independent predictor of response from age grouping (<70 versus ≥70 years), cytogenetics, and blood urea nitrogen in multivariate analysis. Similarly, both chymotrypsin-like and caspase-like activities, but not trypsin-like activity, were predictors of overall survival in univariate analysis (P < 0.0001), but only chymotrypsin-like activity was independent of cytogenetics, age, performance status, blood urea nitrogen, and β2-microglobulin in multivariate Cox regression models. Chymotrypsin-like activity was also a strong independent predictor of survival in patients with intermediate karyotype (n = 124).

Conclusions: Measuring plasma chymotrypsin-like activity may provide a powerful biomarker for risk stratification in patients with AML and advanced-stage MDS, including those with normal karyotype.
A Phase 0 Trial of Riluzole in Patients with Resectable Stage III and IV Melanoma
Authors: Yip Dana , Le Maithao N. , Chan Joseph L.-K. , Lee Jonathan H. , Mehnert Janice A. , Yudd Anthony , Kempf Jeffery , Shih Weichung J. , Chen Suzie , Goydos James S.
Published: May 2009

Purpose: Ectopic expression of GRM1 in murine melanocytes results in transformation into a form of melanoma, and more than 60% of human melanoma samples tested ectopically express GRM1. Stimulation of this receptor in vitro results in up-regulation of activated extracellular signal–regulated kinase (ERK). Furthermore, a xenograft model of melanoma treated with riluzole, an oral GRM1 blocking agent, showed decreased tumor growth compared with the untreated controls. We have now completed a phase 0 trial of riluzole in patients with melanoma.

Experimental Design: Patients enrolled on this trial underwent a pretreatment biopsy, took 200 mg of oral riluzole per day for 14 days, and then underwent resection of their remaining tumor. We compared the levels of pERK and pAKT in the pretreatment and post-treatment samples and assessed the metabolic activity of pretreatment and post-treatment tumors using fluorodeoxyglucose positron emission tomography (FDG-PET) scanning.

Results: We accrued 12 patients and all expressed GRM1. We found a significant decrease in pAKT and/or pERK in post-treatment tumor samples as compared with pretreatment samples in 4 (34%) patients. These four patients had a significant decrease in FDG-PET intensity post-treatment as well. Two other patients had a clinical response with no corresponding metabolic response; five patients had similar pretreatment and post-treatment FDG-PET scan findings; and one patient had progressive disease.

Conclusions: Our data show that glutamate blockade with riluzole can inhibit signaling through the mitogen-activated protein kinase and phosphatidylinositol 3-kinase/AKT pathways and suppress the metabolic activity of melanoma. The ectopic expression of metabotropic glutamate receptors may be important in the pathogenesis of human melanoma, and targeting this pathway may be an effective therapy.
B-Cell Translocation Gene 2 Is Over-Expressed in Peri-Infarct Neurons after Ischaemic Stroke
Authors: Slevin M. , Sanfeliu C. , Turu M.M. , Grau-Olivares M. , Ferrer I. , Boluda S. , Marti-Fabregas J. , Kumar S. , Kumar P. , Krupinski J.
Published: May 2009

Objectives: Recovery from stroke is dependent on the survival of neurons in the dynamic peri-infarcted region. Although several markers of neuronal injury and apoptotic cell death have been described, administration of neuroprotective drugs directed at specific molecules has had limited success. A complete understanding of deregulated genes associated with neuronal death would be beneficial. Our previous microarray studies identified increased expression of a novel protein, the B-cell translocation gene 2 (BTG2), in infarcted regions. Methods: We have used immunohistochemistry and Western blotting to examine the expression and localization of BTG2 in stroked brain tissue and immunofluorescent staining of human fetal brain neurons to determine if oxygen-glucose deprivation affected its expression. Results: We show that BTG2 is strongly expressed in peri-infarcted and infarcted regions of brain tissue, localizing in neuronal nuclei and cytoplasm, whilst being absent or very weakly expressed in normal looking contralateral tissue. Exposure of human fetal brain neurons to oxygen-glucose deprivation also induced BTG2 expression in the cytoplasm and perinuclear regions of cells staining positive for propidium iodide (a marker of nuclear damage). Conclusions: BTG2 may be a modulator of cell survival and differentiation and could help to protect against cell death by inhibition of necrosis and/or apoptotic signalling pathways.
Immunolocalization of DNMT1 and DNMT3a in Salivary Gland Neoplasms
Authors: Cavaliéri Gomes Carolina , da Silveira e Oliveira Carla , Santos Pimenta Luiz Gustavo Garcia , De Marco Luiz , Santiago Gomez Ricardo
Published: May 2009

Objective: Salivary gland neoplasms pathogenesis has not been well established. DNA methylation occurs when methyl groups are added to cytosine nucleotides in specific areas of the gene by the enzyme DNA methyltransferase (DNMT). This chemical modification can alter gene expression without altering DNA sequence. While DNMT3a is mostly involved in de novo methylation, DNMT1 acts as a maintenance methyltransferase. We aimed to investigate the immunoexpression of DNMT3a and DNMT1 in minor salivary gland neoplasms, comparing it with normal tissue. Material: Forty-four formalin-fixed and paraffin-embedded samples of pleomorphic adenoma, adenoid cystic carcinoma, mucoepidermoid carcinoma and polymorphous low-grade adenocarcinoma were included in the study. The DNMT1 and DNMT3a proteins were identified by using a highly sensitive polymer-based system. Results: Positive nuclear and cytoplasmic labeling for DNMT1 was observed in all samples, including the controls. Positive nuclear labeling for DNMT3a was found only in few neoplasms: 1 pleomorphic adenoma (9.0%), 2 adenoid cystic carcinoma (16.6%) and 1 mucoepidermoid (9.0%) cases. Conclusion: Our results were not able to demonstrate a clear correlation between DNMT1 and DNMT3a immunoexpression and salivary gland neoplasms development.
p16ink4a Is a Surrogate Marker for High-Risk and Malignant Cervical Lesions in the Presence of Human Papillomavirus
Authors: Lakshmi S. , Rema P. , Somanathan Thara
Published: May 2009

Objective: To strengthen the role of high-risk human papillomaviruses (HPVs) in the development of cervical cancer, and the association between HPV and the cell cycle inhibitor gene p16ink4a in cervical carcinogenesis. Methods: In this study, the association between p16ink4a and the presence of HPV DNA in cervical lesions was investigated in a total of 177 cervical biopsies classified as benign (n = 42), low-grade squamous intraepithelial lesions (LSIL; n = 34), high-grade squamous intraepithelial lesions (HSIL; n = 48) and invasive cancer (n = 53). p16ink4a expression was analyzed by immunohistochemistry while HPV DNA was localized by in situ hybridization. Results: No significant association was found between p16ink4a expression and HPV presence in benign and LSIL samples. In HSIL and invasive cancer, a significant association was found between p16ink4a overexpression and the presence of HPV DNA. Conclusion: Immunohistochemical p16ink4a expression is associated with HPV infection in HSIL and cervical cancer, suggesting a role of p16 as a biomarker of HPV-associated cervical lesions.
Symmetric Division and Expression of Its Regulatory Gene Numb in Human Cervical Squamous Carcinoma Cells
Authors: Chen Huaizeng , Chen Xiaojun , Ye Feng , Lu Weiguo , Xie Xing
Published: May 2009

Objective: To investigate the mode of cell division and Numb expression in cervical squamous cells. Method: We measured the cleavage angles of cervical squamous epithelia basal cells and Numb protein expression in 50 cases of cervical squamous cell carcinomas (CSCC), 59 cervical intraepithelial neoplasias (CIN) grade II–III, 21 CIN grade I and 48 controls. Results: We found an increase in horizontal divisions in CIN grade I and a decrease in horizontal divisions in CIN grade II–III. Numb expression had a tendency to be stronger from control to CSCC. In CSCC there was a significantly higher expression of mRNA for the Numb-PRRL isoform but not for the Numb-PRRS isoform. Conclusion: Our findings suggest that in the initial stage of CSCC, carcinomatous cells may acquire an increase of symmetric division by altering cleavage orientation and proliferation. Expression of Numb protein in Cervical squamous epithelia and an increased expression of Numb-PRRL isoforms may be among the mechanisms involved in the genesis of CSCC.
Simple Interrupted Percutaneous Suture versus Intradermal Running Suture for Wound Tensile Strength Measurement in Rats: A Technical Note
Authors: Gál P. , Toporcer T. , Vidinský B. , Hudák R. , Živčák J. , Sabo J.
Published: May 2009

Background: In this experimental study, simplicity of measurement and wound tensile strength of wounds fixed by simple interrupted percutaneous suture (SIPS) and intradermal running suture (IRS) were compared. Methods: Twenty-eight male Sprague-Dawley rats were used for the experiment and separated into two groups: SIPS group and IRS group. Under general anesthesia, two parallel full-thickness (4-cm) skin incisions were made on the back of each rat. Wounds in the SIPS group were closed using 4 interrupted percutaneous sutures, whereas wounds in the IRS group were closed by intradermal running suture. Seven animals from each group were sacrificed at 2 and at 5 days after surgery for tensile strength testing. Results: The wound tensile strength of IRS wounds was significantly higher than that of SIPS wounds at 2 days (SIPS = 2.9 ± 0.8 vs. IRS = 3.7 ± 0.9 g/mm2, p < 0.05) and at 5 days (SIPS = 5.6 ± 1.3 vs. IRS = 7.1 ± 1.2 g/mm2, p < 0.01). In addition, the measurement of IRS wounds was easier and faster due to removal of only one suture. Conclusions: These findings highlight the advantages of expanding the use of IRS suturing in experimental studies conducted on rats.
Incidence and Risk Factors of Intracranial Atherosclerotic Stroke: The Northern Manhattan Stroke Study
Authors: Rincon Fred , Sacco Ralph L. , Kranwinkel Grace , Xu Qiu , Paik Myungee C. , Boden-Albala Bernadette , Elkind Mitchell S.V.
Published: May 2009

Background: To assess the prevalence of risk factors as determinants of intracranial atherosclerosis (IATH)-related stroke in a multi-ethnic community-based cohort. Methods: The Northern Manhattan Stroke Study included a population-based incidence study and a nested case-control study. Incident cases of first ischemic stroke were 1:2 when matched to community controls by age, sex, and race/ethnicity. Vascular risk factors were assessed among controls and compared against the following stroke subtypes: IATH, extracranial atherosclerosis (EATH), and non-atherosclerotic (NATH: cardioembolic, lacunar, and cryptogenic). Conditional logistic-regression was used to determine the association between risk factors and stroke subtypes. Results: The crude incidence of IATH was 8/100,000 per year and the relative incidence of IATH was higher than that of EATH in blacks (5.9 vs. 3.2/100,000 per year) and in Hispanics (5.0 vs. 1.7/100,000 per year). The IATH group had a higher prevalence of diabetes mellitus (DM; 67% IATH, 60% EATH, 48% NATH, and 23% controls; p < 0.05 IATH vs. control) and of metabolic syndrome (62% IATH, 40% EATH, 40% NATH, and 35% controls; p < 0.05 IATH vs. control). In multivariate analysis, DM conferred a higher risk for IATH versus NATH stroke (OR, 10.8; 95% CI, 2.0–57 vs. OR, 2.7; 95% CI, 1.9–3.9; p < 0.05) and much lower for EATH (OR, 6.2; 95% CI, 1.2–32). The metabolic syndrome conferred a higher risk for IATH stroke subtype (OR, 4.6; 95% CI, 1.1–18.7) when compared to EATH (OR, 2.3; CI, 0.6–9.1) and NATH (OR, 2.4; CI, 1.7–3.3). Conclusions: DM is a more important determinant for IATH-related stroke than EATH or NATH.
Myocardial Scars Are an Underestimated Cardiovascular Burden in Patients with Internal Carotid Artery Stenosis
Authors: Bruder O. , Göricke S. , Hunold P. , Lowitsch M. , Barkhausen J. , Sabin G.V. , Forsting M. , Fiebach J.B.
Published: May 2009

Background: Patients with internal carotid artery (ICA) stenosis have an increased incidence of coronary heart disease. Evidence about the incidence of clinically silent myocardial infarction (MI) in these patients is limited. Contrast-enhanced cardiac magnetic resonance (CMR) imaging allows for the detection of minor myocardial damage. Objective: We tested whether patients with ICA stenosis exhibit a relevant incidence of silent MI when assessed by CMR. Methods: In a single-center study, 77 consecutive patients (age 68 ± 7 years) with suspected ICA stenosis were imaged prospectively with a combined MRI protocol including T1, T2, diffusion-weighted imaging, fluid-attenuated inversion recovery, and contrast-enhanced MR angiography (CEMRA) imaging of the brain and a short (11 min) CMR protocol with left ventricular function and late gadolinium enhancement imaging. Blinded to any clinical information, two readers evaluated the cardiac and neuroradiologic examinations. Results: Of 154 imaged ICA, 85 presented with stenosis and 17 were occluded. In 7 patients, the suspected ICA stenosis could not be confirmed by CEMRA. In the remaining 70 patients with ICA stenosis, 34.3% had cerebral lesions (15.7% with a homodynamic pattern,18.6% with territorial infarction). CMR detected MI in 29 (41%) patients, whereas ECG and medical history enabled diagnosis in only 7 (10%) patients. Conclusions: ICA stenosis patients have a higher incidence of myocardial scars proving silent MI when detected by contrast-enhanced CMR than clinically expected. Whether the presence and extent of silent MIs detected by CMR affect peri-interventional risk and prognosis of ICA stenosis patients remains to be evaluated in a large patient cohort with long-term follow-up.
Renal Function Predicts Survival in Patients with Acute Ischemic Stroke
Authors: Mostofsky Elizabeth , Wellenius Gregory A. , Noheria Amit , Levitan Emily B. , Burger Mary R. , Schlaug Gottfried , Mittleman Murray A.
Published: May 2009

Background: Among patients with acute ischemic stroke, impaired kidney function has been shown to increase the mortality risk, but the shape of this relationship has not been evaluated in detail. Methods: We estimated the glomerular filtration rate (eGFR) at the time of hospitalization in 1,175 consecutive patients hospitalized with acute ischemic stroke at the Beth Israel Deaconess Medical Center and examined the shape of the association between eGFR and all-cause mortality. Results: There were 508 deaths during a median follow-up of 40.3 months, resulting in a ‘U’-shaped relationship between eGFR and all-cause mortality. The curve was relatively flat between 75 and 110 ml/min/1.73 m2 but increased sharply at lower and higher levels of eGFR (test for nonlinearity: p < 0.0001). Conclusions: Among patients with acute ischemic stroke, a reduced or highly elevated eGFR at hospital admission is associated with a higher mortality rate compared to patients with moderate levels of eGFR.
Stress Influences Brain Enkephalinase, Oxytocinase and Angiotensinase Activities: A New Hypothesis
Authors: Hernández Joaquin , Segarra Ana B. , Ramírez Manuel , Banegas Inmaculada , de Gasparo Marc , Alba Francisco , Vives Francisco , Durán Raquel , Prieto Isabel
Published: May 2009

Brain enkephalin and oxytocin are anxiolytic agents involved in the response mechanism to stress. Degrading enzymes such as enkephalinase and oxytocinase could also be associated with this response. The effect of acute immobilization stress on enkephalinase and oxytocinase activities was determined in the soluble and membrane fractions of the medial prefrontal cortex, hippocampus and amygdala using alanyl- and leucyl-beta-naphthylamide as substrates, the latter in the presence and absence of 20 mML-methionine. No change in aminopeptidase activities was observed in the prefrontal cortex of stressed rats. In contrast, enkephalinase activity decreased in the soluble fraction of the hippocampus but increased in the membrane fraction. In the amygdala, soluble oxytocinase and membrane enkephalinase activities decreased in stressed animals. These results show that acute immobilization stress affects differentially enkephalinase and oxytocinase activities depending on the fraction and brain region analyzed. A reduction in the activity of soluble enkephalinase in the hippocampus and soluble oxytocinase as well as membrane enkephalinase in the amygdala may suggest higher availability/longer action of enkephalin and oxytocin at these locations. This may explain the relative importance of these enzymatic activities in the anxiolytic properties proposed for enkephalins and oxytocin in the hippocampus and amygdala during stress conditions. This interpretation is not applicable to membrane enkephalinase activity in the hippocampus. However, alanyl-beta-naphthylamide hydrolyzing activity not only measures enkephalinase activity, it also reflects the angiotensinase-induced metabolism of angiotensin III to angiotensin IV. Therefore, our results may also mirror an increase in the formation of Ang IV in hippocampus and a decrease in the amygdala in acute stress. In conclusion, aminopeptidase activities in the hippocampus and amygdala may affect enkephalin, oxytocin and angiotensin III metabolism during acute immobilization stress and therefore be involved in the anxiolytic response.
Laparoscopic Removal of a Foreign Body in the Pelvic Cavity through One Port Using a Flexible Cholangioscope
Authors: Kurita Nobuhiro , Shimada Mitsuo , Nakao Toshihiro , Chikakiyo Motoya , Miyatani Tomohiko , Higashijima Jun , Yoshikawa Kozo , Nishioka Masanori , Iwata Takashi
Published: May 2009

Background: More than two trocar ports 12 mm in diameter were usually used to remove a foreign body from the peritoneal cavity using laparoscopic surgery. We designed a method of laparoscopic removal through a single port 5 mm in diameter and a flexible cholangioscope. Patients and Methods: The patient had a ventriculoperitoneal shunt (V-P shunt) catheter implanted for hydrocephalus of unknown cause in his 40s. Endoscopic marsupialization of the third ventricle was performed because of functional failure of the V-P shunt catheter 7 years after the implantation. A falling off of the V-P shunt catheter into the pelvic space was detected, and laparoscopic removal of the V-P shunt catheter was performed. A laparoscope was inserted through a trocar port 5 mm in diameter to confirm the location of the V-P shunt catheter following replacement of the flexible cholangioscope to grasp the catheter. The catheter was held using a snare catheter through the cholangioscope. The V-P shunt catheter was removed by pulling through the trocar port with the flexible cholangioscope. Conclusion: Laparoscopic removal is a good technique for removal of a foreign body in the peritoneal cavity. It enables one-port laparoscopic removal using a flexible cholangioscope.
Early Onset Gastric Cancer No Longer Presents as an Advanced Disease with Ominous Prognosis
Authors: Moreira H. , Pinto-de-Sousa J. , Carneiro F. , Cardoso de Oliveira M. , Pimenta A.
Published: May 2009

Background: This study aimed to characterize the clinicopathological features and prognosis of early onset gastric carcinomas (EOGCs) in a sample of young Portuguese adults. Material and Methods: A subset of 415 patients submitted to gastric resection was divided into 2 groups: A – patients aged 45 years or less (n = 108), and B – patients older than 45 years (n = 307). The groups were compared for several clinicopathological features and the survival of the patients was analyzed by uni- and multivariate analysis. Results: The surgical resection rate was higher in young patients, in whom tumors more frequently displayed an ulcerative shape (p < 0.001) and diffuse/isolated cell-type histology (p < 0.001). In the elderly, the rate of vascular invasion was significantly higher (p = 0.002). Overall, the survival of young patients was higher than in the elderly (p < 0.001). Survival curves were higher in young patients, except for pT4 and pM1 cases. In the multivariate survival analysis, age (p < 0.001), depth of tumor invasion (p < 0.001), nodal status (p < 0.001), vascular invasion (p = 0.011) and distant metastases (p = 0.007) emerged as independent prognostic factors for gastric carcinoma. Conclusion: Young gastric cancer patients no longer present with more advanced disease than the elderly and, overall, survival is better in young patients.