bluuebunny/crossref_metadata_2025 huggingface dataset:
Modulation of muscle sympathetic nerve activity to muscle heating during dynamic exercise
Authors: Cook Jonathan S. , Ray Chester A.
Published: March 2009

Previous studies from our laboratory have demonstrated that altering muscle temperature of the exercising forearm can elicit changes in muscle sympathetic nerve activity (MSNA) during ischemic isometric handgrip. The purpose of the current study was to determine the interactive effect of muscle temperature and blood flow on MSNA responses during dynamic handgrip (DHG). Eight subjects performed two bouts of graded DHG to fatigue followed by 2 min of postexercise muscle ischemia (PEMI). Local heating of the forearm increased muscle temperature from 33.6 ± 0.3 to 38.3 ± 0.5°C ( P < 0.05). Mean arterial pressure and heart rate increased in a linear fashion during graded DHG ( P < 0.05) but were not affected by muscle temperature. MSNA (burst frequency and total activity) at fatigue and PEMI were elevated in all conditions ( P < 0.05). However, MSNA responses were not different between temperature conditions. To ascertain the effect of blood flow, eight additional subjects completed two trials of ischemic DHG under control or warm conditions followed by 2 min of PEMI. MSNA, expressed as burst frequency and total activity, was significantly greater in warm compared with the control trial (Δ14 ± 3 and Δ9 ± 2 bursts/30 s, and Δ1,234 ± 260 and Δ751 ± 199 units/30 s, respectively). This finding supports the concept that muscle heating sensitizes skeletal muscle afferents during muscle contractions and augments MSNA in humans. However, on the basis of these findings, we conclude that muscle blood flow modulates the effect of muscle temperature on MSNA during exercise.
Chromatic Contrast Sensitivity During Optokinetic Nystagmus, Visually Enhanced Vestibulo-ocular Reflex, and Smooth Pursuit Eye Movements
Authors: Schütz Alexander C. , Braun Doris I. , Gegenfurtner Karl R.
Published: March 2009

Recently we showed that sensitivity for chromatic- and high-spatial frequency luminance stimuli is enhanced during smooth-pursuit eye movements (SPEMs). Here we investigated whether this enhancement is a general property of slow eye movements. Besides SPEM there are two other classes of eye movements that operate in a similar range of eye velocities: the optokinetic nystagmus (OKN) is a reflexive pattern of alternating fast and slow eye movements elicited by wide-field visual motion and the vestibulo-ocular reflex (VOR) stabilizes the gaze during head movements. In a natural environment all three classes of eye movements act synergistically to allow clear central vision during self- and object motion. To test whether the same improvement of chromatic sensitivity occurs during all of these eye movements, we measured human detection performance of chromatic and luminance line stimuli during OKN and contrast sensitivity during VOR and SPEM at comparable velocities. For comparison, performance in the same tasks was tested during fixation. During the slow phase of OKN we found a similar enhancement of chromatic detection rate like that during SPEM, whereas no enhancement was observable during VOR. This result indicates similarities between slow-phase OKN and SPEM, which are distinct from VOR.
Genetic control of global gene expression levels in the intestinal mucosa: a human twin study
Authors: Häsler Robert , Begun Alexander , Freitag-Wolf Sandra , Kerick Martin , Mah Nancy , Zvirbliene Aida , Spehlmann Martina E. , von Wurmb-Schwark Nicole , Kupcinskas Limas , Rosenstiel Philip , Schreiber Stefan
Published: March 2009

Phenotypic variation between individuals, such as different mRNA expression levels, is influenced by genetic and nongenetic factors. Although several studies have addressed the interplay between genotypes and expression profiles in various model organisms in the recent years, the detailed and relative contributions of genetic and nongenetic factors in regulating plasticity of gene expression in barrier organs (e.g., skin, gut), which are exposed to continuous environmental challenge, are still poorly understood. Here we systematically monitored the level of genetic control over genomewide mRNA expression profiles in the healthy intestinal mucosa of 10 monozygotic and 10 dizygotic human twin pairs with microarray analyses. Our results, which are supported by real-time PCR and analysis of molecular phylogenetic conservation, indicate that genes associated with energy metabolism and cell and tissue regeneration pathways are under strong genetic control. Conversely, genes associated with immune response seem to be mainly controlled by exogenous factors. Further insights into the relative extent of genetic and nongenetic determinants of transcriptomal profiles and their influence on physiological and pathophysiological mechanisms are crucial to understanding the key role played by gene-environment interactions in health and disease.
Strong Smooth Muscle Differentiation Is Dependent on Myocardin Gene Amplification in Most Human Retroperitoneal Leiomyosarcomas
Authors: Pérot Gaëlle , Derré Josette , Coindre Jean-Michel , Tirode Franck , Lucchesi Carlo , Mariani Odette , Gibault Laure , Guillou Louis , Terrier Philippe , Aurias Alain
Published: March 2009

Myocardin (MYOCD), a serum response factor (SRF) transcriptional cofactor, is essential for cardiac and smooth muscle development and differentiation. We show here by array-based comparative genomic hybridization, fluorescence in situ hybridization, and expression analysis approaches that MYOCD gene is highly amplified and overexpressed in human retroperitoneal leiomyosarcomas (LMS), a very aggressive well-differentiated tumor. MYOCD inactivation by shRNA in a human LMS cell line with MYOCD locus amplification leads to a dramatic decrease of smooth muscle differentiation and strongly reduces cell migration. Moreover, forced MYOCD expression in three undifferentiated sarcoma cell lines and in one liposarcoma cell line confers a strong smooth muscle differentiation phenotype and increased migration abilities. Collectively, these results show that human retroperitoneal LMS differentiation is dependent on MYOCD amplification/overexpression, suggesting that in these well-differentiated LMS, differentiation could be a consequence of an acquired genomic alteration. In this hypothesis, these tumors would not necessarily derive from cells initially committed to smooth muscle differentiation. These data also provide new insights on the cellular origin of these sarcomas and on the complex connections between oncogenesis and differentiation in mesenchymal tumors. [Cancer Res 2009;69(6):2269–78]
Quantitative Phosphoproteomics Reveals a Cluster of Tyrosine Kinases That Mediates Src Invasive Activity in Advanced Colon Carcinoma Cells
Authors: Leroy Cédric , Fialin Camille , Sirvent Audrey , Simon Valérie , Urbach Serge , Poncet Joël , Robert Bruno , Jouin Patrick , Roche Serge
Published: March 2009

The nonreceptor tyrosine kinase Src is frequently overexpressed and/or activated in human colorectal carcinoma (CRC), and its increased activity has been associated with a poor clinical outcome. Src has been implicated in growth and invasion of these cancer cells by still not well-known mechanisms. Here, we addressed Src oncogenic signaling using quantitative phosphoproteomics. Src overexpression increased growth and invasiveness of metastatic SW620 CRC cells. Stable isotope labeling with amino acids in cell culture in combination with liquid chromatography tandem mass spectrometry allowed the identification of 136 proteins which exhibited a significant increase in and/or association with tyrosine phosphorylation upon Src expression. These mainly include signaling, cytoskeleton, and vesicular-associated proteins. Interestingly, Src also phosphorylated a cluster of tyrosine kinases, i.e., the receptors Met and EphA2, the cytoplasmic tyrosine kinase Fak, and pseudo-tyrosine kinase SgK223, which were required for its invasive activity. Similar results were obtained with metastatic Colo205 CRC cells that exhibit high endogenous Src activity. We concluded that Src uses a tyrosine kinases network to promote its invasive activity in CRC and this implicates a reverse signaling via tyrosine kinase receptors. Targeting these tyrosine kinases may be of significant therapeutic value in this cancer. [Cancer Res 2009;69(6):2279–86]
CYP1A1/2 Haplotypes and Lung Cancer and Assessment of Confounding by Population Stratification
Authors: Aldrich Melinda C. , Selvin Steve , Hansen Helen M. , Barcellos Lisa F. , Wrensch Margaret R. , Sison Jennette D. , Kelsey Karl T. , Buffler Patricia A. , Quesenberry Charles P. , Seldin Michael F. , Wiencke John K.
Published: March 2009

Prior studies of lung cancer and CYP1A1/2 in African-American and Latino populations have shown inconsistent results and have not yet investigated the haplotype block structure of CYP1A1/2 or addressed potential population stratification. To investigate haplotypes in the CYP1A1/2 region and lung cancer in African-Americans and Latinos, we conducted a case-control study (1998–2003). African-Americans (n = 535) and Latinos (n = 412) were frequency matched on age, sex, and self-reported race/ethnicity. We used a custom genotyping panel containing 50 single nucleotide polymorphisms in the CYP1A1/2 region and 184 ancestry informative markers selected to have large allele frequency differences between Africans, Europeans, and Amerindians. Latinos exhibited significant haplotype main effects in two blocks even after adjusting for admixture [odds ratio (OR), 2.02; 95% confidence interval (95% CI), 1.28–3.19 and OR, 0.55; 95% CI, 0.36–0.83], but no main effects were found among African-Americans. Adjustment for admixture revealed substantial confounding by population stratification among Latinos but not African-Americans. Among Latinos and African-Americans, interactions between smoking level and haplotypes were not statistically significant. Evidence of population stratification among Latinos underscores the importance of adjusting for admixture in lung cancer association studies, particularly in Latino populations. These results suggest that a variant occurring within the CYP1A2 region may be conferring an increased risk of lung cancer in Latinos. [Cancer Res 2009;69(6):2340–8]
Cetuximab Attenuates Metastasis and u-PAR Expression in Non–Small Cell Lung Cancer: u-PAR and E-Cadherin are Novel Biomarkers of Cetuximab Sensitivity
Authors: Nikolova Dessislava A. , Asangani Irfan A. , Nelson Laura D. , Hughes Dennis P.M. , Siwak Doris R. , Mills Gordon B. , Harms Andrea , Buchholz Erika , Pilz Lothar R. , Manegold Christian , Allgayer Heike
Published: March 2009

Cetuximab, which blocks ligand binding to epidermal growth factor receptor (EGFR), is currently being studied as a novel treatment for non–small cell lung cancer (NSCLC). However, its mechanisms of action toward metastasis, and markers of drug sensitivity, have not been fully elucidated. This study was conducted to (a) determine the effect of Cetuximab on invasion and NSCLC-metastasis; (b) investigate urokinase-type plasminogen activator receptor (u-PAR), a major molecule promoting invasion and metastasis, as a target molecule; (c) delineate molecular mediators of Cetuximab-induced metastasis inhibition; and (d) identify biomarkers of drug sensitivity in NSCLC. Cetuximab treatment resulted in reduced growth and Matrigel invasion of H1395 and A549 NSCLC cell lines, in parallel with reduced u-PAR mRNA and protein. u-PAR down-regulation was brought about by suppressing the binding of JunD and c-Jun to u-PAR promoter motif −190/−171 in vivo, and an inhibition of MAP/ERK kinase signaling. Furthermore, Cetuximab inhibited NSCLC proliferation and metastasis to distant organs in vivo as indicated by the chicken embryo metastasis assay. Low E-cadherin and high u-PAR, but not EGFR, was associated with resistance to Cetuximab in seven NSCLC cell lines. Furthermore, siRNA knockdown of u-PAR led to a resensitization to Cetuximab. Moreover, low E-cadherin and high u-PAR was found in 63% of resected tumor tissues of NSCLC patients progressing under Cetuximab therapy. This is the first study to show u-PAR as a target and marker of sensitivity to Cetuximab, and to delineate novel mechanisms leading to metastasis suppression of NSCLC by Cetuximab. [Cancer Res 2009;69(6):2461–70]
Mitochondrial Dysfunction and Reactive Oxygen Species Imbalance Promote Breast Cancer Cell Motility through a CXCL14-Mediated Mechanism
Authors: Pelicano Helene , Lu Weiqin , Zhou Yan , Zhang Wan , Chen Zhao , Hu Yumin , Huang Peng
Published: March 2009

Although mitochondrial dysfunction and reactive oxygen species (ROS) stress have long been observed in cancer cells, their role in promoting malignant cell behavior remains unclear. Here, we show that perturbation of the mitochondrial respiratory chain in breast cancer cells leads to a generation of subclones of cells with increased ROS, active proliferation, high cellular motility, and invasive behaviors in vitro and in vivo. Gene expression analysis using microarrays revealed that all subclones overexpressed CXCL14, a novel chemokine with undefined function. We further show that CXCL14 expression is up-regulated by ROS through the activator protein-1 signaling pathway and promotes cell motility through elevation of cytosolic Ca2+ by binding to the inositol 1,4,5-trisphosphate receptor on the endoplasmic reticulum. Abrogation of CXCL14 expression using a decoy approach suppressed cell motility and invasion. Our data suggest that mitochondrial dysfunction and ROS stress promote cancer cell motility through a novel pathway mediated by CXCL14. [Cancer Res 2009;69(6):2375–83]
Associations between Selected Biomarkers and Prognosis in a Population-Based Pancreatic Cancer Tissue Microarray
Authors: Takikita Mikiko , Altekruse Sean , Lynch Charles F. , Goodman Mark T. , Hernandez Brenda Y. , Green Mark , Cozen Wendy , Cockburn Myles , Sibug Saber Maria , Topor Marie , Zeruto Chris , Abedi-Ardekani Behnoush , Reichman Marsha E. , Hewitt Stephen M.
Published: March 2009

Pancreatic cancer is the fourth leading cause of cancer death in the United States. Prognostic biomarkers are lacking, and treatment has limited effect on survival. Tissues from Surveillance, Epidemiology, and End Results registries (Iowa, Hawaii, and Los Angeles) were used to build a tissue microarray of 161 pancreatic tumors (113 resections and 48 biopsies). Proportional hazard models adjusted for age, race, sex, stage, time-period of diagnosis, and treatment. Associations were examined between markers (MUC1, MUC2, MUC5AC, synaptophysin, chromogranin, neuron specific enolase, epidermal growth factor receptor, HER2, CD5, CD138, CK5/6, CK19, CK20, and p53) and survival time from diagnosis. After adjusting for covariates, borderline statistically significant associations were seen between expression of each of the three mucins (MUC1, MUC2, and MUC5AC) and shorter survival time. The associations strengthened for 154 (96%) adenocarcinomas, particularly the 120 (75%) well-differentiated to moderately differentiated ductal adenocarcinomas, a tumor type that occurred more often in the cohort among White cases than cases of other racial origin (P < 0.01). For differentiated ductal adenocarcinomas, associations with shorter survival time were seen for expression of all three mucins combined versus other mucin expression patterns (adjusted hazard ratio, 1.8; 95% confidence interval, 1.2–2.6) and for MUC2(+) versus MUC2(−) expression (adjusted hazard ratio, 1.6; 95% confidence interval, 1.1–2.4). Mucin gene expression, particularly MUC2 expression, may have prognostic value for differentiated adenocarcinomas. Tumor histologies differed in this and Japanese cohorts. The tissue microarray is available to evaluate other biomarkers. Tissue-based surveillance can be used to monitor tumor histology in populations and facilitate applied research. [Cancer Res 2009;69(7):2950–5]
Alternate Endpoints for Screening Phase II Studies
Authors: Dhani Neesha , Tu Dongsheng , Sargent Daniel J. , Seymour Lesley , Moore Malcolm J.
Published: March 2009

Phase II trials are screening trials that seek to identify agents with sufficient activity to continue development and those for which further evaluation should be halted. Although definitive phase III trials use progression-free or overall survival to confirm clinical benefit, earlier endpoints are preferable for phase II trials. Traditionally, tumor shrinkage of a predetermined degree (response) has been used as a surrogate of eventual survival benefit based on the observation that high response rates (RR), and particularly complete responses, in the phase II setting resulted in survival benefit in subsequent phase III trials. Recently, some molecularly targeted agents have shown survival and clinical benefit despite very modest RRs in early clinical trials. These observations provide a major conundrum, with concerns of inappropriate termination of development for active agents with low RRs being balanced by concerns of inactive agents being taken to late-phase development with resultant increases in the failure rate of phase III trials. Numerous alternate or complementary endpoints have been explored, incorporating multinomial endpoints (including progression and response), progression-free survival, biomarkers, and, more recently, evaluation of tumor size as a continuous variable. In this review, we discuss the current status of phase II endpoints and present retrospective analyses of two international gastrointestinal cancer studies showing the potential utility of one novel approach. Alternate endpoints, although promising, require additional evaluation and prospective validation before their use as a primary endpoint for phase II trials.
Fluorescence In situ Hybridization Analysis of Circulating Tumor Cells in Metastatic Prostate Cancer
Authors: Leversha Margaret A. , Han Jialian , Asgari Zahra , Danila Daniel C. , Lin Oscar , Gonzalez-Espinoza Rita , Anand Aseem , Lilja Hans , Heller Glenn , Fleisher Martin , Scher Howard I.
Published: March 2009

Purpose: To assess the feasibility of characterizing gene copy number alteration by fluorescence in situ hybridization (FISH) of circulating tumor cells (CTC) isolated using the CellSearch system in patients with progressive castration-resistant metastatic prostate cancer.

Experimental Design: We used probe combinations that included the androgen receptor (AR) and MYC genes for FISH analysis of CTC samples collected from 77 men with castration-resistant metastatic prostate cancer.

Results: High-level chromosomal amplification of AR was detected in 38% and relative gain of MYC in 56% of samples analyzed. No such abnormalities were detected in samples with CTC counts of <10, reflecting ascertainment difficulty in these lower count samples.

Conclusion: The CTC isolated from our patient cohort present a very similar molecular cytogenetic profile to that reported for late-stage tumors and show that FISH analysis of CTC can be a valuable, noninvasive surrogate for routine tumor profiling. That as many as 50% of these patients have substantial amplification of the AR locus indicates that androgen signaling continues to play an important role in late-stage prostate cancer.
Expression of Immunosuppresive B7-H3 Ligand by Hormone-Treated Prostate Cancer Tumors and Metastases
Authors: Chavin Grant , Sheinin Yuri , Crispen Paul L. , Boorjian Stephen A. , Roth Timothy J. , Rangel Laureano , Blute Michael L. , Sebo Thomas J. , Tindall Don J. , Kwon Eugene D. , Karnes R. Jeffrey
Published: March 2009

Purpose: Prostate cancer cells uniformly express the immune cell inhibitory B7-H3 ligand. Enhanced B7-H3 expression correlates with increased disease progression and cancer-specific death after radical prostatectomy (RP).

Experimental Design: To further assess whether B7-H3 expression is hormone regulated and persists as a viable target during (or after) androgen-ablative therapy, we examined B7-H3 ligand expression within primary and metastatic cancer lesions in response to neoadjuvant hormone therapy (NHT) or palliative hormone deprivation. Tumor B7-H3 in RP specimens from men treated with ≥3 months of NHT was compared with B7-H3 in tumors from matched patients who received no therapy before RP. Hormone-treated and untreated metastatic lesions involving bone were also compared for levels of B7-H3 expression.

Results: Of 165 consecutive RP specimens in each cohort studied, sufficient tissues were available for 148 patients (89.7%) treated with NHT versus 127 patients (77.0%) treated with surgery alone. B7-H3 was expressed in 142 (95.9%) tumors from NHT patients compared with 122 (96.0%) tumors from patients treated with surgery alone (P = 0.91). B7-H3 expression intensity in RP specimens was not affected by NHT (P = 0.12). Bone metastases from 11 (32.4%) untreated and 23 (67.6%) androgen-ablated patients revealed that B7-H3 expression increased in response to hormone therapy (P = 0.04) relative to untreated lesions.

Conclusions: Taken together, B7-H3 expression seems to remain stable (or may even increase) in response to hormone therapy. As such, B7-H3 may represent an attractive target to improve treatment of men with high-risk hormone-treated or refractory prostate cancer.
CXCR3 expression is associated with poor survival in breast cancer and promotes metastasis in a murine model
Authors: Ma Xinrong , Norsworthy Kelly , Kundu Namita , Rodgers William H. , Gimotty Phyllis A. , Goloubeva Olga , Lipsky Michael , Li Yanchun , Holt Dawn , Fulton Amy
Published: March 2009

Breast tumor cells express the chemokine receptor CXCR3, which binds the ligands CXCL9, CXCL10, and CXCL11. CXCR3 and other chemokine receptors may mediate tumor metastasis by supporting migration of tumor cells to sites of ligand expression including the lymph nodes, lungs, and bone marrow. We examined the relationship of CXCR3 expression to clinical outcome in 75 women diagnosed with early-stage breast cancer. We detected CXCR3 in malignant epithelium from all tumors. Twelve percent were weakly positive and 64% had moderate levels of CXCR3. Strong CXCR3-positive staining was observed in 24% of tumors. Kaplan-Meier survival curves showed that high CXCR3 expression was associated with poorer overall survival; the unadjusted hazard ratio was 1.56 and it was marginally significant (P = 0.07). When interactions between lymph node status and CXCR3 were considered, the adjusted hazard ratio for CXCR3 was 2.62 (P = 0.02) for women with node-negative disease at diagnosis, whereas the hazard ratio for CXCR3 was not significant for those with node-positive disease. CXCR3 gene silencing inhibited lung colonization and spontaneous lung metastasis from mammary gland–implanted tumors in a murine model. The size or growth rate of the locally growing tumors was not affected. The antimetastatic effect of CXCR3 gene silencing was compromised in mice depleted of Natural Killer cells or with mutations in IFN-γ, suggesting that the role of CXCR3 is not simply to mediate tumor cell trafficking. These studies support the continued examination of CXCR3 as a potential therapeutic target in patients with breast cancer. [Mol Cancer Ther 2009;8(3):490–8]
Potent induction of B- and T-cell immunity against human carcinoembryonic antigen-expressing tumors in human carcinoembryonic antigen transgenic mice mediated by direct lentivector injection
Authors: Loisel-Meyer Severine , Felizardo Tania , Mariotti Jacopo , Mossoba Miriam E. , Foley Jason E. , Kammerer Robert , Mizue Nobuo , Keefe Robert , McCart J. Andrea , Zimmermann Wolfgang , Dropulic Boro , Fowler Daniel H. , Medin Jeffrey A.
Published: March 2009

The applicability of immunotherapy would be dramatically broadened to a greater number of recipients if direct “off-the-shelf” products could be engineered to engender functionally potent immune responses against true “self”-tumor antigens. This would obviate the need for ex vivo culture of dendritic cells or T cells on a patient-by-patient basis, for example. The carcinoembryonic antigen (CEA) is a glycoprotein expressed in normal gut epithelium that is up-regulated in the majority of colon cancers, non-small cell lung cancers, and half of all breast cancers. Such properties make CEA an excellent and important target for cancer immunotherapy. In this study, we show stabilization of 14-day established s.c. mGC4CEA tumors in human CEA (huCEA) transgenic mice following two direct low-dose injections of 0.15 × 106 transducing units of a lentiviral vector (LV) that directs expression of huCEA (LV-huCEA). This stabilization result was reproducible and detailed analyses including antibody assays, multiplex cytokine analyses on unstimulated splenocytes, lymph node cell characterizations, tetramer staining, and immunofluorescence staining of tumor sections showed that this outcome correlated with both a cellular and humoral immune response. Similar tumor outcomes were not seen when mice were vaccinated with a control LV that engineered expression of enGFP only. The long-term potency of this vaccination strategy was also studied and revealed the requirement for maintenance of tumor antigen-specific immunity for efficient tumor control. These data support the use of direct injections of low doses of LV-huCEA for enhancement of tumor immunotherapy directed against CEA. [Mol Cancer Ther 2009;8(3):OF692–11]
Gefitinib (Iressa) represses FOXM1 expression via FOXO3a in breast cancer
Authors: McGovern Ursula B. , Francis Richard E. , Peck Barrie , Guest Stephanie K. , Wang Jun , Myatt Stephen S. , Krol Janna , Kwok Jimmy M-M. , Polychronis Andreas , Coombes R. Charles , Lam Eric W-F.
Published: March 2009

Gefitinib (Iressa) is a specific and effective epidermal growth factor receptor inhibitor. An understanding of the downstream cellular targets of gefitinib will allow the discovery of biomarkers for predicting outcomes and monitoring anti-epidermal growth factor receptor therapies and provide information for overcoming gefitinib resistance. In this study, we investigated the role and regulation of FOXM1 in response to gefitinib treatment in breast cancer. Using the gefitinib-sensitive breast carcinoma cell lines BT474 and SKBR3 as well as the resistant lines MCF-7, MDA-MB-231, and MDA-MB-453, we showed that gefitinib represses the expression of the transcription factor FOXM1 in sensitive, but not resistant, cells. FOXM1 repression by gefitinib is associated with FOXO3a activation and is mediated at the transcriptional level and gene promoter level. These results were verified by immunohistochemical staining of biopsy samples from primary breast cancer patients obtained from a gefitinib neoadjuvant study. We also showed that ectopic expression of an active FOXO3a represses FOXM1 expression, whereas knockdown of FOXO3a expression using small interfering RNA can up-regulate FOXM1 and its downstream targets polo-like kinase, cyclin B1, and CDC25B and rescue sensitive BT474 cells from gefitinib-induced cell proliferative arrest. These results suggest that gefitinib represses FOXM1 expression via FOXO3a in breast cancer. We further showed that overexpression of a wild-type FOXM1 or a constitutively active FOXM1, ΔN-FOXM1, abrogates the cell death induced by gefitinib, indicating that FOXM1 has a functional role in mediating the gefitinib-induced proliferative arrest and in determining sensitivity to gefitinib. In summary, our study defined FOXM1 as a cellular target and marker of gefitinib activity in breast cancer. [Mol Cancer Ther 2009;8(3):582–91]
p31comet Induces Cellular Senescence through p21 Accumulation and Mad2 Disruption
Authors: Yun Miyong , Han Young-Hoon , Yoon Sun Hee , Kim Hee Young , Kim Bu-Yeo , Ju Yeun-Jin , Kang Chang-Mo , Jang Su Hwa , Chung Hee-Yong , Lee Su-Jae , Cho Myung-Haing , Yoon Gyesoon , Park Gil Hong , Kim Sang Hoon , Lee Kee-Ho
Published: March 2009

Functional suppression of spindle checkpoint protein activity results in apoptotic cell death arising from mitotic failure, including defective spindle formation, chromosome missegregation, and premature mitotic exit. The recently identified p31comet protein acts as a spindle checkpoint silencer via communication with the transient Mad2 complex. In the present study, we found that p31comet overexpression led to two distinct phenotypic changes, cellular apoptosis and senescence. Because of a paucity of direct molecular link of spindle checkpoint to cellular senescence, however, the present report focuses on the relationship between abnormal spindle checkpoint formation and p31comet-induced senescence by using susceptible tumor cell lines. p31comet-induced senescence was accompanied by mitotic catastrophe with massive nuclear and chromosomal abnormalities. The progression of the senescence was completely inhibited by the depletion of p21Waf1/Cip1 and partly inhibited by the depletion of the tumor suppressor protein p53. Notably, p21Waf1/Cip1 depletion caused a dramatic phenotypic conversion of p31comet-induced senescence into cell death through mitotic catastrophe, indicating that p21Waf1/Cip1 is a major mediator of p31comet-induced cellular senescence. In contrast to wild-type p31comet, overexpression of a p31 mutant lacking the Mad2 binding region did not cause senescence. Moreover, depletion of Mad2 by small interfering RNA induced senescence. Here, we show that p31comet induces tumor cell senescence by mediating p21Waf1/Cip1 accumulation and Mad2 disruption and that these effects are dependent on a direct interaction of p31comet with Mad2. Our results could be used to control tumor growth. (Mol Cancer Res 2009;7(3):371–82)
Monochorionic Triplet and Monoamniotic Twins Gestation after Intracytoplasmic Sperm Injection and Laser-Assisted Hatching
Authors: Pantos Konstas , Kokkali Georgia , Petroutsou Konstantina , Lekka Kaliopi , Malligiannis Petros , Koratzis Asterios
Published: March 2009

Objective: To report on a patient with a quintuplet pregnancy consisting of a monochorionic triamnionic triplet pregnancy and one case of monoamniotic twins after intracytoplasmic sperm injection (ICSI), laser-assisted hatching, and day 4 embryo transfer. Case Report: A 34-year-old woman who underwent ICSI due to male infertility. She underwent standard controlled ovarian hyperstimulation. Retrieved oocytes were fertilized in vitro, and 3 embryos were transferred on day 4 after laser-assisted hatching. Transvaginal ultrasound revealed a monoamniotic twin pregnancy and a monochorionic-triamniotic triplet pregnancy. A second ultrasound, 1 week later, showed three distinct foci of cardiac motion in one sac, while the sac of the monoamniotic twins had one embryo with heart beat. After extensive counseling with perinatologists about pregnancy complications, the patient elected to performed fetal reduction on the third sac. A healthy male infant was delivered by Caesarean section at 38 weeks of gestation. Conclusions: The exact mechanism of monozygotic multiple gestation is still poorly characterized. Procedures that modify the zona pellucida (e.g. ICSI and assisted embryo hatching) have been suggested as important in the monozygotic multiple gestation hypothesis, yet a definitive relationship between any clinical intervention during in vitro fertilization and the subsequent development of multiple monozygotic gestation remains speculative.
Improving Global Model Precipitation Forecasts over India Using Downscaling and the FSU Superensemble. Part II: Seasonal Climate
Authors: Chakraborty Arindam , Krishnamurti T. N.
Published: March 2009

This study addresses seasonal forecasts of rains over India using the following components: high-resolution rain gauge–based rainfall data covering the years 1987–2001, rain-rate initialization, four global atmosphere–ocean coupled models, a regional downscaling of the multimodel forecasts, and a multimodel superensemble that includes a training and a forecast phase at the high resolution over the internal India domain. The results of monthly and seasonal forecasts of rains for the member models and for the superensemble are presented here. The main findings, assessed via the use of RMS error, anomaly correlation, equitable threat score, and ranked probability skill score, are (i) high forecast skills for the downscaled superensemble-based seasonal forecasts compared to the forecasts from the direct use of large-scale model forecasts were possible; (ii) very high scores for rainfall forecasts have been noted separately for dry and wet years, for different regions over India and especially for heavier rains in excess of 15 mm day−1; and (iii) the superensemble forecast skills exceed that of the benchmark observed climatology. The availability of reliable measures of high-resolution rain gauge–based rainfall was central for this study. Overall, the proposed algorithms, added together, show very promising results for the prediction of monsoon rains on the seasonal time scale.
Vinyl Ester—Glass Hollow Particle Composites: Dynamic Mechanical Properties at High Inclusion Volume Fraction
Authors: Tagliavia G. , Porfiri M. , Gupta N.
Published: March 2009

This study entails the analysis of dynamic mechanical properties of hollow particle filled composites, called syntactic foams. A theoretical model is developed to predict the dynamic mechanical behavior of composite materials containing particle volume fraction up to the packing limit. The modeling approach is based on a differential scheme that extrapolates the properties of infinitely dilute dispersions of hollow particles (microballoons) to high inclusion volume fraction. The model is capable of predicting storage modulus and loss tangent in a wide frequency band of mechanical vibrations. Theoretical predictions are validated with experimental results on 16 compositions of vinyl ester—glass microballoon syntactic foams. Vibration methods are used to assess the dependence of dynamic mechanical properties of syntactic foams on microballoon wall thickness and volume fraction. Theoretical predictions are in close agreement with experimental findings over a wide range of vibration frequencies. Results show that loss tangent generally decreases as the inclusion volume fraction increases and is almost unaffected by microballoon wall thickness and that storage modulus increases with increasing microballoon wall thickness. Therefore, the loss tangent and the storage modulus can be tailored by means of microballoon volume fraction and wall thickness.
Reducing urinary catheter related infections in care homes: a review of the literature
Authors: McNulty Cliodna AM
Published: March 2009

Although much guidance indicates that indwelling urinary catheters should only be used for the management of urinary retention and incontinence as a last resort, approximately 10% of care home residents have them, leading to increased risks of catheter associated infections and mortality. Catheterisation rates and subsequent infections can be reduced through more proactive management of incontinence and toileting, and removal of urinary catheters from residents discharged from hospital. Staffing in care homes should match residents' nursing and continence needs to allow this proactive approach. Audit of care home should and hospital discharge catheterisation rates, combined with feedback and staff training will raise awareness of the benefits of reducing catheterisation rates. Use of the Department of Health's Essential Steps to Safe, Clean Urinary Catheter Care (2006a) will also help to reduce catheter associated infections.
Copy number abnormalities, MYC activity, and the genetic fingerprint of normal B cells mechanistically define the microRNA profile of diffuse large B-cell lymphoma
Authors: Li Cheng , Kim Sang-Woo , Rai Deepak , Bolla Aswani R. , Adhvaryu Siddharth , Kinney Marsha C. , Robetorye Ryan S. , Aguiar Ricardo C. T.
Published: March 2009

MicroRNA (miRNA) deregulation contributes to cancer pathogenesis. However, analysis of miRNAs in diffuse large B-cell lymphoma (DLBCL) has been hindered by a focus on cell lines, limited number of miRNAs examined, and lack of copy number data. To address these restrictions, we investigated genomewide miRNA expression and copy number data in 86 DLBCLs. Permutation analysis showed that 63 miRNAs were recurrently disrupted in DLBCL, including highly expressed oncomirs not previously linked to chromosomal abnormalities. Further, using training and validation tumor groups, we defined a collection of miRNAs that robustly segregates DLBCLs into 3 subsets, which are independent of the cell-of-origin classification, extent of T-cell infiltrate, and tumor site. Instead, these unique miRNA-driven DLBCL subgroups showed markedly different MYC transcriptional activity, which explained the dominance of miRNAs regulated by MYC in their expression signatures. In addition, analysis of miRNA expression patterns of normal B cells and integration of copy number and expression data showed that genomic abnormalities and the genetic fingerprint of nonmalignant cells also contribute to the miRNA profile of DLBCL. In conclusion, we created a comprehensive map of the miRNA genome in DLBCL and, in the process, have uncovered and mechanistically elucidated the basis for additional molecular heterogeneity in this tumor.
Lung ventilation scintigraphy in the assessment of obstructive lung diseases
Authors: Fazzi P. , Albertelli R. , Grana M. , Paggiaro P.L.
Published: March 2009

Educational aims

To describe more recent techniques used to study the regional distribution of ventilation.

To learn the most frequent abnormal patterns of lung ventilation scintigraphy (LVS) in obstructive lung diseases.

To compare the information obtained by this technique with that obtained by traditional lung function tests.

To understand that LVS may provide further information on lung pathophysiology.

LVS is a well known technique that is used to study the distribution of ventilation in pulmonary diseases. However, its application in current practice has been poorly developed, mainly because of methodological problems. Redistribution of convective ventilation is the leading disorder in patients with chronic obstructive pulmonary disease (COPD) and is the target of most treatments, either by drugs or mechanical ventilation. The current availability of a new delivery inhaler (FAI; Medical Products Research, Legnano, Italy) with fine particles allows: 1) fast aerosol administration; 2) high-quality early images; 3) improved visualisation of pulmonary periphery; and 4) wider physiological distribution of the radioaerosol. Several patterns of distribution of pulmonary ventilation can be described in different airway disorders (asthma and COPD). These abnormalities are often too subtle to be picked up by traditional lung function tests, both in the baseline evaluation and after different interventions. Wider use of this technique may increase the knowledge and the management of obstructive lung diseases.
Characterization of the hupSL promoter activity in Nostoc punctiformeATCC 29133
Authors: Holmqvist Marie , Stensjö Karin , Oliveira Paulo , Lindberg Pia , Lindblad Peter
Published: March 2009

Background

In cyanobacteria three enzymes are directly involved in the hydrogen metabolism; a nitrogenase that produces molecular hydrogen, H2, as a by-product of nitrogen fixation, an uptake hydrogenase that recaptures H2 and oxidize it, and a bidirectional hydrogenase that can both oxidize and produce H2.Nostoc punctiforme ATCC 29133 is a filamentous dinitrogen fixing cyanobacterium containing a nitrogenase and an uptake hydrogenase but no bidirectional hydrogenase. Generally, little is known about the transcriptional regulation of the cyanobacterial uptake hydrogenases. In this study gel shift assays showed that NtcA has a specific affinity to a region of the hupSL promoter containing a predicted NtcA binding site. The predicted NtcA binding site is centred at 258.5 bp upstream the transcription start point (tsp). To further investigate the hupSL promoter, truncated versions of the hupSL promoter were fused to either gfp or luxAB, encoding the reporter proteins Green Fluorescent Protein and Luciferase, respectively.

Results

Interestingly, all hupsSL promoter deletion constructs showed heterocyst specific expression. Unexpectedly the shortest promoter fragment, a fragment covering 57 bp upstream and 258 bp downstream the tsp, exhibited the highest promoter activity. Deletion of the NtcA binding site neither affected the expression to any larger extent nor the heterocyst specificity.

Conclusion

Obtained data suggest that the hupSL promoter in N. punctiforme is not strictly dependent on the upstream NtcA cis element and that the shortest promoter fragment (-57 to tsp) is enough for a high and heterocyst specific expression of hupSL. This is highly interesting because it indicates that the information that determines heterocyst specific gene expression might be confined to this short sequence or in the downstream untranslated leader sequence.
In vitromodel for the analysis of synovial fibroblast-mediated degradation of intact cartilage
Authors: Pretzel David , Pohlers Dirk , Weinert Sönke , Kinne Raimund W
Published: March 2009

Introduction

Activated synovial fibroblasts are thought to play a major role in the destruction of cartilage in chronic, inflammatory rheumatoid arthritis (RA). However, profound insight into the pathogenic mechanisms and the impact of synovial fibroblasts in the initial early stages of cartilage destruction is limited. Hence, the present study sought to establish a standardised in vitro model for early cartilage destruction with native, intact cartilage in order to analyse the matrix-degrading capacity of synovial fibroblasts and their influence on cartilage metabolism.

Methods

A standardised model was established by co-culturing bovine cartilage discs with early-passage human synovial fibroblasts for 14 days under continuous stimulation with TNF-α, IL-1β or a combination of TNF-α/IL-1β. To assess cartilage destruction, the co-cultures were analysed by histology, immunohistochemistry, electron microscopy and laser scanning microscopy. In addition, content and/or neosynthesis of the matrix molecules cartilage oligomeric matrix protein (COMP) and collagen II was quantified. Finally, gene and protein expression of matrix-degrading enzymes and pro-inflammatory cytokines were profiled in both synovial fibroblasts and cartilage.

Results

Histological and immunohistological analyses revealed that non-stimulated synovial fibroblasts are capable of demasking/degrading cartilage matrix components (proteoglycans, COMP, collagen) and stimulated synovial fibroblasts clearly augment chondrocyte-mediated, cytokine-induced cartilage destruction. Cytokine stimulation led to an upregulation of tissue-degrading enzymes (aggrecanases I/II, matrix-metalloproteinase (MMP) 1, MMP-3) and pro-inflammatory cytokines (IL-6 and IL-8) in both cartilage and synovial fibroblasts. In general, the activity of tissue-degrading enzymes was consistently higher in co-cultures with synovial fibroblasts than in cartilage monocultures. In addition, stimulated synovial fibroblasts suppressed the synthesis of collagen type II mRNA in cartilage.

Conclusions

The results demonstrate for the first time the capacity of synovial fibroblasts to degrade intact cartilage matrix by disturbing the homeostasis of cartilage via the production of catabolic enzymes/pro-inflammatory cytokines and suppression of anabolic matrix synthesis (i.e., collagen type II). This new in vitro model may closely reflect the complex process of early stage in vivo destruction in RA and help to elucidate the role of synovial fibroblasts and other synovial cells in this process, and the molecular mechanisms involved in cartilage degradation.
Serum levels of soluble receptor for advanced glycation end products and of S100 proteins are associated with inflammatory, autoantibody, and classical risk markers of joint and vascular damage in rheumatoid arthritis
Authors: Chen Yueh-Sheng , Yan Weixing , Geczy Carolyn L , Brown Matthew A , Thomas Ranjeny
Published: March 2009

Introduction

The receptor for advanced glycation end products (RAGE) is a member of the immunoglobulin superfamily of cell surface receptor molecules. High concentrations of three of its putative proinflammatory ligands, S100A8/A9 complex (calprotectin), S100A8, and S100A12, are found in rheumatoid arthritis (RA) serum and synovial fluid. In contrast, soluble RAGE (sRAGE) may prevent proinflammatory effects by acting as a decoy. This study evaluated the serum levels of S100A9, S100A8, S100A12 and sRAGE in RA patients, to determine their relationship to inflammation and joint and vascular damage.

Methods

Serum sRAGE, S100A9, S100A8 and S100A12 levels from 138 patients with established RA and 44 healthy controls were measured by ELISA and compared by unpaired t test. In RA patients, associations with disease activity and severity variables were analyzed by simple and multiple linear regressions.

Results

Serum S100A9, S100A8 and S100A12 levels were correlated in RA patients. S100A9 levels were associated with body mass index (BMI), and with serum levels of S100A8 and S100A12. S100A8 levels were associated with serum levels of S100A9, presence of anti-citrullinated peptide antibodies (ACPA), and rheumatoid factor (RF). S100A12 levels were associated with presence of ACPA, history of diabetes, and serum S100A9 levels. sRAGE levels were negatively associated with serum levels of C-reactive protein (CRP) and high-density lipoprotein (HDL), history of vasculitis, and the presence of the RAGE 82Ser polymorphism.

Conclusions

sRAGE and S100 proteins were associated not just with RA inflammation and autoantibody production, but also with classical vascular risk factors for end-organ damage. Consistent with its role as a RAGE decoy molecule, sRAGE had the opposite effects to S100 proteins in that S100 proteins were associated with autoantibodies and vascular risk, whereas sRAGE was associated with protection against joint and vascular damage. These data suggest that RAGE activity influences co-development of joint and vascular disease in rheumatoid arthritis patients.