bluuebunny/crossref_metadata_2025 huggingface dataset:
Initiation and Development of Lateral Buds in Abiesbalsamea
Authors: Powell G. R.
Published: February 2009

Lateral buds of Abiesbalsamea (L.) Mill, are initiated on shoots within the previous season's vegetative terminal and subterminal buds about 1 week before the latter burst. Initiation coincides with megasporangiate-bud burst. Prophyll primordia form 1 week after bud initiation and cataphyll production continues throughout the shoot-elongation phase. Some lateral buds cease development during, or at the end of, the cataphyll-formation period and become latent. Most lateral buds in the megasporangiate zone of the crown occur on the upper sides of the shoots and may differentiate into megasporangiate or vegetative buds: those in the microsporangiate zone occur largely on the lower sides of the shoots and normally differentiate into microsporangiate buds. Bracts are initiated in megasporangiate buds from late July to October, and during September, ovuliferous scales form at the bases of most bracts. No differentiation is evident within the ovuliferous scales by the end of October. Needles are initiated in vegetative buds from late July to October. In microsporangiate buds microsporophylls form from late July through August. The microsporangia contain sporogenous tissue by early September and angular archesporial cells by October.
Coniferous Stands Characterized with The Weibull Distribution
Authors: Schreuder Hans T. , Swank Wayne T.
Published: February 2009

The Weibull distribution, [Formula: see text], summarized diameter, basal area, surface area, biomass, and crown profile distribution data well for several different ages of white and loblolly pine plantations. The data for diameter, basal area, surface area, and biomass were easily summarized by this one distribution in a theoretically consistent fashion. This is not possible with the normal and the gamma distributions, and the lognormal gives less satisfactory results. The distribution function should prove useful in modeling tree stands since only the parameter values need to be changed over time for the above variables. The change in these parameters may be a good way to characterize and interpret changes in stands over time.
Whole-tree Harvesting Depletes Soil Nutrients
Authors: White Edwin H.
Published: February 2009

This paper reports the effects of whole-tree harvesting of eight cottonwood stands on the soil nutrient pool. The data indicate possible site degradation by depletion of soil reserves of N, P, and K but not Ca and Mg on a range of alluvial site conditions in Alabama. Foresters must establish the rate of nutrient removal in intensive tree cropping systems for a variety of species and sites and develop prescriptions to minimize the impact.
Rooting of Jack Pine (Pinusbanksiana Lamb.) Cuttings
Authors: Zsuffa L.
Published: February 2009

Cuttings from 6- and 10-year-old trees of jack pine rooted under intermittent mist. Well balanced root systems with many fine side roots were produced.The influence of several factors on rooting was studied. Clonal variation in rooting was observed. In general 6- and 10-year-old ortets rooted similarly. The ortets with heavy, long branches rooted better than the ones with light, short branches. Cuttings taken from second-order shoots rooted and survived like those taken from third-order shoots. Cuttings of medium length rooted and survived best.A high percentage of the cuttings of all clones callused. However, a number of callused cuttings later died, possibly due to unfavorable environmental conditions in the propagation house. Survival was highest for clones rooting best.
Nutrient cycles in pine and their adaptation to poor soils
Authors: Miller Hugh G. , Cooper Jean M. , Miller John D. , Pauline Olive J. L.
Published: February 2009

Values for the various fluxes of nitrogen through the tree–soil system were calculated using models derived from measurements of rates of input, accumulation, and transfer in plots of 11-m-tall Pinusnigra var. maritima (Ait.) Melv. that were showing growth responses to varying rates of nitrogen fertilizer. As the result of continuing immobilization in trees and humus, growth of the unfertilized trees was declining and the trees were having to sacrifice older tissues to mobilize sufficient nitrogen for new growth. In the fertilized trees, there was similar mobilization but of excess nitrogen stored during the period that fertilizers were applied. Models of the flux of nitrogen, phosphorus, calcium, and magnesium at optimum growth showed that tight cycling and low rates of immobilization, coupled with effective retention of relatively high atmospheric inputs, probably represents the adaptive mechanism that enables trees to thrive on soils low in potassium and magnesium. However, high rates of immobilization and low rates of atmospheric input relative to tree uptake mean that despite the maintenance of tight cycles, the trees continued to make significant demands on soil reserves of nitrogen, phosphorus, and calcium throughout their life cycle.
Strach storage and radial growth in woody roots of sugar maple
Authors: Wargo Philip M.
Published: February 2009

The timing and pattern of starch storage and the timing of radial root growth in sugar maple, Acersaccharum, were observed with micro- and histo-chemical techniques from May to December. Results of initial monthly observations showed that starch deposits were very low or depleted in May shortly after budbreak but were fairly heavy by mid-July. A starchless ring of xylem was evident on some roots in mid-July and all roots in mid-August but was not visible in early November. Data from weekly observations indicated that the starchless xylem resulted from new tissue development that began in early July after substantial starch storage. The major portion of growth occurred by early September. The new xylem had no visible starch until mid-August. Initially, starch deposits developed in new xylem cells nearest to the previous annual ring. Starch appeared first in the ray parenchyma, next in the xylem fibers adjacent to vessels, and then in xylem fibers more distant from the vessels. By late September, the outer ring of xylem was no longer distinctly different from the older tissue. In contrast with the stem, where radial growth precedes or coincides with carbohydrate storage, substantial storage of carbohydrates preceded radial growth in the roots.
Effect of light intensity during extended photoperiod on growth of amabilis fir, mountain hemlock, and white and Engelmann spruce seedlings
Authors: Arnott J. T.
Published: February 2009

Abiesamabilis (Dougl.) Forbes, Tsugamertensiana (Bong.) Carr., Piceaglauca (Moench) Voss, and Piceaengelmannii Parry seedlings were grown in an outdoor container nursery using one high-pressure sodium vapor lamp to provide a 24-h photoperiod. Eight intensities of lighting were arranged, viz., 220, 80, 40, 20, 12, 8, 5, and 0 lx.Extending the photoperiod and increasing the light intensity had a significant positive effect on the length and weight of seedling shoots and delayed terminal bud set. Root growth response to these treatments was negative, although the differences were rarely significant. Seedlings grown under a light intensity of 220 lx were the largest. The minimum intensity required to produce seedling shoot lengths which were significantly larger than the controls was in the range 20–80 lx.
The pathogenic fungus Caloscyphafulgens in stored conifer seeds in British Columbia and relation of its incidence to ground and squirrel-cache collected cones
Authors: Sutherland Jack R.
Published: February 2009

Stored seeds were assayed for the seed-borne fungus Caloscyphafulgens and the pathogen was found in 0, 25, 3, 0, 0, 0, 29, 16, 14, 0, and 0% of the seed lots of Abiesamabilis, A. grandis, Pseudotsugamenziesii, Pinusponderosa, P. monticola, P. contorta, Piceaglauca, P. engelmannii, P. glauca × P. engelmannii hybrid, Tsugamertensiana, and T. heterophylla, respectively. Within infested seed lots, 0.4 to 22.4% of the seeds were infected. There was no relationship between incidence of infested seed lots and year of cone collection or geographic origin. Overall, squirrel-cache collected cones, especially of Picea spp., had the highest incidence, and slash-picked cones the lowest incidence, of C. fulgens infested seed lots. The percentage of diseased seeds, within infested seed lots, was unrelated to the origin of ground-picked cones.
Electrophysiologic changes in heart failure: focus on pacemaker channelsThis article is one of a selection of papers from the NATO Advanced Research Workshop on Translational Knowledge for Heart Health (published in part 1 of a 2-part Special Issue).
Authors: Sartiani Laura , Stillitano Francesca , Cerbai Elisabetta , Mugelli Alessandro
Published: February 2009

Heart failure is a common clinical syndrome occurring as a result of cardiac overload, injury, and a complex interplay among genetic, neurohormonal, inflammatory, and biochemical factors. Occurrence of arrhythmias in heart failure is largely a consequence of disease-induced electrical remodeling of cardiac myocytes, a phenomenon consisting of alterations of ion channels and the ion-transport function that predispose patients to develop lethal arrhythmias. In most cases, the mechanism is the rapid onset of a ventricular tachyarrhythmia progressing to ventricular fibrillation and hemodynamic compromise. This paper highlights some of the important changes in ion channel expression and function that underlie electrical remodeling of the failing heart. Particular attention will be focused on the presence, features, and pharmacologic modulation of f channels expressed in ventricular cardiac myocytes.
COHERENT ANTI-STOKES RAMAN SCATTERING MICROSCOPY TO MONITOR DRUG DISSOLUTION IN DIFFERENT ORAL PHARMACEUTICAL TABLETS
Authors: JURNA M. , WINDBERGS M. , STRACHAN C. J. , HARTSUIKER L. , OTTO C. , KLEINEBUDDE P. , HEREK J. L. , OFFERHAUS H. L.
Published: February 2009

Coherent anti-Stokes Raman scattering (CARS) microscopy is used to visualize the release of a model drug (theophylline) from a lipid (tripalmitin) based tablet during dissolution. The effects of transformation and dissolution of the drug are imaged in real time. This study reveals that the manufacturing process causes significant differences in the release process: tablets prepared from powder show formation of theophylline monohydrate on the surface which prevents a controlled drug release, whereas solid lipid extrudates did not show formation of monohydrate. This visualization technique can aid future tablet design.
EXTRAOCULAR CELLULAR PHOTOTRANSDUCTION
Authors: ZHU LING , LIU TIMON CHENG-YI , WU MIN , YUAN JIAN-QIN , CHEN TONG-SHENG
Published: February 2009

Photobiomodulation (PBM) is a modulation of monochromatic light or laser irradiation (LI) on biosystems. It is reviewed from the viewpoint of extraocular phototransduction in this paper. It was found that LI can induce extraocular phototransduction, and there may be an exact correspondence relationship of LI at different wavelengths and in different dose zones, and cellular signal transduction pathways. The signal transduction pathways can be classified into two types so that the Gs protein-mediated pathways belong to pathway 1, and the other pathways such as protein kinase Cs -mediated pathways and mitogen-activated protein kinase-mediated pathways belong to pathway 2. Almost all the present pathways found to mediate PBM belong to pathway 2, but there should be a pathway 1-mediated PBM. The previous studies were rather preliminary, and therefore further work should be done.
NON-LINEAR OPTICAL IMAGING OF OBESITY-RELATED HEALTH RISKS: REVIEW
Authors: LE THUC T. , CHENG JI-XIN
Published: February 2009

This review highlights the recent applications of non-linear optical (NLO) microscopy to study obesity-related health risks. A strong emphasis is given to the applications of coherent anti-Stokes Raman scattering (CARS) microscopy where multiple non-linear optical imaging modalities including CARS, sum-frequency generation (SFG), and two-photon fluorescence are employed simultaneously on a single microscope platform. Specific examples on applications of NLO microscopy to study lipid-droplet biology, obesity-cancer relationship, atherosclerosis, and lipid-rich biological structures are discussed.
NON-LINEAR SPECTRAL IMAGING MICROSCOPY STUDIES OF HUMAN HYPERTROPHIC SCAR
Authors: LU KECHENG , ZHUO SHUANGMU , HONG ZHIBIN , CHEN GUANNAN , JIANG XINGSHAN , ZHENG LIQIN , CHEN JIANXIN
Published: February 2009

Skin scar is unique to humans, the major significant negative outcome sustained after thermal injuries, traumatic injuries, and surgical procedures. Hypertrophic scar in human skin is investigated using non-linear spectral imaging microscopy. The high contrast images and spectroscopic intensities of collagen and elastic fibers extracted from the spectral imaging of normal skin tissue, and the normal skin near and far away from the hypertrophic scar tissues in a 10-year-old patient case are obtained. The results show that there are apparent differences in the morphological structure and spectral characteristics of collagen and elastic fibers when comparing the normal skin with the hypertrophic scar tissue. These differences can be good indicators to differentiate the normal skin and hypertrophic scar tissue and demonstrate that non-linear spectral imaging microscopy has potential to noninvasively investigate the pathophysiology of human hypertrophic scar.
PHOTODYNAMIC THERAPY — AN UPDATE ON CLINICAL APPLICATIONS
Authors: HUANG ZHENG , LI LIBO , WANG HONGWEI , WANG XIULI , YUAN KAIHUA , MEYERS ARLEN , YANG LIHAO , HETZEL FRED W.
Published: February 2009

Photodynamic therapy (PDT) has received increased attention since the regulatory approvals of several photosensitizers and light applicators in numerous countries and regions around the world. In recent years, much progress has been seen in basic research as well as clinical application. PDT clinical application has now extended from treating malignant diseases to nonmalignant diseases. This review article will present recent clinical data published in English journals. The data will be organized according to their clinical specialties. The new development and future direction in clinical applications of PDT for the management of both malignant and nonmalignant diseases will be discussed.
Upregulation of voltage-gated Na+ channels by long-term activation of the ghrelin-growth hormone secretagogue receptor in clonal GC somatotropes
Authors: Dominguez Belisario , Felix Ricardo , Monjaraz Eduardo
Published: February 2009

A central question in adenohypophyseal cell physiology concerns the role of transmembrane ionic fluxes in the initiation of the hormone secretion process. In the current report, we investigated the effects of the growth hormone (GH) secretagogues ghrelin and GH-releasing peptide-6 (GHRP-6) on the regulation of the functional expression of voltage-gated Na+ channels using the tumoral somatotrope GC cell line as a model. Cells were cultured under control conditions or in presence of the GH secretagogues (GHS) for 96 h, and Na+ currents ( INa) were characterized in whole cell patch-clamp experiments. GHS treatment significantly increased INa density in a dose-dependent manner. The effects of GHRP-6 were accompanied by an augment in conductance without changes in the kinetics and the voltage dependence of the currents, suggesting an increase in the number of channels in the cell membrane. Sustained inhibition of L-type Ca2+ channel activity decreased INa density and prevented the effects of the GHS, whereas long-term exposure to an L-channel agonist increased INa density and enhanced the actions of GHRP-6, indicating that Ca2+ entry through these channels plays a role in the regulation of Na+ channel expression. Likewise, GHRP-6 failed to enhance Na+ channel expression in the presence of membrane-permeable inhibitors of protein kinases A and C, as well as the Ca2+/calmodulin-dependent kinase II. Conversely, treatment with a cAMP analog or a protein kinase C activator enhanced both basal and GHS-induced secretion of GH measured by enzyme-linked immunoassay, suggesting that GHRP-6 acting through the ghrelin receptor and different signaling pathways enhances Na+ channel membrane expression, which favors hormone release from GC somatotropes.
Conserved Gene Expression Programs Integrate Mammalian Prostate Development and Tumorigenesis
Authors: Pritchard Colin , Mecham Brig , Dumpit Ruth , Coleman Ilsa , Bhattacharjee Madhuchhanda , Chen Qian , Sikes Robert A. , Nelson Peter S.
Published: February 2009

Studies centered at the intersection of embryogenesis and carcinogenesis have identified striking parallels involving signaling pathways that modulate both developmental and neoplastic processes. In the prostate, reciprocal interactions between epithelium and stroma are known to influence neoplasia and also exert morphogenic effects via the urogenital sinus mesenchyme. In this study, we sought to determine molecular relationships between aspects of normal prostate development and prostate carcinogenesis. We first characterized the gene expression program associated with key points of murine prostate organogenesis spanning the initial in utero induction of prostate budding through maturity. We identified a highly reproducible temporal program of gene expression that partitioned according to the broad developmental stages of prostate induction, branching morphogenesis, and secretory differentiation. Comparisons of gene expression profiles of murine prostate cancers arising in the context of genetically engineered alterations in the Pten tumor suppressor and Myc oncogene identified significant associations between the profile of branching morphogenesis and both cancer models. Further, the expression of genes comprising the branching morphogenesis program, such as PRDX4, SLC43A1, and DNMT3A, was significantly altered in human neoplastic prostate epithelium. These results indicate that components of normal developmental processes are active in prostate neoplasia and provide further rationale for exploiting molecular features of organogenesis to understand cancer phenotypes. [Cancer Res 2009;69(5):1739–47]
RASSF1A Mediates p21Cip1/Waf1-Dependent Cell Cycle Arrest and Senescence through Modulation of the Raf-MEK-ERK Pathway and Inhibition of Akt
Authors: Thaler Sonja , Hähnel Patricia S. , Schad Arno , Dammann Reinhard , Schuler Martin
Published: February 2009

Promoter hypermethylation preventing expression of the RAS association domain family 1 isoform A (RASSF1A) gene product is among the most abundant epigenetic deregulations in human cancer. Restoration of RASSF1A inhibits tumor cell growth in vitro and in murine xenograft models. Rassf1a-deficient mice feature increased spontaneous and carcinogen-induced tumor formation. Mechanistically, RASSF1A affects several cellular functions, such as microtubule dynamics, migration, proliferation, and apoptosis; however, its tumor-suppressive mechanism is incompletely understood. To study the functional consequences of RASSF1A expression in human cancer cells, we made use of a doxycycline-inducible expression system and a RASSF1A-deficient lung cancer cell line. We observed that RASSF1A induces cell cycle arrest in G1 phase and senescence in vitro and in tumors established in immunodeficient mice. RASSF1A-mediated growth inhibition was accompanied by the up-regulation of the cyclin-dependent kinase inhibitor p21Cip1/Waf1 and proceeded independently of p53, p14Arf, and p16Ink4a. Loss of p21Cip1/Waf1 or coexpression of the human papilloma virus 16 oncoprotein E7 was found to override RASSF1A-induced cell cycle arrest and senescence. Conditional RASSF1A affected mitogen-activated protein kinase and protein kinase B/Akt signaling to up-regulate p21Cip1/Waf1 and to facilitate its nuclear localization. In summary, RASSF1A can mediate cell cycle arrest and senescence in human cancer cells by p53-independent regulation of p21Cip1/Waf1. [Cancer Res 2009;69(5):1748–57]
PIK3CA Mutations in Colorectal Cancer Are Associated with Clinical Resistance to EGFR-Targeted Monoclonal Antibodies
Authors: Sartore-Bianchi Andrea , Martini Miriam , Molinari Francesca , Veronese Silvio , Nichelatti Michele , Artale Salvatore , Di Nicolantonio Federica , Saletti Piercarlo , De Dosso Sara , Mazzucchelli Luca , Frattini Milo , Siena Salvatore , Bardelli Alberto
Published: February 2009

The monoclonal antibodies (moAb) panitumumab and cetuximab target the epidermal growth factor receptor (EGFR) and have proven valuable for the treatment of metastatic colorectal cancer (mCRC). EGFR-mediated signaling involves two main intracellular cascades: on one side KRAS activates BRAF, which in turn triggers the mitogen-activated protein kinases. On the other, membrane localization of the lipid kinase PIK3CA counteracts PTEN and promotes AKT1 phosphorylation, thereby activating a parallel intracellular axis. Constitutive activation of KRAS bypasses the corresponding signaling cascade and, accordingly, patients with mCRC bearing KRAS mutations are clinically resistant to therapy with panitumumab or cetuximab. We hypothesized that mutations activating PIK3CA could also preclude responsiveness to EGFR-targeted moAbs through a similar mechanism. Here, we present the mutational analysis of PIK3CA and KRAS and evaluation of the PTEN protein status in a cohort of 110 patients with mCRC treated with anti-EGFR moAbs. We observed 15 (13.6%) PIK3CA and 32 (29.0%) KRAS mutations. PIK3CA mutations were significantly associated with clinical resistance to panitumumab or cetuximab; none of the mutated patients achieved objective response (P = 0.038). When only KRAS wild-type tumors were analyzed, the statistical correlation was stronger (P = 0.016). Patients with PIK3CA mutations displayed a worse clinical outcome also in terms of progression-free survival (P = 0.035). Our data indicate that PIK3CA mutations can independently hamper the therapeutic response to panitumumab or cetuximab in mCRC. When the molecular status of the PIK3CA/PTEN and KRAS pathways are concomitantly ascertained, up to 70% of mCRC patients unlikely to respond to EGFR moAbs can be identified. [Cancer Res 2009;69(5):1851–7]
E2F7 Can Regulate Proliferation, Differentiation, and Apoptotic Responses in Human Keratinocytes: Implications for Cutaneous Squamous Cell Carcinoma Formation
Authors: Endo-Munoz Liliana , Dahler Alison , Teakle Ngari , Rickwood Danny , Hazar-Rethinam Mehlika , Abdul-Jabbar Ibtissam , Sommerville Scott , Dickinson Ian , Kaur Pritinder , Paquet-Fifield Sophie , Saunders Nicholas
Published: February 2009

The E2F family of transcription factors plays a crucial role in the regulation of genes involved in cell proliferation, differentiation, and apoptosis. In keratinocytes, the inhibition of E2F is a key step in the control and initiation of squamous differentiation. Because the product of the recently identified E2F7a/E2F7b gene has been shown to repress E2F-regulated promoters, and to be abundant in skin, we examined its role in the epidermis. Our results indicate that E2F7b mRNA expression is selectively associated with proliferation-competent keratinocytes. Moreover, E2F7 was able to antagonize E2F1-induced proliferation and apoptosis. In contrast, although E2F7 was able to inhibit proliferation and initiate differentiation, it was unable to antagonize the differentiation suppression induced by E2F1. These data indicate that E2F7-mediated suppression of proliferation and apoptosis acts through E2F1-dependent pathways, whereas E2F7-induced differentiation acts through an E2F1-independent pathway. These data also suggest that proliferation, differentiation, and survival of primary human keratinocytes can be controlled by the relative ratio of E2F1 to E2F7. Because deregulated proliferation, differentiation, and apoptosis are hallmarks of cancer, we examined the expression levels of E2F1 and E2F7 in cutaneous squamous cell carcinomas (CSCC). We found that both genes were overexpressed in CSCCs compared with normal epidermis. Furthermore, inhibition of E2F7 in a SCC cell line sensitized the cells to UV-induced apoptosis and doxorubicin-induced apoptosis. Combined, these data suggest that the selected disruption of E2F1 and E2F7 in keratinocytes is likely to contribute to CSCC formation and may prove to be a viable therapeutic target. [Cancer Res 2009;69(5):1800–9]
Down-regulation of Vascular Endothelial Growth Factor Receptor 2 Is a Major Molecular Determinant of Proteasome Inhibitor–Mediated Antiangiogenic Action in Endothelial Cells
Authors: Meissner Markus , Reichenbach Gabi , Stein Monika , Hrgovic Igor , Kaufmann Roland , Gille Jens
Published: February 2009

The ubiquitin-proteasome system is the major pathway for intracellular protein degradation in eukaryotic cells. This system controls a wide range of cellular regulatory proteins, including transcription factors and cell cycle regulatory proteins. Recent evidence also established the importance of the proteasome in tumor development, showing antitumor and antiangiogenic actions by using selective inhibitors in vivo. As signaling via the vascular endothelial growth factor receptor 2 (VEGFR2) pathway is critical for angiogenic responses to occur, we explored whether antiangiogenic effects due to proteasome inhibition were partly mediated through decreased endothelial VEGFR2 expression. This study shows that different proteasome inhibitors blocked VEGFR2 expression in a time-dependent and concentration-dependent manner. This blockade was paralleled by the respective inhibition of the formation of capillary-like structures and endothelial cell migration. In contrast, neither tie-2 nor VEGFR1 expression was significantly affected by proteasome inhibitor treatment. The suppressive effects on VEGFR2 expression were not conveyed by increased shedding or a decrease in protein half-life, suggesting that transcriptional mechanisms accounted for the observed effects. In line with this conclusion, proteasome inhibition significantly suppressed VEGFR2 mRNA accumulation. In addition, inhibitor treatment considerably decreased the transcriptional activity of 5′ deletional VEGFR2 promoter gene constructs. Proteasome inhibition–mediated repression was controlled by a GC-rich region that harbored one consensus Sp1-binding site. Subsequent EMSA analyses showed decreased constitutive Sp1-dependent DNA binding in response to proteasome inhibition. In addition, we could show that proteasome inhibitors reduced VEGFR2 mRNA stability. Therefore, VEGFR2 expression may constitute a critical molecular target of proteasome inhibitors that may mediate their antiangiogenic effects in vivo. [Cancer Res 2009;69(5):1976–84]
Tumor-Induced Suppression of CTL Expansion and Subjugation by gp96-Ig Vaccination
Authors: Schreiber Taylor H. , Deyev Vadim V. , Rosenblatt Joseph D. , Podack Eckhard R.
Published: February 2009

Established tumors suppress antitumor immune responses and induce tolerance by incompletely characterized mechanisms, and this phenomenon is an important barrier to tumor immunotherapy. Single vaccination with tumor cells expressing gp96-Ig stimulates robust expansion of tumor-specific CTLs in tumor-naïve mice and this expansion is inhibited by established tumors. Interestingly, frequent vaccinations restore antitumor immune responses in the presence of established tumors. Syngeneic EG7 tumor-bearing mice have heterogeneous responses to frequent vaccination with EG7-gp96-Ig, with 32% complete responders and 68% partial responders. Comparison of responders to nonresponders revealed an inverse correlation between tumor-specific CTL expansion in the peripheral blood and tumor size. To identify immune cells and molecules associated with effective antitumor immune responses, reverse transcription-PCR arrays were performed using cells isolated from the vaccination site. ELISAs, cellular phenotyping, and tumor immunohistochemistry were also performed comparing vaccine responders to nonresponders. These data show that up-regulation of T-bet, RORγt, IFNγ, CCL8, CXCL9, and CXCL10 at the vaccination site are associated with vaccine-induced antitumor immunity. These data correlate with increased CTL expansion in the peripheral blood of responders, increased infiltration of responder tumors by CD8+ cells and interleukin-17+ cells, and decreased infiltration of responder tumors by CD11b+Gr-1+ cells and FoxP3+ cells. Furthermore, serum ELISAs revealed a significant elevation of transforming growth factor-β in nonresponders as compared with responders. Interestingly, CD8+ T cells isolated from responders and nonresponders have equivalent cytotoxic activity in vitro. Taken together, our data suggest that established tumors may escape immunosurveillance by preventing clonal expansion of tumor-specific CTL without inducing anergy. [Cancer Res 2009;69(5):2026–33]
Polychlorinated Biphenyls and Risk of Testicular Germ Cell Tumors
Authors: McGlynn Katherine A. , Quraishi Sabah M. , Graubard Barry I. , Weber Jean-Philippe , Rubertone Mark V. , Erickson Ralph L.
Published: February 2009

Exposure to endocrine-disrupting chemicals, such as polychlorinated biphenyls (PCB), may alter hormonal balance and thereby increase risk of testicular germ cell tumors (TGCT). To study the relationship of PCBs to TGCT, prediagnostic serum samples from 736 cases and 913 controls in the Servicemen's Testicular Tumor Environmental and Endocrine Determinants study were analyzed. Adjusted odds ratios and 95% confidence intervals were estimated using logistic regression. PCB levels were examined in association with all TGCT and, separately, with each histologic type (seminoma and nonseminoma). Risks associated with seven functional groupings of PCBs, as well as sum of PCBs, were also examined. There were significantly decreased risks of TGCT in association with eight PCBs (PCB-118, PCB-138, PCB-153, PCB-156, PCB-163, PCB-170, PCB-180, and PCB-187) and no association with the remaining three (PCB-99, PCB-101, and PCB-183). The same eight congeners were significantly associated with decreased risk of nonseminoma, whereas five (PCB-138, PCB-153, PCB-156, PCB-163, and PCB-170) were associated with decreased risk of seminoma. All functional groupings of PCBs were also associated with decreased risk of TGCT and of nonseminoma, whereas six of the seven functional groups were associated with decreased risk of seminoma. Sum of PCBs was significantly associated with decreased risk of TGCT (Ptrend = 0.006), nonseminoma (Ptrend = 0.007), and seminoma (Ptrend = 0.05). Overall, these data do not support the hypothesis that PCB exposure increases the risk of TGCT. [Cancer Res 2009;69(5):1901–9]
The Complex Relationship between BRCA1 and ERα in Hereditary Breast Cancer
Authors: Gorski Julia J. , Kennedy Richard D. , Hosey Alison M. , Harkin D. Paul
Published: February 2009

Breast cancer 1 (BRCA1) was initially identified as one of the genes conferring genetic predisposition to both breast and ovarian cancer. One of the interesting aspects of BRCA1-linked cancers is the observed specificity for estrogen-responsive tissues such as breast and ovary. Recent advances in our understanding of BRCA1-linked breast cancers have revealed a complex relationship between BRCA1 and estrogen receptor α (ERα) signaling. Estrogen stimulation increases expression of BRCA1 at the mRNA and protein level and conversely BRCA1 functions to both induce ERα mRNA expression and act as a negative regulator of ERα signaling. Here, we review the relationship between BRCA1 and ERα and discuss the use of antiestrogen therapies such as tamoxifen and aromatase inhibitors in the treatment of BRCA1 mutation carriers.
Involvement of FLIP in 2-Methoxyestradiol–Induced Tumor Regression in Transgenic Adenocarcinoma of Mouse Prostate Model
Authors: Ganapathy Manonmani , Ghosh Rita , Jianping Xie , Zhang Xiaoping , Bedolla Roble , Schoolfield John , Yeh I-Tien , Troyer Dean A. , Olumi Aria F. , Kumar Addanki P.
Published: February 2009

Purpose: The purpose of this study is to investigate whether Fas-associated death domain interleukin-1 converting enzyme like inhibitory protein (FLIP) inhibition is a therapeutic target associated with 2-methoxyestradiol (2-ME2)–mediated tumor regression.

Experimental Design: Expression and levels of FLIP were analyzed using (a) real-time PCR and immunoblot analysis in androgen-independent PC-3 cells treated with the newly formulated 2-ME2 and (b) immunohistochemistry in different Gleason pattern human prostate tumors. Transient transfections and chromatin immunoprecipitation (ChIP) assays were used to identify the transcription factors that regulate FLIP. Involvement of FLIP in 2-ME2–induced tumor regression was evaluated in transgenic adenocarcinoma mouse prostate (TRAMP) mice.

Results: High Gleason pattern (5+5) human prostate tumors exhibit significant increase in FLIP compared with low Gleason pattern 3+3 (P = <0.04). 2-ME2 reduced the levels and promoter activity of FLIP (P = 0.001) in PC-3 cells. Transient expression assays show sequences between −503/+242 being sufficient for 2-ME2–induced inhibition of FLIP promoter activity. Cotransfection experiments show that overexpression of Sp1 activated, whereas Sp3 inhibited, Sp1 transactivation of FLIP promoter activity (P = 0.0001). 2-ME2 treatment reduced binding of Sp1 to the FLIP promoter as evidenced by ChIP. Further, levels of FLIP associated with Fas or FADD decreased, whereas cleavage of caspase-8, levels of Bid, and apoptosis increased in response to 2-ME2 treatment in PC-3 cells. Administration of 2-ME2 regressed established prostate tumors in TRAMP mice that were associated with reduced expression of FLIP and Sp1.

Conclusion: Targeting Sp1-mediated FLIP signaling pathway may provide a novel approach for prostate cancer management.
CD8 T-cell Responses against Cyclin B1 in Breast Cancer Patients with Tumors Overexpressing p53
Authors: Sørensen Rikke Bæk , Andersen Rikke Sick , Svane Inge Marie , Engell-Noerregaard Lotte , Hadrup Sine R. , Balslev Eva , Andersen Mads Hald , Straten Per thor
Published: February 2009

Purpose: This study aimed to examine CD8 T-cell reactivity in breast cancer patients against cyclin B1–derived peptides restricted by the human leukocyte antigen (HLA)-A2 molecule.

Experimental Design: Peripheral blood mononuclear cells from 36 breast cancer patients were analyzed by enzyme-linked immunosorbent spot (ELISPOT) for the presence of T cells recognizing the cyclin B1–derived peptides CB9 (AKYLMELTM) and CB-P4 (AKYLMELCC), in addition to modified versions of CB9, CB9L2 (ALYLMELTM) and CB9M2 (AMYLMELTM), both of which display higher affinity to HLA-A2.

Results: Twelve patients harbored a memory CD8 T-cell response against at least one of the peptides; strongest reactivity was detected against the CB9L2 peptide. Because the level of cyclin B1 has been shown to be influenced by the level of p53, which in turn is elevated in cancer cells because of point mutation, we analyzed the level of p53 protein in biopsies from the patients by immune histochemistry. Combined data showed that anti–cyclin B1 reactivity was predominantly detected in patients with tumors characterized by elevated expression of p53. Interestingly, no reactivity was detected against six peptides derived from the p53 protein.

Conclusions: Our data support the notion of cyclin B1 as a prominent target for immunologic recognition in cancer patients harboring p53-mutated cancer cells. Because mutation of p53 is one of the most frequent genetic alterations in human cancers, this suggests that immunotherapy based on targeting of cyclin B1 is broadly applicable in a large proportion of cancer patients.
Dynamic Contrast-Enhanced Magnetic Resonance Imaging As a Biomarker for Prediction of Radiation-Induced Neurocognitive Dysfunction
Authors: Cao Yue , Tsien Christina I. , Sundgren Pia C. , Nagesh Vijaya , Normolle Daniel , Buchtel Henry , Junck Larry , Lawrence Theodore S.
Published: February 2009

Purpose: To determine whether early assessment of cerebral microvessel injury can predict late neurocognitive dysfunction after brain radiation therapy (RT).

Experimental Design: Ten patients who underwent partial brain RT participated in a prospective dynamic contrast-enhanced magnetic resonance imaging (MRI) study. Dynamic contrast-enhanced MRI was acquired prior to, at weeks 3 and 6 during, and 1 and 6 months after RT. Neuropsychological tests were done pre-RT and at the post-RT MRI follow-ups. The correlations between early delayed changes in neurocognitive functions and early changes in vascular variables during RT were analyzed.

Results: No patients had tumor progression up to 6 months after RT. Vascular volumes and blood-brain barrier (BBB) permeability increased significantly in the high-dose regions during RT by 11% and 52% (P < 0.05), respectively, followed by a decrease after RT. Changes in both vascular volume and BBB permeability correlated with the doses accumulated at the time of scans at weeks 3 and 6 during RT and 1 month after RT (P < 0.03). Changes in verbal learning scores 6 months after RT were significantly correlated with changes in vascular volumes of left temporal (P < 0.02) and frontal lobes (P < 0.03), and changes in BBB permeability of left frontal lobes during RT (P < 0.007). A similar correlation was found between recall scores and BBB permeability.

Conclusion: Our data suggest that the early changes in cerebral vasculature may predict delayed alterations in verbal learning and total recall, which are important components of neurocognitive function. Additional studies are required for validation of these findings.
Genetic Signatures of HPV-related and Unrelated Oropharyngeal Carcinoma and Their Prognostic Implications
Authors: Klussmann Jens P. , Mooren Jeroen J. , Lehnen Martin , Claessen Sandra M.H. , Stenner Markus , Huebbers Christian U. , Weissenborn Soenke J. , Wedemeyer Inga , Preuss Simon F. , Straetmans Jos M.J.A.A. , Manni Johannes J. , Hopman Anton H.N. , Speel Ernst-Jan M.
Published: February 2009

Purpose: Patients with human papillomavirus (HPV)-containing oropharyngeal squamous cell carcinomas (OSCC) have a better prognosis than patients with HPV-negative OSCC. This may be attributed to different genetic pathways promoting cancer.

Experimental Design: We used comparative genomic hybridization to identify critical genetic changes in 60 selected OSCC, 28 of which were associated with HPV-16 as determined by HPV-specific PCR and fluorescence in situ hybridization analysis and positive p16INK4A immunostaining. The results were correlated with HPV status and clinical data from patients.

Results: Two thirds of OSCC harbored gain at 3q26.3-qter irrespective of HPV status. In HPV-negative tumors this alteration was associated with advanced tumor stage (P = 0.013). In comparison with HPV-related OSCC, the HPV-negative tumors harbored: (a) a higher number of chromosomal alterations and amplifications (P = 0.03 and 0.039, respectively); (b) significantly more losses at 3p, 5q, 9p, 15q, and 18q, and gains/amplifications at 11q13 (P = 0.002, 0.03; <0.001, 0.02, 0.004, and 0.001, respectively); and (c) less often 16q losses and Xp gains (P = 0.02 and 0.03). Survival analysis revealed a significantly better disease-free survival for HPV-related OSCC (P = 0.02), whereas chromosome amplification was an unfavorable prognostic indicator for disease-free and overall survival (P = 0.01 and 0.05, respectively). Interestingly, 16q loss, predominantly identified in HPV-related OSCC, was a strong indicator of favorable outcome (overall survival, P = 0.008; disease-free survival, P = 0.01) and none of these patients had a tumor recurrence.

Conclusions: Genetic signatures of HPV-related and HPV-unrelated OSCC are different and most likely underlie differences in tumor development and progression. In addition, distinct chromosomal alterations have prognostic significance.
Meta-analysis of Oncogenic Protein Kinase Cι Signaling in Lung Adenocarcinoma
Authors: Erdogan Eda , Klee Eric W. , Thompson E. Aubrey , Fields Alan P.
Published: February 2009

Purpose: Atypical protein kinase Cι (PKCι) is an oncogene in non–small cell lung cancer (NSCLC). Here, we identify four functional gene targets of PKCι in lung adenocarcinoma (LAC), the most prominent form of NSCLC.

Experimental Design: Three independent public domain gene expression data sets were interrogated to identify genes coordinately expressed with PKCι in primary LAC tumors. Results were validated by QPCR in an independent set of primary LAC tumors. RNAi-mediated knockdown of PKCι and the target genes was used to determine whether expression of the identified genes was regulated by PKCι, and whether these target genes play a role in anchorage-independent growth and invasion of LAC cells.

Results: Meta-analysis identified seven genes whose expression correlated with PKCι in primary LAC. Subsequent QPCR analysis confirmed coordinate overexpression of four genes (COPB2, ELF3, RFC4, and PLS1) in an independent set of LAC samples. RNAi-mediated knockdown showed that PKCι regulates expression of all four genes in LAC cells, and that the four PKCι target genes play an important role in the anchorage-independent growth and invasion of LAC cells. Meta-analysis of gene expression data sets from lung squamous cell, breast, colon, prostate, and pancreas carcinomas, as well as glioblastoma, revealed that a subset of PKCι target genes, particularly COPB2 and RFC4, correlate with PKCι expression in many tumor types.

Conclusion: Meta-analysis of public gene expression data are useful in identifying novel gene targets of oncogenic PKCι signaling. Our data indicate that both common and cell type–specific signaling mechanisms contribute to PKCι-dependent transformation.
Ductal Lavage Is an Inefficient Method of Biomarker Measurement in High-Risk Women
Authors: Khan Seema A. , Lankes Heather A. , Patil Deepa B. , Bryk Michele , Hou Nanjiang , Ivancic David , Nayar Ritu , Masood Shahla , Rademaker Alfred
Published: February 2009

Effective methods of serial epithelial sampling to measure breast-specific biomarkers will aid the rapid evaluation of new preventive interventions. We report here a proof-of-principle phase 2 study to assess the utility of ductal lavage (DL) to measure biomarkers of tamoxifen action. We enrolled women with a 5-year breast cancer risk estimate >1.6% or the unaffected breast of women with T1a or T1b breast cancer. After entry DL, participants chose tamoxifen or observation and underwent repeat DL 6 months later. Samples were processed for cytology and immunohistochemistry for estrogen receptor α, Ki-67, and cyclooxygenase-2. Of 182 women recruited, 115 (63%) underwent entry and repeat DL; 85 (47%) had sufficient cells for analysis from ≥1 duct at both time points; in 78 (43%), cells were sufficient from ≥1 matched ducts. Forty-six women chose observation and 39 chose tamoxifen. We observed greater reductions in the tamoxifen group than in the observation group for Ki-67 (adjusted P = 0.03) and estrogen receptor α (adjusted P = 0.07), but not in cyclooxygenase-2 (adjusted P = 0.4) labeling. Cytologic findings showed a trend toward improvement in the tamoxifen group compared with the observation group. Interobserver variability for cytologic diagnosis between two observers showed good agreement (κ = 0.44). Using DL, we observed the expected changes in tamoxifen-related biomarkers; however, poor reproducibility of biomarkers in the observation group, the 53% attrition rate of subjects from recruitment to biomarker analyses, and the expense of DL are significant barriers to the use of this procedure for biomarker assessment over time.
Avoiding Abducens Nerve Palsy during the Percutaneous Balloon Compression Procedure
Authors: Kefalopoulou Zinovia , Markaki Elli , Constantoyannis Constantine
Published: February 2009

Aims: The main purpose of this study is to underline the factors which are likely to increase the risk of a sixth nerve injury after percutaneous balloon compression of the gasserian ganglion. Methods: Medical records and intraoperative radiological images of 79 consecutive patients, who underwent percutaneous balloon compression for treating drugresistant trigeminal neuralgia, were analyzed. Results: Post-operatively, transient palsy of the sixth nerve developed in 2 patients. According to their intraoperative radiographs, the side effect resulted from irregular balloon shape and inflation of the balloon towards the sellar floor. Conclusion: The anatomic position and the shape of the inflated balloon represent key factors in determining the procedure’s benefit or risk of complications.
Thrombolysis in Acute Childhood Stroke: Design and Challenges of the Thrombolysis in Pediatric Stroke Clinical Trial
Authors: Amlie-Lefond Catherine , Chan Anthony K.C. , Kirton Adam , deVeber Gabrielle , Hovinga Collin A. , Ichord Rebecca , Stephens Derek , Zaidat Osama O.
Published: February 2009

Background: Although tissue plasminogen activator (tPA) has revolutionized the treatment of acute ischemic stroke in adults, no thrombolysis trials in childhood stroke have been conducted. Experience in adults cannot be applied to children due to fundamental age-related differences in coagulation systems, stroke pathophysiology and neuropharmacology. Obstacles to acute treatment trials in childhood stroke include delays in diagnosis and minimizing risk in a vulnerable population. Study Design: Thrombolysis in Pediatric Stroke (TIPS) is an international multicenter study to assess the safety of intravenous tPA within 0–3 h and intra-arterial tPA within 3–6 h of onset of arterial ischemic stroke in childhood. Through the International Pediatric Stroke Study, 30 international centers will enroll a total of 48 patients: 24 will be treated with intravenous tPA (0.6, 0.75, 0.9, and 1.0 mg/kg) using the classical dose-finding method, and 24 will be treated with intra-arterial tPA (maximum 0.2, 0.3, 0.4, and 0.5 mg/kg) using a Bayesian dose-finding method. Conclusion: The TIPS trial will be the first clinical trial exploring the safety and feasibility of systemic and local thrombolytic therapy in childhood stroke and the obstacles in conducting such a trial.
Biomarkers in Relation to Cognitive Reserve in Patients with Mild Cognitive Impairment – Proof of Concept
Authors: Rolstad S. , Nordlund A. , Eckerström C. , Gustavsson M.H. , Zetterberg H. , Wallin A.
Published: February 2009

Background: The concept of the cognitive reserve (CR) posits that factors such as education enable compensation for the effect of brain pathology. Consequently, pathology should be more pronounced in individuals with higher CR before becoming clinically apparent. Biomarkers such as total tau (t-tau) and β-amyloid 42 (Aβ42) may be surrogates for pathology in relation to CR in patients with neurodegenerative disease. Objective: To examine the applicability of biomarkers as surrogates for pathology in relation to the CR in patients with mild cognitive impairment (MCI) either converting to dementia or remaining stable at follow-up. Method: Comparisons of baseline t-tau, Aβ42, educational years and global cognition for MCI patients either converting to dementia (n = 57) or remaining stable (n = 91) were made. Patients converting to dementia were grouped on the basis of educational level and compared considering biomarkers and neuropsychological tests. Results: Stable MCI patients were better educated, performed better cognitively, had higher Aβ42 levels and lower levels of t-tau. Converting MCI patients with higher education had lower levels of Aβ42 and performed equally in neuropsychological tests compared to those with lower education. Conclusion: Our results suggest that highly educated MCI patients subsequently converting to dementia display more amyloid pathology.