bluuebunny/crossref_metadata_2025 huggingface dataset:
Waughs and Rumours of Waughs
Authors: Jarrett Bede
Published: August 2007

A wholly new influence over books written on school-life began with Compton Mackenzie and Sinister Street. Previously to that there had indeed been school stories, an endless procession of them, including such masterpieces as Tom Brown's Schooldays and others that like Eric sought to appeal frankly to that innate sentimentalism that lies uncomfortably across the conscience of every English boy. These, however, had culminated in The Hill, The Harrovians, and The Bending of the Twig, which are the three finest probably of that older school of Romanticism, throwing its touching veil of fights, friendship, and laughter over the hardened formalities and dreary littlenesses of normal school-life. These represent, then, the final effort of a style of writing which had become obsolete when dealing with “Olympians,” but was still employed in the happy efforts of grown men to recapture the spirit of boyhood, its inner sanctities and outer brutalities, its sorrows, its heartiness, its monotony, its intrigues as baffling, and to master and pupil as entrancing as the political games that interest and exhaust the diplomats of a larger world.

Then came the first volume of Sinister Street. This must be too well known to all readers of Blackfriars to require much treatment here : but its importance is hardly to be calculated. It opened the eyes of boys to the fascinating study of self-analysis : for it differed in two respects from previous ventures, in that first, it was written by one who had hardly left school, and secondly, it was a very detailed description of the changing attitude of a boy’s mind, its unfolding to good and evil, and thus it interested the reader, as often as not a boy-reader, in watching the changing attitude of his own mind.
Richard Crashaw
Authors: Makepeace John Foster
Published: August 2007

In the year that Charles the First came to the throne, Richard Crashaw was a lad of thirteen at the Charterhouse School. He had lost his mother in his infancy, and his father—the Rev. William Crashaw, a learned theologian, eloquent preacher, indifferent poet and bitter anti-Catholic— had recently died, leaving him with slender resources, but endowed with uncommon abilities and a rare gift for making friends.

He went to Cambridge in 1631 ; was admitted to Pembroke College ; gained a pensionership, having, as he afterwards wrote to a friend, spent all his patrimony in buying books ; and graduated in 1634 when he was twenty-one. In the same year he published a book of Scripture Epigrams in Latin, memorable for the one immortal line, “Nympha pudica Deum vidit, et erubuit,” upon which the skill of many translators, including William Hayley, has been exercised.

To draw a true portrait of Richard Crashaw he must be presented in this University setting. For he was, pre-eminently, in modern phrase, a “‘Varsity man.” The advantage of life at the older universities, it has often been said, has always been found to lie as much in the friendships that common studies foster as in the studies themselves. Crashaw enjoyed both to the full. Clearly, he was a student, and lived austere, laborious days, for he became proficient in the classics, as well as in Hebrew, Italian, and Spanish, besides in acquiring some skill in music and art. And the finest minds at the University were his friends. Of these latter we learn something from the numerous elegies he composed, according to the custom of the day.
Nationality
Authors: Devas Dominic
Published: August 2007

Nationality as an isolating force is new in the world, an artificial, after-Christian growth, dating from the break-up of Christendom at the so-called Reformation in the sixteenth century. Mutual hostility between races—as opposed to conflicts between a prince and his followers of one State, and a prince of another State and his followers—is not new, but a relapse into fore-Christian barbarism, when the cult of local deities, little knowledge of each other, and total inexperience of political union, led men to regard differences of race as natural and insurmountable barriers, when to be a stranger, a man of another race and creed, was—as it has since become—to be necessarily an enemy, so that the same word—hostis—once did for both. This idea of war between races as natural— , says Plato in Book I of his Laws—was gradually checked politically by the rise of the Roman Empire and the permanence of the Imperial idea, and, on the religious side, by Christianity with its doctrine of the unity of God and of the unity of man made to His Image and equal before Him. Thus with the of Plato we may contrast St. Paul’s ‘.There is neither Jew nor Greek : there is neither bond nor free. . . . For you are all one in Christ Jesus’ (Gal. iii. 28).”

Is this a wholly wild and quite untrue presentation of history? Such measure of truth as it may be found to contain will perhaps emerge as we proceed with our enquiry.
The Church in France and the Ruhr
Authors: Gwynn Denis
Published: August 2007

In her decision to seize the Ruhrland, France has committed herself to a course which leaves her practically no possibility of turning back. The gravity of her decision is already appreciated throughout the country, and the Press is busy exhorting all classes of the population to pay no heed to criticism of French policy and to face the trials which she has chosen to encounter with unflinching resolution and with a determination to rely solely upon Frances own resources. This is not the place to discuss the merits or otherwise of the policy which France has adopted. But in order to make clear the extremely serious dangers which it involves for the position of the Church in France if failure in the Ruhr should lead to a violent political reaction towards the Left, it is necessary to indicate briefly the principal factors in the situation.

In the first place it must be stated quite frankly that no one who sympathises with France in the difficulties with which she has been surrounded since the war could well see how the adventure into the Ruhr was to be avoided. The occupation of the Ruhr was Frances last card against Germany, and as all else had failed, France simply had to play it or acquiesce meekly in Germanys triumphant default. It is not fair to say that France is prepared to drive Europe into war rather than undertake to pay for her own reconstruction out of taxes.
Liturgical Hymns
Authors: Britten James
Published: August 2007

Among the few incidents of my ill-spent life which . I hope may be accounted to me for righteousness is the suppression of a hymn-book. At first sight this might appear to somewho do not know what literary crimes are committed under that innocent-seeming namea matter for remorse rather than for pride, for regret rather than for congratulation. But I will venture in justification to apply to myself the response of the prudent Rector of a seminary who, when told by a scrupulous student that he had discovered a new mortal sin, replied : Then, my son, keep it to yourself and never mention it; there are far too many known already. Yet when I add that the hymn-book was to have appeared under the protection of a Cardinal, my daring will seem to demand yet further excuse; so it will be simplest to say what actually happened.

The late Cardinal Vaughan, as is well known, had a burning desire for the conversion of England and, with that object in view, to bring the Church into contact with the people. It was this that impelled him to establish the original Catholic Truth Society for the publication or for dissemination of cheap popular Catholic books; and when, owing to various circumstances, that Society ceased to function, he gladly placed himself at the head of the new organisation established for the same purpose, retaining that position until his death in 1903. Apart from the C.T.S. the Cardinal issued a series of People's Manuals, prepared by himself, which had a very large sale.
Davidson's Transcendental Externalism
Authors: BRIDGES Jason
Published: August 2007

One of the chief aims of Donald Davidson's later work was to show that participation in a certain causal nexus involving two creatures and a shared environment–Davidson calls this nexus “triangulation”–is a metaphysically necessary condition for the acquisition of thought. This doctrine, I suggest, is aptly regarded as a form of what I call transcendental externalism. I extract two arguments for the transcendental‐externalist doctrine from Davidson's writings, and argue that neither succeeds. A central interpretive claim is that the arguments are primarily funded by a particular conception of the nature of non‐human animal life. This conception turns out to be insupportable. The failure of Davidson's arguments presses the question of whether we could ever hope to arrive at far‐reaching claims about the conditions for thought if we deny, as does Davidson, the legitimacy of the naturalistic project in the philosophy of mind.
Précis of Insensitive Semantics
Authors: CAPPELEN Herman , LEPORE Ernie
Published: August 2007

Insensitive Semantics (I) has three components: It defends a positive theory; it presents a methodology for how to distinguish semantic context sensitivity from other kinds of context sensitivity; and finally, it includes chapters critical of other contributors on these issues. In this Précis, we outline each component, but before doing so a few brief ‘big picture’ remarks about the positions defended in is are in order:
Replies
Authors: CAPPELEN Herman , LEPORE Ernie
Published: August 2007

We are in the curious position of disagreeing with Bach about what exactly we disagree about. As we see it, his characterization of our disagreement introduces largely irrelevant terminological issues; and these in turn cover up the fact that he simply hasn't replied to our central criticism. We start with two failed attempts to characterize the disagreement, and then move on to what is central.
Improved Protocols for Microwave-Assisted Cu(I)-Catalyzed Huisgen 1,3-Dipolar Cycloadditions
Authors: Cintas Pedro , Martina Katia , Robaldo Bruna , Garella Davide , Boffa Luisa , Cravotto Giancarlo
Published: August 2007

The Huisgen 1,3-dipolar cycloaddition of azides and acetylenes catalyzed by Cu(I) salts, leading to 1,2,3-triazoles, is one of the most versatile "click reactions". We have developed a series of optimized protocols and new applications of this reaction starting from several substrates, comparing heterogeneous vs homogeneous catalysis, conventional heating vs microwave irradiation or simultaneous microwave/ultrasound irradiation. Both non-conventional techniques strongly promoted the cycloaddition (bromide → azide → triazole), that could be conveniently performed in a one-pot procedure. This was feasible even with such bulky molecules as functionalized β-cyclodextrins (β-CD), starting from 61-O-tosyl-β-CD or from heptakis[6-O-(tert-butyldimethylsilyl)]-21-O-propargyl-β-CD. "Greener" heterogeneous catalysis with charcoal-supported Cu(II) or Cu(I) (prepared under ultrasound) was advantageously employed.
Transfer Hydrogenation of Carbonyl Compounds and Alkenes Catalyzed by Ruthenium(II)-N-Heterocycle Carbene Complexes
Authors: Fekete Marianna , Joó Ferenc
Published: August 2007

The Ru(II)-N-heterocycle carbene complexes [RuCl2(η6-p-cymene)L] (L = 1-butyl-3-methylimid- azol-2-ylidene) and [RuCl(η6-p-cymene)L(pta)]Cl (pta = 1,3,5-triaza-7-phosphaadamantane) showed excellent catalytic activities (with turnover frequencies up to 1116 h-1) in the hydrogen transfer reduction of cinnamaldehyde and several ketones using propan-2-ol/KOH as a H-donor. Similar hydrogenations of trans-stilbene and cyclohexene were characterized by low conversions. The hydrogenation of 4-phenylbut-3-en-2-one and cinnamaldehyde proceeded with moderate selectivities of the formation of the saturated alcohol or of the of C=C hydrogenation (giving saturated ketone or saturated aldehyde). In the case of cinnamaldehyde, the unsaturated alcohol is initially formed; however, subsequent redox isomerization to the saturated aldehyde with the same catalyst diminishes its yield. The hydrogen transfer from formate to 4-phenylbut-3-en-2-one in an aqueous-organic two-phase mixture was also demonstrated.
DESIGN AND IMPLEMENTATION OF A P2P COOPERATIVE PROXY CACHE SYSTEM
Authors: WANG JAMES Z. , BHULAWALA VIPUL
Published: August 2007

In this paper, we design and implement a P2P cooperative proxy caching system based on a novel P2P cooperative proxy caching scheme. To effectively locate the cached web documents, a TTL-based routing protocol is proposed to manage the query and response messages in the P2P cooperative proxy cache system. Furthermore, we design a predict query-route algorithm to improve the TTL-based routing protocol by adding extra information in the query message packets. To select a suitable cache replacement algorithm for the P2P cooperative proxy cache system, three different cache replacement algorithms, LRU, LFU and SIZE, are evaluated using web trace based performance studies on the implemented P2P cooperative proxy cache system. The experimental results show that LRU is an overall better cache replacement algorithm for the P2P proxy cache system although SIZE based cache replacement approach produces slightly better cache hit ratio when cache size is very small. The performance studies also demonstrate that the proposed message routing protocols significantly improve the performance of the P2P cooperative proxy cache system, in terms of cache hit ratio, byte hit ratio, user request latency, and the number of query messages generated in the proxy cache system, compared to the flooding based message routing protocol.
NON-STATIONARY NUMERICAL COVARIANCE ANALYSIS OF LINEAR MULTI DEGREE OF FREEDOM MECHANICAL SYSTEM SUBJECT TO RANDOM INPUTS
Authors: MARANO GIUSEPPE CARLO , ACCIANI GIUSEPPE , CASCELLA LENONARDO GIUSEPPE
Published: August 2007

Structural response of linear multi degree of freedom (MDoF) system subject to random Gaussian dynamic actions is defined by mean of vector and covariance matrix in state space. In case of non-stationary inputs, second-order spectral moments evaluation needs the solution of the so-called Lyapunov matrix differential equation. In this work a numerical scheme for its resolution is proposed, with reference to input processes modeled as linear filtered white noise with time-varying parameters, which is a common situation in amplitude and frequency variable loads. Numerical computational effort is minimized by taking into account symmetry characteristic of state space covariance matrix. As application of the proposed method a multi-storey building is analyzed to obtain reliability associated to maximum inter-storey exceeded over a given acceptable limit. It is assumed to be subject to seismic input described by a amplitude and frequency nonstationary process, by using a generalized non-stationary Kanai Tajimi seismic model. Structure is assumed as a plane shear frame MDoF system. Structural reliability evaluation is referred to "first time out-crossing" and different numerical benchmarks are considered.
Fifty-Five-Year Personal Experience with Human Nutrition Worldwide
Authors: Scrimshaw Nevin S.
Published: August 2007

By 1950 the vitamins had been identified, but little was known of their functions. Beriberi, pellagra, and ariboflavinosis were disappearing, but kwashiorkor and/or marasmus were common in most developing countries. Requirements for protein were still uncertain, and those for essential amino acids or essential fatty acids were unknown. The author's contributions in the field of vitamins began in the 1950s and have been reported in more than 650 publications and in 20 books or monographs. These contributions include establishing the Institute of Nutrition of Central America and Panama, the Department of Nutrition and Food Science at the Massachusetts Institute of Technology, the World Hunger Program of the United Nations University, and the International Nutrition Foundation. His scientific contributions include identification of synergistic interactions of nutrition and infection, use of potassium iodate for fortifying crude moist salt, research in the epidemiology of kwashiorkor and marasmus, development of a successful low-cost protein-rich food for infants and young children, establishment of human protein requirements, and investigation of single-cell protein for food use.
Akt activation and augmented fibronectin production in hyperhexosemia
Authors: Xin Xiping , Chen Shali , Khan Zia A. , Chakrabarti Subrata
Published: August 2007

Dysmetabolic state in diabetes may lead to augmented synthesis of extracellular matrix (ECM) proteins. In the endothelial cells, we have previously demonstrated that glucose-induced fibronectin (FN) production and that of its splice variant, EDB+FN, is regulated by protein kinase B (PKB, also known as Akt). In this study, we investigated the role of Akt1 in ECM protein production in the organs affected by chronic diabetic complications. We studied Akt1/PKBα knockout mice and wild-type control littermates. To avoid confounding effects of systemic insulin, we used 30% galactose feeding to induce hyperhexosemia for 8 wk starting at 6 wk of age. We investigated FN mRNA, EDB+FN mRNA, and transforming growth factor (TGF)-β mRNA expression, Akt phosphorylation, Akt kinase activity, and NF-κB and AP-1 activation in the retina, heart, and kidney. Renal and cardiac tissues were histologically examined. Galactose feeding caused significant upregulation of FN, EDB+FN, and TGF-β in all tissues. FN protein levels paralleled mRNA. Such upregulation were prevented in Akt1-deficient galactose-fed mice. Galactose feeding caused ECM protein deposition in the glomeruli and in the myocardium, which was prevented in the Akt knockout mice. NF-κB and AP-1 activation was pronounced in galactose-fed wild-type mice and prevented in the galactose-fed Akt1/PKBα-deficient group. In the retina and kidney, Ser473 was the predominant site for Akt phosphorylation, whereas in the heart it was Thr308. Parallel experiment in streptozotocin-induced diabetic animals showed similar results. The data from this study indicate that hyperhexosemia-induced Akt/PKB activation may be an important mechanism leading to NF-κB and AP-1 activation and increased ECM protein synthesis in the organs affected by chronic diabetic complications.
Increased susceptibility of aging kidney to ischemic injury: identification of candidate genes changed during aging, but corrected by caloric restriction
Authors: Chen G. , Bridenbaugh E. A. , Akintola A. D. , Catania J. M. , Vaidya V. S. , Bonventre J. V. , Dearman A. C. , Sampson H. W. , Zawieja D. C. , Burghardt R. C. , Parrish A. R.
Published: August 2007

Aging is associated with an increased incidence and severity of acute renal failure. However, the molecular mechanism underlying the increased susceptibility to injury remains undefined. These experiments were designed to investigate the influence of age on the response of the kidney to ischemic injury and to identify candidate genes that may mediate this response. Renal slices prepared from young (5 mo), aged ad libitum (aged-AL; 24 mo), and aged caloric-restricted (aged-CR; 24 mo) male Fischer 344 rats were subjected to ischemic stress (100% N2) for 0–60 min. As assessed by biochemical and histological evaluation, slices from aged-AL rats were more susceptible to injury than young counterparts. Importantly, caloric restriction attenuated the increased susceptibility to injury. In an attempt to identify the molecular pathway(s) underlying this response, microarray analysis was performed on tissue harvested from the same animals used for the viability experiments. RNA was isolated and the corresponding cDNA was hybridized to CodeLink Rat Whole Genome Bioarray slides. Subsequent gene expression analysis was performed using GeneSpring software. Using two-sample t-tests and a twofold cut-off, the expression of 92 genes was changed during aging and attenuated by caloric restriction, including claudin-7, kidney injury molecule-1 (Kim-1), and matrix metalloproteinase-7 (MMP-7). Claudin-7 gene expression peaked at 18 mo; however, increased protein expression in certain tubular epithelial cells was seen at 24 mo. Kim-1 gene expression was not elevated at 8 or 12 mo but was at 18 and 24 mo. However, changes in Kim-1 protein expression were only seen at 24 mo and corresponded to increased urinary levels. Importantly, these changes were attenuated by caloric restriction. MMP-7 gene expression was decreased at 8 mo, but an age-dependent increase was seen at 24 mo. Increased MMP-7 protein expression in tubular epithelial cells at 24 mo was correlated with the gene expression pattern. In summary, we identified genes changed by aging and changes attenuated by caloric restriction. This will facilitate investigation into the molecular mechanism mediating the age-related increase in susceptibility to injury.
Aldosterone promotes proximal tubular cell apoptosis: role of oxidative stress
Authors: Patni Hitesh , Mathew Jayant T. , Luan Liming , Franki Nicholas , Chander Praveen N. , Singhal Pravin C.
Published: August 2007

Aldosterone has attracted significant consideration for its role in the progression of renal injury. Since apoptotic cell loss contributes to the deterioration of renal function, we examined the effect of aldosterone on tubular cell apoptosis. To determine dose and time course effect, human renal proximal tubular (HK2) cells were treated with aldosterone at different doses and for variable time periods followed by evaluation for apoptosis. To determine the role of mineralocorticoid receptors (MR) and oxidative stress, HK2 cells were treated with either vehicle or aldosterone in the presence or absence of spironolactone/antioxidants/free radical scavengers (FRS) followed by evaluation for apoptosis. The presence of MR was evaluated using RT-PCR. Reactive oxygen species (ROS) generation was evaluated using redox-sensitive dyes. Effect of aldosterone was evaluated on dephosphorylation of phospho-Bad and accumulation of cytosolic cytochrome c. Human tubular cells express MR. Aldosterone promotes tubular cell apoptosis in a dose- and time-dependent manner. This effect of aldosterone is mediated through MR and associated with generation of ROS. Antioxidants and FRS partially attenuated the proapoaptotic effect of aldosterone. Aldosterone enhanced dephosphorylation of phospho-Bad and accumulation of cytosolic cytochrome c. We conclude that aldosterone-induced tubular cell apoptosis is mediated through the activation of the mitochondrial pathway and generation of ROS.
Effect of deficiency in SREBP cleavage-activating protein on lipid metabolism during intermittent hypoxia
Authors: Li Jianguo , Nanayakkara Ashika , Jun Jonathan , Savransky Vladimir , Polotsky Vsevolod Y.
Published: August 2007

Obstructive sleep apnea (OSA), a condition leading to intermittent hypoxia (IH) during sleep, has been associated with dyslipidemia, atherosclerosis, and increased cardiovascular mortality. We previously showed in C57BL/6J mice that IH causes hypercholesterolemia and upregulation of sterol regulatory element binding protein (SREBP)-1, a transcription factor of lipid biosynthesis in the liver. The goal of the present study was to provide mechanistic evidence that IH causes hypercholesterolemia via the SREBP-1 pathway. We utilized mice with a conditional knockout of SREBP cleavage-activating protein (SCAP) in the liver ( L-Scap− mice), which exhibit low levels of an active nuclear isoform of SREBP-1 (nSREBP-1). We exposed L-Scap− mice and wild-type (WT) littermates to IH or intermittent air control for 5 days. IH was induced during the 12-h light phase by decreasing FiO2from 20.9% to 5% for a period of 30 s with rapid reoxygenation to 20.9% through the subsequent 30 s. In WT mice, IH increased fasting levels of serum total and HDL cholesterol, serum triglycerides, serum and liver phospholipids, mRNA levels of SREBP-1 and mitochondrial glycerol-3-phosphate acyltransferase (mtGPAT), and protein levels of SCAP, nSREBP-1, and mtGPAT in the liver. In L-Scap− mice, IH did not have any effect on serum and liver lipids, and expression of lipid metabolic genes was not altered. We conclude that hyperlipidemia in response to IH is mediated via the SREBP-1 pathway. Our data suggest that the SREBP-1 pathway could be used as a therapeutic target in patients with both OSA and hyperlipidemia.
Interaction of MLL Amino Terminal Sequences with Menin Is Required for Transformation
Authors: Caslini Corrado , Yang Zhaohai , El-Osta Mohamad , Milne Thomas A. , Slany Robert K. , Hess Jay L.
Published: August 2007

Rearrangements of the mixed lineage leukemia gene MLL are associated with aggressive lymphoid and myeloid leukemias. The resulting MLL fusion proteins enforce high-level expression of HOX genes and the HOX cofactor MEIS1, which is pivotal for leukemogenesis. Both wild-type MLL and MLL fusion proteins interact with the tumor suppressor menin and with the Hoxa9 locus in vivo. Here, we show that MLL sequences between amino acids 5 and 44 are required for interaction with menin and for the transformation of hematopoietic progenitors. Blocking the MLL-menin interaction by the expression of a dominant negative inhibitor composed of amino terminal MLL sequences down-regulates Meis1 expression and inhibits cell proliferation, suggesting that targeting this interaction may be an effective therapeutic strategy for leukemias with MLL rearrangements. [Cancer Res 2007;67(15):7275–83]
p53 Activation in Response to Mitotic Spindle Damage Requires Signaling via BubR1-Mediated Phosphorylation
Authors: Ha Geun-Hyoung , Baek Kwan-Hyuck , Kim Hyun-Soo , Jeong Sook-Jung , Kim Chang-Min , McKeon Frank , Lee Chang-Woo
Published: August 2007

The mitotic spindle checkpoint plays a crucial role in regulating accurate chromosome segregation and preventing the adaptation of multiploid progeny cells. Recent reports have indicated that the induction of p53 by mitotic checkpoint activation is essential for protecting cells from abnormal chromosome ploidization caused by mitotic failure. However, although studies have shown that p53 deficiencies arrest mitosis, compromise apoptosis, and may cause profound aneuploidy, the molecular mechanisms leading to p53 induction following mitotic checkpoint activation remain unknown. Here, we show that the BubR1 mitotic checkpoint kinase interacts with p53 both in vitro and in vivo, with higher levels of interaction in mitotic cells. This interaction contributes to p53 phosphorylation. Silencing of BubR1 expression reduces the phosphorylation and stability of p53, whereas exogenous introduction of BubR1 proteins into BubR1-depleted cells recovers p53 stability. In addition, inhibition of BubR1 expression in the presence of a microtubule inhibitor accelerates chromosomal instability and polyploidy in p53-null cells. These results collectively suggest that p53 activation in response to mitotic spindle damage requires signaling via BubR1-mediated phosphorylation. [Cancer Res 2007;67(15):7155–64]
Targeting Matrix Metalloproteinases and Endothelial Cells with a Fusion Peptide against Tumor
Authors: Zou Yufei , Chen Yahui , Jiang Yongqiang , Gao Jin , Gu Jun
Published: August 2007

Development of novel therapy for patients with tumor is still a challenge at the present time. We designed a fusion peptide (RK5) with two targets as a novel agent against tumor. The fusion peptide RK5 containing the kringle 5 fragment of human plasminogen and a decapeptide (CTTHWGFTLC) was constructed and expressed in yeast. Matrix metalloproteinase (MMP) activity, proliferation, and migration of endothelial cells were examined in vitro, respectively. Angiogenesis, tumor growth, metastasis, and survival time were evaluated in in vivo models. Administration of RK5 was delivered by both protein and gene approach. The results showed that RK5 inhibited the activity of MMP-9 and exhibited more inhibitory effects on proliferation and migration of endothelial cells than that of kringle 5 fragment and decapeptide individually. RK5 also inhibited angiogenesis, tumor growth, and metastasis and increased survival time of mice bearing tumor. In addition, the effectiveness of RK5 could be achieved by both protein and gene delivery. In conclusion, RK5 has potential to inhibit tumor growth and metastasis and to prolong survival time of animals bearing tumor. Therefore, fusion peptide RK5 with two targets provides a new design for the development of antitumor drugs and has potential for clinical application. [Cancer Res 2007;67(15):7295–300]
Tripeptidyl-peptidase II Controls DNA Damage Responses and In vivo γ-Irradiation Resistance of Tumors
Authors: Hong Xu , Lei Lu , Künert Brita , Naredla Rajender , Applequist Steven E. , Grandien Alf , Glas Rickard
Published: August 2007

Cellular responses to γ-irradiation exposure are controlled by phosphatidylinositol 3-kinase–related kinases (PIKK) in the nucleus, and in addition, cytosolic PIKKs may have a role in such responses. Here, we show that the expression of tripeptidyl-peptidase II (TPPII), a high molecular weight cytosolic peptidase, required PIKK signaling and that TPPII was rapidly translocated into the nucleus of γ-irradiated cells. These events were dependent on mammalian target of rapamycin, a cytosolic/mitochondrial PIKK that is activated by γ-irradiation. Lymphoma cells with inhibited expression of TPPII failed to efficiently stabilize p53 and had reduced ability to arrest proliferation in response to γ-irradiation. We observed that TPPII contains a BRCA COOH-terminal–like motif, contained within sequences of several proteins involved in DNA damage signaling pathways, and this motif was important for nuclear translocation of TPPII and stabilization of p53. Novel tripeptide-based inhibitors of TPPII caused complete in vivo tumor regression in mice in response to relatively low doses of γ-irradiation (3–4 Gy/wk). This was observed with established mouse and human tumors of diverse tissue backgrounds, with no tumor regrowth after cancellation of treatment. These TPPII inhibitors had minor effects on tumor growth as single agent and had low cellular toxicity. Our data indicated that TPPII connects signaling by cytosolic/mitochondrial and nuclear PIKK-dependent pathways and that TPPII can be targeted for inhibition of tumor therapy resistance. [Cancer Res 2007;67(15):7165–74]
Decreased Tumor Surveillance after Adoptive T-Cell Therapy
Authors: Matter Matthias , Pavelic Viktor , Pinschewer Daniel D. , Mumprecht Sabine , Eschli Bruno , Giroglou Tsanan , von Laer Dorothee , Ochsenbein Adrian F.
Published: August 2007

The effect of cancer immunotherapy on the endogenous immune response against tumors is largely unknown. Therefore, we studied immune responses against murine tumors expressing the glycoprotein (GP) and/or nucleoprotein of lymphocytic choriomeningitis virus (LCMV) with or without adoptive T-cell therapy. In nontreated animals, CTLs specific for different epitopes as well as LCMV-GP–specific antibodies contributed to tumor surveillance. Adoptive immunotherapy with monoclonal CTLs specific for LCMV-gp33 impaired the endogenous tumor-specific antibody and CTL response by targeting antigen cross-presenting cells. As a consequence and in contrast to expectations, immunotherapy enhanced tumor growth. Thus, for certain immunogenic tumors, a reduction of tumor-specific B- and T-cell responses and enhanced tumor growth may be an unwanted consequence of adoptive immunotherapy. [Cancer Res 2007;67(15):7467–76]
An Improved Bioluminescence Resonance Energy Transfer Strategy for Imaging Intracellular Events in Single Cells and Living Subjects
Authors: De Abhijit , Loening Andreas Markus , Gambhir Sanjiv Sam
Published: August 2007

Bioluminescence resonance energy transfer (BRET) is currently used for monitoring various intracellular events, including protein-protein interactions, in normal and aberrant signal transduction pathways. However, the BRET vectors currently used lack adequate sensitivity for imaging events of interest from both single living cells and small living subjects. Taking advantage of the critical relationship of BRET efficiency and donor quantum efficiency, we report generation of a novel BRET vector by fusing a GFP2 acceptor protein with a novel mutant Renilla luciferase donor selected for higher quantum yield. This new BRET vector shows an overall 5.5-fold improvement in the BRET ratio, thereby greatly enhancing the dynamic range of the BRET signal. This new BRET strategy provides a unique platform to assay protein functions from both single live cells and cells located deep within small living subjects. The imaging utility of the new BRET vector is shown by constructing a sensor using two mammalian target of rapamycin pathway proteins (FKBP12 and FRB) that dimerize only in the presence of rapamycin. This new BRET vector should facilitate high-throughput sensitive BRET assays, including studies in single live cells and small living subjects. Applications will include anticancer therapy screening in cell culture and in small living animals. [Cancer Res 2007;67(15):7175–83]
Krüppel-Like Factor 8 Induces Epithelial to Mesenchymal Transition and Epithelial Cell Invasion
Authors: Wang Xianhui , Zheng Mingzhe , Liu Gang , Xia Weiya , McKeown-Longo Paula J. , Hung Mien-Chie , Zhao Jihe
Published: August 2007

Tumor invasion and metastasis are the main causes of death from cancer. Epithelial to mesenchymal transition (EMT) is a determining step for a cancer cell to progress from a noninvasive to invasive state. Krüppel-like factor 8 (KLF8) plays a key role in oncogenic transformation and is highly overexpressed in several types of invasive human cancer, including breast cancer. To understand the role of KLF8 in regulating the progression of human breast cancer, we first established stable expression of KLF8 in an immortalized normal human breast epithelial cell line. We found that KLF8 strongly induced EMT and enhanced motility and invasiveness in the cells, by analyzing changes in cell morphology and epithelial and mesenchymal marker proteins, and using cell migration and Matrigel invasion assays. Chromatin immunoprecipitations (ChIP), oligonucleotide precipitations, and promoter-reporter assays showed that KLF8 directly bound and repressed the promoter of E-cadherin independent of E boxes in the promoter and Snail expression. Aberrant elevation of KLF8 expression is highly correlated with the decrease in E-cadherin expression in the invasive human breast cancer. Blocking KLF8 expression by RNA interference restored E-cadherin expression in the cancer cells and strongly inhibited the cell invasiveness. This work identifies KLF8 as a novel EMT-regulating transcription factor that opens a new avenue in EMT research and suggests an important role for KLF8 in human breast cancer invasion and metastasis. [Cancer Res 2007;67(15):7184–93]
A Human scFv Antibody against TRAIL Receptor 2 Induces Autophagic Cell Death in Both TRAIL-Sensitive and TRAIL-Resistant Cancer Cells
Authors: Park Kyung-Jin , Lee Seung-Hyun , Kim Tae-In , Lee Hae-Won , Lee Chang-Han , Kim Eun-Hee , Jang Ji-Young , Choi Kyeong Sook , Kwon Myung-Hee , Kim Yong-Sung
Published: August 2007

Tumor necrosis factor (TNF)–related apoptosis-inducing ligand (TRAIL) induces apoptotic cell death in a variety of tumor cells without significant cytotoxicity on normal cells. However, many cancer cells with apoptotic defects are resistant to treatment with TRAIL alone, limiting its potential as an anticancer therapeutic. Here, we report on the tumoricidal activity of a human single-chain fragment variable, HW1, which specifically binds to TRAIL receptor 2 (TR2) without competing with TRAIL for the binding. HW1 treatment as a single agent induces autophagic cell death in a variety of both TRAIL-sensitive and TRAIL-resistant cancer cells, but exhibits much less cytotoxicity on normal cells. The HW1-induced autophagic cell death was inhibited by an autophagy inhibitor, 3-methyladenine, or by RNA interference knockdown of Beclin-1 and Atg7. We also show that the HW1-mediated autophagic cell death occurs predominantly via the c-Jun NH2-terminal kinase pathway in a caspase-independent manner. Analysis of the death-inducing signaling complex induced by HW1 binding to TR2 exhibits the recruitment of TNF receptor–associated death domain and TNF receptor–associated factor 2, but not Fas-associated death domain, caspase-8, or receptor-interacting protein, which is distinct from that induced by TRAIL. Our results reveal a novel TR2-mediated signaling pathway triggering autophagic cell death and provides a new strategy for the elimination of cancer cells, including TRAIL-resistant tumors, through nonapoptotic cell death.[Cancer Res 2007;67(15):7327–34]
Visualizing the Dynamics of EGFR Activity and Antiglioma Therapies In vivo
Authors: Arwert Esther , Hingtgen Shawn , Figueiredo Jose-Luiz , Bergquist Henry , Mahmood Umar , Weissleder Ralph , Shah Khalid
Published: August 2007

Many altered pathways in cancer cells depend on growth factor receptors. In primary malignant gliomas, the amplification/alteration of the epidermal growth factor receptor (EGFR) has been shown to play a significant role in enhancing glioma burden. In an effort to dissect the role of EGFR expression in glioma progression in vivo and evaluate targeted therapies for gliomas, we have genetically engineered glioma cells to visualize the dynamics of EGFR and targeted therapies in real time in vivo. Using engineered lentiviral vectors bearing fusions between EGFR and its exon 2 to 7 deleted variant (EGFRvIII) with green fluorescent protein (GFP) and Renilla luciferase (Rluc), we show that there is a direct correlation between EGFR expression and glioma cell proliferation in the initial stages of glioma progression. To monitor and evaluate EGFR-targeted therapies, we have engineered (a) short hairpin RNAs (shRNA) and (b) clinically used monoclonal antibody, cetuximab. Using EGFR-GFP-Rluc/firefly luciferase (Fluc)-DsRed2 glioma model, we show that both shRNAs and cetuximab result in a considerable reduction in glioma cell proliferation in culture and glioma burden in vivo that can be monitored in real time at a cellular resolution. This study serves as a template to follow the role of growth factor receptor expression in tumor progression and to image therapeutic efficacy of targeted therapies in cancer. [Cancer Res 2007;67(15):7335–42]
Induction of p53-Dependent Senescence by the MDM2 Antagonist Nutlin-3a in Mouse Cells of Fibroblast Origin
Authors: Efeyan Alejo , Ortega-Molina Ana , Velasco-Miguel Susana , Herranz Daniel , Vassilev Lyubomir T. , Serrano Manuel
Published: August 2007

Cellular senescence is emerging as an important in vivo anticancer response elicited by multiple stresses, including currently used chemotherapeutic drugs. Nutlin-3a is a recently discovered small-molecule antagonist of the p53-destabilizing protein murine double minute-2 (MDM2) that induces cell cycle arrest and apoptosis in cancer cells with functional p53. Here, we report that nutlin-3a induces cellular senescence in murine primary fibroblasts, oncogenically transformed fibroblasts, and fibrosarcoma cell lines. No evidence of drug-induced apoptosis was observed in any case. Nutlin-induced senescence was strictly dependent on the presence of functional p53 as revealed by the fact that cells lacking p53 were completely insensitive to the drug, whereas cells lacking the tumor suppressor alternative reading frame product of the CDKN2A locus underwent irreversible cell cycle arrest. Interestingly, irreversibility was achieved in neoplastic cells faster than in their corresponding parental primary cells, suggesting that nutlin-3a and oncogenic signaling cooperate in activating p53. Our current results suggest that senescence could be a major cellular outcome of cancer therapy by antagonists of the p53-MDM2 interaction, such as nutlin-3a. [Cancer Res 2007;67(15):7350–7]
A Growth Hormone Receptor Mutation Impairs Growth Hormone Autofeedback Signaling in Pituitary Tumors
Authors: Asa Sylvia L. , DiGiovanni Rebecca , Jiang Jing , Ward Megan L. , Loesch Kimberly , Yamada Shozo , Sano Toshiaki , Yoshimoto Katsuhiko , Frank Stuart J. , Ezzat Shereen
Published: August 2007

Pituitary tumors are a diverse group of neoplasms that are classified based on clinical manifestations, hormone excess, and histomorphologic features. Those that cause growth hormone (GH) excess and acromegaly are subdivided into morphologic variants that have not yet been shown to have pathogenetic significance or predictive value for therapy and outcome. Here, we identify a selective somatic histidine-to-leucine substitution in codon 49 of the extracellular domain of the GH receptor (GHR) in a morphologic subtype of human GH-producing pituitary tumors that is characterized by the presence of cytoskeletal aggresomes. This GHR mutation significantly impairs glycosylation-mediated receptor processing, maturation, ligand binding, and signaling. Pharmacologic GH antagonism recapitulates the morphologic phenotype of pituitary tumors from which this mutation was identified, inducing the formation of cytoskeletal keratin aggresomes. This novel GHR mutation provides evidence for impaired hormone autofeedback in the pathogenesis of these pituitary tumors. It explains the lack of responsiveness to somatostatin analogue therapy of this tumor type, in contrast to the exquisite sensitivity of tumors that lack aggresomes, and has therapeutic implications for the safety of GH antagonism as a therapeutic modality in acromegaly. [Cancer Res 2007;67(15):7505–11]
ELMO1 and Dock180, a Bipartite Rac1 Guanine Nucleotide Exchange Factor, Promote Human Glioma Cell Invasion
Authors: Jarzynka Michael J. , Hu Bo , Hui Kwok-Min , Bar-Joseph Ifat , Gu Weisong , Hirose Takanori , Haney Lisa B. , Ravichandran Kodi S. , Nishikawa Ryo , Cheng Shi-Yuan
Published: August 2007

A distinct feature of malignant gliomas is the intrinsic ability of single tumor cells to disperse throughout the brain, contributing to the failure of existing therapies to alter the progression and recurrence of these deadly brain tumors. Regrettably, the mechanisms underlying the inherent invasiveness of glioma cells are poorly understood. Here, we report for the first time that engulfment and cell motility 1 (ELMO1) and dedicator of cytokinesis 1 (Dock180), a bipartite Rac1 guanine nucleotide exchange factor (GEF), are evidently linked to the invasive phenotype of glioma cells. Immunohistochemical analysis of primary human glioma specimens showed high expression levels of ELMO1 and Dock180 in actively invading tumor cells in the invasive areas, but not in the central regions of these tumors. Elevated expression of ELMO1 and Dock180 was also found in various human glioma cell lines compared with normal human astrocytes. Inhibition of endogenous ELMO1 and Dock180 expression significantly impeded glioma cell invasion in vitro and in brain tissue slices with a concomitant reduction in Rac1 activation. Conversely, exogenous expression of ELMO1 and Dock180 in glioma cells with low level endogenous expression increased their migratory and invasive capacity in vitro and in brain tissue. These data suggest that the bipartite GEF, ELMO1 and Dock180, play an important role in promoting cancer cell invasion and could be potential therapeutic targets for the treatment of diffuse malignant gliomas. [Cancer Res 2007;67(15):7203–11]
Dual Role of Mitochondrial Reactive Oxygen Species in Hypoxia Signaling: Activation of Nuclear Factor-κB via c-SRC– and Oxidant-Dependent Cell Death
Authors: Lluis Josep M. , Buricchi Francesca , Chiarugi Paola , Morales Albert , Fernandez-Checa José C.
Published: August 2007

Hypoxia is a prominent feature of solid tumor development and is known to stimulate mitochondrial ROS (mROS), which, in turn, can activate hypoxia-inducible transcription factor-1α and nuclear factor-κB (NF-κB). Because NF-κB plays a central role in carcinogenesis, we examined the mechanism of mROS-mediated NF-κB activation and the fate of cancer cells during hypoxia after mitochondrial reduced glutathione (mGSH) depletion. Hypoxia generated mROS in hepatoma (HepG2, H35), neuroblastoma (SH-SY5Y), and colon carcinoma (DLD-1) cells, leading to hypoxia-inducible transcription factor-1α–dependent gene expression and c-Src activation that was prevented in cells expressing a redox-insensitive c-Src mutant (C487A). c-Src stimulation activated NF-κB without IκB-α degradation due to IκB-α tyrosine phosphorylation that was inhibited by rotenone/TTFA or c-Src antagonism. The c-Src–NF-κB signaling contributed to the survival of cells during hypoxia as c-Src inhibition or p65 down-regulation by small interfering RNA–sensitized HepG2 cells to hypoxia-induced cell death. Moreover, selective mGSH depletion resulted in an accelerated and enhanced mROS generation by hypoxia that killed SH-SY5Y and DLD-1 cells without disabling the c-Src–NF-κB pathway. Thus, although mROS promote cell survival by NF-κB activation via c-Src, mROS overgeneration may be exploited to sensitize cancer cells to hypoxia. [Cancer Res 2007;67(15):7368–77]
Interleukin-2 Administration Alters the CD4+FOXP3+ T-Cell Pool and Tumor Trafficking in Patients with Ovarian Carcinoma
Authors: Wei Shuang , Kryczek Ilona , Edwards Robert P. , Zou Linhua , Szeliga Wojciech , Banerjee Mousumi , Cost Marilyn , Cheng Pui , Chang Alfred , Redman Bruce , Herberman Ronald B. , Zou Weiping
Published: August 2007

Interleukin (IL)-2 is used in the immunotherapy of patients with certain cancer and HIV infection. IL-2 treatment reliably results in 16% to 20% objective clinical response rate in cancer patients, with significant durability of responses in selected patients. However, the mechanisms of therapeutic activity in responding versus nonresponding patients remain poorly understood. CD4+CD25+FOXP3+ regulatory T (Treg) cells contribute to immunosuppressive networks in human tumors. We treated 31 ovarian cancer patients with IL-2. We show that administration of IL-2 induces the proliferation of existent Treg cells in patients with ovarian cancer. The potency of Treg cell proliferation is negatively determined by the initial prevalence of Treg cells, suggesting that Treg cells are a factor for self-controlling Treg cell proliferation. After IL-2 cessation, the number of Treg cells more efficiently dropped in clinical responders than nonresponders. Furthermore, IL-2 treatment stimulates chemokine receptor CXCR4 expression on Treg cells, enables Treg cell migration toward chemokine CXCL12 in the tumor microenvironment, and may enforce Treg cell tumor accumulation. Our findings support the concept that administration of IL-2 numerically and functionally affects the Treg cell compartment. These data provide an important insight in evaluating the clinical benefit and therapeutic prediction of IL-2 treatment in patients with cancer. [Cancer Res 2007;67(15):7487–94]