bluuebunny/crossref_metadata_2025 huggingface dataset:
Ghrelin and Growth Hormone Secretagogue Receptor Expression in Mice during Aging
Authors: Sun Yuxiang , Garcia Jose Manuel , Smith Roy G.
Published: December 2006

In well-nourished humans, GH and IGF-I decline during aging, and the responsiveness of the GH axis to exogenous ghrelin is attenuated with age. Intriguingly, the GH/IGF-I axis is rejuvenated by chronic treatment with the ghrelin mimetic MK-0677, resulting in improvements in body composition, suggesting that frail elderly subjects might benefit from treatment with ghrelin and ghrelin mimetics. Mouse models are widely used to study the effects of ghrelin, but the impact of age on the ghrelin pathway is unclear. In this study, total and active ghrelin peptides were measured in plasma, and ghrelin mRNA was quantitated in brain tissue from different aged C57BL/6J mice. Surprisingly, plasma levels of ghrelin peptide slightly increased with age; ghrelin mRNA levels were similar in brains from mice aged 2, 6, 12, and 28 months but higher in mice aged 18 and 24 months. The tissue distribution of Ghsr1a mRNA (ghrelin receptor) was also characterized, and pituitary and brain exhibited the highest levels of expression. In the pituitary gland, the highest concentration of Ghsr1a mRNA was observed at age 1–2 months, it was lower at 6 months, and remained unchanged for up to 30 months of age. This result is consistent with the finding that GH release in response to exogenous ghrelin was not significantly different in mice aged 7–30 months. In the brain, Ghsr1a mRNA levels remained stable during aging. Hence, in C57BL/6J male mice, aging is not associated with changes in circulating ghrelin levels or changes in ghrelin receptor expression in the pituitary gland and brain.
The Low-Density Lipoprotein Class A Module of the Relaxin Receptor (Leucine-Rich Repeat Containing G-Protein Coupled Receptor 7): Its Role in Signaling and Trafficking to the Cell Membrane
Authors: Kern András , Agoulnik Alexander I. , Bryant-Greenwood Gillian D.
Published: December 2006

The relaxin receptor (LGR7, relaxin family peptide receptor 1) is a member of the leucine-rich repeat containing G protein-coupled receptors subgroup C. This and the LGR8 (relaxin family peptide receptor 2) receptor are unique in having a low-density lipoprotein class A (LDL-A) module at their N termini. This study was designed to show the role of the LDL-A in LGR7 expression and function. Point mutants for the conserved cysteines (Cys47 and Cys53) and for calcium binding asparagine (Asp58), a mutant with deleted LDL-A domain and chimeric LGR7 receptor with LGR8 LDL-A all showed no cAMP response to human relaxins H1 or H2. We have shown that their cell surface delivery was uncompromised. The mutation of the putative N-linked glycosylation site (Asn36) decreased cAMP production and reduced cell surface expression to 37% of the wild-type LGR7. All point mutant, chimeric, and wild-type receptor proteins were expressed as the two forms. The immature or precursor form of the receptor was 80 kDa, whereas the mature receptor, delivered to the cell surface was 95 kDa. The glycosylation mutant was also expressed as two forms with appropriately smaller molecular masses. Deletion of the LDL-A module resulted in expression of the mature receptor only. These data suggest that the LDL-A module of LGR7 influences receptor maturation, cell surface expression, and relaxin-activated signal transduction.
Functional Characterization of Polymorphisms in the Kidney Enhancer of the Human Renin Gene
Authors: Itani Hana A. , Liu Xuebo , Pratt J. Howard , Sigmund Curt D.
Published: December 2006

The renin gene is regulated by an enhancer located 2.6 kb upstream of the transcription start site in the mouse and 11 kb upstream in humans. Despite extensive sequence conservation, the mouse renin enhancer is transcriptionally more active than the human renin enhancer. We report that the mechanism accounting for this is a result of sequence variation in the promoter proximal half-site of a retinoic-acid response element present in the enhancer. This sequence difference also prompted us to search for naturally occurring polymorphisms in the renin enhancer among normal and hypertensive human subjects. We sequenced the kidney enhancer from 90 samples derived from the Coriell Polymorphism Discovery Resource and 95 severely hypertensive Caucasian and African-American individuals. A single relatively frequent polymorphism (7, 2, and 7%, respectively in the Coriell, African-American, and Caucasian) was identified in the enhancer, one nucleotide downstream of the promoter distal half-site of the retinoic-acid response element. This variant was transcriptionally silent in transfection assays performed in renin-expressing As4.1 cells, a model of renal juxtaglomerular cells. A singleton polymorphism in the promoter was also identified in a single African-American individual. This polymorphism was located between binding sites for CBF1 and homeobox D10 but was also transcriptionally silent either in the presence or absence of the enhancer. Our study demonstrates the presence of silent polymorphisms in the renin promoter and enhancer, thus underscoring the critical importance of performing functional analyses before initiating expensive clinical studies seeking association between polymorphisms and complex diseases such as hypertension.
A class of risk processes with delayed claims: ruin probability estimates under heavy tail conditions
Authors: Ganesh Ayalvadi , Torrisi Giovanni Luca
Published: December 2006

We consider a class of risk processes with delayed claims, and we provide ruin probability estimates under heavy tail conditions on the claim size distribution.
The upcrossings index and the extremal index
Authors: Ferreira H.
Published: December 2006

For stationary sequences X = {Xn}n≥1 we relate τ, the limiting mean number of exceedances of high levels un by X1,…,Xn, and ν, the limiting mean number of upcrossings of the same level, through the expression θ = (ν/τ)η, where θ is the extremal index of X and η is a new parameter here called the upcrossings index. The upcrossings index is a measure of the clustering of upcrossings of u by variables in X, and the above relation extends the known relation θ = ν/τ, which holds under the mild-oscillation local restriction D″(u) on X. We present a new family of local mixing conditions D̃(k)(u) under which we prove that (a) the intensity of the limiting point process of upcrossings and η can both be computed from the k-variate distributions of X; and (b) the cluster size distributions for the exceedances are asymptotically equivalent to those for the lengths of one run of exceedances or the lengths of several consecutive runs which are separated by at most k − 2 nonexceedances and, except for the last one, each contain at most k − 2 exceedances.
An urn model arising from all-optical networks
Authors: Morrison John A.
Published: December 2006

An occupancy model that has arisen in the investigation of randomized distributed schedules in all-optical networks is considered. The model consists of B initially empty urns, and at stage j of the process dj ≤ B balls are placed in distinct urns with uniform probability. Let Mi(j) denote the number of urns containing i balls at the end of stage j. An explicit expression for the joint factorial moments of M0(j) and M1(j) is obtained. A multivariate generating function for the joint factorial moments of Mi(j), 0 ≤ i ≤ I, is derived (where I is a positive integer). Finally, the case in which the dj, j ≥ 1, are independent, identically distributed random variables is investigated.
Conditioned stable Lévy processes and the Lamperti representation
Authors: Caballero M. E. , Chaumont L.
Published: December 2006

By variously killing a stable Lévy process when it leaves the positive half-line, conditioning it to stay positive, and conditioning it to hit 0 continuously, we obtain three different, positive, self-similar Markov processes which illustrate the three classes described by Lamperti (1972). For each of these processes, we explicitly compute the infinitesimal generator and from this deduce the characteristics of the underlying Lévy process in the Lamperti representation. The proof of this result bears on the behaviour at time 0 of stable Lévy processes before their first passage time across level 0, which we describe here. As an application, for a certain class of Lévy processes we give the law of the minimum before an independent exponential time. This provides the explicit form of the spatial Wiener-Hopf factor at a particular point and the value of the ruin probability for this class of Lévy processes.
Discretionary stopping of one-dimensional Itô diffusions with a staircase reward function
Authors: Bronstein Anne Laure , Hughston Lane P. , Pistorius Martijn R. , Zervos Mihail
Published: December 2006

We consider the problem of optimally stopping a general one-dimensional Itô diffusion X. In particular, we solve the problem that aims at maximising the performance criterion Ex[exp(-∫0τr(Xs)ds)f(Xτ)] over all stopping times τ, where the reward function f can take only a finite number of values and has a ‘staircase’ form. This problem is partly motivated by applications to financial asset pricing. Our results are of an explicit analytic nature and completely characterise the optimal stopping time. Also, it turns out that the problem's value function is not C1, which is due to the fact that the reward function f is not continuous.
Genealogy for supercritical branching processes
Authors: Lagerås Andreas Nordvall , Martin-Löf Anders
Published: December 2006

We study the genealogy of so-called immortal branching processes, i.e. branching processes where each individual upon death is replaced by at least one new individual, and conclude that their marginal distributions are compound geometric. The result also implies that the limiting distributions of properly scaled supercritical branching processes are compound geometric. We exemplify our results with an expression for the marginal distribution for a class of branching processes that have recently appeared in the theory of coalescent processes and continuous stable random trees. The limiting distribution can be expressed in terms of the Fox H-function, and in special cases by the Meijer G-function.
Multitype linear fractional branching processes
Authors: Joffe A. , Letac G.
Published: December 2006

We complete a paper written by Edward Pollak in 1974 on a multitype branching process the generating functions of whose birth law are fractional linear functions with the same denominator. The main tool is a parameterization of these functions adapted using the mean matrix M and an element w of the first quadrant. We use this opportunity to give a self-contained presentation of Pollak's theory.
A four-dimensional random motion at finite speed
Authors: Kolesnik Alexander D.
Published: December 2006

We consider the random motion of a particle that moves with constant finite speed in the space ℝ4 and, at Poisson-distributed times, changes its direction with uniform law on the unit four-sphere. For the particle's position, X(t) = (X1(t), X2(t), X3(t), X4(t)), t > 0, we obtain the explicit forms of the conditional characteristic functions and conditional distributions when the number of changes of directions is fixed. From this we derive the explicit probability law, f(x, t), x ∈ ℝ4, t ≥ 0, of X(t). We also show that, under the Kac condition on the speed of the motion and the intensity of the switching Poisson process, the density, p(x,t), of the absolutely continuous component of f(x,t) tends to the transition density of the four-dimensional Brownian motion with zero drift and infinitesimal variance σ2 = ½.
Characterizations in a random record model with a nonidentically distributed initial record
Authors: Barlevy Gadi , Nagaraja H. N.
Published: December 2006

We consider a sequence, of random length M, of independent, continuous observations Xi, 1 ≤ i ≤ M, where M is geometric, X1 has cumulative distribution function (CDF) G, and Xi, i ≥ 2, have CDF F. Let N be the number of upper records and let Rn, n ≥ 1, be the nth record value. We show that N is independent of F if and only if G(x) = G0(F(x)) for some CDF G0, and that if E(|X2|) is finite then so is E(|Rn|), n ≥ 2, whenever N ≥ n or N = n. We prove that the distribution of N, along with appropriately chosen subsequences of E(Rn), characterize F and G and, along with subsequences of E(Rn - Rn-1), characterize F and G up to a common location shift. We discuss some applications to the identification of the wage offer distribution in job search models.
Some limiting properties of the bounds of the present value function of a life insurance portfolio
Authors: Zhang Yi , Lin Zhengyan , Weng Chengguo
Published: December 2006

Under certain assumptions on the dependence structure of the residual lives of the insured (i.e. their independence, positive association, or negative association), in this paper we establish some laws of large numbers for the convex upper bounds, derived by the technique of comonotonicity, of the present value function of a homogeneous portfolio composed of the whole-life insurance policies.
Navigation in small-world networks: a scale-free continuum model
Authors: Franceschetti Massimo , Meester Ronald
Published: December 2006

The small-world phenomenon, the principle that we are all linked by a short chain of intermediate acquaintances, has been investigated in mathematics and social sciences. It has been shown to be pervasive both in nature and in engineering systems like the World Wide Web. Work of Jon Kleinberg has shown that people, using only local information, are very effective at finding short paths in a network of social contacts. In this paper we argue that the underlying key to finding short paths is scale invariance. In order to appreciate scale invariance we suggest a continuum setting, since true scale invariance happens at all scales, something which cannot be observed in a discrete model. We introduce a random-connection model that is related to continuum percolation, and we prove the existence of a unique scale-free model, among a large class of models, that allows the construction, with high probability, of short paths between pairs of points separated by any distance.
Foster-type criteria for Markov chains on general spaces
Authors: Fralix Brian H.
Published: December 2006

This paper establishes new Foster-type criteria for a Markov chain on a general state space to be Harris recurrent, positive Harris recurrent, or geometrically ergodic. The criteria are based on drift conditions involving stopping times rather than deterministic steps. Meyn and Tweedie (1994) developed similar criteria involving random-sized steps, independent of the Markov chain under study. They also posed an open problem of finding criteria involving stopping times. Our results essentially solve that problem. We also show that the assumption of ψ-irreducibility is not needed when stating our drift conditions for positive Harris recurrence or geometric ergodicity.
Examining Configurations and Firm Performance in a Suboptimal Equifinality Context
Authors: Payne G. Tyge
Published: December 2006

Gresov and Drazin’s (1997) suggestions regarding the study of equifinality are used to investigate the organizational configurations that exists within a suboptimal equifinality context. A suboptimal equifinality situation exists when organizations must satisfy multiple and conflicting functional demands with a restricted set of design options; these restrictions force all organizations to operate suboptimally. Gresov and Drazin (1997) speculate that in such a situation legitimacy claims are made about one function being more important than the other(s) so that an increased quantity of acceptable organizational forms are established. Additionally, they argue that a misfit penalty is likely to exist for organizations that deviate from the preferred configurational design. With data from the medical group industry, these speculations about suboptimal equifinality situations are tested using multivariate regression analyses. Findings show that the configuration type aligning with the preferred functional demand of quality performs relatively higher than those aligning with operational efficiency or attempting to meet both demands. Additionally, the greater the organization’s deviation from that design, the lower the financial performance. In summary, this study demonstrates that it is useful to take a functional equivalence perspective when examining variance in firm performance that is caused by design differences. Such an approach allows a more complete understanding of the fit-performance relationship by explicitly considering three factors: (i) the number of and level of conflict(s) among functional demands placed on the organization, (ii) the structural design options available to the decision maker, and (iii) the trade-offs that exist.
The Antecedents and Performance Implications of Cooperative Exchange Norms
Authors: Bercovitz Janet , Jap Sandy D. , Nickerson Jack A.
Published: December 2006

Our study investigates the antecedents and performance implications of cooperative exchange norms. We argue that, in early relationships, the level of expected cooperative norms in an exchange is the result of a calculative process facilitated by transaction attributes: joint transaction-specific investments and observability. The greater the level of these two exchange attributes, the greater the level of cooperative exchange norms, all else being equal. We further argue that the realized level of cooperative exchange norms can deviate from the expected level because the development of such norms is the result of social processes that management cannot directly and fully control. This gap between realized and expected norms affects exchange performance. Performance suffers when the realized level of cooperative exchange norms falls below the expected level, but overshooting expectations lays a critical groundwork for repeat transactions. The analysis of a survey of 182 collaborative R&D alliances provides initial support for our theory.
Competitive Implications of Interfirm Mobility
Authors: Wezel Filippo Carlo , Cattani Gino , Pennings Johannes M.
Published: December 2006

This paper examines the competitive consequences of interfirm mobility. Because the loss of key members (defined as top decision makers) to competing firms may amount to a replication of a firm’s higher-order routines, we investigate the conditions under which interfirm mobility triggers transfer of routines across organizational boundaries. We examine membership lists pertinent to the Dutch accounting industry to study key member exits and firm dissolutions over the period 1880–1986. We exploit information on the type of membership migration (individual versus collective) and the competitive saliency of the destination firm as inferred from the recipient status (incumbent versus start-up) and its geographic location (same versus different province). The dissolution risk is highest when collective interfirm mobility results in a new venture within the same geographic area. The theoretical implications of this study are discussed.
Microarray-in-a-Tube for Detection of Multiple Viruses
Authors: Liu Quanjun , Bai Yunfei , Ge Qinyu , Zhou Shixin , Wen Tian , Lu Zuhong
Published: December 2006

Background: The detection of multiple viruses is important for pathogenic diagnosis and disease control. Microarray detection is a good method, but requires complex procedures for multiple virus detection.

Methods: We developed a novel PCR assay, the microarray-in-a-tube system, which integrates multiple PCR processes and DNA microarrays for multiple virus detection. A 5 × 5 oligonucleotide microarray for detecting 4 respiratory tract viruses (severe acute respiratory syndrome–associated coronavirus, influenza A virus, influenza B virus, and enterovirus) with inner controls was arranged on the inner surface of a specially designed Eppendorf cap with a flat, optically transparent window.

Results: We were able to perform all detection processes in the encapsulated system without opening the cap. The 4 viruses were successfully amplified by one-step reverse transcription–PCR in the encapsulated tube. After the PCR process, the microarray-in-a-tube was inverted, and the fluorescence-labeled PCR products were directly hybridized on the microarray. Hybridization signals were obtained with an ordinary fluorescent microscope. The sensitivity of the system for virus detection reached 102 copies/μL. With the help of inner controls, the system provided reliable results without false negatives and false positives.

Conclusions: The microarray-in-a-tube system is a rapid, labor-saving tool for multiple virus detection with several advantages, such as convenience, prevention of cross-contamination of the PCR products, and potential for multiple-gene detection.
Stability of Urinary Fractionated Metanephrines and Catecholamines during Collection, Shipment, and Storage of Samples
Authors: Willemsen Jacques J , Ross H Alec , Lenders Jacques WM , Sweep Fred CGJ
Published: December 2006

Background: Measurements of 24-h fractionated urinary metanephrines and catecholamines are used for the diagnosis of pheochromocytoma, but adequate information is needed regarding collection, storage, and shipment conditions.

Methods: Spot urine samples were collected from 8 healthy volunteers. Aliquots were immediately frozen at −20 °C, or acidified to pH 4 and then frozen either directly or after 24 h at room temperature. The remaining urine was left at room temperature for 24 h and then split into one portion that was acidified and one portion that was not. Aliquots were either frozen or allowed to stand at room temperature for an additional 24, 48, 72, 96, and 168 h before freezing. We also tested the efficacy of adding Na2EDTA and Na2S2O5, as an alternative to acidification for preservation of the catecholamines.

Results: No clinically relevant degradation (<5%) was observed for the fractionated metanephrines under any of the storage conditions. In contrast, in ∼50% of the untreated samples catecholamines were partially degraded during the first 24 h at room temperature. Immediate acidification, however, prevented degradation, whereas acidification after 24 h prevented further decay. Addition of Na2EDTA and Na2S2O5 fully prevented degradation of catecholamines during the first 24 h in 4 of 5 cases. In the remaining case, degradation did not exceed 10%.

Conclusion: Preservation of samples for measurements of urinary fractionated metanephrines is not necessary if samples are assayed or frozen within 1 week, which is an important advantage if transport of samples is necessary. In contrast, urinary catecholamines require preservation measures during collection.
Nucleofection Is a Valuable Transfection Method for Transient and Stable Transgene Expression in Adipose Tissue-Derived Stem Cells
Authors: Zaragosi Laure-Emmanuelle , Billon Nathalie , Ailhaud Gérard , Dani Christian
Published: December 2006

Adipose tissue-derived stem cells are a powerful tool for in vitro study of adult stem cell biology. So far, they have not been extensively used for gain or loss of function studies since they are resistant to most common transfection methods. Herein, we tested several classic transfection methods on human multipotent adipose tissue-derived stem (hMADS) cells. Our results showed that lipofectants and calcium phosphate were poorly efficient for transgene delivery in hMADS cells. In contrast, nucleofection, an electroporation-based method that is assumed to target plasmid DNA directly to the cell nucleus, led to a significant transient transgene expression in hMADS cells (up to 76% enhanced green fluorescent protein [EGFP]-positive cells were detected). Furthermore, after selection of hMADS cells that were nucleofected with a selectable plasmid coding for EGFP, stable EGFP expressing clones could be propagated in culture and efficiently induced to differentiate into EGFP-positive adipocytes and osteoblasts. Finally, we verified that nucleofected hMADS cells could produce a functional, transgene-encoded, secreted protein. To this aim, hMADS cells were nucleofected with a plasmid coding for leukemia inhibitory factor (LIF). This protein was detected at high concentrations in supernatants from pCAG-LIF transfected hMADS cells. Moreover, supernatants were able to maintain mouse embryonic stem cells' undifferentiated phenotype, indicating that hMADS cells could secrete a functional LIF protein. Taken together, our data demonstrate that nucleofection allows both transient and stable gene expression in adipose tissue-derived stem cells, without impairing their differentiation potential.
Age-Dependent Increase in Side Population Distribution Within Hematopoiesis: Implications for Our Understanding of the Mechanism of Aging
Authors: Pearce Daniel J. , Anjos-Afonso Fernando , Ridler Christopher M. , Eddaoudi Ayad , Bonnet Dominique
Published: December 2006

It is thought that, as we age, damage to our stem cells may lead to diminished stem cell pool function and, consequently, a reduced organ regeneration potential that contributes to somatic senescence. Stem cells have evolved many antitoxicity mechanisms, and certain mechanisms may be utilized to isolate hematopoietic stem cells. One method exploits the activity of the ATP-binding cassette/G2 transporter to efflux Hoechst 33,342 and results in a stem cell population known as the side population (SP). The SP subset represents a remarkable enrichment for hematopoietic stem cells and provides an opportunity to re-evaluate age-based changes in hematopoietic stem cells. We report here that the frequency of SP cells steadily increases with age, as does the proportion of Lin−/Sca-1+/c-kit+ cells that is capable of Hoechst efflux. Phenotyping, progenitor, and long-term repopulation assays have indicated that SP cells in older mice are still stem cells, albeit with a lower homing efficiency than SP cells from younger mice. Analysis of apoptosis within SP cells has revealed an apoptosis-resistant population in SP cells from old mice. Gene expression analysis has determined that SP cells from old mice have a reduced expression of apoptosis-promoting genes than SP cells from young mice. This increase in SP cells with age seems to be an intrinsic property that may be independent of the age of the microenvironment (niche), and our data might provide some clues as to how this alteration in the proportion of stem/progenitor cells occurs. A possible selection-based mechanism of stem cell pool aging is discussed.

Disclosure of potential conflicts of interest is found at the end of this article.
p66Shc/Notch-3 Interplay Controls Self-Renewal and Hypoxia Survival in Human Stem/Progenitor Cells of the Mammary Gland Expanded In Vitro as Mammospheres
Authors: Sansone Pasquale , Storci Gianluca , Giovannini Catia , Pandolfi Silvia , Pianetti Simona , Taffurelli Mario , Santini Donatella , Ceccarelli Claudio , Chieco Pasquale , Bonafé Massimiliano
Published: December 2006

The comprehension of the basic biology of stem cells is expected to provide a useful insight into the pathogenesis of cancer. In particular, there is evidence that hypoxia promotes stem cell renewal in vitro as well as in vivo. It therefore seems reasonable that stem cell survival and hypoxia response are strictly connected at molecular level. We here report that the 66-kDa isoform of the SHC gene (p66Shc) is induced in a breast cancer cell line by the exposure to hypoxic environment and that it controls the expression of the stem cell regulatory gene Notch-3. Then, we show that p66Shc/Notch-3 interplay modulates self-renewal (by inducing the Notch-ligand Jagged-1) and hypoxia survival (by inducing the hypoxia-survival gene carbonic anhydrase IX) in mammary gland stem/progenitor cells, expanded in vitro as multicellular spheroids (mammospheres). We conclude that mechanisms that regulate stem cell renewal and hypoxia survival are integrated at the level of the p66Shc/Notch3 interplay. Because Notch-3, Jagged-1, and carbonic anhydrase IX are dysregulated in breast cancer, and because p66Shc is an aging-regulating gene, we envision that these data may help in understanding the relationship among aging, cancer, and stem cells.
Significance, detection and markers of disseminated breast cancer cells
Authors: Lacroix Marc
Published: December 2006

The development of distant metastases is the major cause of death from breast cancer. In order to predict and prevent tumour spreading, many attempts are being made to detect small numbers of tumour cells that have shed from the primary lesions and have moved to lymph nodes, blood or bone marrow. This article presents the advantages and the limitations of techniques used for disseminated tumour cells (DTC) detection. DTC markers are listed and the most currently used of them (KRT19, CEACAM5, TACSTD1, MUC1, EGFR, ERBB2, SCGB2A2, SCGB2A1, SCGB1D2, PIP, SBEM, TFF1, TFF3, ANKRD30A, SPDEF, ESR1, SERPINB5 and GABRP) are discussed, notably on the basis of recent data on breast tumour portraits (luminal epithelial-like, basal/myoepithelial-like and ERBB2). The significance of DTC for the prognosis and prediction of response to therapy is examined. DTC viability, the notion of cell dormancy and the concept of breast cancer stem cells are also discussed.
The role of external beam radiotherapy in the treatment of papillary thyroid cancer
Authors: Lee Nancy , Tuttle Michael
Published: December 2006

The role of external beam radiotherapy (EBRT) in treating thyroid cancer has brought forth controversy. Due to various histologic presentations and different natural histories, there is no uniform approach/recommendation among centers and/or authorities regarding the role of EBRT for thyroid cancer. This is particularly true for papillary thyroid carcinoma (PTC) where the clinical course can range from a disease that is cured with simple surgery to an aggressive form of poorly differentiated thyroid cancer with high rates of recurrence/death from disease. In addition, because the majority of the patients with PTC undergo postoperative radioactive iodine (RAI) treatment, the question remains as to what is the exact role of EBRT for PTC in the setting of RAI treatment?

In this issue of Endocrine-Related Cancer, Chow and colleagues identified indications for EBRT and RAI therapy for PTC based on a retrospective review of 1300 patients. The authors concluded that postoperative RAI treatment is indicated in patients with pT2-pT4, pN0-pN1b while postoperative EBRT is recommended for patients with gross residual, positive margin, pT4, pN1b, and lymph nodes > 2 cm disease. Other centers have also published their experience on the value of EBRT for PTC but with different indications. The reasons for the variations from different centers are complex. However, when all published results are taken together, the findings confirm the added value of EBRT to the present management of PTC in a select group of patients, particularly those with high risk features. In this commentary, these issues will be discussed and recommendations regarding the role of EBRT will be given.
Phytoestrogens and breast cancer –promoters or protectors?
Authors: Rice Suman , Whitehead Saffron A
Published: December 2006

The majority of breast cancers are oestrogen dependent and in postmenopausal women the supply of oestrogens in breast tissue is derived from the peripheral conversion of circulating androgens. There is, however, a paradox concerning the epidemiology of breast cancer and the dietary intake of phytoestrogens that bind weakly to oestrogen receptors and initiate oestrogen-dependent transcription. In Eastern countries, such as Japan, the incidence of breast cancer is approximately one-third that of Western countries whilst their high dietary intake of phytoestrogens, mainly in the form of soy products, can produce circulating levels of phytoestrogens that are known experimentally to have oestrogenic effects. Indeed, their weak oestrogenicity has been used to advantage by herbalist medicine to promote soy products as a natural alternative to conventional hormone replacement therapy (HRT). Such usage could increase in light of recent evidence that long-term HRT usage may be associated with an increased risk of breast cancer with a consequent reduction in prescription rates. So, are phytoestrogens safe as a natural alternative to HRT and could they be promoters or protectors of breast cancer? If they are promoters, then we must assume that it is due to their oestrogenic effect. If they are protectors, then other actions of phytoestrogens, including their ability to inhibit enzymes that are responsible for converting androgens and weak oestrogens into oestradiol, must be considered. This paper addresses these questions by reviewing the actions of phytoestrogens on oestrogen receptors and key enzymes that convert androgens to oestrogens in relation to the growth of breast cancer cells. In addition, it compares the experimental and epidemiological evidence pertinent to the potential beneficial or harmful effects of phytoestrogens in relation to the incidence/progression of breast cancer and their efficacy as natural alternatives to conventional HRT.
Tamoxifen-induced ER-α–SRC-3 interaction in HER2 positive human breast cancer; a possible mechanism for ER isoform specific recurrence
Authors: Mc Ilroy Marie , Fleming Fergal J , Buggy Yvonne , Hill Arnold D K , Young Leonie S
Published: December 2006

Differential signalling between the two oestrogen receptor (ER) isoforms in the presence of tamoxifen has been described. We hypothesise that differential recruitment of the steroid receptor co-activator, SRC-3 to ER-α and ER-β may in part explain associations between ER isoforms and response to endocrine treatment. SRC-3 was localised within epithelial cells of breast tumour tissue and was co-localised with ER-α and ER-β, (n = 112). Expression of SRC-3 was found to be positively associated with ER-α (P = 0.0021) and inversely with ER-β (P < 0.0001). Uniquely, this study utilises primary cell cultures derived from patient tumours, thus providing samples not readily available in most molecular model systems. These samples have enabled us to investigate the influence of growth factor pathways on steroid receptor-co-activator interactions. In HER2 (human epidermal growth factor receptor 2) positive primary tumour cell cultures 17β-estradiol induced a decrease in SRC-3, whereas upregulated SRC-3 expression. Furthermore, treatment with tamoxifen-induced SRC-3 recruitment to the ER-oestrogen response element and enhanced interaction between SRC-3 and ER-α, but not ER-β. Knockdown of SRC-3 results in a concomitant loss of expression of the oestrogen target gene pS2. Furthermore, silencing of SRC-3 resensitizes endocrine resistant, HER2 positive cells to the anti-proliferative effects of tamoxifen. The ability of ER-α, but not ER-β to recruit SRC-3 in the presence of tamoxifen may in part explain the differential ER isoform associations with recurrence in human breast cancer.
Differentiated thyroid cancer cell invasion is regulated through epidermal growth factor receptor-dependent activation of matrix metalloproteinase (MMP)-2/gelatinase A
Authors: Yeh Michael W , Rougier Jean-Philippe , Park Jin-Woo , Duh Quan-Yang , Wong Mariwil , Werb Zena , Clark Orlo H
Published: December 2006

Mechanisms of invasion in thyroid cancer remain poorly understood. We hypothesized that signaling via the epidermal growth factor receptor (EGFR) stimulates thyroid cancer cell invasion by altering the expression and cleavage of matrix metalloproteinases (MMPs). Papillary and follicular carcinoma cell lines were treated with EGF, the EGFR tyrosine kinase inhibitor AG1478, and the MMP inhibitors GM-6001 and Col-3. Flow cytometry was used to detect EGFR. In vitro invasion assays, gelatin zymography, and quantitative reverse transcription-PCR were used to assess the changes in invasive behavior and MMP expression and activation. All cell lines were found to overexpress functional EGFR. EGF stimulated invasion by thyroid cancer cells up to sevenfold (P < 0.0001), a process that was antagonized completely by AG1478 and Col-3, partially by GM-6001, but not by the serine protease inhibitor aprotinin. EGF upregulated expression of MMP-9 (2.64- to 8.89-fold, P < 0.0001) and membrane type-1 MMP (MT1-MMP, 1.97- to 2.67-fold, P < 0.0001). This effect was blocked completely by AG1478 and partially by Col-3. The activation of MMP-2 paralleled MT1-MMP expression. We demonstrate that MMPs are critical effectors of invasion in the papillary and follicular thyroid cancer cell lines studied. Invasion is regulated by signaling through EGFR, an effect mediated by augmentation of gelatinase expression and activation. MMP inhibitors and growth factor antagonists may be effective tumoristatic agents for the treatment of aggressive thyroid carcinomas.
Selective spatial upregulation of intratumoral stromal aromatase in breast cancer patients: evidence for imbalance of local estrogen metabolism
Authors: Singer Christian F , Fink-Retter Anneliese , Gschwantler-Kaulich Daphne , Thalhammer Theresia , Hudelist Gernot , Mueller Ruth , Czerwenka Klaus , Kubista Ernst
Published: December 2006

The suppression of local estrogens levels is of key importance in the treatment of ER-positive breast cancer. Essentially all endocrine strategies act by either suppressing estrogen formation or competitively inhibiting receptor-binding in tumor cells. Nevertheless, little is still known about the local expression of aromatase and sulfotransferase which are the key modulators of intra-tumoral estrogen levels. We have performed immunohistochemostry to investigate the expression of aromatase and sulfotransferase in 42 samples obtained directly from malignant breast tumors, and compared it to biopsies obtained from uninvolved tissue in the vicinity of the invasion front, and to distant breast tissue. We found that aromatase was equally detectable in both tumor epithelial and stroma, but was mostly epithelial in non-malignant tissues (P = 0.00008, Fisher’s exact test). Also, aromatase protein expression was significantly more common in tumoral stroma when compared with peritumoral and distant breast stroma (P = 0.00005, and P < 0.00001 respectively). With the notable exception of cystosarcoma phylloides, sulfotransferase protein was detectable only in epithelial tissues, regardless of the location within the diseased breast. However, epithelial sulfotransferase was correlated with epithelial aromatase (r = 0.35461, P = 0.0009, Spearman’s ρ test) and with the epithelial ER status (r = 0.29313, P = 0.005). We have demonstrated a differential aromatase and sulfotransferase protein expression pattern that is dependent on the spatial relation to a malignant breast tumor. Our results indicate a net increase in intratumoral active estrogen levels through increased stromal aromatization, while physiological local inactivation by sulfotransferase activity remains essentially unchanged.
Added value of gastrin receptor scintigraphy in comparison to somatostatin receptor scintigraphy in patients with carcinoids and other neuroendocrine tumours
Authors: Gotthardt Martin , Béhé Martin P , Grass Julia , Bauhofer Artur , Rinke Anja , Schipper Meike L , Kalinowski Marc , Arnold Rudolf , Oyen Wim J G , Behr Thomas M
Published: December 2006

Gastrin receptor scintigraphy (GRS) is a new imaging method primarily developed for the detection of metastases of medullary thyroid carcinoma (MTC). As gastrin-binding CCK2 receptors are also expressed on a variety of other neuroendocrine tumours (NET), we compared GRS to somatostatin receptor scintigraphy (SRS) in patients with NET. SRS and GRS were performed within 21 days in a series of 60 consecutive patients with NET. GRS was directly compared with SRS. If lesions were visible on GRS but not detectable by SRS, other imaging modalities (MRI, CT) and follow-up were used for verification. Of the 60 evaluable patients, 51 had carcinoid tumours, 3 gastrinomas, 2 glucagonomas, 1 insulinoma and 3 paragangliomas. The overall tumour-detection rate was 73.7% for GRS and 82.1% for SRS. In the 11 patients with negative SRS, GRS was positive in 6 (54.5%). Based on the number of tumour sites detected and the degree of uptake, GRS performed better than SRS in 13 patients (21.7%), equivalent images were obtained in 18 cases (30.0%) and SRS performed better in 24 (40.0%) cases. In six of the SRS positive patients, 18 additional sites of tumour involvement could be detected. Overall, GRS detected additional tumour sites in 20% of the patients. Localisation of the primary tumours or their functional status had no influence on the outcome of imaging. GRS should be performed in selected patients as it may provide additional information in patients with NET with equivocal or absent somatostatin uptake.
Microarrays in breast cancer research and clinical practice – the future lies ahead
Authors: Gruvberger-Saal Sofia K , Cunliffe Heather E , Carr Kristen M , Hedenfalk Ingrid A
Published: December 2006

Molecular profiling for classification and prognostic purposes has demonstrated that the genetic signatures of tumors contain information regarding biological properties as well as clinical behavior. This review highlights the progress that has been made in the field of gene expression profiling of human breast cancer. Breast cancer has become one of the most intensely studied human malignancies in the genomic era; several hundred papers over the last few years have investigated various clinical and biological aspects of human breast cancer using high-throughput molecular profiling techniques. Given the grossly heterogeneous nature of the disease and the lack of robust conventional markers for disease prediction, prognosis, and response to treatment, the notion that a transcriptional profile comprising multiple genes, rather than any single gene or other parameter, will be more predictive of tumor behavior is both appealing and reasonable. Promising results have emerged from these studies, correlating gene expression profiles with prognosis, recurrence, metastatic potential, therapeutic response, as well as biological and functional aspects of the disease. Clearly, the integration of genomic approaches into the clinic lies in the near future, but prospective studies based on larger patient cohorts representing the whole spectrum of breast cancer, oncogenic pathway-based studies, attendant care in bioinformatic analyses and validation studies are needed before the full promise of gene expression profiling can be realized in the clinical setting.
Antiproliferative and proapoptotic effects of histone deacetylase inhibitors on gastrointestinal neuroendocrine tumor cells
Authors: Baradari Viola , Huether Alexander , Höpfner Michael , Schuppan Detlef , Scherübl Hans
Published: December 2006

Treatment options of advanced neuroendocrine tumors (NETs) are unsatisfactory. Hence, innovative therapeutic approaches are urgently needed. Inhibition of histone deacetylases (HDACs) is a promising new approach in cancer therapy. While several HDAC inhibitors have already entered clinical trials, the effect of HDAC inhibition on NET has not been investigated. Therefore, we evaluated the antineoplastic effects of three different HDAC inhibitors, trichostatin A (TSA), sodium butyrate (NaB), and MS-275, on growth and apoptosis of the gastrointestinal NET cell lines CM and BON. We could demonstrate that HDAC inhibition dose-dependently inhibited proliferation of both cell lines with IC50 values varying from the millimolar (NaB) to the micromolar (MS-275) and the nanomolar range (TSA). Moreover, HDAC inhibition potently induced apoptosis, which was accompanied by DNA-fragmentation, an up to 12-fold caspase-3 activation and downregulated Bcl-2 expression. Furthermore, HDAC inhibition resulted in cell cycle arrest at the G1–S-transition, which was associated with the suppression of cyclin D1 expression and induction of p21 and p27 expression. For BON cells, we observed an additional block in the G2/M phase, which was aligned with a downregulation of cyclin B1. In addition, combined treatment with MS-275 and somatostatin or the synthetic somatostatin analog octreotide was evaluated. Neither somatostatin nor its stable analog octreotide augmented the antiproliferative effect of MS-275 in NET cells. To conclude, our data show that HDAC inhibition is a promising new approach in the treatment of NET disease, which should be evaluated in clinical studies.
Uptake and antiproliferative effect of molecular iodine in the MCF-7 breast cancer cell line
Authors: Arroyo-Helguera O , Anguiano B , Delgado G , Aceves C
Published: December 2006

This study analyzes the uptake and antiproliferative effect of two different chemical forms of iodine, iodide (I−) and molecular iodine (I2), in MCF-7 cells, which are inducible for the Na+/I− symporter (NIS) and positive for pendrin (PDS). The mouse fibroblast cell line NIH3T3 was used as control. Our results show that in MCF-7 cells, I− uptake is sustained and dependent on NIS, whereas I2 uptake is transient with a maximal peak at 10 min and a final retention of 10% of total uptake. In contrast, no I− was taken up by NIH3T3 cells, and although I2 was captured with the same time pattern as in MCF-7 cells, its uptake was significantly lower, and it was not retained within the cell. The uptake of I2 is independent of NIS, PDS, Na+, and energy, but it is saturable and dependent on protein synthesis, suggesting a facilitated diffusion system. Radioiodine was incorporated into protein and lipid fractions only with I2 treatment. The administration of non-radiolabeled I2 and 6-iodo-5-hydroxy-8,11,14-eicosatrienoic acid (6-iodolactone, an iodinated arachidonic acid), but not KI, significantly inhibited proliferation of MCF-7 cells. Proliferation of NIH3T3 cells was not inhibited by 20 μM I2. In conclusion, these results demonstrate that I2 uptake does not depend on NIS or PDS; they suggest that in mammary cancer cells, I2 is taken up by a facilitated diffusion system and then covalently bound to lipids or proteins that, in turn, inhibit proliferation.
Endocrine therapy resistance can be associated with high estrogen receptor α (ERα) expression and reduced ERα phosphorylation in breast cancer models
Authors: Kuske Barbara , Naughton Catherine , Moore Kate , MacLeod Kenneth G , Miller William R , Clarke Robert , Langdon Simon P , Cameron David A
Published: December 2006

Hormone-dependent estrogen receptor (ER)-positive breast cancer cells may adapt to low estrogen environments such as produced by aromatase inhibitors. In many instances, cells become insensitive to the effects of estrogen but may still retain dependence on ER. We have investigated the expression, function, and activation of ERα in two endocrine-resistant MCF-7 models to identify mechanisms that could contribute to resistance. While MCF-7/LCC1 cells are partially estrogen dependent, MCF-7/LCC9 cells are fully estrogen insensitive and fulvestrant and tamoxifen resistant. In both MCF-7/LCC1 and MCF-7/LCC9 cell lines, high expression of ERα was associated with enhanced binding to the trefoil factor 1 (TFF1) promoter in the absence of estrogen and increased transcription of TFF1 and progesterone receptor. In contrast to the observations derived from hypersensitive and supersensitive models, these cells were truly estrogen independent; nevertheless, removal of ERα by siRNA, or fulvestrant, a specific ER downregulator, inhibited growth indicating dependence on ERα. In the absence of estrogen, neither ERα Ser118 nor Ser167 were phosphorylated as frequently found in other ligand-independent cell line models. Addition of estrogen activated ERα Ser118 in MCF-7 and LCC1 cells but not in LCC9 cells. We suggest that the estrogen-independent growth within these cell lines is accounted for by high levels of ERα expression driving transcription and full estrogen independence explained by lack of ERα activation through Ser118.