bluuebunny/crossref_metadata_2025 huggingface dataset:
Fatalities in swimming pools with lifeguards: USA, 2000–2008
Authors: Pelletier Andrew R , Gilchrist Julie
Published: January 2011

Background

Pools with lifeguards are often perceived to represent a safe environment for swimming.

Objectives

To examine how often fatalities occurred in pools with lifeguards and what factors were common in these incidents.

Design

Case series.

Setting

USA.

Subjects

A case was defined as a death in a swimming pool with at least one lifeguard that occurred in the USA from 2000 to 2008. Potential cases were identified from media accounts using Lexis-Nexis and ProQuest databases. Supplemental information was obtained through internet based searches and review of death certificates.

Results

A total of 140 deaths from 37 states were identified. Of decedents, 109 (78%) were children and 93 (66%) were males. African Americans accounted for 44% of deaths among those of known race. 104 (74%) fatalities occurred during May through August; 85 (61%) deaths occurred in outdoor pools. Sites for fatal incidents included 53 (38%) municipal pools, 21 (15%) community organisations, and 20 (14%) schools. The median swimmer to lifeguard ratio was 13:1. Decedents were initially identified in the pool twice as often by non-lifeguards (eg, other swimmers or bystanders) as by lifeguards.

Conclusions

Deaths in pools with lifeguards are uncommon, but do occur. Groups most affected include children, males, and African Americans. While lifeguards provide an important layer of protection at swimming pools, they alone cannot prevent all drownings. Additional safety measures are needed.
Smoke-free households with children and decreasing rates of paediatric clinical encounters for otitis media in the United States
Authors: Alpert Hillel R , Behm Ilan , Connolly Gregory N , Kabir Zubair
Published: January 2011

Objective

Temporal trends in paediatric encounters for otitis media (OM) were last characterised and observed to be steadily increasing from 1975 to the mid-1990s. The present study uses an ecological design to quantify trends in paediatric encounters for OM concurrent with a period of decline of an important risk factor, secondhand smoke (SHS) exposure among children.

Patients and methods

Annual paediatric ambulatory visit and hospital discharge rates for children ≤6 years with OM as primary diagnosis were computed with nationally representative data for 1993–2006. Percentages of households with children ≤6 years and no-smoking rules were computed using Tobacco Use Supplement to the Current Population Survey data. Average annual percentage changes were determined for covariate-adjusted rates of paediatric encounters for OM using joinpoint analysis.

Results

While percentages of homes with children and no-smoking rules increased by 89% from 45.5% in 1993 to 86.1% in 2006, average annual covariate-adjusted paediatric encounters for OM decreased by 4.6% (95% CI 4.5% to 4.8%) for ambulatory visits and by 9.8% (95% CI 9.1% to 10.6%) for hospital discharges. Coverage by 7-valent pneumococcal conjugate vaccination (PCV7) increased since 2002, while little variation occurred in other potentially associated risk factors.

Conclusions

Paediatric encounter rates for OM decreased steadily over a 13-year period reversing a previously reported long-term increasing trend reported. Reduced SHS exposure, PCV7 coverage since 2002 and other factors may have contributed to the decline. Further research is needed to provide direct estimates of paediatric encounter rates in exposed and unexposed populations for causal inference.
Risk factors for transmission of foot‐and‐mouth disease during an outbreak in southern England in 2007
Authors: Ellis‐Iversen J. , Smith R. P. , Gibbens J. C. , Sharpe C. E. , Dominguez M. , Cook A. J. C.
Published: January 2011

During an outbreak of foot‐and‐mouth disease (FMD) in southern England in 2007, a case‐control study was conducted to identify risk factors for infection and to investigate the relative impact of risk factors on transmission between the infected farms. Seven of the eight case farms in the outbreak and 22 control farms participated. Data were collected via questionnaires and subjected to comparative statistical analysis. Case farms were further classified as primary or secondary according to the likely source of infection during the study. On primary case farms, it was plausible that infection had been introduced directly from the original source. On secondary case farms, FMD infection was more likely to have originated from another infected premises. Calving occurred more frequently on case farms than on control farms during the risk period, and the two primary case farms had a larger proportion of youngstock than the other farms. Secondary case farms (n=5) had a higher composite environmental risk score and a lower biosecurity score than control farms.
Hypoxia. 3. Hypoxia and neurotransmitter synthesis
Authors: Kumar Ganesh K.
Published: January 2011

Central and peripheral neurons as well as neuroendocrine cells express a variety of neurotransmitters/modulators that play critical roles in regulation of physiological systems. The synthesis of several neurotransmitters/modulators is regulated by O2-requiring rate-limiting enzymes. Consequently, hypoxia resulting from perturbations in O2 homeostasis can affect neuronal functions by altering neurotransmitter synthesis. Two broad categories of hypoxia are frequently encountered: continuous hypoxia (CH) and intermittent hypoxia (IH). CH is often seen during high altitude sojourns, whereas IH is experienced in sleep-disordered breathing with recurrent apneas (i.e., brief, repetitive cessations of breathing). This article presents what is currently known on the effects of both forms of hypoxia on neurotransmitter levels and neurotransmitter synthesizing enzymes in the central and peripheral nervous systems.
Using changes in hydrostatic and osmotic pressure to manipulate metabolic function in chondrocytes
Authors: Mizuno Shuichi , Ogawa Rei
Published: January 2011

Articular cartilage has distinct histological depth zones. In each zone, chondrocytes are subject to different hydrostatic (HP) and osmotic pressure (OP) due to weight-bearing and joint-loading. Previous in vitro studies of regeneration and pathophysiology in cartilage have failed to consider the characteristics of histological heterogeneity and the effects of combinations of changes in HP and OP. Thus, we have constructed molecular, biochemical, and histological profiles of anabolic and catabolic molecules produced by chondrocytes from each depth zone isolated from bovine articular cartilage in response to changes in HP and OP. We cultured the chondrocytes with combinations of loading or off-loading of HP at 0–0.5 MPa, 0.5 Hz, and changes in OP of 300–450 mosM over 1 wk, and evaluated mRNA expression and immunohistology of both anabolic and catabolic molecules and amounts of accumulated sulfated glycosaminoglycan. Any changes in HP and OP upregulated mRNA of anabolic and catabolic molecules in surface-, middle-, and deep-zone cells, in descending order of magnitude. Off-loading HP maintained the anabolic and reduced the catabolic mRNA; high OP retained upregulation of catabolic mRNA. These molecular profiles were consistent with immunohistological and biochemical findings. Changes in HP and OP are essential for simulating chondrocyte physiology and useful for manipulating phenotypes.
Neuropeptide Y inhibits cholangiocarcinoma cell growth and invasion
Authors: DeMorrow Sharon , Onori Paolo , Venter Julie , Invernizzi Pietro , Frampton Gabriel , White Mellanie , Franchitto Antonio , Kopriva Shelley , Bernuzzi Francesca , Francis Heather , Coufal Monique , Glaser Shannon , Fava Giammarco , Meng Fanyin , Alvaro Domenico , Carpino Guido , Gaudio Eugenio , Alpini Gianfranco
Published: January 2011

No information exists on the role of neuropeptide Y (NPY) in cholangiocarcinoma growth. Therefore, we evaluated the expression and secretion of NPY and its subsequent effects on cholangiocarcinoma growth and invasion. Cholangiocarcinoma cell lines and nonmalignant cholangiocytes were used to assess NPY mRNA expression and protein secretion. NPY expression was assessed by immunohistochemistry in human liver biopsies. Cell proliferation and migration were evaluated in vitro by MTS assays and matrigel invasion chambers, respectively, after treatment with NPY or a neutralizing NPY antibody. The effect of NPY or NPY depletion on tumor growth was assessed in vivo after treatment with NPY or the neutralizing NPY antibody in a xenograft model of cholangiocarcinoma. NPY secretion was upregulated in cholangiocarcinoma compared with normal cholangiocytes. Administration of exogenous NPY decreased proliferation and cell invasion in all cholangiocarcinoma cell lines studied and reduced tumor cell growth in vivo. In vitro, the effects of NPY on proliferation were blocked by specific inhibitors for NPY receptor Y2, but not Y1 or Y5, and were associated with an increase in intracellular d- myo-inositol 1,4,5-trisphosphate and PKCα activation. Blocking of NPY activity using a neutralizing antibody promoted cholangiocarcinoma growth in vitro and in vivo and increased the invasiveness of cholangiocarcinoma in vitro. Increased NPY immunoreactivity in human tumor tissue occurred predominantly in the center of the tumor, with less expression toward the invasion front of the tumor. We demonstrated that NPY expression is upregulated in cholangiocarcinoma, which exerts local control on tumor cell proliferation and invasion. Modulation of NPY secretion may be important for the management of cholangiocarcinoma.
4F2hc stabilizes GLUT1 protein and increases glucose transport activity
Authors: Ohno Haruya , Nakatsu Yusuke , Sakoda Hideyuki , Kushiyama Akifumi , Ono Hiraku , Fujishiro Midori , Otani Yuichiro , Okubo Hirofumi , Yoneda Masayasu , Fukushima Toshiaki , Tsuchiya Yoshihiro , Kamata Hideaki , Nishimura Fusanori , Kurihara Hiroki , Katagiri Hideki , Oka Yoshitomo , Asano Tomoichiro
Published: January 2011

Glucose transporter 1 (GLUT1) is widely distributed throughout various tissues and contributes to insulin-independent basal glucose uptake. Using a split-ubiquitin membrane yeast two-hybrid system, we newly identified 4F2 heavy chain (4F2hc) as a membrane protein interacting with GLUT1. Though 4F2hc reportedly forms heterodimeric complexes between amino acid transporters, such as LAT1 and LAT2, and regulates amino acid uptake, we investigated the effects of 4F2hc on GLUT1 expression and the associated glucose uptake. First, FLAG-tagged 4F2hc and hemagglutinin-tagged GLUT1 were overexpressed in human embryonic kidney 293 cells and their association was confirmed by coimmunoprecipitation. The green fluorescent protein-tagged 4F2hc and DsRed-tagged GLUT1 showed significant, but incomplete, colocalization at the plasma membrane. In addition, an endogenous association between GLUT1 and 4F2hc was demonstrated using mouse brain tissue and HeLa cells. Interestingly, overexpression of 4F2hc increased the amount of GLUT1 protein in HeLa and HepG2 cells with increased glucose uptake. In contrast, small interfering RNA (siRNA)-mediated 4F2hc gene suppression markedly reduced GLUT1 protein in both cell types, with reduced glucose uptake. While GLUT1 mRNA levels were not affected by overexpression or gene silencing of 4F2hc, GLUT1 degradation after the addition of cycloheximide was significantly suppressed by 4F2hc overexpression and increased by 4F2hc siRNA treatment. Taken together, these observations indicate that 4F2hc is likely to be involved in GLUT1 stabilization and to contribute to the regulation of not only amino acid but also glucose metabolism.
Interfacial sensing by alveolar type II cells: a new concept in lung physiology?
Authors: Ravasio Andrea , Hobi Nina , Bertocchi Cristina , Jesacher Alexander , Dietl Paul , Haller Thomas
Published: January 2011

Alveolar type II (AT II) cells are in close contact with an air-liquid interface (IAL). This contact may be of considerable physiological relevance; however, no data exist to provide a satisfying description of this specific microenvironment. This is mainly due to the experimental difficulty to manipulate and analyze cell-air contacts in a specific way. Therefore, we designed assays to quantify cell viability, Ca2+ changes, and exocytosis in the course of interface contact and miniaturized IAL devices for direct, subcellular, and real-time analyses of cell-interface interactions by fluorescence microscopy or interferometry. The studies demonstrated that the sole presence of an IAL is not sensed by the cells. However, when AT II cells are forced into closer contact with it, they respond promptly with sustained Ca2+ signals and surfactant exocytosis before the occurrence of irreversible cell damage. This points to a paradoxical situation: a potential threat and potent stimulus for the cells. Furthermore, we found that the signalling mechanism underlying sensation of an IAL can be sufficiently explained by mechanical forces. These results demonstrate that the IAL itself can play a major, although so-far neglected, role in lung physiology, particularly in the regulatory mechanisms related with surfactant homeostasis. Moreover, they also support a general new concept of mechanosensation in the lung.
Parabrachial and hypothalamic interaction in sodium appetite
Authors: Dayawansa S. , Peckins S. , Ruch S. , Norgren R.
Published: January 2011

Rats with bilateral lesions of the lateral hypothalamus (LH) fail to exhibit sodium appetite. Lesions of the parabrachial nuclei (PBN) also block salt appetite. The PBN projection to the LH is largely ipsilateral. If these deficits are functionally dependent, damaging the PBN on one side and the LH on the other should also block Na appetite. First, bilateral ibotenic acid lesions of the LH were needed because the electrolytic damage used previously destroyed both cells and axons. The ibotenic LH lesions produced substantial weight loss and eliminated Na appetite. Controls with ipsilateral PBN and LH lesions gained weight and displayed robust sodium appetite. The rats with asymmetric PBN-LH lesions also gained weight, but after sodium depletion consistently failed to increase intake of 0.5 M NaCl. These results dissociate loss of sodium appetite from the classic weight loss after LH damage and prove that Na appetite requires communication between neurons in the LH and the PBN.
Aldosterone stimulates superoxide production in macula densa cells
Authors: Zhu Xiaolong , Manning R. Davis , Lu Deyin , Gomez-Sanchez Celso E. , Fu Yiling , Juncos Luis A. , Liu Ruisheng
Published: January 2011

Two major factors which regulate tubuloglomerular feedback (TGF)-mediated constriction of the afferent arteriole are release of superoxide (O2−) and nitric oxide (NO) by macula densa (MD) cells. MD O2− inactivates NO; however, among the factors that increase MD O2− release, the role of aldosterone is unclear. We hypothesize that aldosterone activates the mineralocorticoid receptor (MR) on MD cells, resulting in increased O2− production due to upregulation of cyclooxygenase-1 (COX-2) and NOX-2, and NOX-4, isoforms of NAD(P)H oxidase. Studies were performed on MMDD1 cells, a renal epithelial cell line with properties of MD cells. RT-PCR and Western blotting confirmed the expression of MR. Aldosterone (10−8 mol/l for 30 min) doubled MMDD1 cell O2− production, and this was completely blocked by MR inhibition with 10−5 mol/l eplerenone. RT-PCR, real-time PCR, and Western blotting demonstrated aldosterone-induced increases in COX-2, NOX-2, and NOX-4 expression. Inhibition of COX-2 (NS398), NADPH oxidase (apocynin), or a combination blocked aldosterone-induced O2− production to the same degree. These data suggest that aldosterone-stimulated MD O2− production is mediated by COX-2 and NADPH oxidase. Next, COX-2 small-interfering RNA (siRNA) specifically decreased COX-2 mRNA without affecting NOX-2 or NOX-4 mRNAs. In the presence of the COX-2 siRNA, the aldosterone-induced increases in COX-2, NOX-2, and NOX-4 mRNAs and O2− production were completely blocked, suggesting that COX-2 causes increased expression of NOX-2 and NOX-4. In conclusion 1) MD cells express MR; 2) aldosterone increases O2− production by activating MR; and 3) aldosterone stimulates COX-2, which further activates NOX-2 and NOX-4 and generates O2−. The resulting balance between O2− and NO in the MD is important in modulating TGF.
Recent advances in distal tubular potassium handling
Authors: Rodan Aylin R. , Cheng Chih-Jen , Huang Chou-Long
Published: January 2011

It is well known that sodium reabsorption and aldosterone play important roles in potassium secretion by the aldosterone-sensitive distal nephron. Sodium- and aldosterone-independent mechanisms also exist. This review focuses on some recent studies that provide novel insights into the sodium- and aldosterone-independent potassium secretion by the aldosterone-sensitive distal nephron. In addition, we discuss a study reporting on the regulation of the mammalian potassium kidney channel ROMK by intracellular and extracellular magnesium, which may be important in the pathogenesis of persistent hypokalemia in patients with concomitant potassium and magnesium deficiency. We also discuss outstanding questions and propose working models for future investigation.
Physiological changes underlying bilateral isometric arm voluntary contractions in healthy humans
Authors: Soteropoulos Demetris S. , Perez Monica A.
Published: January 2011

Many bilateral motor tasks engage simultaneous activation of distal and proximal arm muscles, but little is known about their physiological interactions. Here, we used transcranial magnetic stimulation to examine motor-evoked potentials (MEPs), interhemispheric inhibition at a conditioning-test interval of 10 (IHI10) and 40 ms (IHI40), and short-interval intracortical inhibition (SICI) in the left first dorsal interosseous (FDI) muscle during isometric index finger abduction. The right side remained at rest or performed isometric voluntary contraction with the FDI, biceps or triceps brachii, or the tibialis anterior. Left FDI MEPs were suppressed to a similar extent during contraction of the right FDI and biceps and triceps brachii but remained unchanged during contraction of the right tibialis anterior. IHI10 and IHI40 were decreased during contraction of the right biceps and triceps brachii compared with contraction of the right FDI. SICI was increased during activation of the right biceps and triceps brachii and decreased during activation of the right FDI. The present results indicate that an isometric voluntary contraction with either a distal or a proximal arm muscle, but not a foot dorsiflexor, decreases corticospinal output in a contralateral active finger muscle. Transcallosal inhibitory effects were strong during bilateral activation of distal hand muscles and weak during simultaneous activation of a distal and a proximal arm muscle, whereas GABAergic intracortical activity was modulated in the opposite manner. These findings suggest that in intact humans crossed interactions at the level of the motor cortex involved different physiological mechanisms when bilateral distal hand muscles are active and when a distal and a proximal arm muscle are simultaneously active.
A tingling sanshool derivative excites primary sensory neurons and elicits nocifensive behavior in rats
Authors: Klein Amanda H. , Sawyer Carolyn M. , Zanotto Karen L. , Ivanov Margaret A. , Cheung Susan , Carstens Mirela Iodi , Furrer Stephan , Simons Christopher T. , Slack Jay P. , Carstens E.
Published: January 2011

Szechuan peppers contain hydroxy-α-sanshool that imparts desirable tingling, cooling, and numbing sensations. Hydroxy-α-sanshool activates a subset of sensory dorsal root ganglion (DRG) neurons by inhibiting two-pore potassium channels. We presently investigated if a tingle-evoking sanshool analog, isobutylalkenyl amide (IBA), excites rat DRG neurons and, if so, if these neurons are also activated by agonists of TRPM8, TRPA1, and/or TRPV1. Thirty-four percent of DRG neurons tested responded to IBA, with 29% of them also responding to menthol, 29% to cinnamic aldehyde, 66% to capsaicin, and subsets responding to two or more transient receptor potential (TRP) agonists. IBA-responsive cells had similar size distributions regardless of whether they responded to capsaicin or not; cells only responsive to IBA were larger. Responses to repeated application of IBA at a 5-min interstimulus interval exhibited self-desensitization (tachyphylaxis). Capsaicin did not cross-desensitize responses to IBA to any greater extent than the tachyphylaxis observed with repeated IBA applications. These findings are consistent with psychophysical observations that IBA elicits tingle sensation accompanied by pungency and cooling, with self-desensitization but little cross-desensitization by capsaicin. Intraplantar injection of IBA elicited nocifensive responses (paw licking, shaking-flinching, and guarding) in a dose-related manner similar to the effects of intraplantar capsaicin and serotonin. IBA had no effect on thermal sensitivity but enhanced mechanical sensitivity at the highest dose tested. These observations suggest that IBA elicits an unfamiliar aversive sensation that is expressed behaviorally by the limited response repertoire available to the animal.
Influence of Thermodynamic Effect on Blade Load in a Cavitating Inducer
Authors: Kikuta Kengo , Shimiya Noriyuki , Hashimoto Tomoyuki , Shimagaki Mitsuru , Nanri Hideaki , Yoshida Yoshiki
Published: January 2011

Distribution of the blade load is one of the design parameters for a cavitating inducer. For experimental investigation of the thermodynamic effect on the blade load, we conducted experiments in both cold water and liquid nitrogen. The thermodynamic effect on cavitation notably appears in this cryogenic fluid although it can be disregarded in cold water. In these experiments, the pressure rise along the blade tip was measured. In water, the pressure increased almost linearly from the leading edge to the trailing edge at higher cavitation number. After that, with a decrease of cavitation number, pressure rise occurred only near the trailing edge. On the other hand, in liquid nitrogen, the pressure distribution was similar to that in water at a higher cavitation number, even if the cavitation number as a cavitation parameter decreased. Because the cavitation growth is suppressed by the thermodynamic effect, the distribution of the blade load does not change even at lower cavitation number. By contrast, the pressure distribution in liquid nitrogen has the same tendency as that in water if the cavity length at the blade tip is taken as a cavitation indication. From these results, it was found that the shift of the blade load to the trailing edge depended on the increase of cavity length, and that the distribution of blade load was indicated only by the cavity length independent of the thermodynamic effect.
Evaluating the Effectiveness of a Commercial Portable Air Purifier in Homes with Wood Burning Stoves: A Preliminary Study
Authors: Hart Julie F. , Ward Tony J. , Spear Terry M. , Rossi Richard J. , Holland Nicholas N. , Loushin Brodie G.
Published: January 2011

Wood burning for residential heating is prevalent in the Rocky Mountain regions of the United States. Studies have shown that wood stoves can be a significant source of PM2.5within homes. In this study, the effectiveness of an electrostatic filter portable air purifier was evaluated (1) in a home where a wood stove was the sole heat source and (2) in a home where a wood stove was used as a supplemental heat source. Particle count concentrations in six particle sizes and particle mass concentrations in two particle sizes were measured for ten 12-hour purifier on and ten purifier off trials in each home. Particle count concentrations were reduced by 61–85 percent. Similar reductions were observed in particle mass concentrations. These findings, although limited to one season, suggest that a portable air purifier may effectively reduce indoor particulate matter concentrations associated with wood combustion during home heating.
A de Casteljau Algorithm for q‐Bernstein‐Stancu Polynomials
Authors: Nowak Grzegorz
Published: January 2011

This paper is concerned with a generalization of the q‐Bernstein polynomials and Stancu operators, where the function is evaluated at intervals which are in geometric progression. It is shown that these polynomials can be generated by a de Casteljau algorithm, which is a generalization of that relating to the classical case and q‐Bernstein case.
A Germline Variant in the Interferon Regulatory Factor 4 Gene as a Novel Skin Cancer Risk Locus
Authors: Han Jiali , Qureshi Abrar A. , Nan Hongmei , Zhang Jiangwen , Song Yiqing , Guo Qun , Hunter David J.
Published: January 2011

Genome-wide association studies on pigmentary phenotypes provide a pool of candidate genetic markers for skin cancer risk. The SNPs identified from a genome-wide association study of natural hair color were assessed for associations with the risk of three types of skin cancer simultaneously in a nested case-control study within the Nurses' Health Study [218 melanoma, 285 squamous cell carcinoma (SCC), and 300 basal cell carcinoma (BCC) cases, and 870 common controls]. Along with two known pigmentation loci, MC1R and OCA2, the IRF4 rs12203592 T allele was associated with an increased risk of each type of skin cancer (P value, 6.6 × 10−4 for melanoma, 7.0 × 10−7 for SCC, and 0.04 for BCC). This association was further replicated in additional samples (190 melanoma, 252 SCC, and 634 common controls). The P value in the replication set was 0.03 for melanoma and 4.2 × 10−3 for SCC. The risk of BCC was replicated in an independent set of 213 cases and 718 controls (P value, 0.02). The combined results showed that the association with SCC reached the genome-wide significance level [odds ratio (OR) for additive model = 1.61, 95%CI, 1.36–1.91, P = 3.2 × 10−8]. The OR was 1.49 for melanoma (95%CI, 1.23–1.80; P = 4.5 × 10−5), and 1.32 for BCC (95%CI, 1.11–1.57; P = 1.6×10−3). Given that the T allele was shown previously to be associated with increased expression of IRF4 locus, further studies are warranted to elucidate the role of the IRF4 gene in human pigmentation and skin cancer development. Cancer Res; 71(5); 1533–9. ©2011 AACR.
Paradoxical Relationship between Chromosomal Instability and Survival Outcome in Cancer
Authors: Birkbak Nicolai J. , Eklund Aron C. , Li Qiyuan , McClelland Sarah E. , Endesfelder David , Tan Patrick , Tan Iain B. , Richardson Andrea L. , Szallasi Zoltan , Swanton Charles
Published: January 2011

Chromosomal instability (CIN) is associated with poor prognosis in human cancer. However, in certain animal tumor models elevated CIN negatively impacts upon organism fitness, and is poorly tolerated by cancer cells. To better understand this seemingly contradictory relationship between CIN and cancer cell biological fitness and its relationship with clinical outcome, we applied the CIN70 expression signature, which correlates with DNA-based measures of structural chromosomal complexity and numerical CIN in vivo, to gene expression profiles of 2,125 breast tumors from 13 published cohorts. Tumors with extreme CIN, defined as the highest quartile CIN70 score, were predominantly of the estrogen receptor negative (ER−), basal-like phenotype and displayed the highest chromosomal structural complexity and chromosomal numerical instability. We found that the extreme CIN/ER− tumors were associated with improved prognosis relative to tumors with intermediate CIN70 scores in the third quartile. We also observed this paradoxical relationship between CIN and prognosis in ovarian, gastric, and non–small cell lung cancer, with poorest outcome in tumors with intermediate, rather than extreme, CIN70 scores. These results suggest a nonmonotonic relationship between gene signature expression and HR for survival outcome, which may explain the difficulties encountered in the identification of prognostic expression signatures in ER− breast cancer. Furthermore, the data are consistent with the intolerance of excessive CIN in carcinomas and provide a plausible strategy to define distinct prognostic patient cohorts with ER− breast cancer. Inclusion of a surrogate measurement of CIN may improve cancer risk stratification and future therapeutic approaches. Cancer Res; 71(10); 3447–52. ©2011 AACR.
Oncogenic B-RAF Signaling in Melanoma Impairs the Therapeutic Advantage of Autophagy Inhibition
Authors: Armstrong Jane L. , Corazzari Marco , Martin Shaun , Pagliarini Vittoria , Falasca Laura , Hill David S. , Ellis Nicola , Al Sabah Salim , Redfern Christopher P.F. , Fimia Gian Maria , Piacentini Mauro , Lovat Penny E.
Published: January 2011

Purpose: Metastatic melanoma is characterized by extremely poor survival rates and hence novel therapies are urgently required. The ability of many anticancer drugs to activate autophagy, a lysosomal-mediated catabolic process which usually promotes cell survival, suggests targeting the autophagy pathway may be a novel means to augment therapy.

Experimental Design: Autophagy and apoptosis were assessed in vitro in human melanoma cell lines in response to clinically achievable concentrations of the endoplasmic reticulum (ER) stress-inducing drugs fenretinide or bortezomib, and in vivo using a s.c. xenograft model.

Results: Autophagy was activated in response to fenretinide or bortezomib in B-RAF wild-type cells, shown by increased conversion of LC3 to the autophagic vesicle-associated form (LC3-II) and redistribution to autophagosomes and autolysosomes, increased acidic vesicular organelle formation and autophagic vacuolization. In contrast, autophagy was significantly reduced in B-RAF–mutated melanoma cells, an effect attributed partly to oncogenic B-RAF. Rapamycin treatment was unable to stimulate LC3-II accumulation or redistribution in the presence of mutated B-RAF, indicative of de-regulated mTORC1-dependent autophagy. Knockdown of Beclin-1 or ATG7 sensitized B-RAF wild-type cells to fenretinide- or bortezomib-induced cell death, demonstrating a pro-survival function of autophagy. In addition, autophagy was partially reactivated in B-RAF–mutated cells treated with the BH3 mimetic ABT737 in combination with fenretinide or bortezomib, suggesting autophagy resistance is partly mediated by abrogated Beclin-1 function.

Conclusions: Our findings suggest inhibition of autophagy in combination with ER stress-inducing agents may represent a means by which to harness autophagy for the therapeutic benefit of B-RAF wild-type melanoma. Clin Cancer Res; 17(8); 2216–26. ©2011 AACR.
Increased Vascular Injury Reduces the Degree of Intimal Hyperplasia following Angioplasty in Rabbits
Authors: Fogelstrand Per , Mellander Stefan , Mattsson Erney
Published: January 2011

Background/Aims: Formation of intimal hyperplasia following angioplastic procedures can lead to complications, including restenosis and accelerated atherosclerosis. The vessel wall media is a main source of neointimal cells. However, evidence suggests that there are additional cell sources, such as the adventitia. Here we investigate whether an extensive loss of vascular smooth muscle cells (VSMCs) in the media results in less intimal hyperplasia or if there is compensatory cell recruitment from the adventitia. Methods:A balloon catheter was pulled through the rabbit carotid artery 4 times (major injury) or 2 times (minor injury). Adventitial cells were labeled with 5-bromo-2-deoxyuridine or PKH26. Results:The major injury, but not the minor injury, resulted in a complete loss of VSMCs in large parts of the media and significant leukocyte infiltration. The major injury resulted in less neointima compared with the minor injury. The thinnest neointima was seen at the most injured parts of the media in the major injury group. Cell-tracking experiments showed that the media, but not the adventitia, served as a source of neointimal cells. Conclusion: An augmented angioplastic injury with extensive VSMC loss in rabbits reduced the degree of intimal hyperplasia. No compensatory recruitment of neointimal cells from the adventitia occurred.
Long-Term Ouabain Treatment Impairs Vascular Function in Resistance Arteries
Authors: Wenceslau Camilla Ferreira , Davel Ana Paula , Xavier Fabiano Elias , Rossoni Luciana Venturini
Published: January 2011

Background/Aims: The purpose of this study was to examine the cardiovascular effects of long-term ouabain treatment at different time points. Methods: Systolic blood pressure (SBP) was measured by tail-cuff method in male Wistar rats treated with ouabain (approx. 8.0 µg·day–1) or vehicle for 5, 10 and 20 weeks. Afterwards, vascular function was assessed in mesenteric resistance arteries (MRA) using a wire myograph. ROS production and COX-1 and COX-2, TNF-α, and IL-6 protein expression were investigated. Results: SBP was increased by ouabain treatment up to the 6th week and remained stable until the 20th week. However, noradrenaline-induced contraction increased only in MRA in rats treated with ouabain for 20 weeks. NOS inhibition and endothelium removal increased the noradrenaline response, but to a smaller magnitude in MRA in the ouabain group. Moreover, inhibition of COX-2 or incubation with superoxide dismutase restores noradrenaline-induced contraction in the 20-week ouabain group to control levels. ROS production as well as COX-2, IL-6 and TNF-α protein expression increased in MRA in this group. Conclusion: Although ouabain treatment induced hypertension in all groups, a larger noradrenaline induced contraction was observed over 20 weeks of treatment. This vascular dysfunction was related to COX-2-derived prostanoids and oxidative stress, increased pro- inflammatory cytokines and reduced NO bioavailability.
The GPER1 Agonist G-1 Attenuates Endothelial Cell Proliferation by Inhibiting DNA Synthesis and Accumulating Cells in the S and G2 Phases of the Cell Cycle
Authors: Holm Anders , Baldetorp Bo , Olde Björn , Leeb-Lundberg L.M. Fredrik , Nilsson Bengt-Olof
Published: January 2011

G protein-coupled receptor 30 (GPR30) or G protein-coupled estrogen receptor 1 (GPER1) is expressed in the vasculature, but the importance of vascular GPER1 remains to be clarified. Here we investigate effects of the GPER1 agonist G-1 on endothelial cell proliferation using mouse microvascular endothelial bEnd.3 cells. The bEnd.3 cells express mRNA for GPER1. The bEnd.3 cells expressed both ERα and ERβ immunoreactivities. Treatment with G-1 reduced DNA synthesis and cell number with IC50 values of about 2 µM. GPER1 siRNA prevented G-1-induced attenuation of DNA synthesis. G-1 accumulated cells in S and G2 phases of the cell cycle, suggesting that G-1 blocks transition between G2 and M. G-1 had no effect on DNA synthesis in COS-7 cells only weakly expressing GPER1 mRNA. 17β-Estradiol had no effect on DNA synthesis in physiological concentrations (nM). The ER blocker ICI182780 reduced DNA synthesis with similar potency as G-1. Treatment with the ERK/MAP kinase inhibitor PD98059 had no effect on G-1-induced attenuation of DNA synthesis. G-1- induced antiproliferation was observed not only in bEnd.3 cells but also in human umbilical vein endothelial cells and HMEC-1 endothelial cells. We conclude that the GPER1 agonist G-1 attenuates endothelial cell proliferation via inhibition of DNA synthesis and by accumulation of cells in S and G2.
The Prevalence of Long Spinal Cord Lesions and Anti-Aquaporin 4 Antibodies in Neuromyelitis Optica Patients in Taiwan
Authors: Wang Kai-Chen , Tsai Ching-Piao , Lee Chao-Lin , Chen Shao-Yuan , Chen Shyi-Jou
Published: January 2011

Background and Objective: It was the aim of this study to determine the prevalence of anti-aquaporin 4 antibody (anti-AQP4 Ab) and long spinal cord lesions in neuromyelitis optica (NMO) and multiple sclerosis (MS) patients in Taiwan. Asia has a relatively high rate of NMO compared with MS patients. Anti-AQP4 Ab is an important marker for NMO worldwide, but serological data and clinical profiles of NMO patients in Taiwan have not been reported. Methods: This retrospective study compared the clinical symptoms, demographics, spinal cord lesion length and AQP4 Ab status of 34 patients with NMO with 34 patients diagnosed with conventional MS. Results: Our NMO patients were predominantly middle-aged women (median age 45 years), exhibited many relapses (1.0/year) and displayed a higher Expanded Disability Status Scale score (4.75) than conventional MS patients. NMO patients exhibited long spinal cord lesions as detected by MRI. Forty-one percent of the NMO patients had detectable anti-AQP4 Ab. The Expanded Disability Status Scale score was significantly higher in AQP4 Ab– NMO patients. Conclusion: The prevalence of AQP4 Ab in a Taiwanese NMO group was 41%. Long spinal cord lesions and detection of AQP4 Ab helped to differentiate NMO patients from MS patients. Long spinal cord lesions with the anti-AQP4 Ab test may allow for an earlier diagnosis of NMO and improve therapeutic decisions.
A Case of Well-Tolerated and Safe Deferasirox Administration during the First Trimester of a Spontaneous Pregnancy in an Advanced Maternal Age Thalassemic Patient
Authors: Ricchi Paolo , Costantini Silvia , Spasiano Anna , Di Matola Tiziana , Cinque Patrizia , Prossomariti Luciano
Published: January 2011

In this report, we present a case of a spontaneous pregnancy in a 42-year-old thalassemic woman referred to our Microcythemic Unit while she was on deferasirox (DFX) therapy. The patient had been on treatment with DFX since March 2008 and in March 2009 realized that she was pregnant at 12 weeks of gestation. The conception was spontaneous and the baby was born at full term without complications or malformations.
Ear and Further Anatomic Anomalies in Children Undergoing Stapedotomy
Authors: Kontorinis Georgios , Goetz Friedrich , Lenarz Thomas , Schwab Burkard
Published: January 2011

Purpose: Aim of this study was to examine any stapes fixation-related clinical, intraoperative and high-resolution computed tomography (HRCT) findings in children undergoing stapedotomy. Procedures: Detailed diagnostic assessment was performed in 12 children (18 ears) who underwent stapedotomy within the period 2005–2008. Results: Ear malformations such as malleus-incus anomalies, malleus-epitympanum fixation and absence of the stapedius tendon, small external auditory canal and auricle malformations were clinically and/or intraoperatively diagnosed. HRCT identified a petrous high jugular bulb and malformed inner auditory canal in 4 ears; however, it failed to recognize ossicular fixation. Systemic anomalies, namely kyphoscoliosis, esophageal atresia and finger malformations were identified in 2 children. Summing up, additional anatomic anomalies were found in 8 patients (66.7%). Conclusions: Ear malformations may coexist in children with stapes fixation. Ossicular fixation is not easily recognized with HRCT, and therefore, ossicular mobility should always be tested intraoperatively. In a few cases, pediatric stapes fixation can coexist with systemic dysplasias; in such patients, additional referral to geneticists is recommended.
The GH/IGF-I Axis and Pituitary Function and Size in Adults with Prader-Willi Syndrome
Authors: van Nieuwpoort I.C. , Sinnema M. , Castelijns J.A. , Twisk J.W.R. , Curfs L.M.G. , Drent M.L.
Published: January 2011

Background: In adults with Prader-Willi syndrome (PWS), limited information is available about pituitary function, more specifically the prevalence of growth hormone deficiency (GHD). The aim of this study was to gain more insight into endocrine function in PWS adults, with emphasis on GH secretion. Methods: 15 randomly selected adult PWS individuals were included and 14 healthy brothers and sisters served as a control group. Main outcome measures were IGF-I, IGFBP-3 and peak GH level after a combined GHRH-arginine test. Other pituitary hormone deficits are diagnosed based on serum levels of the concerning hormones. The size of the pituitary gland was measured on MRI images. Results: In PWS adults, IGF-I levels were low and IGFBP-3 levels normal when compared to healthy controls. GHD was diagnosed in 8–38% of the PWS patients, depending on the criteria used. Hypogonadism was present in 87% of the patients. Hypothyroidism and adrenal insufficiency could also be demonstrated. Anterior pituitary size was lower in PWS individuals when compared to healthy controls. Conclusion: In this study, pituitary hormone deficiencies are demonstrated in a considerable number of adults with PWS, hypogonadism and GHD being most prominent. Furthermore, the anterior pituitary is smaller in comparison with healthy controls.
Comparison of Combination Therapy of Olmesartan plus Azelnidipine or Hydrochlorothiazide on Renal and Vascular Damage in SHR/NDmcr-cp Rats
Authors: Nagasu Hajime , Satoh Minoru , Yorimitsu Daisuke , Tomita Naruya , Sasaki Tamaki , Kashihara Naoki
Published: January 2011

Background: Although the recommended target blood pressure for patients with chronic kidney disease is <130/80 mm Hg, this is difficult to achieve by treatment with an angiotensin receptor blocker alone. Addition of either a calcium channel blocker or a diuretic is suggested as second-line medication; however, which combination is most beneficial for target-organ protection remains unknown. Methods: SHR/NDmcr-cp rats were administered no medications (control) or low-dose olmesartan for 2 weeks and then either olmesartan at an increased dose, azelnidipine, or the hydrochlorothiazide for 3 weeks. We assessed oxidative stress in the kidney and aorta, and endothelial function. Results: Urinary protein excretion was lower in all treated rats than in control rats. Oxidative stress caused by activation of NAD(P)H oxidase was observed in the glomeruli and aorta of control rats and was significantly suppressed in the olmesartan/azelnidipine (Olm/Azl) groups. Combination therapy with olmesartan and hydrochlorothiazide (Olm/HCTZ) however failed to suppress oxidative stress. The Olm/Azl groups maintained the endothelial surface layer in the glomeruli and protected endothelial function in the aorta. Conclusion: In an animal model of metabolic syndrome, a combination of Olm/Azl is superior to a combination of Olm/HCTZ in terms of prevention of glomerular and vascular injuries.
Linkage Analysis of Autopsy-Confirmed Familial Alzheimer Disease Supports an Alzheimer Disease Locus in 8q24
Authors: Sillén Anna , Brohede Jesper , Forsell Charlotte , Lilius Lena , Andrade Jorge , Odeberg Jacob , Kimura Toru , Winblad Bengt , Graff Caroline
Published: January 2011

Background/Aims: We have previously reported the results of an extended genome-wide scan of Swedish Alzheimer disease (AD)-affected families; in this paper, we analyzed a subset of these families with autopsy-confirmed AD. Methods: We report the fine-mapping, using both microsatellite markers and single-nucleotide polymorphisms (SNPs), in the observed maximum logarithm of the odds (LOD)-2 unit (LODmax-2) region under the identified linkage peak, linkage analysis of the fine-mapping data with additionally analyzed pedigrees, and association analysis of SNPs selected from candidate genes in the linked interval. The subset was made on the criterion of at least one autopsy-confirmed AD case per family, resulting in 24 families. Results: Linkage analysis of a family subset having at least one autopsy-confirmed AD case showed a significant nonparametric single-point LOD score of 4.4 in 8q24. Fine-mapping under the linkage peak with 10 microsatellite markers yielded an increase in the multipoint (mpt) LOD score from 2.1 to 3.0. SNP genotyping was performed on 21 selected candidate transcripts of the LODmax-2 region. Both family-based association and linkage analysis were performed on extended material from 30 families, resulting in a suggestive linkage at peak marker rs6577853 (mpt LOD score = 2.4). Conclusion: The 8q24 region has been implicated to be involved in AD etiology.
Photodynamic Therapy Combined with Intravitreal Injection of Vascular Endothelial Growth Factor Antibody for Polypoidal Choroidal Vasculopathy
Authors: Moon Sung Woon , Kim Moo Sang , Kim Eung Suk , Yu Seung-Young , Kwak Hyung-Woo
Published: January 2011

Background/Aims: To evaluate the efficacy of photodynamic therapy (PDT) combined with intravitreal injection of anti-vascular-endothelial-growth-factor (anti-VEGF) antibody in patients with polypoidal choroidal vasculopathy (PCV). Methods: Twenty-two eyes of 22 patients with PCV followed for 12 months after combination therapy with PDT and anti-VEGF were retrospectively reviewed. Patients received intravitreal anti-VEGF (1.25 mg bevacizumab or 0.5 mg ranibizumab) within 7 days after PDT. Retreatment with PDT and intravitreal anti-VEGF injections, or with intravitreal anti-VEGF alone, was performed when indicated. The main outcome measures were best-corrected visual acuity (BCVA) and central foveal thickness (CFT). Results: Mean logMAR BCVA was 0.43 at baseline and 0.45, 0.36, 0.30 and 0.28 at 1, 3, 6 and 12 months, respectively, after the initial combination therapy. Mean BCVA was significantly improved at 6 and 12 months after treatment (p < 0.05). Mean CFT was 269.4 µm at baseline and 180.1, 136.7, 127.5 and 139.6 µm at 1, 3, 6 and 12 months, respectively, after the initial combination therapy. CFT decreased significantly throughout the follow-up period. At 12 months, mean BCVA improved by 1.5 lines, and mean CFT decreased by 129.8 µm. Polypoidal lesions disappeared in 7 of the 13 eyes in which indocyanine green angiography was performed at 12 months. No changes in the branching vascular network were observed in any of these 13 eyes. Patients were treated with PDT a mean of 1.3 times and injected with intravitreal anti-VEGF a mean of 3.4 times over the 12-month period. Conclusion: Combined PDT and intravitreal anti-VEGF may improve visual acuity and decrease CFT at 12 months. Large long-term prospective studies are needed to evaluate the efficacy and safety of combination therapy.
Renal Function in Patients Treated with Cinacalcet for Persistent Hyperparathyroidism after Kidney Transplantation
Authors: Henschkowski Jana , Bischoff-Ferrari Heike A. , Wüthrich Rudolf P. , Serra Andreas L.
Published: January 2011

Background and Aim: Cinacalcet effectively reduces calcium in patients with persistent hyperparathyroidism (HPT) after kidney transplantation. We aimed to assess the association of cinacalcet with a decrease in renal function based on a meta-analysis of observational studies in kidney transplant patients with persistent HPT. Method: Meta-analysis of observational studies, no randomized controlled studies were available. We calculated the mean difference between renal function before cinacalcet and at 3 months on cinacalcet treatment for each study. Pooled analyses are based on random effects models. Results: Pooling the studies on kidney transplant patients with persistent HPT (8 studies, n = 115), we found a significant reduction in renal function (p = 0.008). The effect size was 5 µmol/l (p < 0.0001) when pooling the 7 studies where serum creatinine levels were reported. Meta-regression analysis revealed that there was an association between renal function and the amount of calcium reduction under treatment with cinacalcet. A higher delta change in serum calcium levels was associated with a decrease in renal function at 3 months of cinacalcet treatment. Conclusion: Cinacalcet treatment was associated with a decline of renal function in kidney transplant recipients with persistent HPT. Our meta-analysis underscores the need for frequent monitoring of creatinine and calcium levels during cinacalcet treatment.