bluuebunny/crossref_metadata_2025 huggingface dataset:
Coagulopathy of the critically ill equine patient
Authors: Dallap Schaer Barbara L. , Epstein Kira
Published: February 2009

Objective –To review the hemostasis literature relevant to development of coagulopathy in the critically ill equine patient.

Data Source –Original scientific and review articles.

Human Data Synthesis –Inflammation plays a critical role in the activation and amplification of clot formation, as well as the impairment of physiologic anticoagulant mechanisms, and fibrinolysis. Earlier identification of coagulopathy in patients at risk and restoration of physiologic hemostasis may result in better outcome. Development of scoring systems based on information other than coagulation markers alone may better identify patients with subclinical coagulopathy.

Veterinary Data Synthesis –Critically ill equine patients commonly at risk for coagulopathy include those with severe gastrointestinal disease, septic foals, and adults subjected to severe systemic inflammatory response syndrome. Publications provide information regarding coagulation markers helpful for identification of hemostatic dysfunction in specific patient populations, as well as information regarding the influence of coagulopathy on outcome. Data regarding clinically relevant information on therapeutic intervention are lacking.

Conclusions –The relationship between inflammation and endotoxemia and development of coagulopathy is better understood in both human patients and the critically ill equine patient. Prospective clinical trials evaluating clinically relevant and financially feasible approaches to treatment are still needed.
Anti‐apoptotic effects of protein kinase C‐δ and c‐fos in cisplatin‐treated thyroid cells
Authors: Muscella Antonella , Urso Loredana , Calabriso Nadia , Vetrugno Carla , Rochira Alessio , Storelli Carlo , Marsigliante Santo
Published: February 2009

Background and purpose:  We showed previously that cisplatin inititates a signalling pathway mediated by PKC‐δ/extracellular signal‐regulated kinase (ERK), important for maintaining viability in PC Cl3 thyroid cells. The studies described herein examined whether c‐fos was associated with cisplatin resistance and the signalling link between c‐fos and PKC‐δ/ERK.

Experimental approach:  Cells were treated with various pharmacological inhibitors of PKCs and ERK, or were depleted of c‐fos, PKC‐δ, PKC‐ε and caspase‐3 by small interfering RNA (siRNA), then incubated with cisplatin and cytotoxicity assessed.

Key results:  Cisplatin provokes the induction of c‐fos and the activation of conventional PKC‐β, and novel PKC‐δ and ‐ε. The cisplatin‐provoked c‐fos induction was decreased by Gö6976, a PKC‐β inhibitor; by siRNA for PKC‐δ‐ but not that for PKC‐ε or by PD98059, a mitogen‐activated protein kinase/ERK kinase inhibitor. Expression of c‐fos was abolished by GF109203X, an inhibitor of all PKC isoforms, or by PD98059 plus Gö6976 or by PKC‐δ‐siRNA plus Gö6976. When c‐fos expression was blocked by siRNA, cisplatin cytotoxicity was strongly enhanced with increased caspase‐3 activation. In PKC‐δ‐depleted cells treated with cisplatin, caspase‐3 activation was increased and cell viability decreased. In these PKC‐δ‐depleted cells, PD98059 did not affect caspase‐3 activation.

Conclusions and implications:  In PC Cl3 cells, in the cell signalling pathways that lead to cisplatin resistance, PKC‐δ controls ERK activity and, together with PKC‐β, also the induction of c‐fos. Hence, the protective role of c‐fos in thyroid cells has the potential to provide new opportunities for therapeutic intervention.
Interaction between udenafil and tamsulosin in rats: non‐competitive inhibition of tamsulosin metabolism by udenafil via hepatic CYP3A1/2
Authors: Kang HE , Bae SK , Yoo M , Lee DC , Kim YG , Lee MG
Published: February 2009

Background and purpose: Orthostatic hypotension has been observed when PDE 5 (cGMP‐specific phosphodiesterase type 5) inhibitors are co‐administered with α‐adrenoceptor antagonists. Here we assessed the pharmacokinetic and haemodynamic interactions between udenafil and tamsulosin in rats, as both drugs are metabolized via rat hepatic cytochrome P450 3A1/2.

Experimental approach: Interactions between the two drugs were evaluated in rats after simultaneous 1 or 15 min i.v. infusion or after p.o. administration of udenafil (30 mg·kg−1) and/or tamsulosin (1 mg·kg−1).In vitrometabolism of tamsulosin with udenafil was measured to obtain the inhibition constant (Ki) and [I]/Kiratio of udenafil.

Key results: The total area under the plasma concentration–time curve from time zero to time infinity (AUC)s (or AUC0–4h) of tamsulosin were significantly greater after 15 min of i.v. infusion or after oral administration with udenafil, compared with tamsulosin alone. The hepatic first‐pass metabolism of tamsulosin was inhibited by udenafil, and the inhibitionin vitrowas in a non‐competitive mode. The arterial systolic blood pressure was significantly lower at 5, 10 and 60 min after oral co‐administration of the drugs.

Conclusions and implications: The significantly greater AUC of tamsulosin after i.v. and p.o. administration of both drugs may be attributable to non‐competitive inhibition of cytochrome P450 3A1/2‐mediated hepatic tamsulosin metabolism by udenafil. The inhibition was also observed in human liver S9 fractions, suggesting that a reassessment of the oral dosage of tamsulosin is necessary when udenafil and tamsulosin are co‐administered to patients with benign prostatic hyperplasia.
Blood use in the ambulatory setting among elderly in the United States
Authors: Menis Mikhail , Burwen Dale R. , Holness Leslie , Anderson Steven A.
Published: February 2009

BACKGROUND: Our study characterizes blood use in the ambulatory setting by US elderly Medicare beneficiaries during 2001. As the US population ages and delivery of health care in outpatient settings is on the rise, ambulatory blood utilization is expected to increase. There is currently a lack of broad population‐based studies detailing ambulatory blood utilization patterns among the US elderly.

STUDY DESIGN AND METHODS: This descriptive cross‐sectional study of ambulatory blood utilization in institutional outpatient settings used Medicare administrative data (5% sample of enrollees) for calendar year 2001. Blood use was identified by either the presence of recorded blood units or the procedure code(s) for transfusion of whole blood or red blood cells.

RESULTS: Among 1,368,368 elderly Medicare beneficiaries analyzed, 7054 (0.52%) had blood transfusion in institutional outpatient settings, and 34,186 (2.50%) had blood transfusion in the inpatient setting. Of 10,705 institutional outpatient claims with blood use quantified in this cohort of elderly Medicare beneficiaries, the top 10 principal diagnoses using the largest quantities of blood accounted for 66.3% of total blood units transfused. Nine of these 10 principal diagnoses were either for anemias (51%) or neoplasms (13%) and accounted for 64% of total blood units transfused.

CONCLUSION: Our study suggests that most of the ambulatory blood utilization among US elderly is for diagnoses of anemias and neoplasms rather than procedures. Our population‐based study provides valuable information on ambulatory blood utilization patterns which may be used to better understand the reasons for transfusion in the ambulatory setting as blood use is expected to grow.
Heart Rate Turbulence as a Predictor of Cardiac Mortality and Arrhythmic Events in Patients with Dilated Cardiomyopathy: A Prospective Study
Authors: MIWA YOSUKE , IKEDA TAKANORI , SAKAKI KATSURA , MIYAKOSHI MUTSUMI , ISHIGURO HARUHISA , TSUKADA TAKEHIRO , ABE ATSUKO , MERA HISAAKI , YUSU SATORU , YOSHINO HIDEAKI
Published: February 2009

Background: Few studies have described the clinical usefulness of heart rate turbulence (HRT), an autonomic predictor of mortality, in stratifying patients with dilated cardiomyopathy (DCM) at risk of cardiac mortality and arrhythmic events. We prospectively assessed the utility of HRT for risk stratification in patients with ischemic or nonischemic DCM.

Methods: We enrolled 375 consecutive patients with DCM including ischemic (n = 241) and nonischemic causes (n = 134). HRT was measured using an algorithm based on routine 24‐hour Holter electrocardiograms, assessing 2 parameters: turbulence onset (TO) and turbulence slope (TS). HRT was considered positive when both TO was ≥0% and TS was ≤ 2.5 ms/R‐R interval. The primary endpoint was defined as cardiac mortality and the secondary endpoint as occurrence of hemodynamically stable sustained ventricular tachyarrhythmias.

Results: Of patients enrolled, 83 patients (22.1%) were not utilized for HRT assessment because there were too few ventricular premature beats, or for other reasons. Eighty‐one of 292 patients (27.7%) were HRT‐positive. During follow‐up of 445 ± 216 days, 30 patients (10.3%) reached the primary endpoint and 17 patients, the secondary endpoint. The hazard ratio (HR) of patients with an HRT‐positive outcome was 6.4 (95%CI, 3.0–14.1; P < 0.0001) for the primary endpoint and 5.1 (95%CI, 2.8–9.3; P < 0.0001) for combined endpoints. On subanalysis, HRT positivity was significantly associated in both the ischemic and nonischemic DCM patients with both the primary endpoint (HR = 4.9, P = 0.0006 and HR = 12.3, P = 0.002, respectively) and with combined endpoints.

Conclusions: HRT is a powerful risk stratification marker for cardiac mortality and arrhythmic events in patients with DCM whether ischemia is present or not.
Influence of Left Ventricular Lead Location on Outcomes in the COMPANION Study
Authors: SAXON LESLIE A. , OLSHANSKY BRIAN , VOLOSIN KENT , STEINBERG JONATHAN S. , LEE BYRON K. , TOMASSONI GERY , GUARNIERI THOMAS , RAO ANUPAMA , YONG PATRICK , GALLE ELIZABETH , LEIGH JILL , ECKLUND FRED , BRISTOW MICHAEL R.
Published: February 2009

Introduction:
There are no randomized controlled trial data that evaluate mortality and hospitalization rates in cardiac resynchronization therapy (CRT) recipients based on left ventricular (LV) lead location. We analyzed the event‐driven outcomes of mortality and hospitalization as well as functional outcomes including Functional Class, Quality‐of‐Life, and 6‐minute walk distance in 1,520 patients enrolled in the COMPANION study of CRT versus optimal medical therapy.

Methods and Results:
Over a mean follow‐up after implantation of 16.2 months, patients randomized to CRT, regardless of lead location, experienced benefit compared with optimized pharmacologic therapy (OPT), with respect to all‐cause mortality or heart failure hospitalization. All but a posterior location showed benefit with respect to the all‐cause mortality or all‐cause hospitalization outcome. Mortality benefit in CRT‐D patients was indifferent to LV lead position. All functional outcomes including 6‐minute walk distance, Quality‐of‐Life (QOL) and Functional Class improved with CRT, regardless of LV lead location.

Conclusion:
LV lead location was not a major determinant of multiple measures of response to CRT therapy in the COMPANION Trial. While acute data indicate that a left lateral LV lead location results in the most favorable hemodynamic response, these chronic data suggest that positioning an LV lead in an anterior rather than a lateral or posterior LV location has similar benefit.
A Case of Flecainide‐Induced Hyponatremia
Authors: AHMED MUBASHIR , SRA JASBIR , AKHTAR MASOOD , MORTADA M. EYMAN
Published: February 2009

We present a case of flecainide‐induced hyponatremia in a 67‐year‐old woman who was treated for paroxysmal atrial tachycardia. She developed dizziness after starting flecainide and was found to be hyponatremic with a sodium level of 122 mmol/L (decreased from 136 mmol/L). Work‐up failed to reveal other causes of hyponatremia. She was not on diuretics, laxatives, or herbal medications. After discontinuation of flecainide, her symptoms and sodium levels improved. Hyponatremia is a previously unrecognized entity as an adverse effect of flecainide. We will discuss the clinical presentation, lab findings, and a possible explanation for this patient's unusual reaction to flecainide.
Pacing Lead Implantation Without Live Fluoroscopy: Feasibility of Acute Success in the Live Canine Model
Authors: CHOUDHURI INDRAJIT , KRUM DAVID , HARE JOHN , KELLEY JIM , BECKER DAVID , MORTADA M. EYMAN , AKHTAR MASOOD , SRA JASBIR
Published: February 2009

Introduction: Fluoroscopic visualization for transvenous pacing lead placement necessitates lead shielding to minimize radiation exposure. An electromagnetic (EM) navigation system that integrates real‐time intracardiac tracking within an anatomic navigation environment may provide an effective alternative for lead delivery that obviates live fluoroscopy. We assessed feasibility of pacing lead implantation with electromagnetic tracking guided solely by radiographic virtual navigation and compared this to fluoroscopy‐guided implants in a canine model.

Methods: Seven mongrel dogs with normal hearts were randomized to 47 pacing lead placements in the right atrium (RA) or right ventricle (RV) guided by single‐plane fluoroscopy, or an experimental EM navigation system guided by registered fluoroscopic snapshots obtained before implant (EMN). Ability to achieve successful lead delivery acutely was assessed, and pacing parameters as well as fluoroscopy and implant times were measured. Means were compared using a paired t‐test.

Results: All lead delivery attempts were acutely successful. One atrial lead dislodged with EMN, resulting in 46 successful pacing attempts. There was no statistical difference in pacing parameters and time for lead placement between the approaches (EMN vs fluoroscopic navigation [mean ± SD]: RA threshold 1.15 V ± 0.98 V vs 1.95 V ± 0.98 V [P = NS], RV threshold 1.18 V ± 0.58 V vs 1.42 V ± 0.63 V [P = NS], implant time 4:38 ± 2:37 minutes vs 4:44 ± 2:38 minutes [P = NS]). No live fluoroscopy was required for EMN implants.

Conclusion: Pacing lead placement with an EM system guided by preprocedural fluoroscopic views is feasible and comparable to fluoroscopic navigation, and avoids the use of live fluoroscopy.
Emprise et déprise de l'agriculture sur le territoire de l'est de l'Ontario de 1826 à 2006: la dimension environnementale
Authors: WATELET ANNE
Published: February 2009

Les comtés de Stormont, Dundas et Glengarry (SDG), dans l'Est‐ontarien, sont parmi les premiers àêtre colonisés en Ontario, à partir de 1784.

Les établissements se retrouvent au sein de la forêt où les milieux humides sont abondants. Bien que ces derniers aient presque disparu, le paysage est aujourd'hui relativement boisé, ne laissant pas entrevoir les changements radicaux qu'il a subis au cours des deux derniers siècles. L'étude des recensements de 1826 à 2006 révèle la nature et le rythme des changements d'occupation du sol dont on présente ici une première analyse. Quatre phases se dessinent. D'abord l'établissement de l'emprise de l'agriculture sur le territoire jusqu'en 1891, puis l'apogée de l'agriculture, interrompu en 1941 par la déprise agricole. La contraction de l'espace agraire cesse en 1986 alors que débute la quatrième période. L'ouverture des terres à la culture est relativement lente, du fait de conditions biophysiques et socio‐économiques qui se traduisent par l'absence de croissance de la population agricole. Le déboisement et le drainage des terres humides ont entraîné une détérioration des conditions de la vie agricole qui a résulté en une prise en compte des réalités environnementales dans les occupations du sol. Boisés et cultures sont mieux proportionnés aujourd'hui.
Health geography in Canada: where are we headed?
Authors: LUGINAAH ISAAC
Published: February 2009

This paper overviews the emergence of medical/health geography in Canada. The paper discusses the key questions that Canadian health geographers have explored in the past two decades, how these enquiries have featured in the field and how they contribute to the wider discourse of human geography. It also addresses questions on emerging themes and where Canadian health geography will go in the years ahead. With shifting health landscapes in terms of changes in social, political and physical environments, and changes in health care restructuring, Canadian health geographers are entering a new phase of research, teaching and policy. The complexity of the questions that health geographers seek to address means it is necessary to continue to highlight the policy implications of their findings. Health geographers need to emphasize the public agenda through interdisciplinary research and by continuing to work with geographers in other subfields.
Gendered Power in Cultural Contexts: Part I. Immigrant Couples
Authors: MACIEL JOSE A. , VAN PUTTEN ZANETTA , KNUDSON‐MARTIN CARMEN
Published: February 2009

Immigration is a world‐wide phenomenon and practitioners are increasingly called on to work with issues related to it. The authors examine the experience of couples who are immigrants to the United States in regard to gender and power issues. Although the study limited participation to one religious group in order to hold that aspect of culture and gender attitudes constant, the experiences of these couples help to make visible the link between microlevel couple interaction and larger social processes. The results show how the couples manage a delicate balance between the push for gender change and avoiding too much conflict as male power is challenged.
Portuguese Immigrant Families: The Impact of Acculturation
Authors: MORRISON MARIE , JAMES SUSAN
Published: February 2009

Portuguese immigrants to North America represent a large ethnic group with unique family therapy needs. The present study investigates acculturation and the family lives of Portuguese (Azorean) immigrants in Canada. Methods of analytic induction and constant comparison from grounded theory were used to examine transcripts of interviews with 21 Azorean immigrant women and 28 Azorean immigrant men. A model emerged wherein (a) immigration and acculturation act as stressors on the family unit, as described by the categories Process of Change and Family Relationships; (b) family members adopt generation‐ and gender‐specific acculturative strategies, as illustrated by the categories Duas Culturas (Two Cultures) and Falando Portuges (Speaking Portuguese); and (c) as family members acculturate, discords arise and are resolved according to the cultural traits different members have adopted. The categories Discord Resolution and Preocupação (Preoccupations) illustrate this last dynamic. Implications for family therapy with immigrant families include an indication for community‐level interventions, emphasis on confidentiality, awareness of acculturation stress and different acculturative strategies within the family, and aiding the family in the negotiation and integration of a new bicultural reality.
The decrease of expression of ryanodine receptor sub‐type 2 is reversed by gentamycin sulphate in vascular myocytes from mdx mice
Authors: Morel Jean‐Luc , Dabertrand Fabrice , Fritz Nicolas , Henaff Morgana , Mironneau Jean , Macrez Nathalie
Published: February 2009

The mdx mouse, a model of the human Duchenne muscular dystrophy, displays impaired contractile function in skeletal, cardiac and smooth muscles. We explored the possibility that ryanodine receptor (RYR) expression could be altered in vascular muscle. The three RYR sub‐types were expressed in portal vein myocytes. As observed through mRNA and protein levels, RYR2 expression was strongly decreased in mdx myocytes, whereas RYR3 and RYR1 expression were unaltered. The use of antisense oligonucleotide directed against RYR sub‐types indicated that caffeine‐induced Ca2+ response and Ca2+ spark frequency depended on RYR2 and RYR1. In mdx mice, caffeine‐induced Ca2+ responses were decreased in both amplitude and maximal rate of rise, and the frequency of Ca2+ sparks was also strongly decreased. The gentamycin treatment was able to increase both the expression of RYR2 and the caffeine‐induced Ca2+ response to the same level as that observed in wild‐type mice. Taken together, these results confirm that both RYR1 and RYR2 are required for vascular Ca2+ signalling and indicate that inhibition of RYR2 expression may account for the decreased Ca2+ release from the SR in mdx vascular myocytes. Finally, we suggest that gentamycin can restore the Ca2+ signalling in smooth muscle from mdx mice by increasing RYR2 and dystrophin expression. These results may help explain the reduced efficacy of contraction in vascular myocytes of mdx mice and Duchenne muscular dystrophy–afflicted patients. Gentamycin treatment could be a good therapeutic tool to restore the vascular function.
Endostatin induces autophagy in endothelial cells by modulating Beclin 1 and β‐catenin levels
Authors: Nguyen Tri Minh Bui , Subramanian Indira V. , Xiao Xue , Ghosh Goutam , Nguyen Phan , Kelekar Ameeta , Ramakrishnan S.
Published: February 2009

Endostatin is a well‐characterized endogenous inhibitor of angiogenesis that affects cell proliferation and migration by inhibiting integrin and Wnt‐mediated signalling pathways. Here, we show that endothelial cells treated with native and P125A‐endostatin activate autophagy. Because autophagy can either be protective or induce programmed cell death, experiments were carried out to understand the signalling pathways leading to autophagy in endothelial cells. P125A‐endostatin treatment increased the levels of Beclin 1, a crucial molecule in vesicle nucleation and autophagy. The treatment also reduced the levels of Bcl‐2, Bcl‐xL and β‐catenin; however, progressively increasing amounts of Bcl‐2 and Bcl‐xL were found to be complexed with Beclin 1. Increased β‐catenin and Wnt‐mediated signalling reduced Beclin 1 levels and rescued endothelial cells from endostatin‐induced autophagy. Finally, knocking down Beclin 1 levels by RNA interference decreased autophagy and accelerated caspase activation in endostatin‐treated cells. These studies suggest that endothelial cells may initiate autophagy as a survival response to limit the effects of angiogenesis inhibitors. Thus, interfering with autophagy can potentiate the effects of endostatin by promoting a switch to apoptosis.
Healthy human salivary glands contain a DHEA‐sulphate processing intracrine machinery, which is deranged in primary Sjögren's syndrome
Authors: Spaan Michelle , Porola Pauliina , Laine Mikael , Rozman Blaz , Azuma Masayuki , Konttinen Yrjö T.
Published: February 2009

Sjögren's syndrome (SS) patients have low salivary dehydroepiandrosterone (DHEA) and androgen biomarker levels, but high salivary oestrogen levels. The hypothesis was that the healthy glands contain DHEA‐sulphate processing intracrine machinery; the local androgen/oestrogen imbalance suggests that this is disarranged in SS. Indirect immunofluorescence and quantitative real‐time PCR (qRT‐PCR) of steroid sulphatase, sulfotransferase, 3β‐ and 17β‐hydroxysteroid dehydrogenases (3β‐ and 17β‐HSD), 5α‐reductase and aromatase were performed for labial salivary glands of healthy controls and persons with SS. In control acini steroid sulphatase and sulfotransferase immunoreactivities were located in the basolateral cell parts. 3β‐ and 17β‐HSD formed strong, interrupted bands along the basal cell parts. 5α‐reductase was mainly located in acinar cell nuclei and aromatase in the apical cell membrane. All enzymes were more widespread in ducts. In SS, steroid sulphatase was weak and deranged, 3β‐ and 17β‐HSD had lost their strict basal acinar cell localization and 5α‐reductase was mainly found in the cytoplasm of the acinar cells, whereas aromatase showed similar staining in SS and controls. qRT‐PCR of labial salivary glands disclosed all corresponding enzyme mRNAs with the levels of 3β‐HSD in SS being the lowest. Healthy tubuloacinar epithelial cells contain complete intracrine machineries for DHEA(‐sulphate) pro‐hormone processing. These enzymes have in healthy acini an organized architecture, which corresponds with DHEA uptake from the circulation, nuclear site of production of the active dihydrotestosterone (DHT) end product and production of oestrogens into saliva for export to ductal and oral epithelial cells. SS is characterized by low 3β‐HSD levels, which together with impaired subcellular compartmentalization of HSDs and 5α‐reductase may explain the low local DHT and androgen biomarker levels in SS.
Modulation of cardiac ionic homeostasis by 3‐iodothyronamine
Authors: Ghelardoni Sandra , Suffredini Silvia , Frascarelli Sabina , Brogioni Simona , Chiellini Grazia , Ronca‐Testoni Simonetta , Grandy David K. , Scanlan Thomas S. , Cerbai Elisabetta , Zucchi Riccardo
Published: February 2009

3‐iodothyronamine (T1AM) is a novel endogenous relative of thyroid hormone, able to interact with trace amine‐associated receptors, a class of plasma membrane G protein‐coupled receptors, and to produce a negative inotropic and chronotropic effect. In the isolated rat heart 20–25 μM T1AM decreased cardiac contractility, but oxygen consumption and glucose uptake were either unchanged or disproportionately high when compared to mechanical work. In adult rat cardiomyocytes acute exposure to 20 μM T1AM decreased the amplitude and duration of the calcium transient. In patch clamped cardiomyocytes sarcolemmal calcium current density was unchanged while current facilitation by membrane depolarization was abolished consistent with reduced sarcoplasmic reticulum (SR) calcium release. In addition, T1AM decreased transient outward current (Ito) andIK1background current. SR studies involving 20 μM T1AM revealed a significant decrease in ryanodine binding due to reduced Bmax, no significant change in the rate constant of calcium‐induced calcium release, a significant increase in calcium leak measured under conditions promoting channel closure, and no effect on oxalate‐supported calcium uptake. Based on these observations we conclude T1AM affects calcium and potassium homeostasis and suggest its negative inotropic action is due to a diminished pool of SR calcium as a result of increased diastolic leak through the ryanodine receptor, while increased action potential duration is accounted for by inhibition ofItoandIK1currents.
The anti‐angiogenic factor PEDF is present in the human heart and is regulated by anoxia in cardiac myocytes and fibroblasts
Authors: Rychli Kathrin , Kaun Christoph , Hohensinner Philipp J. , Dorfner Adrian J. , Pfaffenberger Stefan , Niessner Alexander , Bauer Michael , Dietl Wolfgang , Podesser Bruno K. , Maurer Gerald , Huber Kurt , Wojta Johann
Published: February 2009

Cardiac diseases such as myocardial infarction and heart failure are among the leading causes of death in western societies. Therapeutic angiogenesis has been suggested as a concept to combat these diseases. The biology of angiogenic factors expressed in the heart such as vascular endothelial growth factor (VEGF) is well studied, whereas data on anti‐angiogenic mediators in the heart are scarce. Here we study the expression of the anti‐angiogenic factor pigment epithelium‐derived factor (PEDF) in the human heart and in human cardiac cells. PEDF expression could be detected in human cardiac tissue on the protein and mRNA levels. PEDF mRNA levels were significantly lower in explanted human ischemic hearts as compared to healthy hearts. Our in vitro experiments showed that human adult cardiac myocytes and fibroblasts constitutively secrete PEDF. In addition to anoxic conditions, cobalt chloride, 2,2′dipyridyl and dimethoxally glycine, which stabilize hypoxia inducible factor‐α decreased PEDF expression. Furthermore we show that PEDF inhibits VEGF‐induced sprouting. We have identified PEDF in healthy and ischemic human hearts and we show that PEDF expression is down‐regulated by low oxygen levels. Therefore, we suggest a role for PEDF in the regulation of angiogenesis in the heart and propose PEDF as a possible therapeutic target in heart disease.
Methods of labeling skin surgical specimens
Authors: Kneilling Manfred , Breuninger Helmut , Kastl Andrea , Heeg Peter , Häfner Hans Martin , Röcken Martin , Möhrle Matthias
Published: February 2009

Background: Accurate pre‐operative or intra‐operative labeling of the skin is often necessary to mark exactly the surgical excision lines. Pre‐operative “unsterile” permanent skin labeling systems are needed for example for vein and sentinel lymph node surgery; here the dyes must resist two surgical skin disinfection procedures. In contrast, excision borders are labeled during surgery using a “sterile” skin marking system.

Methods: Many commercial and non‐commercial pre‐ and intra‐operative skin‐labeling systems are available, such as autologous patient blood, fluorescence triphenylmethane dyes and commercial skin markers. The available skin marking systems have specific advantages and disadvantages. We review the different labeling systems, offering guidelines to help choose a cost‐effective system appropriate for a given surgical procedure.

Results: The Edding® permanent markers 400 und 3000 are well suited for preoperative skin labeling and less expensive than commercial skin labeling systems. Autologous patient blood and eosin are well suited for intra‐operative labeling and are most cost effective. Eosin Y is widely used and well suited for labeling of dark skin, bone, cartilage, and muscle tissue and spares the expense of expensive commercial skin markers.

Conclusion: Knowledge of the many commercial and non‐commercial pre‐ and intra‐operative skin labeling systems and their advantages and disadvantages helps to reduce the use of relatively expensive commercial skin markers.
Can soil seed banks contribute to the restoration of dune slacks under conservation management?
Authors: Plassmann Katharina , Brown Nigel , Jones M. Laurence M. , Edwards‐Jones Gareth
Published: February 2009

Questions: Does the soil seed bank resemble the former early successional stages of a dune slack system more than the established later successional vegetation? Does it have the potential to contribute to the conservation of a highly endangered habitat?

Location: Dune slacks at Newborough Warren, UK.

Methods: The composition of the soil seed bank in two depth layers was determined using the seedling emergence method between March 2004 and April 2005. Long‐term monitoring data on the floristic composition of the established vegetation were obtained from the national conservation agency, and additional monitoring was undertaken in 2003. Floristic composition, seed weights, seed longevity of component species and Ellenberg indicator values were used to compare the seed bank and established vegetation.

Results: The soil seed bank was diverse and contained typical dune slack species, species of early successional stages and species of conservation interest. A comparison between the composition of the seed bank and historical data on the composition of the established vegetation showed that the seed bank reflects earlier successional stages more closely than the current aboveground vegetation. This study increases the scarce information currently available on the seed bank ecology of several species, including two orchid species.

Conclusions: The soil seed bank can be expected to contribute to vegetation change after disturbance. Stimulation of germination from the seed bank through management may contribute to the conservation of both characteristic and threatened species typical of dune slacks.
Landscape metrics indicate differences in patterns and dominant controls of ribbon forests in the Rocky Mountains, USA
Authors: Bekker Matthew F. , Clark Jess T. , Jackson Mark. W.
Published: February 2009

Question: Do landscape metrics reflect differences in dominant factors controlling ribbon forest patterns among sites?

Location: West Flattop Mountain, Glacier National Park, Montana (Flattop); Medicine Bow Mountains, Wyoming (Medicine Bow); Park Range, Colorado (Park Range).

Methods: High‐resolution aerial photography was used to delineate ribbon forest patches, and to calculate landscape metrics to distinguish between long, narrow, regular patterns expected from strong microtopographic control, and smaller, more compact, and variable patterns expected from wind‐snowdrift interactions.

Results: All but two metrics were significantly different (P<0.05) among the three sites. The rank and magnitude of differences indicated that ribbons at Flattop and Park Range are more similar to each other than to those at Medicine Bow. Flattop ribbons were also more elongated, narrower and less variable than those at Park Range, suggesting differences in the type and strength of structural control. Previous research showed that Flattop ribbons occupy regular lithologic ridges, while our observations of ribbons and analysis of geologic maps suggests weaker and less consistent microtopographic control at Park Range, and dominant wind‐snowdrift interactions with little to no microtopographic influence at Medicine Bow.

Conclusions: Landscape metrics indicate differences in pattern among sites that reflect differences in dominant factors influencing ribbon forest development and maintenance. Explanations of ribbon forest dynamics are site‐specific and are more complex than is currently recognized. The sites vary in the level of endogenous versus exogenous control of ribbon patterns, and consequently in the sensitivity of this phenomenon to climate.
Estrogen Metabolism and Breast Cancer
Authors: Parl Fritz F. , Dawling Sheila , Roodi Nady , Crooke Philip S.
Published: February 2009

Oxidative metabolites of estrogens have been implicated in the development of breast cancer, yet relatively little is known about the metabolism of estrogens in the normal breast. We developed an experimental in vitro model of mammary estrogen metabolism in which we combined purified, recombinant phase I enzymes CYP1A1 and CYP1B1 with the phase II enzymes COMT and GSTP1 to determine how 17β‐estradiol (E2) is metabolized. We employed both gas and liquid chromatography with mass spectrometry to measure the parent hormone E2 as well as eight metabolites, that is, the catechol estrogens, methoxyestrogens, and estrogen−GSH conjugates. We used these experimental data to develop an in silico model, which allowed the kinetic simulation of converting E2 into eight metabolites. The simulations showed excellent agreement with experimental results and provided a quantitative assessment of the metabolic interactions. Using rate constants of genetic variants of CYP1A1, CYP1B1, and COMT, the model further allowed examination of the kinetic impact of enzyme polymorphisms on the entire metabolic pathway, including the identification of those haplotypes producing the largest amounts of catechols and quinones. Application of the model to a breast cancer case‐control population defined the estrogen quinone E2‐3,4‐Q as a potential risk factor and identified a subset of women with an increased risk of breast cancer based on their enzyme haplotypes and consequent E2‐3,4‐Q production. Our in silico model integrates diverse types of data and offers the exciting opportunity for researchers to combine metabolic and genetic data in assessing estrogenic exposure in relation to breast cancer risk.
Involvement of Nitric Oxide in Nigrostriatal Dopaminergic System Degeneration
Authors: Di Matteo Vincenzo , Pierucci Massimo , Benigno Arcangelo , Crescimanno Giuseppe , Esposito Ennio , Di Giovanni Giuseppe
Published: February 2009

The present study was undertaken to explore the involvement of nitric oxide (NO) in the 6‐hydroxydopamine (6‐OHDA) experimental model of Parkinson's disease (PD) in rats. The effect of pharmacological manipulation of the NO system was evaluated on striatal dopamine (DA) level decrease produced by the toxin. 7‐nitroindazole (7‐NI, 50 mg/kg i.p.; n= 5) pretreatment significantly restored the striatal DA contents. Conversely, 40 mg/kg i.p. of molsidomine (MOL, n= 5), an NO donor, significantly worsened the neurodegeneration (n= 5) and completely counteracted the neuroprotective effect of 7‐NI (n= 5). Thus, a crucial role for NO in 6‐OHDA induced neurodegeneration is suggested together with a protective benefit for inhibitors of NOS in the treatment of PD.
New Developments in Intracrinology of Human Breast Cancer
Authors: Sasano Hironobu , Nagasaki Shuji , Miki Yasuhiro , Suzuki Takashi
Published: February 2009

Steroid sulfatase (STS) hydrolyses biologically inactive estrogen sulfates to active estrogens, while estrogen sulfotransferase (EST) sulfonates estrogens to estrogen sulfates. Information regarding the expression of STS in human breast carcinoma tissues is still very limited compared to that of aromatase or 17β‐hydroxysteroid dehydrogenases (17β‐HSDs). In our study, EST and STS immunoreactivity was detected in carcinoma cells in 50 and 84 out of 113 breast carcinomas (44.2% and 74.3%, respectively), which was also associated with mRNA levels determined by RT/real‐time PCR. Using microdissection/RT‐PCR analyses, EST mRNA was localized to both carcinoma and intratumoral stromal cells, whereas STS was detected only in carcinoma or parenchymal cells. STS immunoreactivity was positively associated with tumor size. EST immunoreactivity was inversely correlated with tumor size or lymph node status and was significantly associated with a decreased risk of recurrence and improved prognosis. These data suggest that both EST and STS play important roles in the regulation of in situ estrogen production in human breast cancer. In addition, EST is an independent prognostic factor in human breast carcinoma and loss of EST may result in altered estrogen metabolism in hormone‐dependent breast cancer cells. An inhibition of intratumoral STS in the patients with estrogen‐dependent breast carcinoma is also considered to provide more clinical benefits, especially to the patients in which primary source of an availability of intratumoral estrogen is through STS rather than aromatase.
Inhibition of Human Breast Cancer Cell Growth and Enzymatic Activity by a Fermented Nutraceutical
Authors: Marotta F. , Yadav H. , Pathak S. , Minelli E. , Signorelli P. , Lorenzetti A. , Marandola P.
Published: February 2009

Human breast cancer cell lines MCF‐7 (ER‐positive) and Hs578T (ER‐negative) were cultured and one lot incubated for 48 h with 5–50 μg/ml of a fermented phytocompound (MK: Manda‐Koso, Innoshima, Japan). In vitro, it appeared a dose‐dependent decrease of cell viability (5–57%) in MK group in both cell lines (P < 0.001, plateau: 30 μg/ml), decreased beta‐galactosidase activity, enhanced apoptosis, and inversely increased Bax/Bcl2 ratio (P < 0.01) with an upregulation of p53 (P < 0.05). In the in vivo model, Balb‐c mice were inoculated with tumor cells and the treatment group was fed with 20 mg of MK. Tumor weight in MK‐fed group was time‐course reduced by 22% to 51% at 2 and 4 weeks, respectively (P < 0.05) with increased survival (P < 0.05). Tumour tissue of MK‐fed mice showed a downregulated Bcl‐2 with increased Bax/Bcl‐2 ratio, reduced PCNA, and activated caspase 3. Although more studies are ongoing to foster the clinical applicability of MK integrated within a rational chemopreventive and therapeutic strategy, a p53‐mediated mechanism is likely to play a relevant role, besides its reported antioxidant capacity, NK cell activity enhancement, cancer‐cytostatic activity properties.
Estrogen Mediation of Breast Tumor Formation Involves Estrogen Receptor‐Dependent, as Well as Independent, Genotoxic Effects
Authors: Santen Richard , Cavalieri Ercole , Rogan Eleanor , Russo Jose , Guttenplan Joseph , Ingle James , Yue Wei
Published: February 2009

Long‐term exposure to estrogens influences the development of breast cancer in women, but the precise mechanisms involved are not clearly defined. Our working hypothesis is that estrogen modulates this process by two separate processes. One involves the binding of estradiol to estrogen receptor (ER) alpha with stimulation of cell proliferation. Errors in DNA occurring during replication result in fixed mutations when not repaired. The other process results from the formation of genotoxic metabolites of estradiol, which can bind to DNA, cause depurination, and result in mutations. Our collaborative group, funded by a Department of Defense Center of Excellence grant, has examined this hypothesis using a variety of experimental methods. Estradiol and its catechol‐estrogen metabolite 4‐OH‐estradiol causes mutations in cell culture systems and can transform benign MCF‐10F cells, allowing them to cause tumors in SCID mice. We have demonstrated loss of heterozygosity and gains and losses of DNA segments by comparative genomic hybridization methodology. The depurinated estradiol‐guanine and ‐adenine adducts are measurable in MCF‐7 breast cancer cells in culture and in mouse mammary tissue. The double transgenic, alpha estrogen receptor knockout/Wnt‐1 knockin mouse model allows us to dissect out the separate effects of ER‐mediated and ER‐independent actions of estradiol. Knock out of the ER alpha delays the onset of breast tumors in this model, demonstrating a role of receptor‐mediated actions. Oophorectomy delays the onset of tumors and reduces overall incidence, providing evidence for an ER‐independent effect. Taken together, these data suggest that both ER‐dependent and genotoxic ER‐independent effects of estradiol mediate breast cancer development.
Profiling Cancer Stem Cells in Androgen‐Responsive and Refractory Human Prostate Tumor Cell Lines
Authors: Cocciadiferro Letizia , Miceli Vitale , Kang Kyung‐Sun , Polito Lucia M. , Trosko James E. , Carruba Giuseppe
Published: February 2009

In this study, we investigated androgen metabolism in two different human prostate cancer cell lines, the androgen‐responsive LNCaP cells and the nonresponsive PC3 cells. Following 24‐h and 72‐h incubation with either testosterone (T) or androstenedione (Ad) used as precursor, divergent patterns and rates of androgen metabolism were observed. Given the recent interest in the multiple uses of embryonic and adult stem cells for basic and applied research, we compared the expression of three presumptive stem cell markers (Oct‐4, SUZ‐12, and Cripto‐1), along with connexin 43 (Cx43), Cx32, and androgen receptor (AR), used as cell differentiation gene markers. In anchorage‐independent cell growth conditions, the expression levels of candidate markers of cancer stem cells initially increased (days 2–4) but drastically fell thereafter (day 6) in both cell lines. Results of immunocytochemical assay (ICA) largely confirmed those obtained by RT‐PCR. Interestingly, both symmetrical and asymmetrical cell divisions were revealed in PC3 cells using Oct‐4 immunostaining. Our data suggest that both androgen‐responsive and androgen‐nonresponsive prostate tumor cell lines contain a presumptive cancer stem cell population that can be identified using a panel of selected gene markers, including Oct‐4, SUZ‐12, and Cripto‐1.
Targeting Aromatase and Estrogen Signaling in Human Non‐Small Cell Lung Cancer
Authors: Márquez‐Garbán Diana C. , Chen Hsiao‐Wang , Goodglick Lee , Fishbein Michael C. , Pietras Richard J.
Published: February 2009

Lung cancer has become increasingly common in women, and gender differences in the physiology and pathogenesis of the disease have suggested a role for estrogens. In the lung recent data have shown local production of estrogens from androgens via the action of aromatase enzyme and higher levels of estrogen in tumor tissue as compared with surrounding normal lung tissue. High levels of aromatase expression are also maintained in metastases as compared with primary tumors. Consistent with these findings, clinical studies suggest that aromatase expression may be a useful predictive biomarker for prognosis in the management of non‐small cell lung cancer (NSCLC), the most common form of lung malignancy. Low levels of aromatase associate with a higher probability of long‐term survival in older women with early stage NSCLC. Treatment of lung NSCLC xenografts in vivo with an aromatase inhibitor (exemestane) alone or combined with standard cisplatin chemotherapy elicits a significant reduction in tumor progression as compared to paired controls. Further, lung cancer progression is also governed by complex interactions between estrogen and growth factor signaling pathways to stimulate the growth of NSCLC as well as tumor‐associated angiogenesis. We find that combination therapy with the multitargeted growth factor receptor inhibitor vandetanib and the estrogen receptor antagonist fulvestrant inhibit tumor growth more effectively than either treatment administered alone. Thus, incorporation of antiestrogen treatment strategies in standard antitumor therapies for NSCLC may contribute to improved patient outcome, an approach that deserves to be tested in clinical trials.
Clues to Understanding the Oxidation of Estradiol in Humans
Authors: Lahita Robert G. , Schaefer Robert A. , Bradlow H. Leon , Kreek Mary Jeanne
Published: February 2009

Determination of 2‐ and 16α‐hydroxylation of estradiol in patients with a variety of liver disorders using a dynamic method of quantitating the extent of hydroxylation revealed specific and characteristic differences in the metabolic response. Patients with acute or silent variants of hepatitis B had estrogen metabolite patterns that were indistinguishable from those found in the control subjects. Female patients with autoimmune hepatitis (formerly known as lupoid hepatitis), however, showed a moderate significant decrease (P < 0.01) in 2‐hydroxylation as compared with normal controls (mean 16.3 ± 1.9 vs. 33.9 ± 2.5), with no significant change in 16α‐hydroxylation. Male and female subjects with chronic alcoholic cirrhosis were almost devoid of 2‐hydroxylation (mean 2.9 ± 0.5, P < 0.01), but did show a significant increase in 16α‐hydroxylation (P < 0.01). The results, therefore, show that the alterations in patterns of biological oxidation are highly specific and do not reflect a general inability to metabolize estrogens in the cirrhotic patient. However, the results also suggest the possibility that a substantial fraction of 16α‐hydroxylation may occur elsewhere in the body at sites other than in the liver, explaining why this biotransformation pathway is elevated, while the reaction at C‐2 is almost absent in the alcoholic cirrhotic subjects.
Dasymetric Mapping for Estimating Population in Small Areas
Authors: Mennis Jeremy
Published: February 2009

A dasymetric map depicts a statistical surface, most commonly population density, as a set of simply connected regions, such that variation within each region is minimized and the region boundaries approximate the steepest escarpments of the surface. Dasymetric mapping has its roots in early thematic mapping of population, but has recently been taken up by researchers focusing on areal interpolation and population estimation using remote sensing. The process of dasymetric mapping typically involves the disaggregation of population data encoded in choropleth map form using an ancillary spatial data set, most commonly either an area‐class map or satellite image. The functional relationship between the ancillary data and the statistical surface being mapped may be specified a priori by the researcher or estimated using a variety of statistical techniques. Challenges facing dasymetric mapping research include handling spatio‐temporal data and the development of standardized and accessible methods.