bluuebunny/crossref_metadata_2025 huggingface dataset:
Stent Thrombosis in Sweden
Authors: Lagerqvist Bo , Carlsson Jörg , Fröbert Ole , Lindbäck Johan , Scherstén Fredrik , Stenestrand Ulf , James Stefan K.
Published: September 2009

Background—

The objective was to evaluate the role of risk factors and stent type for stent thrombosis (ST) using a large real world registry.

Methods and Results—

We evaluated all consecutive coronary stent implantations in Sweden from May 1, 2005, to June 30, 2007. All cases of ST, documented in the Swedish coronary angiography and angioplasty registry until September 21, 2008, were analyzed. ST was registered in 882 of 73 798 stents. Acute coronary syndromes, insulin-treated diabetes mellitus, smoking, previous coronary intervention, warfarin treatment, small stent diameter, and stenting in restenotic, complex, or bypass graft lesions had the strongest association with ST in the multivariable statistical model. There were considerable differences in the frequency of ST between different stent brands. The overall risk of ST was lower in drug-eluting stents compared with bare metal stents (adjusted risk ratio, 0.79; 99% CI, 0.63 to 0.99). However, from 6 months after stent implantation and onward, the risk for ST was higher in drug-eluting stents compared with bare metal stents (adjusted risk ratio, 2.02; 99% CI, 1.30 to 3.14).

Conclusions—

ST is a multifactor disease, and the incidence varies considerably between patients based on clinical, vessel, and stent characteristics. For drug-eluting stents compared with bare metal stents, the risk pattern was biphasic; initially, bare metal stents demonstrated a higher risk of ST; whereas after the first months, ST risk was higher with drug-eluting stents. Our findings highlight the need for prospective randomized studies with head-to-head comparisons between different stents.
Real-Time 3-Dimensional Echocardiography Provides New Insight Into Mechanisms of Tricuspid Valve Regurgitation in Patients With Hypoplastic Left Heart Syndrome
Authors: Takahashi K. , Inage A. , Rebeyka I.M. , Ross D.B. , Thompson R.B. , Mackie A.S. , Smallhorn J.F.
Published: September 2009

Background—

Tricuspid regurgitation in hypoplastic left heart syndrome has an impact on outcome, but its mechanisms remain unclear.

Methods and Results—

Real-time 3-dimensional echocardiography was performed in 35 patients with hypoplastic left heart syndrome (age, 1 month to 10 years; 10 after first-stage Norwood, 12 after superior cavopulmonary shunt, 13 after Fontan). From the 3-dimensional data set, we marked the annulus in systole and diastole. At mid systole, we marked the location of the papillary muscle tip and point of chordal attachment to the leaflet. We traced the surfaces of the tricuspid valve leaflets and measured the volume of leaflet prolapse, tethering, annular and septal leaflet areas, and papillary muscle position. Seventeen patients had moderate tricuspid regurgitation (prolapse, 7; tethered leaflets, 7) and 18 mild (prolapse, 0; tethered leaflets, 7). Multiple linear regression analysis revealed that moderate tricuspid regurgitation is associated with leaflet tethering and prolapse; that in hypoplastic left heart syndrome with tethered leaflets, the papillary muscle is displaced laterally and the tricuspid annulus is more planar; and that enlargement of the annulus at mid systole, small septal leaflet area, and age affect the degree of prolapse.

Conclusion—

In hypoplastic left heart syndrome, moderate tricuspid regurgitation may be associated with increasing age, geometrical changes of the annulus, leaflet prolapse, lateral papillary muscle displacement, and subsequent leaflet tethering, as well as a smaller septal leaflet.
Engraftment, Differentiation, and Functional Benefits of Autologous Cardiosphere-Derived Cells in Porcine Ischemic Cardiomyopathy
Authors: Johnston Peter V. , Sasano Tetsuo , Mills Kevin , Evers Robert , Lee Shuo-Tsan , Smith Rachel Ruckdeschel , Lardo Albert C. , Lai Shenghan , Steenbergen Charles , Gerstenblith Gary , Lange Richard , Marbán Eduardo
Published: September 2009

Background—

Cardiosphere-derived cells (CDCs) isolated from human endomyocardial biopsies reduce infarct size and improve cardiac function in mice. Safety and efficacy testing in large animals is necessary for clinical translation.

Methods and Results—

Mesenchymal stem cells, which resemble CDCs in size and thrombogenicity, have been associated with infarction after intracoronary infusion. To maximize CDC engraftment while avoiding infarction, we optimized the infusion protocol in 19 healthy pigs. A modified cocktail of CDCs in calcium-free PBS, 100 U/mL of heparin, and 250 μg/mL of nitroglycerin eliminated infusion-related infarction. Subsequent infusion experiments in 17 pigs with postinfarct left ventricular dysfunction showed CDC doses ≥10
7
but <2.5×10
7
result in new myocardial tissue formation without infarction. In a pivotal randomized study, 7 infarcted pigs received 300 000 CDCs/kg (≈10
7
total) and 7 received placebo (vehicle alone). Cardiac magnetic resonance imaging 8 weeks later showed CDC treatment decreased relative infarct size (19.2% to 14.2% of left ventricle infarcted,
P
=0.01), whereas placebo did not (17.7% to 15.3%,
P
=0.22). End-diastolic volume increased in placebo, but not in CDC-treated animals. Hemodynamically, the rate of pressure change (dP/dt) maximum and dP/dt minimum were significantly better with CDC infusion. There was no difference between groups in the ability to induce ventricular tachycardia, nor was there any tumor or ectopic tissue formation.

Conclusions—

Intracoronary delivery of CDCs in a preclinical model of postinfarct left ventricular dysfunction results in formation of new cardiac tissue, reduces relative infarct size, attenuates adverse remodeling, and improves hemodynamics. The evidence of efficacy without obvious safety concerns at 8 weeks of follow-up motivates human studies in patients after myocardial infarction and in chronic ischemic cardiomyopathy.
Cardiomyocyte-Specific Overexpression of Human Stem Cell Factor Improves Cardiac Function and Survival After Myocardial Infarction in Mice
Authors: Xiang Fu-Li , Lu Xiangru , Hammoud Lamis , Zhu Ping , Chidiac Peter , Robbins Jeffrey , Feng Qingping
Published: September 2009

Background—

Soluble stem cell factor (SCF) has been shown to mobilize bone marrow stem cells and improve cardiac repair after myocardial infarction (MI). However, the effect of membrane-associated SCF on cardiac remodeling after MI is not known. The present study investigated the effects of cardiomyocyte-specific overexpression of the membrane-associated isoform of human SCF (hSCF) on cardiac function after MI.

Methods and Results—

A novel mouse model with tetracycline-inducible and cardiac-specific overexpression of membrane-associated hSCF was generated. MI was induced by left coronary artery ligation. Thirty-day mortality after MI was decreased in hSCF/tetracycline transactivator (tTA) compared with wild-type mice. In vivo cardiac function was significantly improved in hSCF/tTA mice at 5 and 30 days after MI compared with wild-type mice. Endothelial progenitor cell recruitment and capillary density were increased and myocardial apoptosis was decreased in the peri-infarct area of hSCF/tTA mice. Myocyte size was decreased in hSCF/tTA mice 30 days after MI compared with WT mice. Furthermore, hSCF overexpression promoted de novo angiogenesis as assessed by matrigel implantation into the left ventricular myocardium.

Conclusions—

Cardiomyocyte-specific overexpression of hSCF improves myocardial function and survival after MI. These beneficial effects of hSCF may result from increases in endothelial progenitor cell recruitment and neovascularization and decreases in myocardial apoptosis and cardiac remodeling.
Renal Impairment of Pure Autonomic Failure
Authors: Garland Emily M. , Gamboa Alfredo , Okamoto Luis , Raj Satish R. , Black Bonnie K. , Davis Thomas L. , Biaggioni Italo , Robertson David
Published: September 2009

Supine hypertension is difficult to manage in patients with pure autonomic failure (PAF), because treatment can worsen orthostatic hypotension. Supine hypertension in PAF has been associated with left ventricular hypertrophy, but end organ damage in the kidney has not been assessed. We reviewed hemodynamic and laboratory data of 64 male patients with PAF who were 69±11 (mean±SD) years old. Systolic blood pressure fell by 67±40 mm Hg within 10 minutes of standing, with an inappropriately low 13±11-bpm increase in heart rate. Plasma norepinephrine levels were below normal (0.62±0.32 nmol/L supine and 1.28±1.25 nmol/L standing). A control data set of 75 men (67±12 years) was obtained from a deidentified version of the Vanderbilt University Medical Center electronic medical chart database. Compared with controls, PAF patients had lower hemoglobin (8.3±0.9 versus 9.3±0.8 mmol/L;
P
<0.001), packed cell volume (0.40±0.04 versus 0.45±0.04;
P
<0.001), and red blood cell count (4.4±0.5×10
12
versus 5.0±0.5×10
12
cells/L;
P
<0.001). Serum creatinine and blood urea nitrogen levels were elevated in patients. Forty-eight percent of patients with PAF had supine hypertension (supine systolic blood pressure: ≥150 mm Hg). Serum creatinine was higher in patients with supine hypertension (133±44 versus 106±27 μmol/L;
P
=0.021) and estimated glomerular filtration rate was lower (57±22 versus 70±20 mL/min per 1.73 m
2
;
P
=0.022) compared with patients who did not have supine hypertension. These findings may indicate that renal function is diminished in PAF in association with supine hypertension.
Arterial Wave Reflections During the Menstrual Cycle of Healthy Women
Authors: Papaioannou Theodore G. , Stamatelopoulos Kimon S. , Georgiopoulos George , Vlachopoulos Charalambos , Georgiou Stellios , Lykka Marita , Lambrinoudaki Irene , Papamichael Christos M. , Stefanadis Christodoulos I.
Published: September 2009

Increased wave reflection is an independent factor associated with cardiovascular diseases, risk, and mortality. The influence of the menstrual cycle on wave reflections and particularly on the reproducibility of their measurement has never been examined. The aim of the present study was to examine the reproducibility and variability of wave reflection indices in premenopausal healthy women during their menstrual cycle. Thirty-two women were examined at 3 phases of their menstrual cycle: days 1 to 2 (menstrual phase), days 6 to 14 (late follicular), and days 4 to 7 after ovulation (early luteal phase). Applanation tonometry of the radial artery and aortic pulse wave analysis were performed for the calculation of augmentation pressure, augmentation index, and timing of reflected waves. Reproducibility of these measures was evaluated by intraclass correlation coefficient and Bland-Altman analysis, whereas ANOVA was performed to assess their variability during the menstrual cycle. The SD of augmentation index differences between repeated measurements within the menstrual cycle ranged from 7.6% to 9.9%. Bland-Altman analysis indicated no evidence of systemic bias and no trend for the reproducibility of measurements to vary with their underlying mean value. Intraclass correlation coefficient indicated a moderate reproducibility of augmentation index and augmentation pressure (>0.80) and a rather low reproducibility for timing of reflected waves (0.43). Mean augmentation pressure, augmentation index, and timing of reflected waves did not vary significantly during the menstrual cycle (ANOVA). Measurement of wave reflections at the same phase of the menstrual cycle or statistical adjustment could be suggested for optimal study design and data interpretation.
Potassium Inhibits Dietary Salt-Induced Transforming Growth Factor-β Production
Authors: Ying Wei-Zhong , Aaron Kristal , Wang Pei-Xuan , Sanders Paul W.
Published: September 2009

Human and animal studies demonstrate an untoward effect of excess dietary NaCl (salt) intake on cardiovascular function and life span. The endothelium in particular augments the production of transforming growth factor (TGF)-β, a fibrogenic growth factor, in response to excess dietary salt intake. This study explored the initiating mechanism that regulates salt-induced endothelial cell production of TGF-β. Male Sprague-Dawley rats were given diets containing different amounts of NaCl and potassium for 4 days. A bioassay for TGF-β demonstrated increased (35.2%) amounts of active TGF-β in the medium of aortic ring segments from rats on the high-salt diet compared with rats maintained on a 0.3% NaCl diet. Inhibition of the large-conductance, calcium-activated potassium channel inhibited dietary salt-induced vascular production of TGF-β but did not affect production of TGF-β by ring segments from rats on the low-salt diet. Immunohistochemical and Western analyses demonstrated the α subunit of the calcium-activated potassium channel in endothelial cells. Increasing medium [K
+
] inhibited production of dietary salt-induced vascular production levels of total and active TGF-β but did not alter TGF-β production by aortic rings from rats on the 0.3% NaCl diet. Increasing dietary potassium content decreased urinary active TGF-β in animals receiving the high-salt diet but did not change urinary active TGF-β in animals receiving the low-salt diet. The findings demonstrated an interesting interaction between the dietary intake of potassium and excess NaCl and further showed the fundamental role of the endothelial calcium-activated potassium channel in the vascular response to excess salt intake.
Recurrent Febrile Infections and Neutropenia in a Child With Silent Celiac Disease
Authors: Leonardi Salvatore , Vitaliti Giovanna , La Rosa Mario
Published: September 2009

Backgound. Today it is known that celiac disease (CD) may be silent or present with atypical extraintestinal symptoms (silent/atypical CD). Case report. The authors report the case of a 3-year-old child in whom the diagnosis of CD was made after several admissions to hospital for recurrent febrile infections and persistently moderate neutropenia. After the beginning of a gluten-free diet, we observed a remission of febrile infections and an increase in neutrophil count until it reached normal ranges. Conclusions. We suggest that recurrent febrile infections and moderate neutropenia be included in the diagnostic workup for atypical/silent CD in the general population.
The Effects of Initially Misclassified Data on the Effectiveness of Discriminant Function Analysis and Finite Mixture Modeling
Authors: Holden Jocelyn E. , Kelley Ken
Published: September 2009

Classification procedures are common and useful in behavioral, educational, social, and managerial research. Supervised classification techniques such as discriminant function analysis assume training data are perfectly classified when estimating parameters or classifying. In contrast, unsupervised classification techniques such as finite mixture models (FMM) do not require, or even use if available, knowledge of group status to estimate parameters or classifying. This study investigates the impact of two types of misclassification errors on the classification accuracy of discriminant function analysis (both linear [LDA] and quadratic [QDA]) and FMM for two groups with a single predictor. Analytic and Monte Carlo results are provided for a variety of misclassification scenarios to investigate the performance of the two methods. Discriminant function techniques recovered the highest overall percentages of correctly classified data, whereas FMM captured higher percentages of the smaller group when group sizes are unequal. LDA marginally outperformed QDA under misclassified conditions.
Poor Neighbourhoods and Poor Services: Evidence on the ‘Rationing’ of Environmental Service Provision to Deprived Neighbourhoods
Authors: Hastings Annette
Published: September 2009

There is growing political concern that poor neighbourhoods often receive inadequate public services, although little research to date which explores how and why this may be the case. Earlier research on environmental services has suggested that poor neighbourhoods do not tend to get levels of this service which are proportionate to their needs. This paper extends an earlier analysis to explore the processes by which this underprovision occurs. It is argued that three ‘rationing’ processes are central to explanations: institutional rationing which describes a systemic bias against fully meeting the needs of poor neighbourhoods; reactive rationing in which service practices and standards are varied between neighbourhoods; and political rationing where service levels and standards are sensitive to variations in the political resources of neighbourhoods. The analysis reveals how these rationing processes produce levels of environmental maintenance in poor neighbourhoods which are insufficient to address needs.
Time-for-Money Exchanges Between Older and Younger Generations in Swedish Families
Authors: Lennartsson Carin , Silverstein Merril , Fritzell Johan
Published: September 2009

Despite the maturation of welfare states, family solidarity continues to be strong and a growing body of research has shown that substantial financial transfers are passed from older to younger generations within the family. At the same time, family solidarity in terms of instrumental and social support is found to be mutual. This study examines eventual reciprocity in time-for-money exchanges, by combining two large-scale Swedish representative longitudinal studies. It analyzes how earlier social contacts (time) are related to financial transfers (money) and to what extent social class and gender differentials are visible. The findings indicate that parents provide economic transfers if they have more frequent social contact with any of their children, and that these time investments pay off for children who were of higher social class origins. In contrast, no support for gender-specific patterns is found. In conclusion, family solidarity seems to have different bases in different social strata.
Mechanical Properties of Polypropylene Filled with Electron Beam Modified Surface-treated Titanium Dioxide Nanoparticles
Authors: Madani M.
Published: September 2009

An irradiation grafting method was applied for the surface modification of titanium dioxide nanoparticles so that the latter can be added to polymeric materials for improving their mechanical performance. The modification of filler has been developed by coating this with an acrylate monomer, trimethylol propane triacrylate (TMPTA), followed by electron beam irradiation. Nanocomposites of polypropylene with different loading of unmodified and modified nano-fillers were prepared. The mechanical properties including modulus and tensile strength of the resultant nanocomposites are discussed as a function of filler loading. The composites with treated filler showed enhanced mechanical properties. It was found that the reinforcing effects of the treated nanoparticles on the polymer matrix could be fully brought into play at a rather low filler loading in comparison to conventional particulate-filled composites. Unlike the approaches for manufacturing of the other types of nanocomposites, including intercalation polymerization, the current technique is characterized by many advantages, such as simple, low cost, easy to control, and having broader applicability. Finally, experimental results were compared with theoretical predictions.
Behavior of Kevlar/Epoxy Composite Plates Under Ballistic Impact
Authors: Kumar Sunil , Durga Shankar Gupta  , Singh Inderdeep , Sharma Apurbba
Published: September 2009

This study investigates the ballistic response of laminated composite plates using numerical simulations. Numerical simulations were carried out to determine the ballistic response of thick Kevlar/epoxy composite plates, commonly used in body armor. These plates were impacted at velocities between 100 and 1000 m/s. The numerical parametric study of ballistic impact caused by cylindrical projectile is undertaken to obtain an estimate for the ballistic limit velocity, energy absorbed by the plate, and the contact duration. The effect of mass and diameter of the projectile on ballistic limit velocity was also studied. The results obtained hereby are in good agreement with the experimental data presented by other researchers.
Flexural and Morphological Properties of Poly(Methyl Methacrylate)/ Hydroxyapatite Composites: Effects of Planetary Ball Mill Grinding Time
Authors: Tham W.L. , Chow W.S. , Mohd Ishak Z.A.
Published: September 2009

Poly(methyl methacrylate) (PMMA)/hydroxyapatite (HA) composites have potential applications in bone cement, prosthesis, and dental implant. In this study, PMMA containing 5 wt% of HA is prepared using polymerization followed by compression molding. PMMA and HA powder are ground using planetary ball milling. The grinding time takes from 30 to 120 min. The effects of the grinding time and particle size of the PMMA/HA powder on the flexural properties and morphology of the composites are investigated. The structure patterns of PMMA/5HA are characterized using X-Ray diffraction (XRD). No new phase is observed in the XRD pattern with the different sizes of PMMA/HA powder. This indicates that planetary milling solely reduces the size of PMMA/ HA powder. However, it does not modify the structure of PMMA and HA. A reduction of ~40% in the particle diameter is observed in both PMMA and PMMA/5HA powder after subjected to planetary milling for 60 min. For the planetary ball mill-ground PMMA/HA powder, the flexural modulus of the respective PMMA composites is slightly increased. Planetary milling can increase the volume of fine particles in the composites specimens, which results in a more homogeneous distribution of HA and a reduction of void contents in PMMA matrix. The reduction in void content is observed on the fractured surface of PMMA composites through field emission scanning electron microscopy.
Bone Cell Responses of Titanium Blasted with Bioactive Glass Particles
Authors: Choi Chang-Rak , Yu Hye-Sun , Kim Chul-Hwan , Lee Jae-Hoon , Oh Chung-Hun , Kim Hae-Won , Lee Hae-Hyoung
Published: September 2009

Surface modification of Ti-based metals is an important issue in improving the bone cell responses and bone-implant integration. Blasting Ti with granules (mostly alumina) is commonly used to prepare a clean surface and provide a level of roughness. In this study, glass granules with a bioactive composition were used as the blasting source to improve the surface bioactivity and biocompatibility of a Ti substrate. Bioactive glass particles with a composition of 70SiO 2 · 25CaO · 5P2O5 were prepared using a sol—gel method. A Ti disc was blasted with glass particles using a dental blasting unit (BG-Ti). A Ti disc blasted with commercial spherical-shaped glass (G-Ti) and a disc without blasting (Ti) were also prepared for comparison. The blasted Ti contained a large number of glass particles after the blasting process. The surface roughness of the samples in ascending order was G-Ti>BG-Ti>Ti. Murine-derived preosteoblasts (MC3T3-E1) were seeded on the samples, and the cell growth, differentiation, and mineralization behaviors were observed. The osteoblastic cells attached well and spread actively over all the sample groups with extensive cytoskeletal processes. The level of cell growth on the BG-Ti showed a continual increase with culturing up to 7 days, showing good cell viability. However, there was no significant difference (ANOVA, p<0.05) with respect to the G-Ti and Ti groups. In particular, the alkaline phosphatase (AP) activity of the cells was significantly higher on the BG-Ti than on the other groups after culturing for 14 days. Moreover, the mineralization behavior of the cells, as assessed by Alizarin S Red, was superior on the BG-Ti to that observed on the other groups after culturing for 14 and 28 days. Overall, the blasting of Ti with a bioactive glass composition is considered beneficial for producing substrates with enhanced osteogenic potential.
Montmorillonite-Alginate Nanocomposites as a Drug Delivery System: Intercalation and In Vitro Release of Vitamin B1 and Vitamin B6
Authors: Kevadiya Bhavesh D. , Joshi Ghanshyam V. , Patel Hasmukh A. , Ingole Pravin G. , Mody Haresh M. , Bajaj Hari C.
Published: September 2009

Sustained intestinal delivery of thiamine hydrochloride (Vitamin B 1; VB1) and pyridoxine hydrochloride (Vitamin B6; VB6) seems to be a feasible alternative to existing therapy. The vitamins (VB1/VB6) intercalated in montmorillonite (MMT) and intercalated VB1/VB6-MMT hybrid is further used for synthesis of VB1/VB6-MMT-alginate nanocomposite beads by gelation method and in vitro release in the intestinal environment. The structure and surface morphology of the synthesized VB1/VB 6-MMT hybrid, VB1/VB6-alginate and VB1/VB 6-MMT-alginate nanocomposite beads were characterized by XRD, FT-IR, TGA and SEM. In vitro release experiments revealed that the VB1/VB 6 releases suddenly from VB1/VB6-MMT hybrid and is pH dependent. The controlled release of VB1/VB6 from VB1/VB6-MMT-alginate nanocomposite beads was observed to be controlled as compared to their release from VB1/VB 6-MMT hybrid and VB1/VB6-alginate beads.
Polysulfone-based Ionomers for Fuel Cell Applications
Authors: Iojoiu Cristina , Sanchez Jean-Yves
Published: September 2009

This paper is a review that is focused on ionomers based on aromatic polysulfone backbone and intended to be used in proton exchange membrane fuel cells or in direct methanol fuel cells. Emphasis is placed on the different chemical routes to prepare the ionomers. Special attention is given to the impact of the ionomer structure on the conductivity performance and on the dimensional stability of the membranes at high temperatures.
The web could spell catastrophe
Authors: Hagerty Bill
Published: September 2009

Mort Zuckerman, property billionaire, media magnate and intellectual giant, says papers need a new business model — and customers must pay
Fear and loathing in tabloid Britain
Authors: Mullin Chris
Published: September 2009

Of course politicians were culpable but for years there have been media lies about MPs. So have we lost all sense of proportion?
FOI: Whitehall strikes back
Authors: Hayes Jeremy
Published: September 2009

The idea of the public’s “right to know” is fading fast as politicians and officials try to limit freedom of information
Media ineptitude? We’ve been framed
Authors: Simpson Daniel
Published: September 2009

As views that frame stories shape their message, “free market”and “growth” propagandists control the news, believes one commentator
Congrats BJR, (but do try harder)
Authors: Letts Quentin
Published: September 2009

A distinguished critic reviews 20 years of British Journalism Review and concludes it’s OK, but that Derek Birdsall is brilliant
Scientific and logistical challenges in designing the CONTROL trial: recombinant factor VIIa in severe trauma patients with refractory bleeding
Authors: Dutton Richard , Hauser Carl , Boffard Kenneth , Dimsitts Jeannett , Bernard Gordon , Holcomb John , Leppäniemi Ari , Tortella Bartholemew , Bouillon Bertil , CONTROL Steering Committee
Published: September 2009

Background Clinical research in trauma patients poses multiple challenges in study design. These reflect the heterogeneity of injury and treatment, the paucity of acceptable study endpoints aside from mortality, and the difficulties inherent in obtaining informed consent in acutely ill populations. A current example of this problem is the study of recombinant factor VIIa (rFVIIa), which has attracted considerable interest as a systemic procoagulant agent for use in trauma patients with exsanguinating hemorrhage.

Purpose To report on the implementation of an international trial — CONTROL — intended to assess the efficacy and safety of rFVIIa in trauma, and discuss trauma research study design in light of this experience.

Methods The CONTROL trial international steering committee confronted a number of barriers in the design of the CONTROL trial. They addressed methodologies for (1) standardizing entry criteria for trauma patients suffering inherently heterogeneous injuries, (2) obtaining informed consent in an acutely injured population with altered levels of consciousness, (3) avoiding futile care, while recruiting subjects with incompletely diagnosed injuries, (4) standardizing trauma intensive care across different investigating sites and countries, and (5) establishing study endpoints that were both clinically relevant and convincing to regulatory authorities. The resulting study methodology is reported.

Results The CONTROL trial began active recruitment in October 2005, and was halted on June 11, 2008 because the observed mortality in the 576 enrolled patients was so far below expectations that the study would lack sufficient statistical power at the planned number of subjects to demonstrate a benefit. The utility of the endpoints selected for study will not be known until completion of data analysis. Limitations Any clinical trial in trauma patients must cope with the urgency of care required, issues of patient heterogeneity, standardization of care across multiple centers, and the difficulties of obtaining informed consent.

Conclusion Research in acutely hemorrhaging trauma patients presents numerous scientific and ethical challenges. The methodology of the CONTROL study is presented as an example of how some of these challenges can be approached and managed, and of the pitfalls that may arise. Clinical Trials 2009; 6: 467—479. http://ctj.sagepub.com
Transforming growth factor-β–induced protein (TGFBIp/β ig-h3) activates platelets and promotes thrombogenesis
Authors: Kim Ha-Jeong , Kim Pan-Kyung , Bae Sang Mun , Son Hye-Nam , Thoudam Debraj Singh , Kim Jung-Eun , Lee Byung-Heon , Park Rang-Woon , Kim In-San
Published: September 2009

Transforming growth factor-β–induced protein (TGFBIp)/βig-h3 is a 68-kDa extracellular matrix protein that is functionally associated with the adhesion, migration, proliferation, and differentiation of various cells. The presence of TGFBIp in platelets led us to study the role of this protein in the regulation of platelet functions. Upon activation, platelet TGFBIp was released and associated with the platelets. TGFBIp mediates not only the adhesion and spread of platelets but also activates them, resulting in phosphatidylserine exposure, α-granule secretion, and increased integrin affinity. The fasciclin 1 domains of TGFBIp are mainly responsible for the activation of platelets. TGFBIp promotes thrombus formation on type I fibrillar collagen under flow conditions in vitro and induces pulmonary embolism in mice. Moreover, transgenic mice, which have approximately a 1.7-fold greater blood TGFBIp concentration, are significantly more susceptible to collagen- and epinephrine-induced pulmonary embolism than wild-type mice. These results suggest that TGFBIp, a human platelet protein, plays important roles in platelet activation and thrombus formation. Our findings will increase our understanding of the novel mechanism of platelet activation, contributing to a better understanding of thrombotic pathways and the development of new antithrombotic therapies.
BMS-214662 induces mitochondrial apoptosis in chronic myeloid leukemia (CML) stem/progenitor cells, including CD34+38− cells, through activation of protein kinase Cβ
Authors: Pellicano Francesca , Copland Mhairi , Jorgensen Heather G. , Mountford Joanne , Leber Brian , Holyoake Tessa L.
Published: September 2009

Chronic myeloid leukemia (CML) is a hematopoietic stem cell disorder maintained by cancer stem cells. To target this population, we investigated the mechanism of action of BMS-214662, developed as a farnesyl transferase inhibitor (FTI) and unique in inducing apoptosis in these cells. By contrast, a related congener and equally effective FTI, BMS-225975 does not induce apoptosis, indicating a novel mechanism of action. BMS-214662 significantly and selectively induced apoptosis in primitive CD34+38− CML compared with normal cells. Apoptosis proceeded via the intrinsic pathway: Bax conformational changes, loss of mitochondrial membrane potential, generation of reactive oxygen species, release of cytochrome c, and caspase-9/3 activation were noted. Up-regulation of protein kinase Cβ (PKCβ), down-regulation of E2F1, and phosphorylation of cyclin A–associated cyclin-dependent kinase 2 preceded these changes. Cotreatment of CML CD34+ and CD34+38− cells with PKC modulators, bryostatin-1, or hispidin markedly decreased these early events and the subsequent apoptosis. None of these events was elicited by BMS-214662 in normal CD34+ cells or by BMS-225975 in CML CD34+ cells. These data suggest that BMS-214662 selectively elicits a latent apoptotic pathway in CML stem cells that is initiated by up-regulation of PKCβ and mediated by Bax activation, providing a molecular framework for development of novel therapeutics.
Rac1 activation inhibits E-cadherin-mediated adherens junctions via binding to IQGAP1 in pancreatic carcinoma cells
Authors: Hage Beatrix , Meinel Katrin , Baum Iris , Giehl Klaudia , Menke Andre
Published: September 2009

Background

Monomeric GTPases of the Rho family control a variety of cellular functions including actin cytoskeleton organisation, cell migration and cell adhesion. Defects in these regulatory processes are involved in tumour progression and metastasis. The development of metastatic carcinoma is accompanied by deregulation of adherens junctions, which are composed of E-cadherin/β- and α-catenin complexes.

Results

Here, we show that the activity of the monomeric GTPase Rac1 contributes to inhibition of E-cadherin-mediated cell-cell adhesion in pancreatic carcinoma cells. Stable expression of constitutively active Rac1(V12) reduced the amount of E-cadherin on protein level in PANC-1 pancreatic carcinoma cells, whereas expression of dominant negative Rac1(N17) resulted in an increased amount of E-cadherin. Extraction of proteins associated with the actin cytoskeleton as well as coimmunoprecipitation analyses demonstrated markedly decreased amounts of E-cadherin/catenin complexes in Rac1(V12)-expressing cells, but increased amounts of functional E-cadherin/catenin complexes in cells expressing Rac1(N17). Cell aggregation and migration assays revealed, that cells containing less E-cadherin due to expression of Rac1(V12), exhibited reduced cell-cell adhesion and increased cell motility. The Rac/Cdc42 effector protein IQGAP1 has been implicated in regulating cell-cell adhesion. Coimmunoprecipitation studies showed a decrease in the association between IQGAP1 and β-catenin in Rac1(V12)-expressing PANC-1 cells and an association of IQGAP1 with Rac1(V12). Elevated association of IQGAP1 with the E-cadherin adhesion complex via β-catenin correlated with increased intercellular adhesion of PANC-1 cells.

Conclusion

These results indicate that active Rac1 destabilises E-cadherin-mediated cell-cell adhesion in pancreatic carcinoma cells by interacting with IQGAP1 which is associated with a disassembly of E-cadherin-mediated adherens junctions. Inhibition of Rac1 activity induced increased E-cadherin-mediated cellular adhesion.
A practical approach for the validation of sterility, endotoxin and potency testing of bone marrow mononucleated cells used in cardiac regeneration in compliance with good manufacturing practice
Authors: Soncin Sabrina , Lo Cicero Viviana , Astori Giuseppe , Soldati Gianni , Gola Mauro , Sürder Daniel , Moccetti Tiziano
Published: September 2009

Background

Main scope of the EU and FDA regulations is to establish a classification criterion for advanced therapy medicinal products (ATMP). Regulations require that ATMPs must be prepared under good manufacturing practice (GMP). We have validated a commercial system for the determination of bacterial endotoxins in compliance with EU Pharmacopoeia 2.6.14, the sterility testing in compliance with EU Pharmacopoeia 2.6.1 and a potency assay in an ATMP constituted of mononucleated cells used in cardiac regeneration.

Methods

For the potency assay, cells were placed in the upper part of a modified Boyden chamber containing Endocult Basal Medium with supplements and transmigrated cells were scored. The invasion index was expressed as the ratio between the numbers of invading cells relative to cell migration through a control insert membrane.

For endotoxins, we used a commercially available system based on the kinetic chromogenic LAL-test. Validation of sterility was performed by direct inoculation of TSB and FTM media with the cell product following Eu Ph 2.6.1 guideline.

Results and discussion

The calculated MVD and endotoxin limit were 780× and 39 EU/ml respectively. The 1:10 and 1:100 dilutions were selected for the validation. For sterility, all the FTM cultures were positive after 3 days. For TSB cultures, Mycetes and B. subtilis were positive after 5 and 3 days respectively. The detection limit was 1-10 colonies.

A total of four invasion assay were performed: the calculated invasion index was 28.89 ± 16.82% (mean ± SD).

Conclusion

We have validated a strategy for endotoxin, sterility and potency testing in an ATMP used in cardiac regeneration. Unlike pharmaceutical products, many stem-cell-based products may originate in hospitals where personnel are unfamiliar with the applicable regulations. As new ATMPs are developed, the regulatory framework is likely to evolve. Meanwhile, existing regulations provide an appropriate structure for ensuring the safety and efficacy of the next generation of ATMPs. Personnel must be adequately trained on relevant methods and their application to stem-cell-based products.
The formation of cysteine-linked dimers of BST-2/tetherin is important for inhibition of HIV-1 virus release but not for sensitivity to Vpu
Authors: Andrew Amy J , Miyagi Eri , Kao Sandra , Strebel Klaus
Published: September 2009

Background

The Human Immunodeficiency virus type 1 (HIV-1) Vpu protein enhances virus release from infected cells and induces proteasomal degradation of CD4. Recent work identified BST-2/CD317 as a host factor that inhibits HIV-1 virus release in a Vpu sensitive manner. A current working model proposes that BST-2 inhibits virus release by tethering viral particles to the cell surface thereby triggering their subsequent endocytosis.

Results

Here we defined structural properties of BST-2 required for inhibition of virus release and for sensitivity to Vpu. We found that BST-2 is modified by N-linked glycosylation at two sites in the extracellular domain. However, N-linked glycosylation was not important for inhibition of HIV-1 virus release nor did it affect surface expression or sensitivity to Vpu. Rodent BST-2 was previously found to form cysteine-linked dimers. Analysis of single, double, or triple cysteine mutants revealed that any one of three cysteine residues present in the BST-2 extracellular domain was sufficient for BST-2 dimerization, for inhibition of virus release, and sensitivity to Vpu. In contrast, BST-2 lacking all three cysteines in its ectodomain was unable to inhibit release of wild type or Vpu-deficient HIV-1 virions. This defect was not caused by a gross defect in BST-2 trafficking as the mutant protein was expressed at the cell surface of transfected 293T cells and was down-modulated by Vpu similar to wild type BST-2.

Conclusion

While BST-2 glycosylation was functionally irrelevant, formation of cysteine-linked dimers appeared to be important for inhibition of virus release. However lack of dimerization did not prevent surface expression or Vpu sensitivity of BST-2, suggesting Vpu sensitivity and inhibition of virus release are separable properties of BST-2.
Genotype distribution of cervical human papillomavirus DNA in women with cervical lesions in Bioko, Equatorial Guinea
Authors: García-Espinosa Benjamín , Nieto-Bona MaPaz , Rueda Sonsoles , Silva-Sánchez Luís Fernando , Piernas-Morales MaConcepción , Carro-Campos Patricia , Cortés-Lambea Luís , Moro-Rodríguez Ernesto
Published: September 2009

Background

The HVP vaccine is a useful tool for preventing cervical cancer. The purpose of this study is to determine the most frequent HPV genotypes in Equatorial Guinea in order to develop future vaccination strategies to apply in this country.

Methods

A campaign against cervical cancer was carried out in the area on a total of 1,680 women. 26 of the women, following cytological screening, were treated surgically with a loop electrosurgical excision procedure (LEEP). Cases were studied histologically and were genotyped from paraffin blocks by applying a commercial kit that recognized 35 HPV types.

Results

Cytological diagnoses included 17 HSIL, 1 LSIL, 5 ASC-H and 3 AGUS. Histological diagnosis resulted in 3 cases of microinvasive squamous cell carcinoma stage IA of FIGO, 9 CIN-3, 8 CIN-2, 2 CIN-1, 3 flat condylomas and mild dysplasia of the endocervical epithelium. Fifteen of twenty-five cases genotyped were positive for HPV (60%). HPV 16 and 33 were identified in four cases each, HPV 58 in two other cases, and HPV 18, 31, 52, and 82 in one case, with one HPV 16 and 58 coinfection.

Conclusion

The frequency of HPV types in the African area varies in comparison to other regions, particularly in Europe and USA. Vaccination against the five most common HPV types (16, 33, 58, 18, and 31) should be considered in the geographic region of West Africa and specifically in Equatorial Guinea.
Steroid Receptor RNA Activator Protein (SRAP): a potential new prognostic marker for estrogen receptor-positive/node-negative/younger breast cancer patients
Authors: Yan Yi , Skliris George P , Penner Carla , Chooniedass-Kothari Shilpa , Cooper Charlton , Nugent Zoann , Blanchard Anne , Watson Peter H , Myal Yvonne , Murphy Leigh C , Leygue Etienne
Published: September 2009

Introduction

The steroid receptor RNA activator is a functional RNA suspected to participate in the mechanisms underlying breast tumor progression. This RNA is also able to encode for a protein, Steroid Receptor RNA Activator Protein (SRAP), whose exact function remains to be determined. Our aim was to assess, in a large breast cancer cohort, whether levels of this protein could be associated with outcome or established clinical parameters.

Methods

Following antibody validation, SRAP expression was assessed by tissue-microarray (TMA) analysis of 372 breast tumors. Clinical follow-up and parameters such as steroid receptor and node status were available for all the corresponding cases. Immunohistochemical scores were independently determined by three investigators and averaged. Statistical analyses were performed using standard univariate and multivariate tests.

Results

SRAP levels were significantly (Mann-Whitney rank sum test, P < 0.05) higher in estrogen receptor-alpha positive (ER+, n = 271), in progesterone receptor positive (PR+, n = 257) and in older patients (age > 64 years, n = 182). When considering ER+ tumors, PR+ tumors, or younger patients (≤ 64 years), cases with high SRAP expression had a significantly (Mantel-Cox test, P < 0.05) worse breast cancer specific survival (BCSS) than those with low SRAP levels. SRAP also appeared as a very powerful indicator of poor prognostic for BCSS in the subset of ER+, node negative and young breast cancer patients (Cox regression analysis, n = 60, BCSS Hazard Ratio = 8.61, P < 0.006).

Conclusions

Our data suggest that SRAP levels might provide additional information on potential risk of recurrence and negative outcome in a specific set of patients with otherwise good prognosis when considering only estrogen receptor and nodal status.
Nebulized antithrombin limits bacterial outgrowth and lung injury in Streptococcus pneumoniae pneumonia in rats
Authors: Hofstra Jorrit J , Cornet Alexander D , de Rooy Bart F , Vlaar Alexander P , van der Poll Tom , Levi Marcel , Zaat Sebastian AJ , Schultz Marcus J
Published: September 2009

Introduction

Disturbed alveolar fibrin turnover is a cardinal feature of severe pneumonia. Clinical studies suggest that natural inhibitors of coagulation exert lung-protective effects via anticoagulant and possibly also anti-inflammatory pathways. Intravenous infusion of the natural anticoagulants increases the risk of bleeding. Local administration may allow for higher treatment dosages and increased local efficacy while at the same time reducing the risk of bleeding. We evaluated the effect of nebulized anticoagulants on pulmonary coagulopathy and inflammation in a rat model of Streptococcus pneumoniae pneumonia.

Methods

In this randomized controlled in vivo laboratory study rats were challenged intratracheally with S. pneumoniae, inducing pneumonia, and randomized to treatment with normal saline (placebo), recombinant human activated protein C (rh-APC), plasma-derived antithrombin (AT), heparin or danaparoid, by means of nebulization.

Results

S. pneumoniae infection increased pulmonary levels of thrombin-antithrombin complexes and fibrin degradation products. All nebulized anticoagulants significantly limited pulmonary coagulopathy. None of the agents except danaparoid resulted in changes in systemic coagulopathy. Treatment with plasma-derived AT reduced outgrowth of S. pneumoniae and histopathologic damage in lungs. In vitro experiments confirmed outgrowth was reduced in bronchoalveolar lavage fluid (BALF) from rats treated with plasma-derived AT compared with placebo. Neutralizing of cationic components in BALF diminished the inhibitory effects on bacterial outgrowth of BALF, suggesting a role for cationic antimicrobial proteins.

Conclusions

Nebulization of anticoagulants attenuates pulmonary coagulopathy during S. pneumoniae pneumonia in rats while only danaparoid affects systemic coagulation. Nebulized plasma-derived AT reduces bacterial outgrowth and exerts significant lung-protective effects.
Amphiregulin and Epiregulin mRNA Expression in Primary Tumors Predicts Outcome in Metastatic Colorectal Cancer Treated With Cetuximab
Authors: Jacobs Bart , De Roock Wendy , Piessevaux Hubert , Van Oirbeek Robin , Biesmans Bart , De Schutter Jef , Fieuws Steffen , Vandesompele Jo , Peeters Marc , Van Laethem Jean-Luc , Humblet Yves , Pénault-Llorca Frederique , De Hertogh Gert , Laurent-Puig Pierre , Van Cutsem Eric , Tejpar Sabine
Published: September 2009

Purpose

To study the power of the epidermal growth factor receptor (EGFR) epiregulin (EREG) and amphiregulin (AREG) ligands' expression in primary tumors to predict the outcome in patients with chemorefractory metastatic colorectal cancer (cmCRC) treated with the combination of cetuximab and irinotecan.

Patients and Methods

Gene expression measurements and KRAS mutation analysis were performed on archival formalin-fixed paraffin-embedded primary tumors of 220 cmCRC patients. Response was measured using RECIST (Response Evaluation Criteria in Solid Tumors) criteria. The relation between ligand expression levels and outcome was evaluated using logistic regression for response and Cox regression for survival data. Receiver operating characteristics analysis was performed for response and survival data. CIs for the performance indices were obtained with a nonparametric bootstrap procedure. Findings were externally validated on a series of 67 samples treated in a similar setting.

Results

In KRAS wild type (WT) patients, there was a significant association between log-transformed ligand expression and response for EREG (odds ratio for objective response, 1.90; 95% CI, 1.27 to 2.83; P = .0005; concordance index [c-index], 0.681) and for AREG (odds ratio for objective response, 1.862; 95% CI, 1.22 to 2.72; P = .0017; c-index, 0.673). In a Cox regression model, dichotomized ligand expression was significantly associated with progression-free survival (PFS) and overall survival (OS). EREG PFS hazard ratio (HR) was 0.41 (95% CI, 0.274 to 0.609; P < .001; time-dependent c-index [Cτ index], 0.640), and AREG PFS HR was 0.43 (95% CI, 0.29 to 0.64; P < .001; Cτ index, 0.627). EREG OS HR was 0.42 (95% CI, 0.28 to 0.63; P < .0001; Cτ index, 0.639), and AREG OS HR was 0.40 (95% CI, 0.27 to 0.64; P < .0001; Cτ index, 0.625). There was no predictive power of ligand expression in patients with KRAS mutation.

Conclusion

Expression of EGFR ligands in primary tumors significantly predicts outcome in KRAS WT cmCRC treated with cetuximab and irinotecan.