bluuebunny/crossref_metadata_2025 huggingface dataset:
Blood puncture as a nondestructive sampling tool to obtain DNA in frogs: comparison of protocols and survival analysis
Authors: MENDOZA A. M. , GARCÍA‐RAMÍREZ J. C. , CÁRDENAS‐HENAO H.
Published: January 2012

In molecular biology studies of Anura, nondestructive methods to obtain genetic material are needed as alternatives to toe clipping. This work evaluates a nondestructive method for sampling DNA from blood puncture, comparing the performance of three different extraction protocols (Qiagen Kit, Salting‐out and Chelex). We collected 134 individuals of Eleutherodactylus johnstonei, extracting blood via puncture of the medial vein using commercial‐grade glucometer lancets. We extracted 100–1880 ng DNA, finding no differences between the extraction protocols. We compared the quality of the resulting DNA through amplification and sequencing of the 16S mitochondrial gene. Amplification was successful for the three extraction protocols, although Chelex showed better performance, making it the most recommendable protocol for extraction of DNA from blood. The resulting sequences corresponded to those registered in the GenBank for this species. Additionally, we found no significant differences in survival or weight change between the individuals that were manipulated and a control group (mean survival 66.7% treated, 62.9% untreated). Data reveal that blood samples obtained by puncture are a convenient alternative to other tissues (phalange, buccal swab, liver) that have traditionally been used as DNA sources for anurans. The technique is applicable to small and large species, covering most anuran diversity, provides enough DNA for many genetic applications and produces no noticeable effect on the survival or performance, given that it does not affect the motor parts or the dexterity of the animals.
Late‐evening snack with branched‐chain amino acids improves liver function after radiofrequency ablation for hepatocellular carcinoma
Authors: Morihara Daisuke , Iwata Kaoru , Hanano Takayuki , Kunimoto Hideo , Kuno Shizuka , Fukunaga Atsushi , Yotsumoto Kaoru , Takata Kazuhide , Tanaka Takashi , Sakurai Kunitoshi , Iwashita Hideyuki , Ueda Shu‐ichi , Hirano Genryu , Yokoyama Keiji , Nakane Hidetoshi , Nishizawa Shinya , Yoshikane Makoto , Anan Akira , Takeyama Yasuaki , Kakumitsu Shigeru , Kitamura Yuji , Sakamoto Masaharu , Irie Makoto , Shakado Satoshi , Sohda Tetsuro , Watanabe Hiroshi , Sakisaka Shotaro
Published: January 2012

Aim:  This prospective study was designed to examine whether consumption of a branched‐chain amino acid (BCAA)‐enriched nutrient mixture as a late‐evening snack (LES) helps maintain and/or improve liver functioning in liver cirrhosis (LC) patients who have undergone radiofrequency ablation (RFA) for hepatocellular carcinoma (HCC).

Methods:  An equal number (10) of 30 LC patients who had undergone RFA for HCC was randomly assigned to a standard diet group (control) group, a morning BCAA (M‐BCAA) administration group, or a LES with BCAA (LES‐BCAA) administration group. Liver function testing was performed and Child–Pugh scores (CPS) calculated for each group to assess the improvement at 1, 4 and 12 weeks post‐RFA.

Results:  Compared to the control and M‐BCAA groups, the LES‐BCAA group experienced a rapid and significant improvement in albumin and total serum bilirubin levels and in CPS that began during the initial post‐RFA period. These results indicate that LES with BCAA supplementation significantly improved the CPS of the LES‐BCAA group at 4 and 12 weeks post‐RFA. Although no patients experienced serious adverse effects, two patients who had been diagnosed with diabetes mellitus before undergoing RFA required blood sugar management to improve glycemic control and one subject withdrew due to supplement‐induced vomiting.

Conclusion:  LES with BCAA supplementation significantly and rapidly improves liver functioning and CPS in LC patients who have undergone RFA for HCC. Control of blood sugar levels is necessary when calorie‐containing BCAA is administrated to LC patients with impaired glucose tolerance.
Does Audit Market Concentration Harm the Quality of Audited Earnings? Evidence from Audit Markets in 42 Countries*
Authors: Francis Jere R. , Michas Paul N. , Seavey Scott E.
Published: January 2012

Audit regulators around the world have expressed concern over market dominance by Big 4 accounting firms and the potential adverse effect it may have on the quality of audited financial statements. We use cross‐country variation in the audit market structure of 42 countries to examine two separate aspects of Big 4 dominance: (1) Big 4 market concentration as a group relative to non–Big 4 auditors; and (2) concentration within the Big 4 group in which one or more of the Big 4 firms is dominant relative to the other Big 4 firms. We find that in countries where the Big 4 (as a group) conduct more listed company audits, both Big 4 and non–Big 4 clients have higher quality audited earnings compared to clients in countries with smaller Big 4 market shares. In contrast, in countries where there is a greater concentration within the Big 4 group, we find that Big 4 clients have lower quality audited earnings compared to countries with more evenly distributed market shares among the Big 4. Thus concentration within the Big 4 group appears to be detrimental to audit quality in a country and of legitimate concern to regulators and policymakers. However, Big 4 dominance per se does not appear to harm audit quality and is in fact associated with higher earnings quality, after controlling for other country characteristics that potentially affect earnings quality.
Output Information Based Iterative Learning Control Law Design With Experimental Verification
Authors: Hladowski Lukasz , Galkowski Krzysztof , Cai Zhonglun , Rogers Eric , Freeman Chris T. , Lewin Paul L.
Published: January 2012

This paper considers iterative learning control law design using the theory of linear repetitive processes. This setting enables trial-to-trial error convergence and along-the-trial performance to be considered simultaneously in the design. It is also shown that this design extends naturally to include robustness to unmodeled plant dynamics. The results from experimental application of these laws to a gantry robot performing a pick and place operation are given, together with a discussion of the positioning of this approach relative to alternatives and possible further research.
Optimal Sliding Mode Cascade Control for Stabilization of Underactuated Nonlinear Systems
Authors: Muske Kenneth R. , Ashrafiuon Hashem , Nersesov Sergey , Nikkhah Mehdi
Published: January 2012

This paper presents an optimal sliding mode cascade control for stabilization of a class of underactuated nonlinear mechanical systems. A discrete-time, nonlinear model predictive control structure is used to optimally select and update the parameters of the sliding mode control surfaces at specified intervals in order to achieve a desired performance objective. The determination of these surface parameters is subject to constraints that arise from the stability conditions imposed by the sliding mode control law and the physical limits on the system such as input saturation. Nominal stability of the optimal cascade control structure is demonstrated and its robust performance is illustrated using an experimental rotary inverted pendulum system.
A New Mechanistic Approach for Micro End Milling Force Modeling
Authors: Jun Martin B. G. , Goo Chanseo , Malekian Mohammad , Park Simon
Published: January 2012

This paper investigates the mechanistic modeling of micro end milling forces, with consideration of the effects of plowing, elastic recovery, effective rake angle, and flank face rubbing. Two different mechanistic models are developed for shearing- and plowing-dominant regimes. Micro end milling experiments are conducted to validate the model for Aluminum 6061; and, the model appropriately predicts force profiles for a wide range of feed rates, and prediction of the root mean square (RMS) values of the resultant forces is, on average, within a 12% error. The study of the model shows that plowing and rubbing force contributions are significant, especially at low feed rates. The edge radius is found to have a significant effect on plowing and rubbing force components and the effective rake angle, which indicates that it is important to maintain a low edge radius to reduce micro end milling forces.
The Interactions of Microhole Sidewall With Plasma induced by Femtosecond Laser Ablation in High-Aspect-Ratio Microholes
Authors: Wu Benxin , Tao Sha , Lei Shuting
Published: January 2012

High-aspect-ratio microholes have many important applications, but their drilling is very challenging. Femtosecond (fs) laser ablation provides a potential solution, but involves many complicated physical processes that have not been well understood, which have hindered its practical application. One of these is that the plasma induced by laser ablation at the hole bottom will transfer some of its energy to the hole sidewall as it expands in the microhole. The plasma–sidewall interaction has been rarely studied in literature, and it is still not clear if or not the energy transferred from the plasma is sufficient to cause significant material removal from the sidewall. Direct time-resolved observations are extremely difficult due to the small temporal/spatial scales and the spatial constraint inside the hole, while the sidewall characterization after laser ablation is difficult to distinguish between the possible material removal due to plasma energy transfer and that due to direct laser energy absorption by the sidewall. In this paper, a physics-based model is applied as the investigation tool to study the plasma–sidewall interaction in fs laser drilling of high-aspect-ratio microholes. It has been found that for the studied conditions the energy transferred from the plasma is not sufficient to cause significant material removal from the sidewall through any thermally induced phase change process.
Workpiece Thermal Distortion in Minimum Quantity Lubrication Deep Hole Drilling—Finite Element Modeling and Experimental Validation
Authors: Tai Bruce L. , Jessop Andrew J. , Stephenson David A. , Shih Albert J.
Published: January 2012

This paper presents the three dimensional (3-D) finite element analysis (FEA) to predict the workpiece thermal distortion in drilling multiple deep-holes under minimum quantity lubrication (MQL) condition. Heat sources on the drilling hole bottom surface (HBS) and hole wall surface (HWS) are first determined by the inverse heat transfer method. A 3-D heat carrier consisting of shell elements to carry the HWS heat flux and solid elements to carry the HBS heat flux has been developed to conduct the heat to the workpiece during the drilling simulation. A thermal–elastic coupled FEA was applied to calculate the workpiece thermal distortion based on the temperature distribution. The concept of the heat carrier was validated by comparing the temperature calculation with an existing 2-D advection model. The 3-D thermal distortion was validated experimentally on an aluminum workpiece with four deep-holes drilled sequentially. The measured distortion on the reference point was 61 μm, which matches within uncertainty the FEA predicted distortion of 51 μm.
In situ fabrication of blue ceramic coatings on wrought Al Alloy 2024 by plasma electrolytic oxidation
Authors: Wang Zhijiang , Nie Xueyuan , Hu Henry , Hussein Riyad O.
Published: January 2012

In situ formation of ceramic coatings on 2024 Al alloy with a blue color was successfully achieved using a plasma electrolytic oxidation process working at atmospheric pressure. This novel blue ceramic coating overcomes the shortcomings of surface treatments resulting from conventional dyeing processes by depositing organic dyes into the porous structure of anodic film, which has poor resistance to abrasion and rapid fading when exposed to sunlight. X-ray diffraction, scanning electron microscopy, and energy dispersive spectroscopy were employed to characterize the microstructure of the blue ceramic coating. The fabricated ceramic coating was composed of CoAl2O4, α-Al2O3, and γ-Al2O3. By controlling the working parameters, the distribution of the CoAl2O4 phase on the surface can be adjusted, and plays a key role in the appearance of the coating. Electrochemical testing, thermal cycling method, and pin-on-disk sliding wear testing were employed to evaluate corrosion, thermal cycling, and wear resistance of the ceramic coatings. The results indicate that the blue ceramic coating has a similar polarization resistance to that of conventional anodic film and can significantly enhance the corrosion resistance of aluminum alloy. There are no destructive horizontal cracks observed within the blue ceramic coating when subjected to 120 times of thermal cycling, which heats the samples up to 573 K and followed by submersion in water at room temperature for 10 min. Compared with the aluminum substrate as well as a conventional anodic film coated aluminum sample, the wear resistance of the blue ceramic coating coated sample was significantly increased while the coefficient of friction was decreased from 0.34 to 0.14.
Integration of a thin film III–V edge emitting laser and a polymer microring resonator on an SiO2/Si substrate
Authors: Palit Sabarni , Royal Matthew , Jokerst Nan , Kirch Jeremy , Mawst Luke
Published: January 2012

The planar integration of on-chip laser sources with optical sensing elements, waveguide optical interconnect, and detectors enables the implementation of portable, efficient chip-scale systems, with applications in areas that include medical, environmental, biological, and chemical sensing systems. In this paper, the planar integration of a thin film strain compensated In0.2Ga0.8As/GaAs single quantum well laser with a tapered polymer (SU-8-2002) waveguide and overlap coupled SU-8-2002 microring resonator is achieved on an SiO2/Si platform. Two laser/waveguide configurations, one with the tapered waveguide separated from the laser front facet and the other with the waveguide overlapping the front facet, were demonstrated, and this laser/waveguide structure was then integrated with a polymer microring resonator on SiO2/Si. Lasing operation for the integrated system was verified through power-current characteristics and spectra at the through and drop ports of the microring resonator. This integrated system forms an essential component toward realizing planar chip-scale optical sensing systems.
Homoepitaxial N-polar GaN layers and HEMT structures grown by rf-plasma assisted molecular beam epitaxy
Authors: Storm D. F. , Meyer D. J. , Katzer D. S. , Binari S. C. , Paskova Tanya , Preble E. A. , Evans K. R. , Zhou Lin , Smith David J.
Published: January 2012

The authors have investigated the growth and structural and electrical properties of homoepitaxial GaN layers and GaN/AlGaN heterostructures grown on free-standing, hydride vapor phase epitaxy grown, N-polar GaN:Fe substrates by rf-plasma molecular beam epitaxy. Secondary-ion mass spectroscopic analysis of unintentionally doped and Be-doped N-polar GaN layers indicate that oxygen is the dominant impurity in all layers and is largely insensitive to growth temperature in the range investigated (675 °C < TS < 760 °C). Transmission electron microscopy (TEM) indicates that threading dislocations are generated at the regrowth interface in these samples; in contrast to homoepitaxial growth on Ga-polar GaN, and that the density of threading dislocations diminishes as the growth temperature increases. However, examination by TEM indicates that threading dislocations are not generated at the regrowth interface of samples subjected to pregrowth substrate surface cleaning by gallium deposition and desorption and subsequent growth of ultrathin (15 Å) initial AlN layers. N-polar GaN/AlGaN heterostructures grown on Be-doped homoepitaxial N-polar GaN buffers exhibit low buffer leakage and Hall mobilities up to 1680 cm2/Vs at sheet densities of 1.3 × 1013 cm−2. High electron mobility transistors have been fabricated on these structures; drain current densities over 700 mA/mm and breakdown voltages as high as 70 V have been measured.
Determination of Drimarene Blue X-BLN at a glassy carbon electrode by differential pulse voltammetry
Authors: Radi Abd-Elgawad M. , Mostafa Mohammed R. , Elshafey Reda M. , Hegazy Talaat A.
Published: January 2012

The electrochemical oxidation behaviour of Drimarene Blue X-BLN (DB) has been investigated in phosphate buffers (pH 2.54–10.18) by cyclic and differential pulse voltammetry (DPV) at a glassy carbon electrode (GCE). The oxidation of DB dye generated well-defined pH-dependent two pairs of quasi-reversible anodic-cathodic peak couples. DB exhibited the second redox couple over the entire pH range, while the first redox couple disappeared for pH ≥ 6.70. The redox processes were adsorption-controlled. An electroanalytical method was developed for the determination of DB in phosphate buffer solution (pH 2.85) as supporting electrolyte using DPV. The anodic current heights varied linearly with DB concentrations in the ranges 2 × 10–6–3 × 10–5 and 6 × 10–6–3 × 10–5 mol l–1 with limits of detection (LOD) of 8.7 × 10–7 and 5.7 × 10–7 mol l–1 and limits of quantification (LOQ) of 2.9 × 10–6 and 1.9 × 10–6 mol l–1 for the first and second anodic peaks, respectively. Validation parameters, such as accuracy, precision and recovery were evaluated. The proposed method was successfully applied to the determination of DB in tap water and the analytical results compared well with those obtained by the spectrophotometric method.
Effect of Acupuncture on Perception Threshold: A Randomised Controlled Trial
Authors: Wu Shuang , Yamaguchi Hidenori , Shibutani Koh
Published: January 2012

Objective

To determine the current perception thresholds (CPTs) for arm and mental foramen areas to enable a quantitative evaluation of the effectiveness of acupuncture treatment.

Methods

Ninety-eight volunteers enrolled as participants and were randomly assigned to one of three groups: an acupuncture group (34 subjects); a sham acupuncture group (32 subjects) and a waiting group (32 subjects). CPTs for the arm and mental foramen areas were determined before and after acupuncture at LI4 Hegu of the left hand. A Neurometer CPT was used to evaluate the perception threshold at the homolateral mental foramen and arm. For further exploration, thresholds of homolateral and contralateral mental foramina were determined before and after acupuncture in a subgroup of 13 participants in the acupuncture group.

Results

Acupuncture at LI4 increased the perception thresholds of the left mental foramen in the acupuncture group significantly (p<0.01). The increases were significantly greater than in the control group for all frequencies, and significantly greater than sham acupuncture for 250 Hz and 5 Hz. In the subgroup, only the CPTs at 5 Hz increased significantly for the contralateral mental foramen.

Conclusions

Acupuncture at LI4 increases the perception threshold in the mandibular area, but not in the arm. This finding is probably related to the analgesic effect of acupuncture.
Eosinophils in the oesophageal mucosa: clinical, pathological and epidemiological relevance in children: a cohort study
Authors: Cohen Marta C , Rao Prithviraj , Thomson Mike , Al-Adnani Mudher
Published: January 2012

Objectives

Eosinophilic oesophagitis (EO) shows eosinophilic infiltration of the mucosa and can present with symptoms indistinguishable from gastrooesophageal reflux disease (GORD). The authors describe the clinical, endoscopic and histopathological features of all cases of histological EO presenting during 2007–2008 with a 2-year follow-up. The incidence of paediatric EO and the features of a subgroup with features of both GORD and EO (‘overlap’ syndrome (OS)) are described.

Design

Biopsies with an average of 15 eosinophils/high-power field (HPF) were reviewed in the cohort. OS was suggested when there was coexistence of clinical and histological features of EO and GORD (abnormal pH study), which improved with proton pump inhibitors.

Setting

Tertiary care.

Participants

All cases with ≥15 eosinophils/HPF entered the study.

Primary outcome measures

Patients with EO had an average of 15 eosinophils/HPF.

Secondary outcome measures

Other histological features of EO included microabscesses, dilated intercellular spaces, basal cell hyperplasia, papillary elongation, etc.

Results

24 cases of EO were identified, 13 men and 11 women. The incidence of paediatric oesophageal eosinophilia in the region was 9/100 000 children. 11 of the 24 patients (46%) presented with some form of allergy, six with poor feeding/food aversion, five with dysphagia and four with vomiting. After follow-up, 56.5% were confirmed to have EO, 30.5% responded to treatment for GORD and were categorised as OS, 9% developed eosinophilic gastroenteritis and 4% did not have further upper gastrointestinal symptoms.

Conclusions

Accurate diagnosis of EO, especially the differentiation from GORD, requires appropriate clinicopathological correlation. A significant proportion of patients with eosinophilia in the mucosa also have GORD (OS). These patients improve after treating the underlying GORD. The study was registered as a Service Evaluation with the Trust (number SE74).
Cancer risk with folic acid supplements: a systematic review and meta-analysis
Authors: Wien Tale Norbye , Pike Eva , Wisløff Torbjørn , Staff Annetine , Smeland Sigbjørn , Klemp Marianne
Published: January 2012

Objective

To explore if there is an increased cancer risk associated with folic acid supplements given orally.

Design

Systematic review and meta-analysis of controlled studies of folic acid supplementation in humans reporting cancer incidence and/or cancer mortality. Studies on folic acid fortification of foods were not included.

Data sources

Cochrane Library, Medline, Embase and Centre of Reviews and Dissemination, clinical trial registries and hand-searching of key journals.

Results

From 4104 potential references, 19 studies contributed data to our meta-analyses, including 12 randomised controlled trials (RCTs). Meta-analysis of the 10 RCTs reporting overall cancer incidence (N=38 233) gave an RR of developing cancer in patients randomised to folic acid supplements of 1.07 (95% CI 1.00 to 1.14) compared to controls. Overall cancer incidence was not reported in the seven observational studies. Meta-analyses of six RCTs reporting prostate cancer incidence showed an RR of prostate cancer of 1.24 (95% CI 1.03 to 1.49) for the men receiving folic acid compared to controls. No significant difference in cancer incidence was shown between groups receiving folic acid and placebo/control group, for any other cancer type. Total cancer mortality was reported in six RCTs, and a meta-analysis of these did not show any significant difference in cancer mortality in folic acid supplemented groups compared to controls (RR 1.09, 95% CI 0.90 to 1.30). None of the observational studies addressed mortality.

Conclusions

A meta-analysis of 10 RCTs showed a borderline significant increase in frequency of overall cancer in the folic acid group compared to controls. Overall cancer incidence was not reported in the seven observational studies. Prostate cancer was the only cancer type found to be increased after folic acid supplementation (meta-analyses of six RCTs). Prospective studies of cancer development in populations where food is fortified with folic acid could indicate whether fortification similar to supplementation moderately increases prostate cancer risk.
Ethnic differences in disease presentation of uterine cancer in New Zealand women
Authors: Firestone Ridvan Tupai , Ellison-Loschmann Lis , Shelling Andrew N , Ekeroma Alec , Ikenasio-Thorpe Bettina A , Pearce Neil , Jeffreys Mona
Published: January 2012

Objectives

Little is known about the ethnic differences in disease presentation of uterine cancer in New Zealand women. The objectives of this study were two-fold: (1) to estimate the incidence and mortality of uterine cancer among women in New Zealand and (2) to examine the association of ethnicity and socioeconomic status with tumour stage and grade, at presentation of uterine cancer.

Methods

Retrospective survey of cancer cases identified from the New Zealand Cancer Registry. The authors analysed all 3203 uterine cancer cases registered with the New Zealand Cancer Registry during the period 1 January 1997 to 31 December 2006. Ethnic groups were defined based on the self-identified ethnicity recorded on the cancer registry: Ma–ori, Pacific and non-M–aori non-Pacific women. Socioeconomic status was categorised as quintiles of the New Zealand Deprivation Index 2006. The mortality to incidence ratio was used as a measure of prognosis. Logistic regression was used to estimate age, ethnic and deprivation adjusted odds ratios (ORs) and 95% confidence intervals (CIs).

Results

Pacific and M–aori women have higher incidence (32.4 and 17.7 per 100 000 women, respectively) and mortality rates of uterine cancer (12.1 and 7.4 per 100 000 women, respectively). Women in the most deprived areas are more likely to present with an advanced stage of uterine cancer (OR 1.64, 95% CI 1.09–2.48). M–aori and Pacific women are less likely to present with well-differentiated tumours (OR 0.69, 95% CI 0.52–0.92 and OR 0.72, 95% CI 0.52–0.99, respectively).

Conclusions

M–aori and Pacific women, and those from lower socioeconomic areas, are more likely to present with advanced uterine cancer.
COUNTING SINGULARITIES VIA FITTING IDEALS
Authors: JORGE-PÉREZ V. H. , MIRANDA A. J. , SAIA M. J.
Published: January 2012

The stable singularities of differential map germs constitute the main source of studying the geometric and topological behavior of these maps. In particular, one interesting problem is to find formulae which allow us to count the isolated stable singularities which appear in the discriminant of a stable deformation of a finitely determined map germ. Mond and Pellikaan showed how the Fitting ideals are related to such singularities and obtain a formula to count the number of ordinary triple points in map germs from ℂ2to ℂ3, in terms of the Fitting ideals associated with the discriminant.

In this article we consider map germs from (ℂn+m, 0) to (ℂm, 0), and obtain results to count the number of isolated singularities by means of the dimension of some associated algebras to the Fitting ideals. First in Corollary 4.5 we provide a way to compute the total sum of these singularities. In Proposition 4.9, for m = 3 we show how to compute the number of ordinary triple points. In Corollary 4.10 and with f of co-rank one, we show a way to compute the number of points formed by the intersection between a germ of a cuspidal edge and a germ of a plane.

Furthermore, we show in some examples how to calculate the number of isolated singularities using these results.
SOME RESULTS ON FUNDAMENTAL GROUPS AND BETTI NUMBERS OF FINSLER MANIFOLDS
Authors: SHEN YIBING , ZHAO WEI
Published: January 2012

In this paper the relationship between the Ricci curvature and the fundamental groups of Finsler manifolds are studied. We give an estimate of the first Betti number of a compact Finsler manifold. Two finiteness theorems for fundamental groups of compact Finsler manifolds are proved. Moreover, the growth of fundamental groups of Finsler manifolds with almost-nonnegative Ricci curvature are considered.
Lipid rafts are disrupted in mildly inflamed intestinal microenvironments without overt disruption of the epithelial barrier
Authors: Bowie Rachel V. , Donatello Simona , Lyes Clíona , Owens Mark B. , Babina Irina S. , Hudson Lance , Walsh Shaun V. , O'Donoghue Diarmuid P. , Amu Sylvie , Barry Sean P. , Fallon Padraic G. , Hopkins Ann M.
Published: January 2012

Intestinal epithelial barrier disruption is a feature of inflammatory bowel disease (IBD), but whether barrier disruption precedes or merely accompanies inflammation remains controversial. Tight junction (TJ) adhesion complexes control epithelial barrier integrity. Since some TJ proteins reside in cholesterol-enriched regions of the cell membrane termed lipid rafts, we sought to elucidate the relationship between rafts and intestinal epithelial barrier function. Lipid rafts were isolated from Caco-2 intestinal epithelial cells primed with the proinflammatory cytokine interferon-γ (IFN-γ) or treated with methyl-β-cyclodextrin as a positive control for raft disruption. Rafts were also isolated from the ilea of mice in which colitis had been induced in conjunction with in vivo intestinal permeability measurements, and lastly from intestinal biopsies of ulcerative colitis (UC) patients with predominantly mild or quiescent disease. Raft distribution was analyzed by measuring activity of the raft-associated enzyme alkaline phosphatase and by performing Western blot analysis for flotillin-1. Epithelial barrier integrity was estimated by measuring transepithelial resistance in cytokine-treated cells or in vivo permeability to fluorescent dextran in colitic mice. Raft and nonraft fractions were analyzed by Western blotting for the TJ proteins occludin and zonula occludens-1 (ZO-1). Our results revealed that lipid rafts were disrupted in IFN-γ-treated cells, in the ilea of mice with subclinical colitis, and in UC patients with quiescent inflammation. This was not associated with a clear pattern of occludin or ZO-1 relocalization from raft to nonraft fractions. Significantly, a time-course study in colitic mice revealed that disruption of lipid rafts preceded the onset of increased intestinal permeability. Our data suggest for the first time that lipid raft disruption occurs early in the inflammatory cascade in murine and human colitis and, we speculate, may contribute to subsequent disruption of epithelial barrier function.
Nicotine suppresses hyperexcitability of colonic sensory neurons and visceral hypersensivity in mouse model of colonic inflammation
Authors: Abdrakhmanova Galya R. , Kang Minho , Imad Damaj M. , Akbarali Hamid I.
Published: January 2012

Recently, we reported that nicotine in vitro at a low 1-μM concentration suppresses hyperexcitability of colonic dorsal root ganglia (DRG; L1-L2) neurons in the dextran sodium sulfate (DSS)-induced mouse model of acute colonic inflammation ( 1 ). Here we show that multiple action potential firing in colonic DRG neurons persisted at least for 3 wk post-DSS administration while the inflammatory signs were diminished. Similar to that in DSS-induced acute colitis, bath-applied nicotine (1 μM) gradually reduced regenerative multiple-spike action potentials in colonic DRG neurons to a single action potential in 3 wk post-DSS neurons. Nicotine (1 μM) shifted the activation curve for tetrodotoxin (TTX)-resistant sodium currents in inflamed colonic DRG neurons (voltage of half-activation changed from −37 to −32 mV) but did not affect TTX-sensitive currents in control colonic DRG neurons. Further, subcutaneous nicotine administration (2 mg/kg b.i.d.) in DSS-treated C57Bl/J6 male mice resulted in suppression of hyperexcitability of colonic DRG (L1-L2) neurons and the number of abdominal constrictions in response to intraperitoneal injection of 0.6% acetic acid. Collectively, the data suggest that neuronal nicotinic acetylcholine receptor-mediated suppression of hyperexcitability of colonic DRG neurons attenuates reduction of visceral hypersensitivity in DSS mouse model of colonic inflammation.
Handling objects in old age: forces and moments acting on the object
Authors: Parikh Pranav J. , Cole Kelly J.
Published: January 2012

We measured the external moments and digit-tip force directions acting on a freely moveable object while it was grasped and manipulated by old (OA) and young (YA) adults. Participants performed a grasp and lift task and a precision orientation (key-slot) task with a precision (thumb-finger) grip. During the grasp-lift task the OA group misaligned their thumb and finger contacts and produced greater grip force, greater external moments on the object around its roll axis, and oriented force vectors differently compared with the YA group. During the key-slot task, the OA group was more variable in digit-tip force directions and performed the key-slot task more slowly. With practice the OA group aligned their digits, reduced their grip force, and minimized external moments on the object, clearly demonstrating that the nervous system monitored and actively manipulated one or more variables related to object tilt. This was true even for the grip-lift task, a task for which no instructions regarding object orientation were given and which could tolerate modest amounts of object tilt without interfering with task goals. Although the OA group performed the key-slot task faster with experience, they remained slower than the YA group. We conclude that with old age comes a reduced ability to control the forces and moments applied to objects during precision grasp and manipulation. This may contribute to the ubiquitous slowing and deteriorating manual dexterity in healthy aging.
Erythropoietin administration acutely stimulates resting energy expenditure in healthy young men
Authors: Christensen Britt , Vendelbo Mikkel H. , Krusenstjerna-Hafstrøm Thomas , Madsen Michael , Pedersen Steen B. , Jessen Niels , Møller Niels , Jørgensen Jens Otto L.
Published: January 2012

Treatment with recombinant human erythropoietin (rHuEpo) improves insulin sensitivity in patients with end-stage renal disease, and animal studies indicate that Epo increases fat oxidation. However, the metabolic effects of rHuEpo have never been experimentally studied in healthy humans. The aim was to investigate the effects of an acute rHuEpo bolus on substrate metabolism and insulin sensitivity in healthy young men. Ten healthy young men were studied in a single-blinded, randomized crossover design with a 2-wk washout period receiving 400 IU/kg rHuEpo or placebo. Substrate metabolism was evaluated by indirect calorimetry and tracer infusions, and insulin sensitivity by a hyperinsulinemic euglycemic clamp; and PCR and Western blotting measured protein expression and content, respectively. Resting energy expenditure (REE) increased significantly after rHuEpo [basal: 1,863.3 ± 67.2 (kcal/day) (placebo) vs. 2,041.6 ± 81.2 (rHuEpo), P < 0.001; clamp: 1,903.9 ± 68.3 (placebo) vs. 2,015.7 ± 114.4 (rHuEpo), P = 0.03], but the increase could not be explained by changes in mRNA levels of uncoupling protein 2 or 3. Fat oxidation in the basal state tended to be higher after rHuEpo but could not be explained by changes in mRNA levels of CPT1 and PPARα or AMPK and ACC protein phosphorylation. Insulin-stimulated glucose disposal, glucose metabolism, and whole body and forearm protein metabolism did not change significantly in response to rHuEpo. In conclusion, a single injection of rHuEpo acutely increases REE in healthy human subjects. This calorigenic effect is not accompanied by distinct alterations in the pattern of substrate metabolism or insulin sensitivity.
LRIG1 Modulates Cancer Cell Sensitivity to Smac Mimetics by Regulating TNFα Expression and Receptor Tyrosine Kinase Signaling
Authors: Bai Longchuan , McEachern Donna , Yang Chao-Yie , Lu Jianfeng , Sun Haiying , Wang Shaomeng
Published: January 2012

Smac mimetics block inhibitor of apoptosis proteins to trigger TNFα-dependent apoptosis in cancer cells. However, only a small subset of cancer cells seem to be sensitive to Smac mimetics and even sensitive cells can develop resistance. Herein, we elucidated mechanisms underlying the intrinsic and acquired resistance of cancer cells to Smac mimetics. In vitro and in vivo investigations revealed that the expression of the cell surface protein LRIG1, a negative regulator of receptor tyrosine kinases (RTK), is downregulated in resistant derivatives of breast cancer cells sensitive to Smac mimetics. RNA interference–mediated downregulation of LRIG1 markedly attenuated the growth inhibitory activity of the Smac mimetic SM-164 in drug-sensitive breast and ovarian cancer cells. Furthermore, LRIG1 downregulation attenuated TNFα gene expression induced by Smac mimetics and increased the activity of multiple RTKs, including c-Met and Ron. The multitargeted tyrosine kinase inhibitors Crizotinib and GSK1363089 greatly enhanced the anticancer activity of SM-164 in all resistant cell derivatives, with the combination of SM-164 and GSK1363089 also completely inhibiting the outgrowth of resistant tumors in vivo. Together, our findings show that both upregulation of RTK signaling and attenuated TNFα expression caused by LRIG1 downregulation confers resistance to Smac mimetics, with implications for a rational combination strategy. Cancer Res; 72(5); 1229–38. ©2012 AACR.
Phase II Efficacy and Pharmacogenomic Study of Selumetinib (AZD6244; ARRY-142886) in Iodine-131 Refractory Papillary Thyroid Carcinoma with or without Follicular Elements
Authors: Hayes D. Neil , Lucas Amy S. , Tanvetyanon Tawee , Krzyzanowska Monika K. , Chung Christine H. , Murphy Barbara A. , Gilbert Jill , Mehra Ranee , Moore Dominic T. , Sheikh Arif , Hoskins Janelle , Hayward Michele C. , Zhao Ni , O'Connor Wendi , Weck Karen E. , Cohen Roger B. , Cohen Ezra E.W.
Published: January 2012

Purpose: A multicenter, open-label, phase II trial was conducted to evaluate the efficacy, safety, and tolerability of selumetinib in iodine-refractory papillary thyroid cancer (IRPTC).

Experimental Design: Patients with advanced IRPTC with or without follicular elements and documented disease progression within the preceding 12 months were eligible to receive selumetinib at a dose of 100 mg twice daily. The primary endpoint was objective response rate using Response Evaluation Criteria in Solid Tumors. Secondary endpoints were safety, overall survival, and progression-free survival (PFS). Tumor genotype including mutations in BRAF, NRAS, and HRAS was assessed.

Results: Best responses in 32 evaluable patients out of 39 enrolled were 1 partial response (3%), 21 stable disease (54%), and 11 progressive disease (28%). Disease stability maintenance occurred for 16 weeks in 49%, 24 weeks in 36%. Median PFS was 32 weeks. BRAF V600E mutants (12 of 26 evaluated, 46%) had a longer median PFS compared with patients with BRAF wild-type (WT) tumors (33 versus 11 weeks, respectively, HR = 0.6, not significant, P = 0.3). The most common adverse events and grades 3 to 4 toxicities included rash, fatigue, diarrhea, and peripheral edema. Two pulmonary deaths occurred in the study and were judged unlikely to be related to the study drug.

Conclusions: Selumetinib was well tolerated but the study was negative with regard to the primary outcome. Secondary analyses suggest that future studies of selumetinib and other mitogen-activated protein (MAP)/extracellular signal-regulated kinase (ERK; MEK) inhibitors in IRPTC should consider BRAF V600E mutation status in the trial design based on differential trends in outcome. Clin Cancer Res; 18(7); 2056–65. ©2012 AACR.
Lenalidomide-Induced Immunomodulation in Multiple Myeloma: Impact on Vaccines and Antitumor Responses
Authors: Noonan Kimberly , Rudraraju Lakshmi , Ferguson Anna , Emerling Amy , Pasetti Marcela F. , Huff Carol A. , Borrello Ivan
Published: January 2012

Purpose: To show that the immunomodulatory drug lenalidomide can be used in patients with relapsed multiple myeloma to augment vaccine responses.

Experimental Design: Early phase clinical trial of patients with multiple myeloma who received at least one prior therapy. Patients were treated with single-agent lenalidomide and randomized to receive two vaccinations with pneumococcal 7-valent conjugate vaccine (PCV) on different schedules. Cohort A received the first PCV vaccination prior to the initiation of lenalidomide and the second vaccination while on lenalidomide. Cohort B received both vaccinations while on lenalidomide.

Results: PCV-specific humoral and cellular responses were greater in cohort B than A and were more pronounced in the bone marrow than the blood, suggesting that maximal vaccine efficacy was achieved when both vaccines were administered concomitantly with lenalidomide. Patients with a clinical myeloma response showed evidence of a tumor-specific immune response with increases in myeloma-specific IFN-γ+ T cells and reductions in Th-17 cells.

Conclusions: This is the first clinical evidence showing that lenalidomide augments vaccine responses and endogenous antitumor immunity in patients and as such may serve as an adjuvant for cancer and possibly infectious vaccines. Clin Cancer Res; 18(5); 1426–34. ©2012 AACR.
Molecular and Cellular Effects of NEDD8-Activating Enzyme Inhibition in Myeloma
Authors: McMillin Douglas W. , Jacobs Hannah M. , Delmore Jake E. , Buon Leutz , Hunter Zachary R. , Monrose Val , Yu Jie , Smith Peter G. , Richardson Paul G. , Anderson Kenneth C. , Treon Steven P. , Kung Andrew L. , Mitsiades Constantine S.
Published: January 2012

The NEDD8-activating enzyme is upstream of the 20S proteasome in the ubiquitin/proteasome pathway and catalyzes the first step in the neddylation pathway. NEDD8 modification of cullins is required for ubiquitination of cullin-ring ligases that regulate degradation of a distinct subset of proteins. The more targeted impact of NEDD8-activating enzyme on protein degradation prompted us to study MLN4924, an investigational NEDD8-activating enzyme inhibitor, in preclinical multiple myeloma models. In vitro treatment with MLN4924 led to dose-dependent decrease of viability (EC50 = 25–150 nmol/L) in a panel of human multiple myeloma cell lines. MLN4924 was similarly active against a bortezomib-resistant ANBL-6 subline and its bortezomib-sensitive parental cells. MLN4924 had submicromolar activity (EC50 values <500 nmol/L) against primary CD138+ multiple myeloma patient cells and exhibited at least additive effect when combined with dexamethasone, doxorubicin, and bortezomib against MM.1S cells. The bortezomib-induced compensatory upregulation of transcripts for ubiquitin/proteasome was not observed with MLN4924 treatment, suggesting distinct functional roles of NEDD8-activating enzyme versus 20S proteasome. MLN4924 was well tolerated at doses up to 60 mg/kg 2× daily and significantly reduced tumor burden in both a subcutaneous and an orthotopic mouse model of multiple myeloma. These studies provide the framework for the clinical investigation of MLN4924 in multiple myeloma. Mol Cancer Ther; 11(4); 942–51. ©2012 AACR.
Effects of Proton Pump Inhibitors on Pediatric Inflammatory Esophagogastric Polyps
Authors: Choi Kyong Eun , Kim Mi Jin , Lee Ji Hyuk , Lee Jong Seung , Lee Jee Hyun , Choe Yon Ho
Published: January 2012

Background: The aim of this study was to investigate the effects of proton pump inhibitors on symptomatic inflammatory esophagogastric polyps (IEPs) in a pediatric cohort and to determine the optimal duration of treatment. Methods: The 11 patients with IEPs were managed with lansoprazole. Follow-up endoscopies were performed at 2 and 6 months after the start of medication. Medication was discontinued when the clinical symptoms completely resolved and the polyp size was reduced by more than 50% compared to the initial size. Results:The initial polyp size was 13.7 ± 3.3 mm. After 2 months of medication, the polyp size was reduced to 8.0 ± 5.8 mm. At 6 months, the polyp size was 4.7 ± 2.2 mm. The mean duration of medication was 4.8 ± 2.1 months. The duration of medication and the change in the polyp size appeared to have a linear correlation (p < 0.001). According to the formula used to calculate polyp size, the optimal duration of treatment was more than 7 months for complete resolution of the polyps. Conclusions: Proton pump inhibitor was effective for the treatment of IEPs. About 5 months of lansoprazole was adequate to treat IEPs in children. The optimal duration for complete resolution of the polyp might be more than 7 months.
The Effects of Adiponectin and Leptin on Human Endothelial Cell Proliferation: A Live-Cell Study
Authors: Álvarez Granada , Visitación Bartolomé M. , Miana María , Jurado-López Raquel , Martín Ruben , Zuluaga Pilar , Martinez-Martinez Ernesto , Nieto M. Luisa , Alvarez-Sala Luis A. , Millán Jesús , Lahera Vicente , Cachofeiro Victoria
Published: January 2012

The effect of adiponectin and leptin on the proliferation of the human microvascular endothelial cell line (HMEC-1) was studied in the absence or presence of fetal bovine serum (FBS). The participation of extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinase/Akt (PI-3K/Akt) pathways in this effect were evaluated. We studied the effect of both adipokines on the motility, mitosis, proliferation and cell death processes of HMEC-1 cells using live-cell imaging techniques. Adiponectin but not leptin further increased the proliferative effect induced by FBS on HMEC-1. This effect seems to be the consequence of an increase in the mitotic index in adiponectin-treated cells when compared to untreated ones. The presence of either the mitogen-activated protein kinase (MAPK) inhibitor (PD98059), or PI-3K inhibitor (LY294002), reduced the effect of adiponectin in a dose-dependent manner. Neither adipokine was able to affect HMEC-1 proliferation in FBS-free conditions. Duration of mitosis, cell motility and the cell death process were similar in all conditions. These data suggest that adiponectin and leptin exert different effects on endothelial cell function. Adiponectin was able to potentiate proliferation of HMEC-1. This effect involves the activation of both PI3-K/Akt and ERK/MAPK pathways. However, it seems to exert minimal effects on HMEC-1 function in the case of leptin.
A Risk-Adapted Strategy of Adjuvant Paclitaxel/Carboplatin in Early-Stage Ovarian Cancer: Time-Dependent Effect of 4 versus 6 Cycles on Outcome
Authors: Bamias Aristotle , Bamia Christina , Karadimou Alexandra , Soupos Nikolaos , Zagouri Flora , Rodolakis Alexandros , Haidopoulos Dimitrios , Vlahos George , Thomakos Nikos , Antsaklis Aristidis , Dimopoulos Meletios A.
Published: January 2012

Objective: We investigated the efficacy of risk-adapted adjuvant paclitaxel/carboplatin chemotherapy in early-stage ovarian carcinoma. Methods: Fifty-three patients were treated according to the risk of relapse: patients with stages IA or IB or with grade 1 (low risk) received 4 cycles of paclitaxel and carboplatin; patients with IC/IIA and grade 2 or 3 (high risk) received 6 cycles of chemotherapy. The outcome was compared with that of 95 patients who were all treated with 4 cycles. Results: Median follow-up was 88, 113 and 42 months for the whole cohort, non-risk-adapted and risk-adapted treatment, respectively. Five-year relapse-free and disease-specific survival was 86 and 93% for the whole population, 96 and 97% for low-risk and 81 and 91% for high-risk patients. Risk classification was the only significant prognostic factor for relapse-free (p = 0.011) and disease-specific survival (p = 0.039). Among high-risk patients, the administration of 6 cycles was associated with a significantly lower relapse rate after censoring events, which occurred beyond 2 years (3 vs. 18%; p = 0.013), but this difference was diminished at 5 years (23 vs. 25%; p = 0.797). Conclusions: Six cycles of chemotherapy reduced the risk of relapse within 2 years, but the benefit from two additional cycles beyond this time is questionable.