bluuebunny/crossref_metadata_2025 huggingface dataset:
Travel for treatment: students' perspective on medical tourism
Authors: Reddy Sumanth G. , York Valerie K. , Brannon Laura A.
Published: January 2010

Medical tourism is the act of travelling abroad for health care. The theory of planned behaviour was used to investigate the medical tourism beliefs of 336 undergraduate students in a basic psychology course at a large midwestern American university. Students did not have positive intentions for mere willingness to seek more information about travelling to a developing country to receive medical treatment. An educational intervention is necessary to help promote travel for medical treatment. The intervention may include educating people on the availability of quality health care, highly trained competent doctors and the ability to vacation and see another country. Copyright © 2010 John Wiley & Sons, Ltd.
Septoplasty retards midfacial growth in a rabbit model
Authors: Wong Kevin K. , Filatov Sergei , Kibblewhite Douglas J.
Published: January 2010

Objectives/Hypothesis:

Septoplasty in the prepubertal age group is a controversial operation because of concerns regarding retardation of subsequent midfacial growth. This study uses a validated rabbit snout model to determine if septoplasty results in midfacial growth retardation, and if the retardation can be averted by replacing resected cartilage with a porous polyethylene implant.

Study Design:

Prospective controlled animal study.

Methods:

Sixteen 9‐week‐old New Zealand white rabbits were divided into four groups: two control and two experimental. The experimental groups both underwent conservative submucoperichondrial surgical excision of a portion of their septum, leaving a generous dorsal and caudal septal strut for nasal support similar to what would be done clinically in humans. One of the experimental groups had replacement of the resected area by a rigid porous polyethylene graft, and in the other experimental group the resected area was left empty. Cephalometric measurements were performed on radiographs taken at the time of implantation, and at sacrifice when the rabbits were mature at 24 weeks of age.

Results:

There were significant disturbances in snout growth between experimental and control groups, and no significant differences between the two experimental groups or between the two control groups.

Conclusions:

These data demonstrate midfacial growth retardation after conservative septoplasty in young rabbits, and that this retardation could not be averted by replacing the resected cartilage with a porous polyethylene graft. These data are in agreement with human studies. Laryngoscope, 2010
Spray cryotherapy for the treatment of glottic and subglottic stenosis
Authors: Krimsky William S. , Rodrigues Michael P. , Malayaman Navara , Sarkar Saiyad
Published: January 2010

Objectives/Hypothesis:

Functional partial occlusion of the glottic and subglottic areas by stenosis and strictures is challenging to manage despite a variety of surgical and endoluminal approaches that are prone to complications and inconsistent outcomes. We report here the first three human cases of glottic and subglottic narrowing treated with spray cryotherapy alone or in combination with balloon dilation.

Study Design:

Institutional review board‐approved clinical human trial.

Methods:

A 42‐year‐old female with idiopathic subglottic strictures, a 74‐year‐old female with glottic strictures and vocal cord stenosis following neck radiation, and a 33‐year‐old female with strictures from a previous tracheal stent were treated by four cycles of a 5‐second cryotherapy spray alone or with balloon dilation. The effects of treatment were observed up to 6 months, 12 weeks, and 9 months, respectively.

Results:

In all cases, patency of the stenosed areas was achieved with minimal bleeding and at least some degree of normalization of the glottic and subglottic mucosa. Airway patency and laryngeal functions were restored without complications.

Conclusions:

The use of spray cryotherapy alone or in conjunction with balloon dilation is a promising and effective therapeutic approach to treating glottic and subglottic narrowing. Laryngoscope, 2010
New Thermo‐Sensitive Graft Copolymers Based on a Poly(N‐isopropylacrylamide) Backbone and Functional Polyoxazoline Grafts with Random and Diblock Structure
Authors: Rueda Juan Carlos , Zschoche Stefan , Komber Hartmut , Krahl Franziska , Arndt Karl‐Friedrich , Voit Brigitte
Published: January 2010

New thermo‐sensitive functionalized graft copolymers characterized by a poly(N‐isopropylacrylamide) backbone and grafts containing 2‐ethyl‐2‐oxazoline and 2‐(2‐methoxycarbonylethyl)‐2‐oxazoline units were synthesized. The conformation transition temperatures of the graft copolymers could be modified by variation of the molar composition in the side chain, by different side chain structure (random distribution of both oxazolines vs. diblock structure) and by hydrolysis of the methylester to the acid form. Graft copolymers with long functional oxazoline side chains allowed the stabilization of aggregates above the phase transition temperature of the backbone until the LCST of the side chain. The temperature window allowing for the formation of stable aggregates was widened with acid functions in the corona.

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Correlation between Morphology, Water Uptake, and Proton Conductivity in Radiation‐Grafted Proton‐Exchange Membranes
Authors: Balog Sandor , Gasser Urs , Mortensen Kell , Gubler Lorenz , Scherer Guenther G. , Ben youcef Hicham
Published: January 2010

An SANS investigation of hydrated proton exchange membranes is presented. Our membranes were synthesized by radiation‐induced grafting of ETFE with styrene in the presence of a crosslinker, followed by sulfonation of the styrene. The contrast variation method was used to understand the relationship between morphology, water uptake, and proton conductivity. The membranes are separated into two phases. The amorphous phase hosts the water and swells upon hydration, swelling being inversely proportional to the degree of crosslinking. Hydration and proton conductivity exhibit linear dependence on swelling. Proton conductivity and volumetric fraction of water are related by a power law, indicating a percolated network of finely dispersed aqueous pores in the hydrophilic domains.

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Folic Acid Modified Poly(lactide‐co‐glycolide) Nanoparticles, Layer‐by‐Layer Surface Engineered for Targeted Delivery
Authors: Zhou Jie , Romero Gabriela , Rojas Elena , Moya Sergio , Ma Lie , Gao Changyou
Published: January 2010

The layer‐by‐layer (LbL) assembly technique was applied for the surface modification of biodegradable poly(lactide‐co‐glycolide) nanoparticles (NPs), employing poly(acrylic acid) (PAA), and polyethylenimine (PEI) as building blocks. Amino terminated poly(ethylene glycol) (PEG) and folate decorated PEG (PEG‐FA) were grafted onto the multilayers via condensation between carboxylic groups and amine groups from PEG or PEG‐FA. The LbL assembly and the covalent functionalization were monitored by means of ζ‐potential measurements and the quartz crystal microbalance with dissipation technique (QCM‐D). Protein adsorption after incubation of the NPs in culture medium containing optionally the serum proteins was investigated and related to cellular uptake. Experiments on cellular uptake showed that after PEGylation the uptake ratio of the NPs decreased significantly, but became three times larger when PEG‐FA was grafted on the NPs instead of the PEG.

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Modulatory effects of 5Hz rTMS over the primary somatosensory cortex in focal dystonia—An fMRI‐TMS study
Authors: Schneider Susanne A. , Pleger Burkhard , Draganski Bogdan , Cordivari Carla , Rothwell John C. , Bhatia Kailash P. , Dolan Ray J.
Published: January 2010

Dystonia is associated with impaired somatosensory ability. The electrophysiological method of repetitive transcranial magnetic stimulation (rTMS) can be used for noninvasive stimulation of the human cortex and can alter cortical excitability and associated behavior. Among others, rTMS can alter/improve somatosensory discrimation abilities, as shown in healthy controls. We applied 5Hz‐rTMS over the left primary somatosensory cortex (S1) in 5 patients with right‐sided writer's dystonia and 5 controls. We studied rTMS effects on tactile discrimination accuracy and concomitant rTMS‐induced changes in hemodynamic activity measured by functional magnetic resonance imaging (fMRI). Before rTMS, patients performed worse on the discrimination task than controls even though fMRI showed greater task‐related activation bilaterally in the basal ganglia (BG). In controls, rTMS led to improved discrimination; fMRI revealed this was associated with increased activity of the stimulated S1, bilateral premotor cortex and BG. In dystonia patients, rTMS had no effect on discrimination; fMRI showed similar cortical effects to controls except for no effects in BG. Improved discrimination after rTMS in controls is linked to enhanced activation of S1 and BG. Failure of rTMS to increase BG activation in dystonia may be associated with the lack of effect on sensory discrimination in this group and may reflect impaired processing in BG‐S1 connections. Alternatively, the increased BG activation seen in the baseline state without rTMS may reflect a compensatory strategy that saturates a BG contribution to this task. © 2010 Movement Disorder Society
Portable duplex ultrasonography: A diagnostic and decision‐making tool in reconstructive microsurgery
Authors: Gravvanis Andreas , Karakitsos Dimitrios , Dimitriou Vasilios , Zogogiannis Ioannis , Katsikeris Nick , Karabinis Andreas , Tsoutsos Dimosthenis
Published: January 2010

Unidirectional Doppler is a common diagnostic tool by the Reconstructive Microsurgeons; however, it may generate false signals and surely provides less imaging data as compared to duplex ultrasonography. We have reviewed the use of Portable Duplex Ultrasonography (PDU) in 16 patients who underwent complex soft‐tissue/bone reconstruction, aiming to determine its role in the design and management of free tissue transfer. According to our data, there were modifications either of the surgical plan and/or of patient's management, based on PDU findings, in 10 out of 16 patients (62.5%). The use of ultrasound directed to subtle modifications in three patients (19%), but to significant changes of the surgical plan in four patients (25%). Also, the use of ultrasound improved significantly the postoperative management in three patients (19%). Thus, significant impact of PDU in patient's treatment was recorded in 44% of cases. Portable ultrasound represents generally available method for preoperative, intraoperative, and postoperative diagnosis and decision‐making in free tissue transfer, hence could replace in the near future the unidirectional Doppler in the hands of Microsurgeons. © 2010 Wiley‐Liss, Inc. Microsurgery 30:348–353, 2010.
Impact of dietary polyphenols on human platelet function – A critical review of controlled dietary intervention studies
Authors: Ostertag Luisa M. , O'Kennedy Niamh , Kroon Paul A. , Duthie Garry G. , de Roos Baukje
Published: January 2010

Cardiovascular disease is a chronic disease influenced by many factors, with activated blood platelets being one of them. Platelets play a central role in the formation of plaques within blood vessels, contributing to early inflammatory events. Consumption of diets rich in plant‐based products protects against the development of cardiovascular disease. Polyphenols, which are secondary plant metabolites found in a wide range of foodstuffs and beverages, may be partially responsible for these effects. Their protective properties include inhibitory effects on platelet functionin vitroandin vivo. However, the bioavailability of many polyphenols is poor and it is unclear whether sufficient quantities can be obtained by dietary means to exert protective effects. Consequently, this review summarizes 25 well‐controlled human intervention studies examining the effect of polyphenol‐rich diets on platelet function. These studies report a huge variety of research methods, study designs, and study subjects, resulting in controversial assertions. One consistent finding is that cocoa‐related products, however, have platelet‐inhibiting effects when consumed in moderate amounts. To assess whether other classes of dietary polyphenols, or their metabolites, also beneficially affect platelet function requires more well‐controlled intervention studies as well as the adoption of more uniform methods to assess platelet aggregation and activation.
Preparation and properties of new high performance maleimide‐triazine resins for resin transfer molding
Authors: Guan Qingbao , Gu Aijuan , Liang Guozheng , Zhou Cheng , Yuan Li
Published: January 2010

High performance matrix is the key base for preparing advanced composites via resin transfer molding (RTM). A novel high performance modified maleimide‐triazine (BT) resin system (coded as MBT) for RTM was developed, which is made of 4,4′‐bismaleimidodi‐ phenylmethane, o,o'‐diallylbisphenol A, 2,2′‐bis (4‐cyanatophenyl) isopropylidene, and hyperbranched polysiloxane (HBPSi). The effects of HBPSi on the processing and performance parameters of MBT system are evaluated. Results show that the processing characteristics of the MBT system are greatly dependent on the content of HBPSi in the system, while three MBT resins developed in this paper have significantly better integrated properties than BT resin. For example, compared to original BT resin, MBT resins have enlarged pot life (>8 hr) and good reactivity; more interestingly, cured MBT resins exhibit better dielectric properties and moisture resistance; in addition, MBT resins with suitable content of HBPSi have improved flexural and impact strengths as well as outstanding thermal property, suggesting that MBT system is the right kind of matrices with great potentiality for fabricating advanced structural and functional composites via RTM technique. Copyright © 2010 John Wiley & Sons, Ltd.
ReSASC: A resampling‐based algorithm to determine differential protein expression from spectral count data
Authors: Little Kristina M. , Lee Jae K. , Ley Klaus
Published: January 2010

Label‐free methods for MS/MS quantification of protein expression are becoming more prevalent as instrument sensitivity increases. Spectral counts (SCs) are commonly used, readily obtained, and increase linearly with protein abundance; however, a statistical framework has been lacking. To accommodate the highly non‐normal distribution of SCs, we developed ReSASC (resampling‐based significance analysis for spectral counts), which evaluates differential expression between two conditions by pooling similarly expressed proteins and sampling from this pool to create permutation‐based synthetic sets of SCs for each protein. At a set confidence level and corresponding p‐value cutoff, ReSASC defines a new p‐value, p′, as the number of synthetic SC sets with p>pcutoff divided by the total number of sets. We have applied ReSASC to two published SC data sets and found that ReSASC compares favorably with existing methods while being easy to operate and requiring only standard computing resources.
The role of PI 3‐kinase p110β in AKT signally, cell survival, and proliferation in human prostate cancer cells
Authors: Hill Karen M. , Kalifa Sara , Das Jharna R. , Bhatti Tahira , Gay Martha , Williams Danielle , Taliferro‐Smith LaTonia , De Marzo Angelo M.
Published: January 2010

BACKGROUND

Class IA PI 3‐kinases produce phosphatidylinositol (3,4,5)‐trisphosphate (PIP3). PIP3 is bound by AKT which facilities its activation by PDK1. Activated AKT promotes cell survival and stimulates cell proliferation. Class IA PI 3‐kinases are heterodimers consisting of a regulatory subunit p85 and a catalytic subunit p110. The p110α isoform has been shown to be mutated in a number of tumor types. A number of recent studies suggest that the p110β isoform may be functionally relevant in prostate cancer. In this study we extend this work to include the examination of the expression and functional properties of p110α and p110β in three different prostate cancer cell lines, DU145, LNCaP, PC3, as well as the non‐tumorigenic but immortalized RWPE1 prostate epithelial cell line.

METHODS

Western blot analysis was used to measure protein expression and quantitative real‐time PCR was used to measure mRNA levels. After targeted knockdown using isoform‐specific siRNAs to reduce PI 3‐kinase p110α or p110β isoform expression, we measured downstream signally events such as phosphorylation of AKT, ERK 1/2, PDK, and FOXO, as well as biological consequences such as changes in apoptosis, and alterations in cell cycle progression.

RESULTS

In all three prostate cancer cell lines examined, targeted knockdown of p110β, and not p110α, resulted in significantly reduced AKT, PDK, and FOXO phosphorylation. While knockdown of either p110 isoform resulted in an increase in apoptosis and a cell cycle arrest in G1 in the remaining non‐apoptotic cells, these effects were much more pronounced with knockdown of p110β.

CONCLUSIONS

Our results support the concept that p110β appears to be the predominant functional class I PI 3‐kinase isoform in prostate cancer cells. Prostate 70: 755–764, 2010. © 2010 Wiley‐Liss, Inc.
Prostate‐specific antigen (PSA) is activated by KLK2 in prostate cancer ex vivo models and in prostate‐targeted PSA/KLK2 double transgenic mice
Authors: Williams Simon A. , Xu Yi , De Marzo Angelo M. , Isaacs John T. , Denmeade Samuel R.
Published: January 2010

BACKGROUND

Prostate‐specific antigen (PSA) is a serine protease secreted as a zymogen. Previously, cell‐free biochemical studies have identified various kallikreins (KLK) as candidate activating proteases. In this study, KLK2‐mediated activation of PSA in cell‐based in vitro, xenograft, and transgenic models was evaluated.

METHODS

Du145‐derived PSA‐ or KLK2‐expressing clones were coincubated in vitro and in vivo to evaluate KLK2‐induced PSA activity. While mice possess orthologs of KLK4‐15, they do not have functional orthologs of PSA or KLK2. Therefore, transgenic animals expressing PSA or both PSA and KLK2 were generated to assess orthotopic PSA activation.

RESULTS

PSA is activated by KLK2 when the cells are physically in contact, and through co‐conditioned media. In vivo, the free (inactive PSA) to total (active + inactive PSA) ratio in the blood is decreased when PSA and KLK2‐expressing cells are co‐inoculated subcutaneously, suggesting increased active PSA. Additionally, double‐transgenic mice expressing both genes in the prostate produce more active PSA compared to single transgenic animals. A longitudinal evaluation over a 2‐year period demonstrated no morphologic changes (i.e., no PIN or prostate cancer) due to PSA or PSA/KLK2 double transgene expression relative to non‐transgenic mice.

CONCLUSIONS

These data demonstrate, with biologically relevant models, that KLK2 is the protease responsible for activating PSA. While PSA is involved in the processing and release of a number of important growth factors, our results suggest that active PSA is not sufficient to induce the development of prostate cancer or prostate cancer precursors in aging PSA transgenic mice. Prostate 70: 788–796, 2010. © 2010 Wiley‐Liss, Inc.
Should Routine Analysis of the MEN1 Gene be Performed in all Patients with Primary Hyperparathyroidism Under 40 Years of Age?
Authors: Skandarajah Anita , Barlier Anne , Morlet‐Barlat Nathalie , Sebag Frederic , Enjalbert Alain , Conte‐Devolx Bernard , Henry Jean‐François
Published: January 2010

Background

Familial hyperparathyroidism, especially Multiple Endocrine Neoplasia Type 1, is more likely to present with primary hyperparathyroidism (1° HPT) at a young age, mandating bilateral exploration of the parathyroid glands. However, the majority of young patients will not be gene carriers or have a family history. Recent evidence suggests that young adults under 40, in whom there is no suspicion of family history, can be managed with the same pre‐ and perioperative strategy as used for sporadic primary HPT of any age. Our aim was to evaluate the prevalence of mutations in the MEN1 gene in young adults under 40 who present with apparent sporadic 1° HPT.

Methods

A retrospective review was undertaken of all patients who underwent surgery for 1° HPT between 1993 and 2004. From a total of 1253 patients, 87 (6.2%) were under the age of 40. Thirty‐three patients provided informed consent to a detailed personal and family history, physical examination, and genetic analysis of the MEN1 gene. Twelve patients were subsequently excluded as they were known gene carriers prior to surgery (10 MEN1 and 2 MEN2A patients). Twenty‐one patients underwent genetic analysis.

Results

Of the 21 patients who consented to genetic analysis, the mean age was 30.8 years (range = 18–39 years with 43% younger than 30). These patients had no suspicious family or personal histories suggestive of a MEN phenotype. Fifteen patients presented with symptomatic hypercalcemia. All 21 patients underwent parathyroid surgery by conventional cervicotomy (12) or endoscopic parathyroidectomy in cases (9) where the parathyroid gland was localized preoperatively. Nineteen patients (91%) had uniglandular disease. Surgical cure was achieved in all patients. Of the 21 patients, only one patient (4.7%) was found to have a MEN1 gene mutation (exon 3, at codon 190, c;680_681delGGinsC). This patient was found to have double adenomas at surgery with subsequent histological confirmation. The overall prevalence of MEN1 mutation in all patients under 40 was 13%.

Conclusion

Although young age is often the only criterion to suspect MEN1, our results do not support routine MEN1 analysis in patients under 40. We propose that these patients be managed with the same preoperative and surgical approach as those presenting with sporadic 1° HPT of any age.
Effects of acarbose on ruminal fermentation, blood metabolites and microbial profile involved in ruminal acidosis in lactating cows fed a high-carbohydrate ration
Authors: Blanch Marta , Calsamiglia Sergio , Devant Maria , Bach Alex
Published: January 2010

The objective was to evaluate the effects of an inhibitor of α-amylase and glucosidase (acarbose, Pfizer Limited, Corby, UK) on ruminal fermentation, blood metabolism and microbial profile in dairy cows in a 2×2 cross-overexperiment. Eight Holstein cows fitted with rumen cannulas (milk yield, 24·3±2·35 kg/d, body weight, 622±54 kg, days in milk, 183±67, 5 multiparous and 3 primiparous) were used. Treatments were: control (no additive, CTR) and α-amylase and glucosidase inhibitor (0·75 g acarbose-premix/cow per d, AMI). Animals were given ad-libitum access to a high non-fibre carbohydrate (NFC) partial mixed ration (PMR) containing 17·6% crude protein, 28·3% neutral detergent fibre, and 46·5% NFC in the dry matter and supplementary concentrate during milking. Blood samples were taken to determine blood glucose, insulin and urea within the first hour after the morning feeding on two separate days in each period. Samples of ruminal contents were collected during 3 d in each period at 0, 4 and 8 h after feeding to determine volatile fatty acid and ammonia-N concentrations and to quantify protozoa,Streptococcus bovisandMegasphaera elsdenii. Rumen pH was recorded electronically at 22-min intervals during 6 d in each period. Results were analysed using a mixed-effects model. Cows on AMI treatment spent less time with ruminal pH <5·6 compared with cows in the CTR group (3·74 and 6·52±0·704 h/d, respectively). Cows in the AMI group had greater daily average pH compared with those in the CTR group (6·05 and 5·92±0·042, respectively). AMI animals tended (P=0·09) to have lowerStr. bovistoMeg. elsdeniiratio than CTR (4·09 and 26·8±12·0, respectively). These results indicate that dietary supplementation with acarbose in dairy cattle fed high-production rations may be effective in reducing the time for which rumen pH is suboptimal, with no negative effects on ruminal fermentation and blood metabolites.
On the tidal prism–channel area relations
Authors: D'Alpaos Andrea , Lanzoni Stefano , Marani Marco , Rinaldo Andrea
Published: January 2010

We verify the broad applicability of tidal prism cross‐sectional area relationships, originally proposed to relate the total water volume entering a lagoon during a characteristic tidal cycle (the tidal prism) to the size of its inlet, to arbitrary sheltered cross sections within a tidal network. We suggest, with reasonable approximation defining a statistical tendency rather than a pointwise equivalence, that the regime of tidal channels may be anywhere related to local landscape‐forming prisms embedded in a characteristic spring tide oscillation. The importance of the proposed extension stems from its potential for quantitative predictions of the long‐term morphological evolution of whole tidal landforms, in response to forcings affecting tidal prisms. This is the case, in particular, for alterations of relative mean sea levels possibly driven by climate change. Various 1‐D and 2‐D morphodynamic and hydrodynamic models are employed to evaluate peak flow rates, bottom shear stresses, and the ensuing local tidal prisms. One‐dimensional morphodynamic models describing both the longitudinal and cross‐sectional evolution of tidal channels are used to verify the validity of the relationship for sheltered sections. Relevant hydrodynamic features determined through accurate 2‐D numerical models are compared with those obtained through time‐invariant equivalents, defining a mean watershed by an energy landscape from averaged free surface gradients. Empirical evidence gathered within the lagoon of Venice (Italy) supports the proposed extension. We conclude that the geomorphic law relating tidal prisms to channel cross‐sectional areas anywhere within a tidal landscape is a valuable tool for studies on long‐term tidal geomorphology.
Regular Aerobic Exercise, Without Weight Loss, Improves Endothelium‐dependent Vasodilation in Overweight and Obese Adults
Authors: Mestek Michael L. , Westby Christian M. , Van Guilder Gary P. , Greiner Jared J. , Stauffer Brian L. , DeSouza Christopher A.
Published: January 2010

Lifestyle modification in the form of weight reduction by caloric restriction alone or in combination with regular aerobic exercise significantly improves endothelium‐dependent vasodilation in overweight and obese adults. We determined whether regular aerobic exercise, independent of weight loss, improves endothelium‐dependent vasodilation in overweight and obese adults. Twenty overweight and obese adults (age 53 ± 1 years; BMI: 30.2 ± 0.8 kg/m2) were studied before and after a 3‐month aerobic exercise training intervention. Forearm blood flow (FBF) responses were determined (via plethysmography) in response to intra‐arterial infusion of acetylcholine and sodium nitroprusside. There were no changes in body mass or composition with the intervention. FBF responses to acetylcholine were ∼35% higher (P < 0.01) after (4.1 ± 0.9 to 14.7 ± 4.3 ml/100 ml tissue/min) compared with before (4.2 ± 0.8 to 11.0 ± 3 ml/100 ml tissue/min) exercise training. FBF responses to sodium nitroprusside were unchanged. These results indicate that regular aerobic exercise improves endothelium‐dependent vasodilation in overweight and obese adults, independent of changes in body mass or composition.
miR‐519d Overexpression Is Associated With Human Obesity
Authors: Martinelli Rosanna , Nardelli Carmela , Pilone Vincenzo , Buonomo Tonia , Liguori Rosario , Castanò Ilenia , Buono Pasqualina , Masone Stefania , Persico Giovanni , Forestieri Pietro , Pastore Lucio , Sacchetti Lucia
Published: January 2010

Obesity is a consequence of imbalance of food intake and energy expenditure that results in storage of energy as fat, primarily in adipose tissue. MicroRNAs are non‐coding RNAs that regulate gene expression in metabolic pathways and they are also involved in fat‐cell development. The aim of this study was to evaluate whether microRNA dysfunction contributes to obesity. We analyzed, by microarray, the expression profile of 1,458 microRNAs in subcutaneous adipose tissue (SAT) from nondiabetic severely obese (n = 20) and nonobese adults (n = 8). Among 42 differently expressed microRNAs, we confirmed by reverse‐transcription PCR (RT‐PCR) that miR‐519d was overexpressed whereas the protein levels of peroxisome proliferator‐activated receptor‐α (PPARA) (a predicted miR 519d target) were lower, at western analysis, in severely obese vs. nonobese subjects. We also show that miR‐519d specifically and dose‐dependently suppressed translation of the PPARA protein, and increased lipid accumulation during preadipocyte differentiation. Because PPARA plays a central role in fatty acid homeostasis, and in the transcriptional regulation of genes that are necessary for maintenance of the redox balance during the oxidative catabolism of fatty acids, we suggest that PPARA loss and miR‐519d overexpression could be associated with metabolic imbalance and subsequent adipocyte hypertrophy in SAT during obesity.
Effects of Heritability, Shared Environment, and Nonshared Intrauterine Conditions on Child and Adolescent BMI
Authors: Salsberry Pamela J. , Reagan Patricia B.
Published: January 2010

Heritability studies of BMI, based upon twin samples, have identified genetic and shared environmental components of BMI, but have been largely silent about the nonshared environmental factors. Intrauterine factors have been identified as having significant long‐term effects on BMI and may be a critical source of nonshared environmental influence. Extant studies based on samples of either unrelated individuals or twins cannot separate the effects of genetics, shared environments, and nonshared intrauterine conditions because the one lacks variation in the degree of relatedness and the other has insufficient variation in intrauterine conditions. This study improves upon these prior studies by using a large, sibling‐based sample to examine heritability, shared environmental, and nonshared intrauterine influences on BMI during two age periods in childhood (6–8 years; 12–14 years). The primary interest was in determining the effects of the intrauterine environment on BMI as a component of the nonshared environment and in determining whether there were sex‐specific differences in heritability and/or in the intrauterine factors. These were estimated using regression‐based techniques introduced by DeFries and Fulker. Heritability of BMI was estimated to be 0.20–0.28 at 6–8 years and 0.46–0.61 at 12–14 years. Differences in heritability were found at 12–14 years between same‐sex as compared to mixed‐sex pairs. The shared environmental effect was significant at 6–8 years but insignificant at 12–14 years. Differences in birth weight were significant in all groups at 6–8 years suggesting long‐term effects of the nonshared intrauterine environment; at 12–14 years, birth weight was no longer significant for girls.
Endothelial Progenitor Cell Function, Apoptosis, and Telomere Length in Overweight/Obese Humans
Authors: MacEneaney Owen J. , Kushner Erich J. , Westby Christian M. , Cech Jennifer N. , Greiner Jared J. , Stauffer Brian L. , DeSouza Christopher A.
Published: January 2010

Excess adiposity is associated with increased cardiovascular morbidity and mortality. Endothelial progenitor cells (EPCs) play an important role in vascular repair. We tested the hypothesis that increased adiposity is associated with EPC dysfunction, characterized by diminished capacity to release angiogenic cytokines, increased apoptotic susceptibility, reduced cell migration, and shorter telomere length. A total of 67 middle‐aged and older adults (42–67 years) were studied: 25 normal weight (normal weight; BMI: 18.5–24.9 kg/m2) and 42 overweight/obese (overweight/obese; BMI: 25.0–34.9 kg/m2). Cells with phenotypic EPC characteristics were isolated from peripheral blood. EPC release of vascular endothelial growth factor (VEGF) and granulocyte colony–stimulating factor (G‐CSF) was determined in the absence and presence of phytohemagglutinin (10 µg/ml). Intracellular active caspase‐3 and cytochrome c concentrations were determined by immunoassay. Migratory activity of EPCs in response to VEGF (2 ng/ml) and stromal cell–derived factor‐1α (SDF‐1α; 10 ng/ml) was determined by Boyden chamber. Telomere length was assessed by Southern hybridization. Phytohemagglutinin‐stimulated release of VEGF (90.6 ± 7.6 vs. 127.2 ± 11.6 pg/ml) and G‐CSF (896.1 ± 77.4 vs. 1,176.3 ± 126.3 pg/ml) was ∼25% lower (P < 0.05) in EPCs from overweight/obese vs. normal weight subjects. Staurosporine induced a ∼30% greater (P < 0.05) increase in active caspase‐3 in EPCs from overweight/obese (2.8 ± 0.2 ng/ml) compared with normal weight (2.2 ± 0.2) subjects. There were no significant differences in EPC migration to either VEGF or SDF‐1α. Telomere length did not differ between groups. These results indicate that increased adiposity adversely affects the ability of EPCs to release proangiogenic cytokines and resist apoptosis, potentially compromising their reparative potential.
Son Is Essential for Nuclear Speckle Organization and Cell Cycle Progression
Authors: Sharma Alok , Takata Hideaki , Shibahara Kei-ichi , Bubulya Athanasios , Bubulya Paula A.
Published: January 2010

Subnuclear organization and spatiotemporal regulation of pre-mRNA processing factors is essential for the production of mature protein-coding mRNAs. We have discovered that a large protein called Son has a novel role in maintaining proper nuclear organization of pre-mRNA processing factors in nuclear speckles. The primary sequence of Son contains a concentrated region of multiple unique tandem repeat motifs that may support a role for Son as a scaffolding protein for RNA processing factors in nuclear speckles. We used RNA interference (RNAi) approaches and high-resolution microscopy techniques to study the functions of Son in the context of intact cells. Although Son precisely colocalizes with pre-mRNA splicing factors in nuclear speckles, its depletion by RNAi leads to cell cycle arrest in metaphase and causes dramatic disorganization of small nuclear ribonuclear protein and serine-arginine rich protein splicing factors during interphase. Here, we propose that Son is essential for appropriate subnuclear organization of pre-mRNA splicing factors and for promoting normal cell cycle progression.
Nuclear Translocation of β-Actin Is Involved in Transcriptional Regulation during Macrophage Differentiation of HL-60 Cells
Authors: Xu Yong Zhong , Thuraisingam Thusanth , de Lima Morais David Anderson , Rola-Pleszczynski Marek , Radzioch Danuta
Published: January 2010

Studies have shown that nuclear translocation of actin occurs under certain conditions of cellular stress; however, the functional significance of actin import remains unclear. Here, we demonstrate that during the phorbol 12-myristate 13-acetate (PMA)-induced differentiation of HL-60 cells toward macrophages, β-actin translocates from the cytoplasm to the nucleus and that this process is dramatically inhibited by pretreatment with p38 mitogen-activated protein kinase inhibitors. Using chromatin immunoprecipitation-on-chip assays, the genome-wide maps of β-actin binding to gene promoters in response to PMA treatment is analyzed in HL-60 cells. A gene ontology-based analysis shows that the identified genes belong to a broad spectrum of functional categories such as cell growth and differentiation, signal transduction, response to external stimulus, ion channel activity, and immune response. We also demonstrate a correlation between β-actin occupancy and the recruitment of RNA polymerase II at six selected target genes, and β-actin knockdown decreases the mRNA expression levels of these target genes induced by PMA. We further show that nuclear β-actin is required for PMA-induced transactivation of one target gene, solute carrier family 11 member 1, which is important for macrophage activation. Our data provide novel evidence that nuclear accumulation of β-actin is involved in transcriptional regulation during macrophage-like differentiation of HL-60 cells.
Enhanced H2AX Phosphorylation, DNA Replication Fork Arrest, and Cell Death in the Absence of Chk1
Authors: Gagou Mary E. , Zuazua-Villar Pedro , Meuth Mark
Published: January 2010

H2AX phosphorylation at serine 139 (γH2AX) is a sensitive indicator of both DNA damage and DNA replication stress. Here we show that γH2AX formation is greatly enhanced in response to replication inhibitors but not ionizing radiation in HCT116 or SW480 cells depleted of Chk1. Although H2AX phosphorylation precedes the induction of apoptosis in such cells, our results suggest that cells containing γH2AX are not committed to death. γH2AX foci in these cells largely colocalize with RPA foci and their formation is dependent upon the essential replication helicase cofactor Cdc45, suggesting that H2AX phosphorylation occurs at sites of stalled forks. However Chk1-depleted cells released from replication inhibitors retain γH2AX foci and do not appear to resume replicative DNA synthesis. BrdU incorporation only occurs in a minority of Chk1-depleted cells containing γH2AX foci after release from thymidine arrest and, in cells incorporating BrdU, DNA synthesis does not occur at sites of γH2AX foci. Furthermore activated ATM and Chk2 persist in these cells. We propose that the γH2AX foci in Chk1-depleted cells may represent sites of persistent replication fork damage or abandonment that are unable to resume DNA synthesis but do not play a direct role in the Chk1 suppressed death pathway.
Intracellular Targeting Signals and Lipid Specificity Determinants of the ALA/ALIS P4-ATPase Complex Reside in the Catalytic ALA α-Subunit
Authors: López-Marqués Rosa L. , Poulsen Lisbeth R. , Hanisch Susanne , Meffert Katharina , Buch-Pedersen Morten J. , Jakobsen Mia K. , Pomorski Thomas Günther , Palmgren Michael G.
Published: January 2010

Members of the P4subfamily of P-type ATPases are believed to catalyze flipping of phospholipids across cellular membranes, in this way contributing to vesicle biogenesis in the secretory and endocytic pathways. P4-ATPases form heteromeric complexes with Cdc50-like proteins, and it has been suggested that these act as β-subunits in the P4-ATPase transport machinery. In this work, we investigated the role of Cdc50-like β-subunits of P4-ATPases for targeting and function of P4-ATPase catalytic α-subunits. We show that the Arabidopsis P4-ATPases ALA2 and ALA3 gain functionality when coexpressed with any of three different ALIS Cdc50-like β-subunits. However, the final cellular destination of P4-ATPases as well as their lipid substrate specificity are independent of the nature of the ALIS β-subunit they were allowed to interact with.
Global Up-Regulation of Microtubule Dynamics and Polarity Reversal during Regeneration of an Axon from a Dendrite
Authors: Stone Michelle C. , Nguyen Michelle M. , Tao Juan , Allender Dana L. , Rolls Melissa M.
Published: January 2010

Axon regeneration is crucial for recovery after trauma to the nervous system. For neurons to recover from complete axon removal they must respecify a dendrite as an axon: a complete reversal of polarity. We show that Drosophila neurons in vivo can convert a dendrite to a regenerating axon and that this process involves rebuilding the entire neuronal microtubule cytoskeleton. Two major microtubule rearrangements are specifically induced by axon and not dendrite removal: 1) 10-fold up-regulation of the number of growing microtubules and 2) microtubule polarity reversal. After one dendrite reverses its microtubules, it initiates tip growth and takes on morphological and molecular characteristics of an axon. Only neurons with a single dendrite that reverses polarity are able to initiate tip growth, and normal microtubule plus-end dynamics are required to initiate this growth. In addition, we find that JNK signaling is required for both the up-regulation of microtubule dynamics and microtubule polarity reversal initiated by axon injury. We conclude that regulation of microtubule dynamics and polarity in response to JNK signaling is key to initiating regeneration of an axon from a dendrite.
Functional embedding for the classification of gene expression profiles
Authors: Wu Ping-Shi , Müller Hans-Georg
Published: January 2010

Motivation: Low sample size n high-dimensional large p data with n≪p are commonly encountered in genomics and statistical genetics. Ill-conditioning of the variance-covariance matrix for such data renders the traditional multivariate data analytical approaches unattractive. On the other side, functional data analysis (FDA) approaches are designed for infinite-dimensional data and therefore may have potential for the analysis of large p data. We herein propose a functional embedding (FEM) technique, which exploits the interface between multivariate and functional data, aiming at borrowing strength across the sample through FDA techniques in order to resolve the difficulties caused by the high dimension p.

Results: Using pairwise dissimilarities among predictor variables, one obtains a univariate configuration of these covariates. This is interpreted as variable ordination that defines the domain of a suitable function space, thus leading to the FEM of the high-dimensional data. The embedding may then be followed by functional logistic regression for the classification of high-dimensional multivariate data as an example for downstream analysis. The resulting functional classification is evaluated on several published gene expression array datasets and a mass spectrometric data, and is shown to compare favorably with various methods that have been employed previously for the classification of these high-dimensional gene expression profiles.

Availability: The implementation of FEM and Classification via Functional Embedding (CFEM) as described in this article was done with the PACE package written in Matlab. The latest version of PACE is publicly accessible at http://anson.ucdavis.edu/∼mueller/data/programs.html. An example MATLAB script for FEM is available at http://www.lehigh.edu/∼psw205/psw205.html

Contact:  psw205@lehigh.edu; mueller@wald.ucdavis.edu
Comparative evaluation of a mixed-fisheries effort-management system based on the Faroe Islands example
Authors: Baudron Alan , Ulrich Clara , Nielsen J. Rasmus , Boje Jesper
Published: January 2010

Baudron, A., Ulrich, C., Nielsen, J. R., and Boje, J. 2010. Comparative evaluation of a mixed-fisheries effort-management system based on the Faroe Islands example. – ICES Journal of Marine Science, 67: 1036–1050. Total allowable catch (TAC) management has in many fisheries, especially mixed fisheries, failed to meet conservation objectives. For instance, for the Faroe Plateau mixed demersal fisheries, the TAC system failed to achieve the objective of an average annual fishing mortality of 0.45 for the three gadoid stocks cod (Gadus morhua), haddock (Melanogrammus aeglefinus), and saithe (Pollachius virens). Therefore, in 1996, an effort-regulation system with individual transferable effort quotas was introduced to manage the fisheries. Experience has shown that effort management without additional stock-specific measures may not be appropriate for such fisheries. A management strategy evaluation model was developed to compare an effort-management system based on the Faroese example with a TAC system as currently applied in EU fisheries. Results show that when stocks are considered in isolation, a total allowable effort system does not necessarily perform better than a TAC one. It depends on stock status and dynamics, the level of uncertainty, and the reactivity of the system to changes in scientific advice. When the stocks are considered together in mixed fisheries, effort management seems, however, to be appropriate, and interannual flexibility of the system appears to be the best compromise between short- and long-term objectives, as well as between biological sustainability and economic return.
Rapid Detection of Phytophthora ramorum and P. kernoviae by Two-Minute DNA Extraction Followed by Isothermal Amplification and Amplicon Detection by Generic Lateral Flow Device
Authors: Tomlinson J. A. , Dickinson M. J. , Boonham N.
Published: January 2010

A method for nucleic-acid-based detection of pathogens in plant material has been developed which comprises a simple and rapid method for extracting DNA on the nitrocellulose membranes of lateral-flow devices, loop-mediated isothermal amplification (LAMP) of target DNA using labeled primers, and detection of the generically labeled amplification products by a sandwich immunoassay in a lateral-flow-device format. Each of these steps can be performed without specialist equipment and is suitable for on-site use, and a result can be obtained in just over an hour. A LAMP assay for the detection of plant DNA (cytochrome oxidase gene) can be used in conjunction with pathogen-specific assays to confirm negative results. The use of this method is demonstrated for the detection of Phytophthora ramorum, the causal agent of sudden oak death and dieback/leaf blight in a range of tree, shrub, and herbaceous species, and the recently described pathogen P. kernoviae.
Group provisioning limits sharing conflict among nestlings in joint-nesting Taiwan yuhinas
Authors: Shen Sheng-Feng , Chen Hsueh-Chen , Vehrencamp Sandra L. , Yuan Hsiao-Wei
Published: January 2010

Offspring often compete over limited available resources. Such sibling competition may be detrimental to parents both because it entails wasted expenditure and because it allows stronger offspring to obtain a disproportionate share of resources. We studied nestling conflict over food and its resolution in a joint-nesting species of bird, the Taiwan yuhina (Yuhina brunneiceps). We show that adult yuhinas coordinate their feeding visits, and that this coordination limits competition among nestlings, leading to a ‘fairer’ division of resources. Transponder identification and video-recording systems were used to observe adult feeding and nestling begging behaviours. We found that: (i) yuhinas feed nestlings more often in large parties than in small parties; (ii) feeding events occurred non-randomly in bouts of very short intervals; and (iii) food distribution among nestlings was more evenly distributed, and fewer nestlings begged, during large-party feeding bouts compared with small-party feeding bouts. To our knowledge, this is the first study in a cooperative breeding species showing that adults can influence food allocation and competition among nestlings by coordinating their feeding visits. Our results confirm the hypothesis that the monopolizability of food affects the intensity of sibling competition, and highlight the importance of understanding the temporal strategies of food delivery.