bluuebunny/crossref_metadata_2025 huggingface dataset:
Transfer function characteristics of the neural and peripheral arterial baroreflex arcs at rest and during postexercise muscle ischemia in humans
Authors: Ogoh Shigehiko , Fisher James P. , Young Colin N. , Raven Peter B. , Fadel Paul J.
Published: March 2009

Previous studies have demonstrated an increase in the arterial baroreflex (ABR) control of muscle sympathetic nerve activity (MSNA) during isolated activation of the muscle metaboreflex with postexercise muscle ischemia (PEMI). However, the increased ABR-MSNA control does not appear to manifest in an enhancement in the ABR control of arterial blood pressure (BP), suggesting alterations in the transduction of MSNA into a peripheral vascular response and a subsequent ABR-mediated change in BP. Thus we examined the operating gains of the neural and peripheral arcs of the ABR and their interactive relationship at rest and during muscle metaboreflex activation. In nine healthy subjects, graded isolation of the muscle metaboreflex was achieved by PEMI following isometric handgrip performed at 15% and 30% maximal voluntary contraction (MVC). To obtain the sensitivities of the ABR neural and peripheral arcs, the transfer function gain from BP to MSNA and MSNA to femoral vascular conductance, respectively, was analyzed. No changes from rest were observed in the ABR neural or peripheral arcs during PEMI after 15% MVC handgrip. However, PEMI following 30% MVC handgrip increased the low frequency (LF) transfer function gain between BP and MSNA (ABR neural arc; +58 ± 28%, P = 0.036), whereas the LF gain between MSNA and femoral vascular conductance (ABR peripheral arc) was decreased from rest (−36 ± 8%, P = 0.017). These findings suggest that during high-intensity muscle metaboreflex activation an increased ABR gain of the neural arc appears to offset an attenuation of the peripheral arc gain to help maintain the overall ABR control of systemic BP.
Air pollution and cardiac remodeling: a role for RhoA/Rho-kinase
Authors: Ying Zhekang , Yue Peibin , Xu Xiaohua , Zhong Mianhua , Sun Qinghua , Mikolaj Michael , Wang Aixia , Brook Robert D. , Chen Lung Chi , Rajagopalan Sanjay
Published: March 2009

Exposure to ambient air pollution has been associated with increases in blood pressure. We have previously demonstrated activation of the Rho/Rho kinase pathway in experimental hypertension in rats. In this investigation, we evaluated the effects of particulate matter of <2.5 μm (PM2.5) exposure on cardiovascular responses and remodeling and tested the effect of Rho kinase inhibition on these effects. C57BL/6 mice were exposed to concentrated ambient PM2.5 or filtered air for 12 wk followed by a 14-day ANG II infusion in conjunction with fasudil, a Rho kinase antagonist, or placebo treatment. Blood pressure was monitored, followed by analysis of vascular function and ventricular remodeling indexes. PM2.5 exposure potentiated ANG II-induced hypertension, and this effect was abolished by fasudil treatment. Cardiac and vascular RhoA activation was enhanced by PM2.5 exposure along with increased expression of the guanine exchange factors (GEFs) PDZ-RhoGEF and p115 RhoGEF in PM2.5-exposed mice. Parallel with increased RhoA activation, PM2.5 exposure increased ANG II-induced cardiac hypertrophy and collagen deposition, with these increases being normalized by fasudil treatment. In conclusion, PM2.5 potentiates cardiac remodeling in response to ANG II through RhoA/Rho kinase-dependent mechanisms. These findings have implications for the chronic cardiovascular health effects of air pollution.
Paraplegia increased cardiac NGF content, sympathetic tonus, and the susceptibility to ischemia-induced ventricular tachycardia in conscious rats
Authors: Lujan Heidi L. , Chen Ying , DiCarlo Stephen E.
Published: March 2009

Midthoracic spinal cord injury is associated with ventricular arrhythmias that are mediated, in part, by enhanced cardiac sympathetic activity. Furthermore, it is well known that sympathetic neurons have a lifelong requirement for nerve growth factor (NGF). NGF is a neurotrophin that supports the survival and differentiation of sympathetic neurons and enhances target innervation. Therefore, we tested the hypothesis that paraplegia is associated with an increased cardiac NGF content, sympathetic tonus, and susceptibility to ischemia-induced ventricular tachyarrhythmias. Intact and paraplegic (6–9 wk posttransection, T5 spinal cord transection) rats were instrumented with a radiotelemetry device for recording arterial pressure, temperature, and ECG, and a snare was placed around the left main coronary artery. Following recovery, the susceptibility to ventricular arrhythmias (coronary artery occlusion) was determined in intact and paraplegic rats. In additional groups of matched intact and paraplegic rats, cardiac nerve growth factor content (ELISA) and cardiac sympathetic tonus were determined. Paraplegia, compared with intact, increased cardiac nerve growth factor content (2,146 ± 286 vs. 180 ± 36 pg/ml, P < 0.05) and cardiac sympathetic tonus (154 ± 4 vs. 68 ± 4 beats/min, P < 0.05) and decreased the ventricular arrhythmia threshold (3.6 ± 0.2 vs. 4.9 ± 0.2 min, P < 0.05). Thus altered autonomic behavior increases the susceptibility to ventricular arrhythmias in paraplegic rats.
Thyroid hormone analog 3,5-diiodothyropropionic acid promotes healthy vasculature in the adult myocardium independent of thyroid effects on cardiac function
Authors: Liu Yingheng , Wang Dajun , Redetzke Rebecca A. , Sherer Benjamin A. , Gerdes A. Martin
Published: March 2009

Patients with hypothyroidism are at a higher risk for coronary vascular disease. Patients with diabetes and related vascular complications also have an increased incidence of low thyroid function. While thyroid hormones (THs) may be key regulators of a healthy vasculature, potential undesirable side effects hinder their use in the treatment of vascular disorders. TH analogs such as 3,5-diiodothyropropionic acid (DITPA) may provide a safer treatment option. However, the relative potency of DITPA on vascular growth, cardiac function, and metabolism is poorly understood. We hypothesized that the vascular growth-promoting effects of DITPA can be obtained with a minimum effect on cardiac function. Thyroidectomized Sprague-Dawley rats were given slow-release pellets with either thyroxine (T4, 2.7 or 5.2 mg) or DITPA (80 mg) for 6 wk and were compared with placebo. Heart mass, body mass, body temperature, serum THs, cardiac function (echocardiograms and hemodynamics), and myocardial arteriolar density were determined. Hypothyroidism led to reductions in cardiac function, heart mass, body temperature, and myocardial arterioles. High-dose T4 prevented arteriolar loss and the development of hypothyroidism. Low-dose T4 partially prevented the reduction in cardiac function but had minimal effects on arteriolar loss. In contrast, DITPA treatment prevented myocardial arteriolar loss but not the progression of hypothyroid-induced changes in cardiac function. The results suggested that DITPA can promote a healthy vasculature independently from its thyroid-related metabolic effects. Drugs in this class may provide new therapeutic options for patients with vascular disease.
Regulation of nitrite transport in red blood cells by hemoglobin oxygen fractional saturation
Authors: Vitturi Dario A. , Teng Xinjun , Toledo José C. , Matalon Sadis , Lancaster Jack R. , Patel Rakesh P.
Published: March 2009

Allosteric regulation of nitrite reduction by deoxyhemoglobin has been proposed to mediate nitric oxide (NO) formation during hypoxia. Nitrite is predominantly an anion at physiological pH, raising questions about the mechanism by which it enters the red blood cell (RBC) and whether this is regulated and coupled to deoxyhemoglobin-mediated reduction. We tested the hypothesis that nitrite transport by RBCs is regulated by fractional saturation. Using human RBCs, nitrite consumption was faster at lower fractional saturations, consistent with faster reactions with deoxyheme. A membrane-based regulation was suggested by slower nitrite consumption with intact versus lysed RBCs. Interestingly, upon nitrite addition, intracellular nitrite concentrations attained a steady state that, despite increased rates of consumption, did not change with decreasing oxygen tensions, suggesting a deoxygenation-sensitive step that either increases nitrite import or decreases the rate of nitrite export. A role for anion exchanger (AE)-1 in the control of nitrite export was suggested by increased intracellular nitrite concentrations in RBCs treated with DIDS. Moreover, deoxygenation decreased steady-state levels of intracellular nitrite in AE-1-inhibited RBCs. Based on these data, we propose a model in which deoxyhemoglobin binding to AE-1 inhibits nitrite export under low oxygen tensions allowing for the coupling between deoxygenation and nitrite reduction to NO along the arterial-to-venous gradient.
Voltage-independent calcium channels mediate lipopolysaccharide-induced hyporeactivity to endothelin-1 in the rat aorta
Authors: El-Awady Mohammed S. H. , Smirnov Sergey V. , Watson Malcolm L.
Published: March 2009

The roles of intracellular calcium concentration ([Ca2+]i) and Ca2+sensitization in lipopolysaccharide (LPS)-induced vascular smooth muscle (VSM) hyporesponsiveness are incompletely understood. To investigate these roles, contraction responses to endothelin-1 (ET-1) and 80 mM KCl; relaxation responses to nifedipine; the expression levels of mRNAs of ET-1 and its receptors (ETAor ETB); the expression levels of protein kinase C (PKC) and phosphorylation of Rho kinase (ROKα), CPI-17, and myosin phosphatase target subunit-1 (MYPT1); and changes in aortic VSM cell [Ca2+]iwere measured in LPS-treated aortic rings from male Wistar rats (250–300 g). LPS (10 μg/ml, 20 h) decreased contraction induced by ET-1 (0.3–100 nM) or 80 mM KCl. LPS-induced hypocontractility was not observed in the absence of external Ca2+, but LPS-treated aorta remained hypocontractile on subsequent stepwise restoration of extracellular Ca2+(0.01–10 mM). Vascular relaxation to nifedipine; mRNA expression levels of ET-1, ETA, or ETB; protein expression levels of PKC; and phosphorylation levels of ROKα, CPI-17, and MYPT1 were not affected by LPS. In isolated aortic VSM cells, ET-1 caused a transient initial increase in [Ca2+]i, followed by a maintained tonic increase in [Ca2+]i, which was decreased by LPS pretreatment and was dependent on external Ca2+. Subsequent restoration of extracellular Ca2+increased [Ca2+]i, but this increase was lower in the LPS-treated group. This difference in response to extracellular Ca2+addition was not affected by diltiazem, but was abolished by SKF-96365. Therefore, LPS induces hyporeactivity to ET-1 in rat aorta that depends on external Ca2+influx through non-voltage-operated Ca2+channels, but not on ET-1 receptor expression or Ca2+sensitization.
A severe deficiency of coagulation factor VIIa results in attenuation of the asthmatic response in mice
Authors: Shinagawa Kazuhiko , Ploplis Victoria A. , Castellino Francis J.
Published: March 2009

Eosinophil counts in the bronchoalveolar lavage fluid of wild-type (WT) mice increased after ovalbumin (OVA) challenge, a response that was diminished in comparably challenged low-expressing coagulation factor VII (FVIItTA/tTA) mice. Levels of T helper type 2 (Th2) cytokines, IL-4, IL-5, and IL-13, and eosinophil-attracting chemokines, eotaxin and RANTES, were also lower in the OVA-challenged FVIItTA/tTAmice. Eosinophils purified from low-FVII mice underwent apoptosis at a faster rate compared with WT eosinophils, and eosinophil migration in response to eotaxin was reduced in eosinophils obtained from FVIItTA/tTAmice. Airway hyperresponsiveness and mucous layer thickness were reduced in OVA-treated FVIItTA/tTAmice, and addition of exogenous coagulation factor X (FX) enhanced mucin production in human epithelial NCI-H292 cells. Correspondingly, incubation of FX with NCI-H292 cells resulted in activated (a) FX production, suggesting that the components required for FX activation were present on NCI-H292 cells. These results demonstrate that FVIIa functions in the asthmatic response to an allergen by stimulating lung eosinophilia, airway hyperresponsiveness, and mucin production, this latter effect through its ability to activate FX in conjunction with tissue factor.
An Iterative Algorithm Combining Viscosity Method with Parallel Method for a Generalized Equilibrium Problem and Strict Pseudocontractions
Authors: Peng Jian-Wen , Liou Yeong-Cheng , Yao Jen-Chih
Published: March 2009

We introduce a new approximation scheme combining the viscosity method with parallel method for finding a common element of the set of solutions of a generalized equilibrium problem and the set of fixed points of a family of finitely strict pseudocontractions. We obtain a strong convergence theorem for the sequences generated by these processes in Hilbert spaces. Based on this result, we also get some new and interesting results. The results in this paper extend and improve some well-known results in the literature.
Segmentation of Visitors to a Heritage Site Using High-resolution Time-space Data
Authors: Tchetchik Anat , Fleischer Aliza , Shoval Noam
Published: March 2009

A novel method of data collection based on high-resolution time-space data recorded by global positioning system units was used to segment visitors to the Old City of Acre heritage site in Israel. This technique of accurately tracking the temporal and spatial behavior of visitors carrying the global positioning system units overcomes the well-known limitations of traditional data collection methods. Discrete-choice methods and a system of censored regressions were used to identify the decision-making process at each stage of the visit. The segmentation of the visitors was based on these decisions. It was found that different attributes associate the visitors with different segments at each level. Although the findings might be site specific, the data collection and the segmentation method could be used by tourism planners and decision makers at other sites.
Classification of pain in cancer patients – a systematic literature review
Authors: Knudsen AK , Aass N , Fainsinger R , Caraceni A , Klepstad P , Jordhøy M , Hjermstad MJ , Kaasa S
Published: March 2009

One of the aims of the European Palliative Care Research Collaborative (EPCRC) is to achieve consensus on a classification system for cancer pain. We performed a systematic literature review to identify existing classification systems and domains/items used to classify cancer patients with pain. In a systematic search in the databases Medline and Embase, covering 1986–2006, 692 hits were obtained. 92 papers were evaluated to address pain classification. Six standardised classification systems were identified; three of them systematically developed and partially validated. Both pain characteristics and patient characteristics relevant for cancer pain classification were included in the classification systems. All but one of the standardised systems aim at predicting treatment response or adequacy of treatment. Several domains and items used to describe cancer pain but not formally described as part of a classification system were also identified and systematised. The existing approaches to pain classification in cancer patients are different, mostly not thoroughly validated, and none is widely applied. An internationally accepted classification system for cancer pain could improve research and cancer pain management. This systematic review suggests a need for developing an international consensus on how to classify pain in cancer patients.
Nature, race, and parks: past research and future directions for geographic research
Authors: Byrne Jason , Wolch Jennifer
Published: March 2009

Geographic research on parks has been wide-ranging but has seldom examined how and why people use parks, leaving these questions to leisure science, which privileges socio-demographic variables over urban socio-spatial explanations (eg, historical, political-economic, and location factors). This article examines recent geographic perspectives on park use, drawing upon environmental justice, cultural landscape, and political ecology paradigms to redirect our attention from park users to a more critical appreciation of the historical, socio-ecological, and political-economic processes that operate through, and in turn shape, park spaces and park-going behaviors. We challenge partial, user-orientated approaches and suggest new directions for geographic research on parks.
South Africa
Authors: Buhlungu Sakhela
Published: March 2009

This article explores the ways in which a form of intellectual engagement has gone beyond merely studying society and sought to influence processes of change by engaging with actors outside disciplinary scholarship and the academy. In South Africa, the broad subdiscipline of labor studies provides probably the best illustration of this engagement, which Burawoy has termed public sociology. The article traces the emergence and growth of public sociology, initially from the position of relative privilege in the ivory tower and later to more direct forms of engagement with the new publics that emerged in the antiapartheid struggle. The discussion explains why the labor movement became the focal point of public sociology in South Africa. Finally, the article argues that the advent of democracy led to a growing assertiveness among the antiapartheid movements, including labor. Not only did this alter the terms on which public sociology was undertaken, it also resulted in a decline of public sociology inherited from the antiapartheid struggle.
US National Breastfeeding Monitoring and Surveillance: Current Status and Recommendations
Authors: Chapman Donna J. , Pérez-Escamilla Rafael
Published: March 2009

Eleven federally funded datasets assessing breastfeeding behaviors in the United States (Early Childhood Longitudinal Survey, Infant Feeding Practices Survey II, National Health and Nutrition Examination Survey, National Immunization Survey, National Survey of Children's Health, National Survey of Early Childhood Health, National Survey of Family Growth, Pediatric Nutrition Surveillance System, Pregnancy Nutrition Surveillance System, Pregnancy Risk Assessment Monitoring Survey, and WIC Participant and Program Characteristics) were reviewed to evaluate breastfeeding variables (initiation, duration and exclusivity) and determine whether relevant breastfeeding determinants were collected to evaluate breastfeeding practices from a health disparities perspective. The datasets used inconsistent breastfeeding definitions, limited ethnic descriptors, and varied regarding availability of relevant determinants. Multiple datasets collect breastfeeding data, but a coordinated US breastfeeding monitoring and surveillance system does not exist. Suggestions to improve this system include: standardizing breastfeeding definitions, expanding ethnic/racial descriptors, collecting additional relevant variables, and reducing recall periods. J Hum Lact. 25(2):139-150.
Fentanyl Transdermal Analgesia During Pregnancy and Lactation
Authors: Cohen Ronald S.
Published: March 2009

This report describes an infant who was born to a mother with chronic pain treated with fentanyl 100 µg/h transdermal patch throughout her pregnancy and during lactation. On day of life 27, when the baby was feeding and gaining weight on maternal milk, samples of the baby's blood and maternal milk were sent for analysis. The mother's milk fentanyl level was 6.4 ng/ mL. The infant's blood fentanyl level was undetectable. This preliminary report suggests that fentanyl transdermal patch treatment might be a viable option for managing chronic pain during lactation. J Hum Lact. 25(3):359-361.
Sex differences in child and adolescent mortality in the Nordic countries, 1981—2000
Authors: Gissler Mika , Rahkonen Ossi , Mortensen Laust , Arntzen Annett , Cnattingius Sven , Nybo Andersen Anne-Marie , Hemminki Elina
Published: March 2009

Aims: Unlike the situation for infant and adult mortality, there are only a few studies on child and adolescent mortality. The purpose of this study was to examine sex differences in child and adolescent mortality by age and cohort in four Nordic countries over a 20-year period. Methods: Data on all live-born children were received from national population-covering birth registries from 1981 to 2000 (Denmark, n = 1,184,926; Finland, n = 841,470 (from 1987 to 2000); Norway, n = 1,090,127; and Sweden, n = 1,961,911). Data on mortality and causes of death until the age of 20 years were received from the national cause-of-death registers. Results: The overall mortality rates were higher in Denmark and Norway than in Finland and Sweden, among both boys and girls, and the difference between countries increased over time. In all countries, boys had higher mortality rates than girls. Overall, the sex difference was larger in Denmark and Norway (36% and 33% higher, respectively) than in Finland and Sweden (both 24%). The sex differences were more pronounced for deaths of those aged 5—19 years than for those aged 0—4 years. Twelve per cent of all deaths among boys and 10% of those among girls were due to external causes, mainly unintentional injury deaths or intentional deaths. For children and adolescents aged 5—19 years, the corresponding figures were 43% for boys and 35% for girls. Conclusions: Boys have excess mortality, but the sex difference is lower in countries with lower mortality. Boys' excess mortality was only partly accounted for by deaths from external causes. Avoidable deaths and causes of death need further investigation.
Clerics die, doctors survive: A note on death risks among highly educated professionals
Authors: Erikson Robert , Torssander Jenny
Published: March 2009

Aims: Mortality is strongly associated with education. We present relative death risks of men and women in 12 educational/ occupational groups in Sweden today, with a focus on individuals with higher education. Methods: Results from Cox regressions are reported for 12 educational groups with special emphasis on those with professional education, e.g. clerics, physicians, people with medical PhDs, and university teachers. The study is based on register data of the total Swedish population in the age group of 30—64 (n = 3,734,660). Results: There is a considerable variation in mortality between educational groups. Men with compulsory education run a risk that is more than three times higher than that of professors outside medicine, and other educational groups fall in between. Medical doctors and physicians have relatively low death risks compared to those with compulsory education — less than 50% among men and less than 60% among women — although professors in medicine deviate by having higher risks than their colleagues in other subjects. Those with a theological exam show higher risks of dying during the follow-up period compared to others of a similar educational level. Professors outside medicine experience the lowest death risks of all identified groups. Conclusions: Men and women with a professional education have comparatively low death risks, particularly low among medical doctors and university employees, while the clergy seems to experience relatively higher death risks than others with a similar level of education. These patterns may reflect the effects of education as well as the selection of men and women to higher education.
Robust Damage Metric in Terms of Magnitude and Phase for Impedance-based Structural Health Monitoring
Authors: Xianhua Liu  , Paurobally Roshun
Published: March 2009

Structural health monitoring has been a research interest for its great potential for life safety and economic benefits for decades. Structural vibration impedance by way of piezoceramic patch excitation and sensoring offers a local damage detection technique and has caused a wide research interest. The commonly used damage index in this method is the rate of change of the real part of the measured electromechanical impedance. This paper studied several damage indices constructed by the real and imaginary parts or magnitude and phase. It theoretically deducted and concluded that the damage index in terms of changes in real part is in fact not a properly defined index and is physically obscure; on the other hand, indices in terms of the change in magnitude and phase were shown to be physically clear and properly defined. It further verified that a new damage index in terms of the whole complex-valued impedance turned out to be the elegant combination of the magnitude and phase damage indices in the form of their Euclidean norm. Experimental result further demonstrated that the damage index in terms of real part was largely influenced by the phase, whereas the whole complex impedance offered an optimized and robust damage metric.
Near-field Sub-band Beamforming for Damage Detection in Bridges
Authors: Medda Alessio , DeBrunner Victor
Published: March 2009

Traditional structural health monitoring techniques based on the vibration response of bridge structures are limited because of several factors — including a poorly formed aggregate system model, very low SNR, and unrealistic boundary conditions. Many times, these methods rely on global parameters to describe the dynamic behavior of local structural elements. In this paper, we proposed a novel efficient SHM technique that employs the use of compactly supported sub-band space/ frequency and time/frequency analysis using local vibration characteristics. To overcome the problem of the low-error sensitivity of features extracted from vibration signals, a near-field adaptive beamforming approach was used. This technique allows the sensor array to `scan' local portions of the structure, resulting in accurate spatial selectivity on the array and high signal-to-noise ratio for any given scan direction. Moreover, the sensor array was in direct contact with the vibrating structure, and thus the measured source is in the near-field of the array. Therefore, compensation of the sensor output was needed. Sub-band analysis and adaptive beamforming were integrated in a wavelet packet sub-band framework. We utilized the 3D energy map of the optimized sub-band signals for any given scan direction and for each sub-band center frequency as the damage detection feature. We validated our method using classical finite elements approximation to the dynamic behavior of the system. The comparison of a simulated undamaged simply supported beam with a damaged equivalent revealed that damage is localized in frequency and aligned with the direction of the simulated damage. The energy signature comparison after the beamforming stage validates these results, localizing the damage in areas of high probability around the direction of the damage. Automatic analysis of the energy comparison map was possible using a pseudo density estimation of the damage location, allowing for an automatic damage detection procedure. The focus of our research was aimed toward the adaptation of this damage detection method to real highway bridges.
microRNA 184 regulates expression of NFAT1 in umbilical cord blood CD4+ T cells
Authors: Weitzel R. Patrick , Lesniewski Mathew L. , Haviernik Peter , Kadereit Suzanne , Leahy Patrick , Greco Nicholas J. , Laughlin Mary J.
Published: March 2009

The reduced expression of nuclear factor of activated T cells-1 (NFAT1) protein in umbilical cord blood (UCB)–derived CD4+ T cells and the corresponding reduction in inflammatory cytokine secretion after stimulation in part underlies their phenotypic differences from adult blood (AB) CD4+ T cells. This muted response may contribute to the lower incidence and severity of high-grade acute graft-versus-host disease (aGVHD) exhibited by UCB grafts. Here we provide evidence that a specific microRNA, miR-184, inhibits NFAT1 protein expression elicited by UCB CD4+ T cells. Endogenous expression of miR-184 in UCB is 58.4-fold higher compared with AB CD4+ T cells, and miR-184 blocks production of NFAT1 protein through its complementary target sequence on the NFATc2 mRNA without transcript degradation. Furthermore, its negative effects on NFAT1 protein and downstream interleukin-2 (IL-2) transcription are reversed through antisense blocking in UCB and can be replicated via exogenous transfection of precursor miR-184 into AB CD4+ T cells. Our findings reveal a previously uncharacterized role for miR-184 in UCB CD4+ T cells and a novel function for microRNA in the early adaptive immune response.
Efficient construction of producer cell lines for a SIN lentiviral vector for SCID-X1 gene therapy by concatemeric array transfection
Authors: Throm Robert E. , Ouma Annastasia A. , Zhou Sheng , Chandrasekaran Anantharaman , Lockey Timothy , Greene Michael , De Ravin Suk See , Moayeri Morvarid , Malech Harry L. , Sorrentino Brian P. , Gray John T.
Published: March 2009

Retroviral vectors containing internal promoters, chromatin insulators, and self-inactivating (SIN) long terminal repeats (LTRs) may have significantly reduced genotoxicity relative to the conventional retroviral vectors used in recent, otherwise successful clinical trials. Large-scale production of such vectors is problematic, however, as the introduction of SIN vectors into packaging cells cannot be accomplished with the traditional method of viral transduction. We have derived a set of packaging cell lines for HIV-based lentiviral vectors and developed a novel concatemeric array transfection technique for the introduction of SIN vector genomes devoid of enhancer and promoter sequences in the LTR. We used this method to derive a producer cell clone for a SIN lentiviral vector expressing green fluorescent protein, which when grown in a bioreactor generated more than 20 L of supernatant with titers above 107 transducing units (TU) per milliliter. Further refinement of our technique enabled the rapid generation of whole populations of stably transformed cells that produced similar titers. Finally, we describe the construction of an insulated, SIN lentiviral vector encoding the human interleukin 2 receptor common γ chain (IL2RG) gene and the efficient derivation of cloned producer cells that generate supernatants with titers greater than 5 × 107 TU/mL and that are suitable for use in a clinical trial for X-linked severe combined immunodeficiency (SCID-X1).
Efficient yeast ChIP-Seq using multiplex short-read DNA sequencing
Authors: Lefrançois Philippe , Euskirchen Ghia M , Auerbach Raymond K , Rozowsky Joel , Gibson Theodore , Yellman Christopher M , Gerstein Mark , Snyder Michael
Published: March 2009

Background

Short-read high-throughput DNA sequencing technologies provide new tools to answer biological questions. However, high cost and low throughput limit their widespread use, particularly in organisms with smaller genomes such as S. cerevisiae. Although ChIP-Seq in mammalian cell lines is replacing array-based ChIP-chip as the standard for transcription factor binding studies, ChIP-Seq in yeast is still underutilized compared to ChIP-chip. We developed a multiplex barcoding system that allows simultaneous sequencing and analysis of multiple samples using Illumina's platform. We applied this method to analyze the chromosomal distributions of three yeast DNA binding proteins (Ste12, Cse4 and RNA PolII) and a reference sample (input DNA) in a single experiment and demonstrate its utility for rapid and accurate results at reduced costs.

Results

We developed a barcoding ChIP-Seq method for the concurrent analysis of transcription factor binding sites in yeast. Our multiplex strategy generated high quality data that was indistinguishable from data obtained with non-barcoded libraries. None of the barcoded adapters induced differences relative to a non-barcoded adapter when applied to the same DNA sample. We used this method to map the binding sites for Cse4, Ste12 and Pol II throughout the yeast genome and we found 148 binding targets for Cse4, 823 targets for Ste12 and 2508 targets for PolII. Cse4 was strongly bound to all yeast centromeres as expected and the remaining non-centromeric targets correspond to highly expressed genes in rich media. The presence of Cse4 non-centromeric binding sites was not reported previously.

Conclusion

We designed a multiplex short-read DNA sequencing method to perform efficient ChIP-Seq in yeast and other small genome model organisms. This method produces accurate results with higher throughput and reduced cost. Given constant improvements in high-throughput sequencing technologies, increasing multiplexing will be possible to further decrease costs per sample and to accelerate the completion of large consortium projects such as modENCODE.
Preferential uptake of the non steroid anti-inflammatory drug diclofenac into inflamed tissues after a single oral dose in rats
Authors: Schweitzer A , Hasler-Nguyen N , Zijlstra J
Published: March 2009

Background

Diclofenac is a nonsteroidal anti-inflammatory drug which is available as prescription (RX) and over-the-counter (OTC) medication for the systemic and topical treatment of painful and inflammatory conditions such as arthritis and back pain. This study was undertaken to investigate the distribution and retention of diclofenac and/or its metabolites in inflamed tissues, using the carrageenan-induced inflammation model and quantitative whole body autoradiography in rats.

Methods

[14C]diclofenac sodium was administrated as a single 2 mg/kg oral dose 1 h after injection of carrageenan into one front and one hind footpads and subcutaneously into the dorsum of the neck of rats. A control animal received saline injections. Three carrageenan-treated rats and one control rat were sacrificed at 1, 4, 8, and 24 h after [14C]diclofenac sodium administration (total of 4 rats/time point). The carcasses were immediately snap-frozen and prepared for cryosectioning. Lengthwise whole-body sections (40 μm thick), including all major tissues, were obtained from different levels across the body. The tissue concentrations of total radiolabeled components were determined using quantitative autoradioluminography.

Results

The radioactivity patterns demonstrated that diclofenac and/or its metabolites preferentially distributed into the inflamed tissues. In unharmed tissues the distribution was similar in control and treated animals. The exposure, based on the areas under the tissue concentration versus time (AUC0-tlast), was 26 and 53 fold higher in the inflamed neck and inflamed footpads of treated animals than in control rats; the exposures in unharmed tissues were similar in the treated and control rats, and the AUC0-tlast was 17 fold higher in the inflamed paws than in the non inflamed footpads of the carrageenan-treated rats. The higher exposure in the inflamed tissues may be explained partly to the fact that the elimination of total radiolabeled components from inflamed tissues (t1/2 = 6 h) appeared lower than from the corresponding unharmed tissues (t1/2 = 2 h).

Conclusion

This animal study demonstrated that diclofenac and/or its metabolites were rapidly and preferentially taken up and retained in inflamed tissues. Although there were theoretical considerations that mildly acidic NSAID may show some preferential distribution in inflamed tissues there was no clear experimental proof for diclofenac until the present study.
De novo complex intra chromosomal rearrangement after ICSI: characterisation by BACs micro array-CGH
Authors: Kasakyan Serdar , Lohmann Laurence , Aboura Azeddine , Quimsiyeh Mazin , Menezo Yves , Tachdjian Gerard , Benkhalifa Moncef
Published: March 2009

Background

In routine Assisted Reproductive Technology (ART) men with severe oligozoospermia or azoospermia should be informed about the risk of de novo congenital or chromosomal abnormalities in ICSI program. Also the benefits of preimplantation or prenatal genetic diagnosis practice need to be explained to the couple.

Methods

From a routine ICSI attempt, using ejaculated sperm from male with severe oligozoospermia and having normal karyotype, a 30 years old pregnant woman was referred to prenatal diagnosis in the 17th week for bichorionic biamniotic twin gestation. Amniocentesis was performed because of the detection of an increased foetal nuchal translucency for one of the fetus by the sonographic examination during the 12th week of gestation (WG). Chromosome and DNA studies of the fetus were realized on cultured amniocytes

Results

Conventional, molecular cytogenetic and microarray CGH experiments allowed us to conclude that the fetus had a de novo pericentromeric inversion associated with a duplication of the 9p22.1-p24 chromosomal region, 46,XY,invdup(9)(p22.1p24) [arrCGH 9p22.1p24 (RP11-130C19 → RP11-87O1)x3]. As containing the critical 9p22 region, our case is in coincidence with the general phenotype features of the partial trisomy 9p syndrome with major growth retardation, microcephaly and microretrognathia.

Conclusion

This de novo complex chromosome rearrangement illustrates the possible risk of chromosome or gene defects in ICSI program and the contribution of array-CGH for mapping rapidly de novo chromosomal imbalance.
Effects of HIV-1 infection in vitro on transendothelial migration by monocytes and monocyte-derived macrophages
Authors: Westhorpe Clare L V , Zhou Jingling , Webster Nicole L , Kalionis Bill , Lewin Sharon R , Jaworowski Anthony , Muller William A , Crowe Suzanne M
Published: March 2009

Monocytes constitutively migrate from the bloodstream across the vascular endothelium for systemic immune surveillance and maintenance of macrophage populations. They also perform reverse transendothelial migration (TEM) across the endothelium, which is required for entry of tissue monocytes/macrophages into the lymphatics or back into the bloodstream. We have modeled these processes previously using HUVEC monolayers grown on three-dimensional collagen matrices. The aim of the present study was to determine whether HIV-1 infection of monocytes/macrophages in vitro affects TEM. Purified primary human monocytes and monocyte-derived macrophages (MDM) expressed important TEM proteins such as CD62L, CD18, PECAM-1, CCR2, and CCR8. Purified monocytes underwent efficient forward and reverse TEM across HUVEC, and this function was maintained by MDM after up to 15 days of culture. Monocytes exposed to HIV-1 for 2 days had unaltered forward or reverse TEM. However, HIV-1 infection of MDM for 7 days decreased reverse TEM by an average of 66.5% compared with mock-infected MDM (n=9 independent donors; P=0.004), without affecting forward TEM. Decreased reverse TEM by HIV-infected MDM required viral RT and was not a result of alterations in surface expression of CCR8 or p-glycoprotein or a general impairment in mobility, as assessed by migration toward fMLP. This study indicates that HIV-1 infection of macrophages reduces their capacity to emigrate from the subendothelial extracellular matrix in vitro, which could result in defective cell-mediated immune responses to infections and promote establishment of viral reservoirs of HIV in tissue macrophages in vivo.
Qualitative and Quantitative Assessment of Taste and Smell Changes in Patients Undergoing Chemotherapy for Breast Cancer or Gynecologic Malignancies
Authors: Steinbach Silke , Hummel Thomas , Böhner Christina , Berktold Sabina , Hundt Walter , Kriner Monika , Heinrich Petra , Sommer Harald , Hanusch Claus , Prechtl Anita , Schmidt Burghart , Bauerfeind Ingo , Seck Katharina , Jacobs Volker R. , Schmalfeldt Barbara , Harbeck Nadia
Published: March 2009

Purpose

Smell and taste changes during chemotherapy are significant complaints of cancer patients. Loss of olfactory/gustatory function can lead to malnutrition, weight loss, and possibly a prolonged morbidity of chemotherapy-induced adverse effects, decreased quality of life, poor compliance, and even decreased therapy response. This prospective study comprehensively investigated, to our knowledge for the first time, smell and taste changes in a cohort of 87 patients undergoing chemotherapy for breast cancer or gynecologic malignancies.

Patients and Methods

Olfactory function was tested using Sniffin' Sticks (Burghart; Wedel, Germany) and gustatory function was tested using taste strips before, during, and immediately and 3 months after chemotherapy.

Results

Olfactory and gustatory function significantly decreased during chemotherapy and recovered almost completely 3 months after chemotherapy. Scores of odor thresholds were affected more than those of discrimination or identification. The olfactory function of older patients was affected more than that of younger patients. There was no difference in the olfactory function during chemotherapy with respect to the chemotherapeutic agent or initial diagnosis (breast or ovarian cancer). Regarding taste, scores of salty taste were affected more than scores of sweet, sour, or bitter taste. The gustatory function did not differ significantly during chemotherapy with respect to age or diagnosis but did differ with respect to the chemotherapeutic agent. Taxane-based chemotherapy caused the most severe disorders.

Conclusion

Chemotherapy has a significant but transient effect on olfactory and gustatory function, possibly causing reduced appetite, a low energy intake, and weight loss. Additional spices and flavoring may compensate for this diminished chemosensory function, enhancing patient compliance and quality of life.
Is the ISCP serving us well? A survey of UK trainees
Authors: Pothier DD , Giddings C
Published: March 2009

In August 2008 a compulsory annual fee of £125 was imposed on surgical trainees to support the ongoing costs of the Joint Committee on Surgical Training (JCST). This included the costs of running the Intercollegiate Surgical Curriculum Programme (ISCP) and contributing towards the Orthopaedic Competence Assessment Programme (OCAP). Fees are to be paid by all trainees using ISCP or OCAP surgical curricula from August 2008. This excludes Calman trainees as long as they do not use the electronic portfolio. Calman trainees are, however, able to use the e-logbook without charge.
The new MRCS: changes from 2008
Authors: Ward David , Parker Michael
Published: March 2009

In the 1990s the MRCS examination succeeded the original FRCS examination because of changes in surgical training. Since then several factors have caused the surgical royal colleges to review the MRCS: the introduction of run-through and core surgical training instead of basic surgical training (BST); earlier specialisation; less exposure to the range of surgical specialties; and the development of the Intercollegiate Surgical Curriculum Programme (ISCP).
From nepotism to speed-dating: selection of specialty trainees in surgery
Authors: Blencowe N , Bloor J , Sheffield J , Hollowood A
Published: March 2009

When Sainsbury's recruit checkout assistants, candidates complete a detailed application form and attend a two-part interview lasting up to one-and-a-half hours. The majority of interviews for recruitment to specialty training in 2007 lasted 30 minutes. Is 30 minutes long enough to make an assessment about whether a junior doctor possesses the skills necessary to become a consultant surgeon?
An academic foundation programme in trauma and orthopaedic surgery
Authors: Smith CM , Cooper L , Dutton T , Costa ML
Published: March 2009

There have been substantial recent changes to the structure of clinical academic training. In its 2004 white paper, Science and innovation: working towards a ten-year investment framework, the government issued a call for improvements in clinical research in the NHS, to ensure that scientific advances would translate to genuine improvements in patients' care. In response to this, the UK Clinical Research Collaboration was set up to enhance the partnership between government, industry and medical sectors. They identified three current major problems in clinical academic training: lack of a clear entry route and career structure; lack of flexibility in job content and location; and a shortage of suitably structured posts on training completion. In 2004, 10% of academic posts were unfilled and there were 23% fewer junior academic staff than three years previously.
Using unembalmed cadavers in surgical training
Authors: Mahadevan Vishy
Published: March 2009

Until 1995 the scope of anatomical teaching at the College was more or less determined by and tailored to the requirements of the FRCS and FDS primary examinations. To that end, the anatomy department of the College was staffed by medically qualified anatomists, a prosector and anatomy technicians. The relatively simple facilities in the department of the time included anatomy demonstration rooms, seminar rooms, a prosectorium in which donated cadavers (embalmed in formalin) were dissected by the prosector and a storage facility for the prosected specimens. Intensive courses in anatomy and other basic surgical sciences were conducted at frequent intervals and these popular courses were attended by junior surgical trainees preparing for the FRCS primary.
Launch of the ATLS® 8th edition
Authors: Roden Rosalind
Published: March 2009

On 27 March 2009 The Royal College of Surgeons of England will host the launch of the eighth edition of the Advanced Trauma Life Support® (ATLS®) programme in the UK. ATLS® has just entered its 21st year of life in the UK. The courses are run in 120 regional centres and in 2008 trained over 4,200 providers. ATLS® represents the gold standard for those involved in the early management of major trauma. The programme currently has over 600 active instructors from a wide range of specialties, including general surgery, emergency medicine, trauma and orthopaedics, anaesthesiology and radiology. More than anything ATLS® promotes a common language among those of us who work together to care for trauma victims.
The new IMOrth: the candidate's perspective
Authors: Bains Manjinder , Sheriteh Zahra , Ryan Fiona S
Published: March 2009

The format of the Intercollegiate Membership in Orthodontics (IMOrth) exam has recently changed dramatically and the first sitting took place in June 2008. As the first cohort of candidates to go through the new format, we felt it would be helpful to share our experiences.
Eagle Project phase 2: clinical skills unit
Authors: Thomas WEG
Published: March 2009

Following the outstanding success of phase 1 of the Eagle Project, the Wolfson Surgical Skills Centre, it is with great excitement and anticipation that the College launches phase 2, the clinical skills unit. The completion of this very ambitious second phase of the multimillion-pound project is a tribute to teamworking and has only been possible through close cooperation between the College, generous benefactors and industry. The impact that this new unit will have is enormous. It will provide flexible state-of-the-art facilities that will allow much stronger engagement with course development and course delivery.
Oral and maxillofacial surgery – future uncertain?
Authors: Martin Ian
Published: March 2009

Oral and maxillofacial surgery (OMFS) occupies the confluence of the two great professions of medicine and dentistry. It has developed as a consequence of cross-fertilisation of ideas between surgery and dentistry, from which patients have benefited. While the last decade witnessed OMFS becoming firmly established as one of the nine SAC-recognised surgical specialties, only in the last couple of years has the dental specialty of oral surgery became disarticulated from OMFS.
Facing the future: the FGDP(UK) facial aesthetics masterclass
Authors: Hook Melanie
Published: March 2009

The Faculty of General Dental Practice UK (FGDP(UK)) is proud to announce its innovative facial aesthetics masterclass, due to be launched in July 2009. It is the only facial aesthetics course for dentists in the UK that combines instruction in the performance of practical procedures with dissection exercises on fresh, unembalmed cadaver heads (see also article on page 80).