bluuebunny/crossref_metadata_2025 huggingface dataset:
R5 human immunodeficiency virus type 1 with efficient DC-SIGN use is not selected for early after birth in vertically infected children
Authors: Borggren Marie , Navér Lars , Casper Charlotte , Ehrnst Anneka , Jansson Marianne
Published: December 2012

The binding of human immunodeficiency virus (HIV) to C-type lectin receptors may result in either enhanced trans-infection of T-cells or virus degradation. We have investigated the efficacy of HIV-1 utilization of DC-SIGN, a C-type lectin receptor, in the setting of intrauterine or intrapartum mother-to-child transmission (MTCT). Viruses isolated from HIV-1-infected mothers at delivery and from their vertically infected children both shortly after birth and later during the progression of the disease were analysed for their use of DC-SIGN, binding and ability to trans-infect. DC-SIGN use of a child’s earlier virus isolate tended to be reduced as compared with that of the corresponding maternal isolate. Furthermore, the children’s later isolate displayed enhanced DC-SIGN utilization compared with that of the corresponding earlier virus. These results were also supported in head-to-head competition assays and suggest that HIV-1 variants displaying efficient DC-SIGN use are not selected for during intrauterine or intrapartum MTCT. However, viruses with increased DC-SIGN use may evolve later in paediatric HIV-1 infections.
Upregulation of special AT-rich-binding protein 1 by Epstein–Barr virus latent membrane protein 1 in human nasopharyngeal cells and nasopharyngeal cancer
Authors: Endo Kazuhira , Shackelford Julia , Aga Mitsuharu , Yoshizaki Tomokazu , Pagano Joseph S.
Published: December 2012

A global regulator of chromatin remodelling and gene expression, special AT-rich-binding protein 1 (SATB1) has been implicated in promotion of growth and metastasis of a number of cancers. Here, we demonstrate that the principal oncogene of Epstein–Barr virus (EBV), latent membrane protein 1 (LMP1) upregulates SATB1 RNA and protein expression in human nasopharyngeal cell lines. Silencing of endogenously expressed SATB1 with specific short hairpin RNA decreases cell proliferation and resistance to apoptosis induced by growth factor withdrawal. Additionally, we provide evidence that LMP1-mediated expression of Survivin, a multifunctional protein involved in promoting cell growth and survival, is mediated at least in part by SATB1 in human nasopharyngeal cells. Finally, we show that SATB1 protein levels are elevated in tissue samples from patients with nasopharyngeal carcinoma (NPC), and are directly correlated with the expression of LMP1. Taken together, our results suggest that SATB1 functions as a pro-metastatic effector of LMP1 signalling in EBV-positive NPC.
Neutralization of porcine endogenous retrovirus by antibodies against the membrane-proximal external region of the transmembrane envelope protein
Authors: Waechter Alexander , Eschricht Magdalena , Denner Joachim
Published: December 2012

Immunization of different species including goats, rats, hamsters and guinea pigs with the recombinant ectodomain of the transmembrane envelope (TM) protein p15E of porcine endogenous retrovirus (PERV) has been shown to result in the production of virus-neutralizing antibodies. The sera recognize two groups of epitopes, one located in the fusion peptide-proximal region (FPPR) and the second in the membrane-proximal external region (MPER) of p15E. Most interestingly, the epitopes in the MPER are similar to epitopes in the TM protein gp41 of human immunodeficiency virus type 1 (HIV-1) recognized by mAbs 2F5 and 4E10, which broadly neutralize HIV-1. To study which epitope and which antibody population are involved in the process of neutralization of PERV, this study generated a new antiserum in a goat using an elongated ectodomain of p15E. The immune serum neutralized PERV at a higher titre and recognized broader epitopes in the FPPR and MPER of p15E. For the first time, antibody subpopulations were isolated from this serum using affinity chromatography with immobilized proteins and peptides corresponding to the FPPR and MPER of p15E. Only the affinity-purified antibodies specifically binding the MPER neutralized PERV, indicating that, as in the case of HIV-1, the MPER is an important target of neutralizing activity.
Somatic neurofibromatosis type 1 (NF1) inactivation characterizes NF1-associated pilocytic astrocytoma
Authors: Gutmann David H. , McLellan Michael D. , Hussain Ibrahim , Wallis John W. , Fulton Lucinda L. , Fulton Robert S. , Magrini Vincent , Demeter Ryan , Wylie Todd , Kandoth Cyriac , Leonard Jeffrey R. , Guha Abhijit , Miller Christopher A. , Ding Li , Mardis Elaine R.
Published: December 2012

Low-grade brain tumors (pilocytic astrocytomas) arising in the neurofibromatosis type 1 (NF1) inherited cancer predisposition syndrome are hypothesized to result from a combination of germline and acquired somaticNF1tumor suppressor gene mutations. However, genetically engineered mice (GEM) in which mono-allelic germlineNf1gene loss is coupled with bi-allelic somatic (glial progenitor cell)Nf1gene inactivation develop brain tumors that do not fully recapitulate the neuropathological features of the human condition. These observations raise the intriguing possibility that, while loss of neurofibromin function is necessary for NF1-associated low-grade astrocytoma development, additional genetic changes may be required for full penetrance of the human brain tumor phenotype. To identify these potential cooperating genetic mutations, we performed whole-genome sequencing (WGS) analysis of three NF1-associated pilocytic astrocytoma (PA) tumors. We found that the mechanism of somaticNF1loss was different in each tumor (frameshift mutation, loss of heterozygosity, and methylation). In addition, tumor purity analysis revealed that these tumors had a high proportion of stromal cells, such that only 50%–60% of cells in the tumor mass exhibited somaticNF1loss. Importantly, we identified no additional recurrent pathogenic somatic mutations, supporting a model in which neuroglial progenitor cellNF1loss is likely sufficient for PA formation in cooperation with a proper stromal environment.
Alteration of the Alkaloid Profile in Genetically Modified Tobacco Reveals a Role of Methylenetetrahydrofolate Reductase in Nicotine N-Demethylation
Authors: Hung Chiu-Yueh , Fan Longjiang , Kittur Farooqahmed S. , Sun Kehan , Qiu Jie , Tang She , Holliday Bronwyn M. , Xiao Bingguang , Burkey Kent O. , Bush Lowell P. , Conkling Mark A. , Roje Sanja , Xie Jiahua
Published: December 2012

Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme of the tetrahydrofolate (THF)-mediated one-carbon (C1) metabolic network. This enzyme catalyzes the reduction of 5,10-methylene-THF to 5-methyl-THF. The latter donates its methyl group to homocysteine, forming methionine, which is then used for the synthesis of S-adenosyl-methionine, a universal methyl donor for numerous methylation reactions, to produce primary and secondary metabolites. Here, we demonstrate that manipulating tobacco (Nicotiana tabacum) MTHFR gene (NtMTHFR1) expression dramatically alters the alkaloid profile in transgenic tobacco plants by negatively regulating the expression of a secondary metabolic pathway nicotine N-demethylase gene, CYP82E4. Quantitative real-time polymerase chain reaction and alkaloid analyses revealed that reducing NtMTHFR expression by RNA interference dramatically induced CYP82E4 expression, resulting in higher nicotine-to-nornicotine conversion rates. Conversely, overexpressing NtMTHFR1 suppressed CYP82E4 expression, leading to lower nicotine-to-nornicotine conversion rates. However, the reduced expression of NtMTHFR did not affect the methionine and S-adenosyl-methionine levels in the knockdown lines. Our finding reveals a new regulatory role of NtMTHFR1 in nicotine N-demethylation and suggests that the negative regulation of CYP82E4 expression may serve to recruit methyl groups from nicotine into the C1 pool under C1-deficient conditions.
Experience with the use of hydroxychloroquine for the treatment of lupus erythematosus
Authors: Momose Yoko , Arai Satoru , Eto Hikaru , Kishimoto Mitsumasa , Okada Masato
Published: December 2012

Abstract

Hydroxychloroquine (HCQ) is generally used to treat systemic lupus erythematosus (SLE) in Western countries. However, chloroquine retinopathy became a problem in Japan, and chloroquine has never been used since then. Even now HCQ remains non‐approved. Therefore, the Japanese Hydroxychloroquine Study Group has been organized, and activities have started to have HCQ approved within Japan. In the present study, we investigated the effectiveness of HCQ against the skin manifestations of lupus erythematosus. There were seven patients, all female, and they consisted of four patients with SLE (skin lesion type: discoid lupus erythematosus [DLE] in three, subacute cutaneous lupus erythematosus in one and lupus erythematosus profundus in one), two patients with cutaneous lupus erythematosus (both DLE), and one patient with a combination of SLE and dermatomyositis. HCQ was effective in three patients and ineffective in the two patients. We could not judge the efficacy of HCQ in the other two patients. There were no adverse effects in any of the patients. Efficacy was exhibited against telangiectasia and erythema. HCQ is also an effective and safe treatment for Japanese patients, and it is hoped that it will be approved for use in Japan very soon.
Hospital Use of Agency‐Employed Supplemental Nurses and Patient Mortality and Failure to Rescue
Authors: Aiken Linda H. , Shang Jingjing , Xue Ying , Sloane Douglas M.
Published: December 2012

Objective

To determine the association between the use of agency‐employed supplemental registered nurses (SRNs) to staff hospitals and patient mortality and failure to rescue (FTR).

Data Sources

Primary survey data from 40,356 registered nurses in 665 hospitals in four states in 2006 were linked with American Hospital Association and inpatient mortality data from state agencies for approximately 1.3 million patients.

Study Design

Logistic regression models were used to examine the association between SRN use and 30‐day in‐hospital mortality and FTR, controlling for patient and hospital characteristics, nurse staffing, the proportion of nurses with bachelor's degrees, and quality of the work environment.

Principal Findings

Before controlling for multiple nurse characteristics of hospitals, higher proportions of agency‐employed SRNs in hospitals appeared to be associated with higher mortality (OR = 1.06) and FTR (OR = 1.05). Hospitals with higher proportions of SRNs have poorer work environments, however, and the significant relationships between SRNs and mortality outcomes were rendered insignificant when work environments were taken into account.

Conclusions

Higher use of SRNs does not appear to have deleterious consequences for patient mortality and may alleviate nurse staffing problems that could produce higher mortality.
Comparison of early biomarker strategies with the Heart Foundation of Australia/Cardiac Society of Australia and New Zealand guidelines for risk stratification of emergency department patients with chest pain
Authors: Cullen Louise , Parsonage William A , Greenslade Jaimi , Lamanna Arvin , Hammett Christopher J , Than Martin , Ungerer Jacobus PJ , Chu Kevin , O'Kane Shanen , Brown Anthony FT
Published: December 2012

Objectives

To compare cardiac risk stratification using a 0 and 2 h point‐of‐care (POC) cardiac troponin (cTn), 0 and 2 h POC multi‐biomarkers against the 0 and 6 h laboratory cTn reference standard.

Methods

A prospective observational study of ED patients presenting with chest pain was performed. Patients were risk stratified and treated as per the Heart Foundation of Australia/Cardiac Society of Australia and New Zealand (HF‐A/CS‐ANZ) guidelines using the 0 and 6 h laboratory cTn (T6). Patients were further stratified using a 0 and 2 h POC cTn (T2) plus 0 and 2 h POC multi‐biomarkers (cTn, creatine kinase‐MB, myoglobin) (M2). The T6, T2 and M2 strategies were compared using the 30‐day major adverse cardiac events as the primary outcome.

Results

Seven hundred and four patients (median age 54 years, male 62.1%) were enrolled. Using the T6 reference standard, 2%, 61% and 37% were stratified as low, intermediate and high risk, respectively. The 30‐day event rates were 0%, 3.5% and 28.6%, respectively. The T2 strategy stratified 1.5%, 57% and 41% as low, intermediate and high risk, respectively, with 30‐day event rates of 0%, 4.2% and 24.8%, respectively. The M2 strategy resulted in significantly different risk distribution with 1%, 40% and 59% stratified as low, intermediate and high risk, respectively, with 30‐day event rates of 0%, 3.9% and 18.8%, respectively.

Conclusion

Using a 2 h POC cTn‐only biomarker strategy with the HF‐A/CS‐ANZ guidelines accurately identified a population at intermediate risk of 30‐day events in whom further objective testing might be accelerated, allowing subsequent early discharge of the majority of this cohort. Within 2 h of presentation a high risk population could be identified in whom early management, including admission, was required.
A primer for clinical researchers in the emergency department: Part V: How to describe data and basic medical statistics
Authors: Donath Susan , Davidson Andrew , Babl Franz E
Published: December 2012

In this series we address key topics for clinicians who conduct research as part of their work in the ED. In this section we will address important statistical concepts for clinical researchers and readers of clinical research publications. We use practical clinical examples of how to describe clinical data for presentation and publication, and explain key statistical concepts and tests clinical researchers will likely use for the majority of ED datasets.
Potential of triptolide in reproductive management of the house rat,Rattus rattus
Authors: SINGLA Neena , KAUR Gaganpreet , BABBAR Bhupinder Kaur , SANDHU Bhupinder Singh
Published: December 2012

Mature and healthy male house rats,Rattus rattus(n= 160) were fed on bait (cracked wheat: powdered sugar, 98:2) containing different concentrations of triptolide (0.1, 0.05, 0.025 and 0%) for 7 and 14 days in no‐choice and bi‐choice feeding tests in the laboratory. The objective of the study was to record the antifertility affects of triptolide after 30 and 60 days of termination of treatment. Results revealed no significant effect of triptolide treatment on weights of testis, epididymis, seminal vesicles and prostate gland of rats. Overall, sperm motility, live sperm count, sperm density and sperm morphology in the cauda epididymal fluid were found to differ significantly (P≤ 0.05) between untreated and treated groups of rats. The major effect of triptolide on sperm morphology was in the form of sperm head tail separation, which was up to 56.0% in rats treated for 14 days in no‐choice and autopsied after 30 days. A significant effect (P≤ 0.05) of triptolide treatment was observed on the histomorphology of the testis, which included a dose‐dependent decrease in diameter of seminiferous tubules, thickness of germinal epithelium and numbers of various spermatogenic cells. Cell associations in the seminiferous epithelial cycle were poorly developed in rats ingesting medium (4.7–5.1 mg/100 g bw) and high doses (6.9–7.2 mg/100 g bw) of triptolide than rats ingesting low doses (1.8–2.3 mg/100 g bw) and untreated rats. The cell stages affected had not recovered fully within the 60 day period following triptolide withdrawal. The present study suggests the potential of triptolide in reproductive management ofRattus rattus.
Optimizing the size of the area surveyed for monitoring a Eurasian lynx (Lynx lynx) population in the Swiss Alps by means of photographic capture–recapture
Authors: ZIMMERMANN Fridolin , BREITENMOSER‐WÜRSTEN Christine , MOLINARI‐JOBIN Anja , BREITENMOSER Urs
Published: December 2012

We studied the influence of surveyed area size on density estimates by means of camera‐trapping in a low‐density felid population (1–2 individuals/100 km2). We applied non‐spatial capture–recapture (CR) and spatial CR (SCR) models for Eurasian lynx during winter 2005/2006 in the northwestern Swiss Alps by sampling an area divided into 5 nested plots ranging from 65 to 760 km2. CR model density estimates (95% CI) for models M0 and Mh decreased from 2.61 (1.55–3.68) and 3.6 (1.62–5.57) independent lynx/100 km2, respectively, in the smallest to 1.20 (1.04–1.35) and 1.26 (0.89–1.63) independent lynx/100 km2, respectively, in the largest area surveyed. SCR model density estimates also decreased with increasing sampling area but not significantly. High individual range overlaps in relatively small areas (the edge effect) is the most plausible reason for this positive bias in the CR models. Our results confirm that SCR models are much more robust to changes in trap array size than CR models, thus avoiding overestimation of density in smaller areas. However, when a study is concerned with monitoring population changes, large spatial efforts (area surveyed ≥760 km2) are required to obtain reliable and precise density estimates with these population densities and recapture rates.
A method to study slug predation on seedlings in the field
Authors: Honek A. , Saska P. , Martinkova Z. , Koprdova S.
Published: December 2012

Dandelion is a commonAsteraceaespecies that populates disturbed sites and gaps within swards where it becomes an important competitor of grasses. The natural control against dandelion includes seedling predation, with slugs, particularlyArion lusitanicus, being the most important in the Czech Republic. However, the study of slug seedling consumption is difficult because naturally established seedlings are not always available. Therefore, we developed a method of exposing laboratory‐grown seedlings as bait for slug predation. Dandelion seeds were sown in plastic cups containing a bottom layer of moist substrate. The emerged seedlings were thinned to 20 and displayed in an open area. During 2008–2010, the seedling baits were placed at 15 sites at 1 month intervals throughout the dandelion vegetative season, in parallel with plasticine baits that monitored slug feeding activity. For each 1 month interval, seedling survival was observed for a period of 8 days, and the estimated time to death was calculated; the percentage of surviving seedlings was then recorded. This method of seedling presentation demonstrated that local and temporal variation in seedling survival is correlated with slug feeding activity. The advantage of this technique is that the seedlings in baits may be presented at any place and time, as required by the experimental design; however, we found that the estimated time to seedling death was shorter for the exposed baits than for the naturally established seedlings. The method is suitable for the study of plant species other than dandelion, and also for aims other than the study of spatiotemporal trends in seedling consumption by slugs.
The unconventional geminivirus Beet curly top Iran virus: satisfying Koch's postulates and determining vector and host range
Authors: Soleimani R. , Matic S. , Taheri H. , Behjatnia S. A. A. , Vecchiati M. , Izadpanah K. , Accotto G. P.
Published: December 2012

Beet curly top Iran virus (BCTIRV) is a geminivirus with unusual genomic organisation, recently reported in Iran, infecting sugarbeet and a few other plant species. Although three BCTIRV sequences have been reported, demonstration that BCTIRV DNA is the causal agent of the disease was missing. A full‐length genomic DNA was obtained from symptomatic leaves of sugarbeet collected in the Sivand area of Iran, and its nucleotide sequence was determined (BCTIRV‐Siv, 2845 nt). To satisfy Koch's postulates, an infectivity assay was developed by inserting a 1.4‐mer of BCTIRV‐Siv DNA in Agrobacterium tumefaciens and using it in agroinoculation experiments. The cloned viral DNA was capable of infecting sugarbeets, reproducing the leaf curling and vein enations observed in the field. These results demonstrate that the single DNA component of BCTIRV is sufficient for infectivity. Host range studies indicated that some economically important crops can be affected, such as spinach, tomato and sweet pepper, as well as important laboratory plants including Nicotiana benthamiana, Arabidopsis thaliana and Jimson weed. Circulifer haematoceps, the dominant leafhopper species present in sugarbeet fields in Iran, was successfully used to transmit the disease. The availability of an infectious clone will facilitate extended host range studies, to determine the potential risks to other crops, as well as genetic studies on this unusual member of the family Geminiviridae.
Antagonistic effect of flumazenil after midazolam sedation on arterial‐cardiac baroreflex
Authors: UEDA K. , OGAWA Y. , AOKI K. , HIROSE N. , GOKAN D. , KATO J. , OGAWA S. , IWASAKI K.
Published: December 2012

Background

Flumazenil is generally administered to antagonise the sedative effect of midazolam. However, although flumazenil completely antagonises the sedative effect of midazolam, a few effects remain unantagonised. Hence, it is unclear whether flumazenil restores the attenuation of the arterial‐cardiac baroreflex (i.e. arterial‐heart rate reflex) induced by midazolam. We investigated the antagonistic effect of flumazenil administered after midazolam on cardiac baroreflex, to reveal whether complete recovery from midazolam‐induced sedation by flumazenil administration is accompanied by restoration of midazolam's attenuating effects on the cardiac baroreflex.

Method

Twelve healthy male subjects received midazolam followed by flumazenil until complete recovery from midazolam sedation. Before and during midazolam sedation, and after flumazenil administration, cardiac baroreflex function was assessed by sequence analysis and transfer function analysis between spontaneous oscillations in systolic arterial pressure and R‐R interval.

Results

During midazolam sedation, defined by an Observer's Assessment of Alertness/Sedation scale score of 3, BIS value decreased significantly. Simultaneously, the baroreflex indices of the two analyses decreased significantly compared with baseline, suggesting attenuated cardiac baroreflex function. With complete recovery from midazolam sedation by flumazenil, indicated by an Observer's Assessment of Alertness/Sedation scale score of 5, BIS values returned to the baseline level. Simultaneously, cardiac baroreflex indices also returned to baseline levels.

Conclusion

The present results suggest that complete recovery from midazolam sedation by flumazenil is accompanied by restoration of the attenuated cardiac baroreflex function induced by midazolam.
Using citizen‐based survey data to determine densities of large mammals: a case study from Mana Pools National Park, Zimbabwe
Authors: Dunham Kevin M. , du Toit Andrew J.
Published: December 2012

Since 1993, members of the national wildlife society have undertaken annual surveys of large mammals in the Zambezi alluvial woodlands of Mana Pools National Park, Zimbabwe. Data are collected along 36 systematically‐arranged transects. We provide the first thorough assessment of the data from any survey within this long‐term project. The transect data from 2011 were analysed withDISTANCEsoftware to assess if the data were suitable for determining the densities of large mammals using distance sampling techniques. Successful application of distance sampling depended on observers using printed, large‐scale, georeferenced satellite images onto which they mapped the location of animal groups detected. The assumptions of the distance sampling were well met and thus the 2011 survey provided reliable estimates of the densities of nine species of common large mammals on the Zambezi alluvium during the late dry season. Estimated density in this dry‐season concentration area varied from 3.6 km−2for kudu, to 204 km−2for impala. The precision of the estimates ranged from a coefficient of variation of 7.9% for elephant, to 25.5% for buffalo. For elephant, warthog and baboon, the morning and afternoon densities differed significantly.
Fetal Membrane Biomarker Network Diversity and Disease Functions Induced by Intra‐amniotic Pathogens
Authors: Bhat Geeta , Peltier Morgan R. , Syed Tariq Ali , Drobek Cayce O. , Saade George , Menon Ramkumar
Published: December 2012

Problem

Intra‐amniotic pathogens and by‐products activate innate immune responses encompassing multitudes of signaling molecules and pathways that can result in spontaneous preterm birth (PTB). This study investigates fetal membrane response to bacterial stimulation using a bioinformatics approach.

Method of study

Dysregulated biomarker (IL1‐β, IL‐2, IL‐8, IL‐10, and TNF‐α) data from fetal membranes at term stimulated with Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma hominis, E. coli, Group B Streptococci, Polyporhans gingivalis, or Gardnerella vaginalis with 50% (v/v) amniotic fluid (AF) were analyzed by Ingenuity Pathway Analysis.

Results

In racially stratified analysis, networks representing late‐stage immune inflammation were seen in African‐Americans in AF absence. Inflammation was dominant in AF presence as well. In Caucasians, late‐stage immune response was dominant with AF, but not in its absence.

Conclusions

Fetal membrane biofunctions in response to bacteria reflect early‐ and late‐stage innate immune defenses that vary based on the presence of AF and subject race.
Barriers to the 4‐h rule: What causes delays for gynaecology patients in the emergency department?
Authors: Pakmehr Sahar , Petersen Rodney W. , Quinlivan Julie A.
Published: December 2012

Objectives

To explore factors that led to noncompliance with the 4‐h rule for gynaecology patients in a general emergency department.

Methods

A cross‐sectional cohort study was performed at a general emergency department. The files of all female patients aged from birth to 100 years presenting from 1 January 2009 to 31 December 2010 were screened. Those patient's files where a coded gynaecological diagnosis was made were reviewed. A time flow analysis was then undertaken of 580 consecutive files to evaluate barriers to admission or discharge of patients within the 4‐h period. A further 300 files were audited to determine whether suboptimal management by emergency department staff contributed towards delays.

Results

There were 134 438 presentations to the emergency department, of which 2968 were gynaecology presentations (2.2%). The overall compliance with the 4‐h rule was 66%. Patients with acute triage status, who were pregnant or who were eventually admitted, were more likely to be managed in compliance with the 4‐h rule. The main barriers to compliance were incomplete examinations by emergency department staff; waiting for ultrasound examinations and blood test results; delays waiting for specialty review; and delays caused by initial review by surgical teams.

Conclusion

Specific barriers to compliance with the 4‐h rule can be identified in gynaecology patients. Strategies specific to overcome these barriers can be developed to improve compliance.
Enteroaggregative Escherichia coli quantification in children stool samples using quantitative PCR
Authors: Lima Ila Fernanda Nunes , Quetz Josiane da Silva , Guerrant Richard Littleton , Nataro James P. , Houpt Eric R. , Lima Aldo Angelo Moreira , Havt Alexandre
Published: December 2012

Enteroaggregative Escherichia coli (EAEC) is a common cause of infectious diarrhea, especially in children living in poor‐resource countries. In this article, we present a SYBR Green‐based real‐time polymerase chain reaction (qPCR) method for quantitative detection of EAEC in DNA directly extracted from human stool samples. To test the proposed qPCR system, we examined specificity, sensitivity, repeatability, and also the degree of DNA extraction efficiency using EAEC strain 042 spiked into EAEC‐free stool sample. The specificity of this assay was proved using six strains of EAEC, seven strains of other E. coli types, and one strain of Shigella. The detection limit of qPCR was 67 CFU/reaction. In naturally infected stool samples, we found EAEC in quantities varying from 6.7 × 105 to 2 × 109 CFU/g of feces. We could not detect any reduction after stool DNA extraction for the amounts of 107 and 106 CFU/mL of spiked EAEC. This qPCR assay is simple, rapid, reproducible, sensitive, specific, and allows rapid EAEC quantification to be used in a variety of further EAEC studies. This new quantitative method provides a relatively simple means to quantify EAEC, which will likely be key to understanding the pathophysiology and impact of EAEC infection.
Erythropoietin doping in cycling: lack of evidence for efficacy and a negative risk–benefit
Authors: Heuberger Jules A. A. C. , Cohen Tervaert Joost M. , Schepers Femke M. L. , Vliegenthart Adriaan D. B. , Rotmans Joris I. , Daniels Johannes M. A. , Burggraaf Jacobus , Cohen Adam F.
Published: December 2012

Imagine a medicine that is expected to have very limited effects based upon knowledge of its pharmacology and (patho)physiology and that is studied in the wrong population, with low‐quality studies that use a surrogate end‐point that relates to the clinical end‐point in a partial manner at most. Such a medicine would surely not be recommended. The use of recombinant human erythropoietin (rHuEPO) to enhance performance in cycling is very common. A qualitative systematic review of the available literature was performed to examine the evidence for the ergogenic properties of this drug, which is normally used to treat anaemia in chronic renal failure patients. The results of this literature search show that there is no scientific basis from which to conclude that rHuEPO has performance‐enhancing properties in elite cyclists. The reported studies have many shortcomings regarding translation of the results to professional cycling endurance performance. Additionally, the possibly harmful side‐effects have not been adequately researched for this population but appear to be worrying, at least. The use of rHuEPO in cycling is rife but scientifically unsupported by evidence, and its use in sports is medical malpractice. What its use would have been, if the involved team physicians had been trained in clinical pharmacology and had investigated this properly, remains a matter of speculation. A single well‐controlled trial in athletes in real‐life circumstances would give a better indication of the real advantages and risk factors of rHuEPO use, but it would be an oversimplification to suggest that this would eradicate its use.
Increased leucocyte apoptosis in transfused β‐thalassaemia patients
Authors: Walter Patrick B. , Porter John , Evans Patricia , Kwiatkowski Janet L. , Neufeld Ellis J. , Coates Thomas , Giardina Patricia J. , Grady Robert W. , Vichinsky Elliott , Olivieri Nancy , Trachtenberg Felicia , Alberti Daniele , Fung Ellen , Ames Bruce , Higa Annie , Harmatz Paul ,   Thalassemia Clinical Research Network
Published: December 2012

This exploratory study assessed apoptosis in peripheral blood leucocytes (PBL) from β‐thalassaemia patients receiving chronic transfusions and chelation therapy (deferasirox or deferoxamine) at baseline, 1, 6, and 12 months. At baseline, thalassaemic PBLs presented 50% greater levels of Bax (BAX), 75% higher caspase‐3/7, 48% higher caspase‐8 and 88% higher caspase‐9 activities and 428% more nucleosomal DNA fragmentation than control subjects. Only neutrophils correlated significantly with apoptotic markers. Previously, we showed that over the treatment year, hepatic iron declined; we now show that the ratio of Bax/Bcl‐2 (BCL2), (−27·3%/year), and caspase‐9 activity (−13·3%/year) declined in both treatment groups, suggesting that chelation decreases body iron and indicators of PBL apoptosis.
What happens to health professionals when the ill patient is the health care system? Understanding the experience of practising medicine in the Romanian socio‐cultural context
Authors: Spânu Florina , Băban Adriana , Bria Mara , Dumitrascu Dan L.
Published: December 2012

Objectives

Our aims were to investigate the sources of work strain and stress, and the way in which they are experienced by Romanian health professionals in a work context shaped by the ongoing 20 years long reform of the national health care system.

Design

An exploratory, qualitative design was used to investigate medical professionals’ perceptions of stress and work strain.

Methods

Twenty eight interviews and two focus groups were conducted with 38 physicians, residents and nurses, between the age of 26 and 53. A semi‐structured interview guide was used for data gathering and the major themes were identified using thematic analysis of the transcripts.

Results

Three themes emerged in the analysis: governance and health system management, scarcity of resources, and health system reputation. Health professionals described the image of a suffering health system, exhausted by an inconsistent management plan, underfunded and understaffed; a system that is a constant source of discontent, bitterness and doubts for them and their patients.

Conclusions

Romanian health professionals’ experiences reveal a health care system which after 20 years of reform managed to shape a learned helplessness culture within the medical community and drive a large proportion of its workforce across the borders.

Statement of contribution

What is already known on this subject? Cultural, political, and socio‐economic factors influence the work context and shape the employees’ working experience.

What does this study add? We describe Romanian health professionals’ (HPs) relationship with the national health system, the work environment it creates, and its impact on the quality of their work experience. We draw on HPs’ experience to understand how the socio‐economic and political context shapes a culture of learned helplessness and blame against the health system, within the medical community, and the exodus of Romanian HPs searching for working opportunities abroad.
Protein phosphatase 1 coordinates CFTR‐dependent airway epithelial HCO3− secretion by reciprocal regulation of apical and basolateral membrane Cl−‐HCO3− exchangers
Authors: Garnett James P , Hickman Emma , Tunkamnerdthai Orathai , Cuthbert Alan W , Gray Michael A
Published: December 2012

Background and Purpose

Our recent studies on human airway serous‐like Calu‐3 cells showed that cAMP agonists stimulated a HCO3− rich secretion containing up to 80 mM HCO3−. This alkaline secretion relied on a coordinated switch in the activity of distinct Cl−‐HCO3− anion exchangers (AE) located at different regions of the cell. At the apical membrane, cAMP agonists activated the electroneutral AE pendrin (SLC26A4), together with cystic fibrosis transmembrane conductance regulator (CFTR), while at the basolateral membrane the agonists inhibited AE2 (SLC4A2). However, the underlying mechanism(s) that orchestrates this cAMP‐dependent switch in AE activity has not been elucidated.

Experimental Approach

Apical and basolateral Cl−‐HCO3− exchange was assessed by measuring Cl−‐dependent changes in intracellular pH (pHi).

Key Results

We show that protein phosphatase 1 (PP1), together with CFTR, play central roles in this reciprocal regulation of AE activity. Activation of pendrin by cAMP agonists, but not inhibition of the basolateral exchanger, was protein kinase A‐dependent. Knocking down CFTR expression, or blocking its activity with GlyH‐101, led to incomplete inhibition of the basolateral AE by cAMP, supporting a role for CFTR in this process. Addition of the PP1/2A inhibitor, okadaic acid, but not the PP2A specific inhibitor fostreicin, mimicked the effect of cAMP stimulation. Furthermore, okadaic acid‐treated Calu‐3 monolayers produced a more alkaline fluid than untreated cells, which was comparable with that produced by cAMP stimulation.

Conclusions and Implications

These results identify PP1 as a novel regulator of AE activity which, in concert with CFTR, coordinates events at both apical and basolateral membranes, crucial for efficient HCO3− secretion from Calu‐3 cells.
Children's Social Relationships With Current and Near‐Future Robots
Authors: Kahn Peter H. , Gary Heather E. , Shen Solace
Published: December 2012

abstract

Children will come of age with increasingly sophisticated social robots, which mimic both animal and human form. Will children develop social and even moral relationships with these robots? In this article, we review some of the research that suggests that the answer is yes. We propose that through the creation of social robots, a new ontological category is emerging, one that does not map onto humans, animals, or artifacts. We raise the concern that because these robots can be conceptualized as both social entities and objects, children might dominate them and reify a master–servant relationship, and that in such ways, this could lead to detrimental developmental outcomes, even as the robots benefit children in other ways. We also call on the developmental field to recognize the exponential growth of technological systems in children's lives, and to be future oriented to remain relevant.
Childhood Poverty, Chronic Stress, Self‐Regulation, and Coping
Authors: Evans Gary W. , Kim Pilyoung
Published: December 2012

abstract

Poverty is a powerful factor that can alter lifetime developmental trajectories in cognitive, socioemotional, and physical health outcomes. Most explanatory work on the underlying psychological processes of how poverty affects development has focused on parental investment and parenting practices, principally responsiveness. Our primary objective in this article was to describe a third, complementary pathway—chronic stress and coping—that may also prove helpful in understanding the developmental impacts of early childhood poverty throughout life. Disadvantaged children are more likely than their wealthier peers to confront a wide array of physical stressors (e.g., substandard housing, chaotic environments) and psychosocial stressors (e.g., family turmoil, separation from adult caregivers). As exposure to stressors accumulates, physiological response systems that are designed to handle relatively infrequent, acute environmental demands are overwhelmed. Chronic cumulative stressors also disrupt the self‐regulatory processes that help children cope with external demands.
Extra‐pancreatic effects of incretin‐based therapies: potential benefit for cardiovascular‐risk management in type 2 diabetes
Authors: van Genugten R. E. , Möller‐Goede D. L. , van Raalte D. H. , Diamant M.
Published: December 2012

Development of cardiovascular disease is one of the major complications of type 2 diabetes mellitus (T2DM). The chronic hyperglycaemic state is often accompanied by dyslipidaemia, hypertension, low‐grade systemic inflammation and oxidative stress which collectively result in a high risk of micro‐ and macrovascular complications. Current glucose‐lowering agents do not sufficiently address fore‐mentioned macrovascular‐risk factors. Recently, new therapeutic agents were introduced, based on the incretin hormone glucagon‐like peptide‐1 (GLP‐1), that is, the GLP‐1 receptor agonists (GLP‐1RA) and dipeptidyl‐peptidase 4 (DPP‐4) inhibitors. Beside its effect on pancreatic insulin secretion, GLP‐1 exerts several extra‐pancreatic effects such as slowing down gastric emptying, promoting satiety and reducing food intake and weight loss. Also, GLP‐1 and GLP‐1RA were shown to improve cardiovascular‐risk profiles, by reducing body fat content, blood pressure, circulating lipids and inflammatory markers in patients with T2DM. This review summarizes the presently known evidence with regard to extra‐pancreatic effects of the incretin‐based agents, focusing on the actions that improve the cardiovascular‐risk profile. We present available data from clinical trials of at least 24 week duration, but also findings from small‐sized clinical ‘proof of principle’ studies. We conclude that GLP‐1 RA and to a lesser extent DPP‐4 inhibitors are promising agents with regard to their effects on body weight, blood pressure and lipids, which collectively ameliorate the cardiovascular‐risk profile and as such may have added value in the treatment of T2DM. However, large‐sized long‐term outcome studies are warranted to show the true added value of these agents in the treatment of patients with T2DM.
Changes in cortical oscillations linked to multisensory modulation of nociception
Authors: Mancini Flavia , Longo Matthew R. , Canzoneri Elisa , Vallar Giuseppe , Haggard Patrick
Published: December 2012

Pain can be modulated by several contextual factors. For example, simply viewing one's own body can reduce pain, suggesting that the visual context may influence the processing of nociceptive stimuli. We studied changes in electroencephalographic (EEG) oscillatory activity related to visual modulation of nociception, comparing cortical oscillations during innocuous or noxious contact heat, while participants viewed either their own hand or a neutral object at the same location. Viewing the body compared with viewing the object reduced the intensity ratings of noxious stimuli, but not of innocuous heat. Time–frequency analysis of EEG data revealed that noxious, as opposed to warm, stimulation was associated with reduced beta (15–25 Hz) power. Classically, such decreases in oscillatory power indicate increases in sensory cortical activation. These event‐related oscillatory changes were moreover modulated by the visual context; viewing one's own body increased noxious stimulation‐induced beta oscillatory activity bilaterally, relative to viewing a neutral object, possibly indicating inhibition of cortical nociceptive processing. These results demonstrate that visual–nociceptive interactions involve changes in sensorimotor EEG rhythms.
Variability in life‐history and ecological traits is a buffer against extinction in mammals
Authors: González‐Suárez Manuela , Revilla Eloy
Published: December 2012

Anthropogenic degradation of the world's ecosystems is leading to a widespread and accelerating loss of biodiversity. However, not all species respond equally to existing threats, raising the question: what makes a species more vulnerable to extinction? We propose that higher intraspecific variability may reduce the risk of extinction, as different individuals and populations within a species may respond differently to occurring threats. Supporting this prediction, our results show that mammalian species with more variable adult body masses, litter sizes, sexual maturity ages and population densities are less vulnerable to extinction. Our findings reveal the role of local variation among populations, particularly of large mammals, as a buffering mechanism against extinction, and emphasise the importance of considering trait variation in comparative analyses and conservation management.
The incidence and implications of clouds for cloud forest plant water relations
Authors: Goldsmith Gregory R. , Matzke Nicholas J. , Dawson Todd E.
Published: December 2012

Although clouds are the most recognisable and defining feature of tropical montane cloud forests, little research has focussed on how clouds affect plant functioning. We used satellite and ground‐based observations to study cloud and leaf wetting patterns in contrasting tropical montane and pre‐montane cloud forests. We then studied the consequences of leaf wetting for the direct uptake of water accumulated on leaf surfaces into the leaves themselves. During the dry season, the montane forest experienced higher precipitation, cloud cover and leaf wetting events of longer duration than the pre‐montane forest. Leaf wetting events resulted in foliar water uptake in all species studied. The capacity for foliar water uptake differed significantly between the montane and pre‐montane forest plant communities, as well as among species within a forest. Our results indicate that foliar water uptake is common in these forest plants and improves plant water status during the dry season.
Seizures in patients with Alzheimer’s disease or vascular dementia: A population‐based nested case–control analysis
Authors: Imfeld Patrick , Bodmer Michael , Schuerch Markus , Jick Susan S. , Meier Christoph R.
Published: December 2012

Purpose:  Patients with Alzheimer’s disease (AD) have an increased risk of developing seizures or epilepsy. Little is known about the role of risk factors and about the risk of developing seizures/epilepsy in patients with vascular dementia (VD). The aim of this study was to assess incidence rates (IRs) of seizures/epilepsy in patients with AD, VD, or without dementia, and to identify potential risk factors of seizures or epilepsy.

Methods:  We conducted a follow‐up study with a nested case–control analysis using the United Kingdom–based General Practice Research Database (GPRD). We identified patients aged ≥65 years with an incident diagnosis of AD or VD between 1998 and 2008 and a matched comparison group of dementia‐free patients. Conditional logistic regression was used to estimate the odds ratio (OR) with a 95% confidence interval (CI) of developing seizures/epilepsy in patients with AD or VD, stratified by age at onset and duration of dementia as well as by use of antidementia drugs.

Key Findings:  Among 7,086 cases with AD, 4,438 with VD, and 11,524 matched dementia‐free patients, we identified 180 cases with an incident diagnosis of seizures/epilepsy. The IRs of epilepsy/seizures for patients with AD or VD were 5.6/1,000 person‐years (py) (95% CI 4.6–6.9) and 7.5/1,000 py (95% CI 5.7–9.7), respectively, and 0.8/1,000 py (95% CI 0.6–1.1) in the dementia‐free group. In the nested case–control analysis, patients with longer standing (≥3 years) AD had a slightly higher risk of developing seizures or epilepsy than those with a shorter disease duration, whereas in patients with VD the contrary was observed.

Significance:  Seizures or epilepsy were substantially more common in patients with AD and VD than in dementia‐free patients. The role of disease duration as a risk factor for seizures/epilepsy seems to differ between AD and VD.