Rate and associated factors of transient ischemic attack misdiagnosis.
The goal of this study was to investigate the rate and associated factors of Transient Ischemic Attack (TIA) misdiagnosis. We retrospectively analyzed consecutive patients with an initial diagnosis of TIA in the emergency department (ED) in a 9-month period. All hospitalized TIA patients were evaluated by a neurologist within 24 h and had at least one hospital discharge follow-up visit within three months. Patients' clinical data and neuroimaging were reviewed. The final diagnosis was independently verified by two stroke neurologists. Out of 276 patients with the initial diagnosis of TIA, 254 patients (mean age 68.7 ± 15.4 years, 40.9% male, 25.2% final diagnosis of TIA) were included in the analysis. Twenty-four patients (9.4%) were referred to our rapid-access TIA clinic. The rate of TIA misdiagnosis among TIA clinic referred patients was 45.8%. Among the 230 patients in inpatient setting, the rate of TIA misdiagnosis was 60.0%. A hospital discharge diagnosis of TIA was observed in 54.3% of hospitalized patients; however, only 24.8% had the final diagnosis of TIA. Among hospitalized patients, the univariate analysis suggests a significant difference (P < .05) between the two groups (correctly versus misdiagnosed patients) in terms of hospital discharge diagnosis, final diagnosis, history of diabetes mellitus, and coronary artery disease. In regression model hospital discharge diagnosis (P < .001), final diagnosis (P < .001), and diabetes mellitus (P = .018) retained independent association with TIA misdiagnosis. Our study indicates a high rate of TIA misdiagnosis in the emergency department, hospital, and outpatient clinics.


N-ras mutations in human cutaneous melanoma from sun-exposed body sites.
In 7 of 37 patients with cutaneous melanoma, mutations in the N-ras gene were found. The primary tumors of these seven patients were exclusively localized on body sites continuously exposed to sunlight. Moreover, the ras mutations were all at or near dipyrimidine sites known to be targets of UV damage. Two primary tumors were biclonal with respect to ras mutation. An active role for UV irradiation in induction of the mutations is suggested.


NKCC transport mediates the insulinotropic effects of taurine and other small neutral amino acids.
Despite its high concentration in pancreatic islets of Langerhans and broad range of antihyperglycemic effects, the route facilitating the import of dietary taurine into pancreatic β-cell and mechanisms underlying its insulinotropic activity are unclear. We therefore studied the impact of taurine on beta-cell function, alongside that of other small neutral amino acids, L-alanine and L-proline. Pharmacological profiling of insulin secretion was conducted using clonal BRIN BD11 β-cells, the impact of taurine on the metabolic fate of glucose carbons was assessed using NMR and the findings were verified by real-time imaging of Ca2+ dynamics in the cytosol of primary mouse and human islet beta-cells. In our hands, taurine, alanine and proline induced secretory responses that were dependent on the plasma membrane depolarisation, import of Ca2+, homeostasis of K+ and Na+ as well as on cell glycolytic and oxidative metabolism. Taurine shifted the balance between the oxidation and anaplerosis towards the latter, in BRIN BD11 beta-cells. Furthermore, the amino acid signalling was significantly attenuated by inhibition of Na+-K+-Cl- symporter (NKCC). These data suggest that taurine, like L-alanine and L-proline, acutely induces glucose-dependent insulin-secretory responses by modulating electrogenic Na+ transport, with potential role of intracellular K+ and Cl- in the signal transduction. The acute action delineated would be consistent with antidiabetic potential of dietary taurine supplementation.


Nitrosative stress and transcription.
Low NO concentrations synthesized by constitutively expressed NO synthases act on several signaling pathways activating transcription factors (TF), such as NF-kappaB or AP-1, and thereby influence gene expression. In contrast, during inflammatory reactions the inducible NO synthase produces NO for prolonged periods of time. The resulting nitrosative stress directly affects redox-sensitive TF like NF-kappaB, AP-1, Oct-1, c-Myb, or zinc finger-containing TF, but also additional mechanisms have been identified. Nitrosative stress in some cases induces expression of TF (AP-1, p53), indirectly modulates activity or stability of TF (HIF-1, p53) or their inhibitors (NF-kappaB), or modulates accessibility of promoters via increased DNA methylation or histone deacetylation. Depending on the promoter the result is induced, increased, decreased or even totally inhibited expression of various target genes. In unstimulated cells nitrosative stress increases NF-kappaB- or AP-1-dependent transcription, while in activated cells nitrosative stress rather abolishes NF-kappaB- or AP-1-dependent transcription. Sometimes the oxygen concentration also is of prime importance, since under normoxic conditions nitrosative stress activates HIF-1-dependent transcription, while under hypoxic conditions nitrosative stress leads to inhibition of HIF-1-dependent transcription. This review summarizes what is known about effects of physiological NO levels as well as of nitrosative stress on transcription.


Biomarkers, interventions and healthy ageing.
Population ageing is probably the single most important healthcare challenge the developed and developing world will face in the 21(st) century. This is because the later part of the human life course is marked by the emergence of a wide spectrum of pathological impairments which increase morbidity and reduce quality of life. The processes driving these increases in mortality and morbidity are often conceptualised as highly complex and multi-causal. Indeed, it has been suggested that there is no human 'ageing process', only distinct, disease-specific mechanisms of pathology. However, humans are not the only organisms within the biosphere to show ageing and the use of cross-species approaches has demonstrated that common ageing processes exist and allowed some of the common genetic pathways controlling them to be identified. Mutants in these pathways either delay or accelerate the development of late life diseases giving rise to extended healthy lives or progerias, respectively. These advances in fundamental understanding open opportunities for a more detailed investigation of the key causal mechanisms underlying ageing and the exploitation of that knowledge for improved interventions in later life.


MDR1 and ERCC1 expression predict outcome of patients with locally advanced bladder cancer receiving adjuvant chemotherapy.
The role of adjuvant chemotherapy in patients with locally advanced bladder cancer still remains to be defined. We hypothesized that assessing the gene expression of the chemotherapy response modifiers multidrug resistance gene 1 (MDR1) and excision repair cross-complementing 1 (ERCC1) may help identify the group of patients benefiting from cisplatin-based adjuvant chemotherapy. Formalin-fixed paraffin-embedded tumor samples from 108 patients with locally advanced bladder cancer, who had been enrolled in AUO-AB05/95, a phase 3 trial randomizing a maximum of three courses of adjuvant cisplatin and methotrexate (CM) versus methotrexate, vinblastine, epirubicin, and cisplatin (M-VEC), were included in the study. Tumor cells were retrieved by laser-captured microdissection and analyzed for MDR1 and ERCC1 expression using a quantitative real-time reverse transcription-polymerase chain reaction assay. Gene expression levels were correlated with clinical outcomes by multivariate Cox proportional hazards regression analysis. Expressions of MDR1 and ERCC1 were independently associated with overall progression-free survival (P = .001, relative risk = 2.9 and P = .01, relative risk = 2.24, respectively). The correlation of high MDR1 expression with inferior outcome was stronger in patients receiving M-VEC, whereas ERCC1 analysis performed equally in the CM and M-VEC groups. High MDR1 and ERCC1 gene expressions are associated with inferior outcome after cisplatin-based adjuvant chemotherapy for locally advanced bladder cancer. Prospective studies are warranted to define a role for MDR1 and ERCC1 analysis in individualizing multimodality treatment in locally advanced bladder cancer.


Efficient selection of DARPins with sub-nanomolar affinities using SRP phage display.
There is an ever-increasing demand to select specific, high-affinity binding molecules against targets of biomedical interest. The success of such selections depends strongly on the design and functional diversity of the library of binding molecules employed, and on the performance of the selection strategy. We recently developed SRP phage display that employs the cotranslational signal recognition particle (SRP) pathway for the translocation of proteins to the periplasm. This system allows efficient filamentous phage display of highly stable and fast-folding proteins, such as designed ankyrin repeat proteins (DARPins) that are virtually refractory to conventional phage display employing the post-translational Sec pathway. DARPins comprise a novel class of binding molecules suitable to complement or even replace antibodies in many biotechnological or biomedical applications. So far, all DARPins have been selected by ribosome display. Here, we harnessed SRP phage display to generate a phage DARPin library containing more than 10(10) individual members. We were able to select well behaved and highly specific DARPins against a broad range of target proteins having affinities as low as 100 pM directly from this library, without affinity maturation. We describe efficient selection on the Fc domain of human IgG, TNFalpha, ErbB1 (EGFR), ErbB2 (HER2) and ErbB4 (HER4) as examples. Thus, SRP phage display makes filamentous phage display accessible for DARPins, allowing, for example, selection under harsh conditions or on whole cells. We envision that the use of SRP phage display will be beneficial for other libraries of stable and fast-folding proteins.


Knowledge and attitudes of Indian surgeons regarding professional medical writing support.
The number of scientific papers published from India each year is low. We tried to determine how often Indian surgeons publish in biomedical journals, challenges preventing them from publishing more often, their awareness of the international guidelines that acknowledge professional medical writing as an ethical service, and their willingness to hire medical writers for their publications. A convenience sample of 100 of the surgeons attending a national conference were explained about the survey and asked to complete a survey questionnaire. All results were expressed as absolute frequencies and percentages. Of the 81 who responded, 43 (53%) had never published. Only 31/81 (38%) had published in the last 5 years. The challenges preventing more frequent publication were reported to be the lack of support for carrying out statistical analyses (58%), lack of time (41%), lack of sufficient data (38%), and lack of support for literature search (37%). Twenty-two (27%) surgeons were aware that they could hire a professional medical writer (PMW), 26 (32%) knew that the International Committee of Medical Journal Editors allowed PMWs to contribute to articles published in the biomedical journals, 75 (93%) were interested in hiring medical writers or to consider it in the future. Only 6 (7.4%) showed no such interest. There was poor awareness and great interest in hiring PMWs. It might be beneficial to increase the awareness of clinicians about the scientific expertise and communication skills of PMWs and how they can add value to biomedical publications.


Lung Recruitment Strategies During High Frequency Oscillatory Ventilation in Preterm Lambs.
Background: High frequency oscillatory ventilation (HFOV) is considered a lung protective ventilation mode in preterm infants only if lung volume is optimized. However, whilst a "high lung volume strategy" is advocated for HFOV in preterm infants this strategy is not precisely defined. It is not known to what extent lung recruitment should be pursued to provide lung protection. In this study we aimed to determine the relationship between the magnitude of lung volume optimization and its effect on gas exchange and lung injury in preterm lambs. Methods: 36 surfactant-deficient 124-127 d lambs commenced HFOV immediately following a sustained inflation at birth and were allocated to either (1) no recruitment (low lung volume; LLV), (2) medium- (MLV), or (3) high lung volume (HLV) recruitment strategy. Gas exchange and lung volume changes over time were measured. Lung injury was analyzed by post mortem pressure-volume curves, alveolar protein leakage, gene expression, and histological injury score. Results: More animals in the LLV developed a pneumothorax compared to both recruitment groups. Gas exchange was superior in both recruitment groups compared to LLV. Total lung capacity tended to be lower in the LLV group. Other parameters of lung injury were not different. Conclusions: Lung recruitment during HFOV optimizes gas exchange but has only modest effects on lung injury in a preterm animal model. In the HLV group aiming at a more extensive lung recruitment gas exchange was better without affecting lung injury.


Cleavage of RIPK1 by caspase-8 is crucial for limiting apoptosis and necroptosis.
The aspartate-specific cysteine protease caspase-8 suppresses necroptotic cell death mediated by RIPK3 and MLKL. Indeed, mice that lack caspase-8 die in a RIPK3- and MLKL-dependent manner during embryogenesis1-3. In humans, caspase-8 deficiency is associated with immunodeficiency4 or very early onset inflammatory bowel disease5. The substrates that are cleaved by caspase-8 to prevent necroptosis in vivo have not been defined. Here we show that knock-in mice that express catalytically inactive caspase-8(C362A) die as embryos owing to MLKL-dependent necroptosis, similar to caspase-8-deficient mice. Thus, caspase-8 must cleave itself, other proteins or both to inhibit necroptosis. Mice that express caspase-8(D212A/D218A/D225A/D387A), which cannot cleave itself, were viable, as were mice that express c-FLIP or CYLD proteins that had been mutated to prevent cleavage by caspase-8. By contrast, mice that express RIPK1(D325A), in which the caspase-8 cleavage site Asp325 had been mutated, died mid-gestation. Embryonic lethality was prevented by inactivation of RIPK1, loss of TNFR1, or loss of both MLKL and the caspase-8 adaptor FADD, but not by loss of MLKL alone. Thus, RIPK1(D325A) appears to trigger cell death mediated by TNF, the kinase activity of RIPK1 and FADD-caspase-8. Accordingly, dying endothelial cells that contain cleaved caspase-3 were abnormally abundant in yolk sacs of Ripk1D325A/D325A embryos. Heterozygous Ripk1D325A/+ cells and mice were viable, but were also more susceptible to TNF-induced cell death than were wild-type cells or mice. Our data show that Asp325 of RIPK1 is essential for limiting aberrant cell death in response to TNF, consistent with the idea that cleavage of RIPK1 by caspase-8 is a mechanism for dismantling death-inducing complexes.


Circular motion geometry using minimal data.
Circular motion or single axis motion is widely used in computer vision and graphics for 3D model acquisition. This paper describes a new and simple method for recovering the geometry of uncalibrated circular motion from a minimal set of only two points in four images. This problem has been previously solved using nonminimal data either by computing the fundamental matrix and trifocal tensor in three images or by fitting conics to tracked points in five or more images. It is first established that two sets of tracked points in different images under circular motion for two distinct space points are related by a homography. Then, we compute a plane homography from a minimal two points in four images. After that, we show that the unique pair of complex conjugate eigenvectors of this homography are the image of the circular points of the parallel planes of the circular motion. Subsequently, all other motion and structure parameters are computed from this homography in a straighforward manner. The experiments on real image sequences demonstrate the simplicity, accuracy, and robustness of the new method.


Treatment of patients with chronic obstructive pulmonary disease as practised in a defined Hong Kong community: a cross-sectional pilot survey.
To examine the characteristics of chronic obstructive pulmonary disease patients of the Kwai-Tsing area, Hong Kong, and the chronic treatments they received. DESIGN. Cross-sectional survey. Four clinic settings in Hong Kong: Respiratory Specialist Clinic, Princess Margaret Hospital (group 1); Medical Specialist Clinics, Princess Margaret Hospital (group 2); General Outpatient Clinics, Princess Margaret Hospital (group 3); South Kwai Chung Chest Clinic, Department of Health (group 4). Thirty physician-diagnosed chronic obstructive pulmonary disease patients in each of the above groups with post-bronchodilator 1-second forced expiratory volume/forced vital capacity ratios of less than 70% predicted values, who had been followed up at any of the participating clinics for at least 6 months. There were 111 male and nine female patients. The median age was 72.5 years and 79% had at least one medical co-morbidity. The mean duration of their chronic obstructive pulmonary disease was 9.8 years, and their mean post-bronchodilator 1-second forced expiratory volumes were 45% (for males) and 58% (for females) of predicted values. There were significantly fewer stage I and more stage IV patients in group 1. Influenza vaccination coverage within the previous 1 year was 54% and did not differ significantly between groups. Chronic obstructive pulmonary disease education was given significantly more often to group 1 patients. Short-acting beta agonists were used to treat all patients but long-acting bronchodilators and pulmonary rehabilitation were used almost exclusively in group 1. Overall, long-acting bronchodilators and pulmonary rehabilitation were offered to 16% and 5%, respectively, of those for whom these were indicated (according to international guidelines). In general there was insufficient education and under-treatment for chronic obstructive pulmonary disease patients. Management of such patients warrants improvements by way of increased accessibility to structured education programmes, pulmonary rehabilitation programmes, long-acting bronchodilator drugs, and respiratory specialist care.


The N400 Effect during Speaker-Switch-Towards a Conversational Approach of Measuring Neural Correlates of Language.
Language occurs naturally in conversations. However, the study of the neural underpinnings of language has mainly taken place in single individuals using controlled language material. The interactive elements of a conversation (e.g., turn-taking) are often not part of neurolinguistic setups. The prime reason is the difficulty to combine open unrestricted conversations with the requirements of neuroimaging. It is necessary to find a trade-off between the naturalness of a conversation and the restrictions imposed by neuroscientific methods to allow for ecologically more valid studies. Here, we make an attempt to study the effects of a conversational element, namely turn-taking, on linguistic neural correlates, specifically the N400 effect. We focus on the physiological aspect of turn-taking, the speaker-switch, and its effect on the detectability of the N400 effect. The N400 event-related potential reflects expectation violations in a semantic context; the N400 effect describes the difference of the N400 amplitude between semantically expected and unexpected items. Sentences with semantically congruent and incongruent final words were presented in two turn-taking modes: (1) reading aloud first part of the sentence and listening to speaker-switch for the final word, and (2) listening to first part of the sentence and speaker-switch for the final word. A significant N400 effect was found for both turn-taking modes, which was not influenced by the mode itself. However, the mode significantly affected the P200, which was increased for the reading aloud mode compared to the listening mode. Our results show that an N400 effect can be detected during a speaker-switch. Speech articulation (reading aloud) before the analyzed sentence fragment did also not impede the N400 effect detection for the final word. The speaker-switch, however, seems to influence earlier components of the electroencephalogram, related to processing of salient stimuli. We conclude that the N400 can effectively be used to study neural correlates of language in conversational approaches including speaker-switches.


Atrial fibrillation and the athletic heart.
Endurance exercise, despite a plethora of proven health benefits, is increasingly recognized as a potential cause of lone atrial fibrillation. Moderate exercise reduces all-cause mortality and protects against developing atrial fibrillation. However, more intense exercise regimes confer modest incremental health benefits, induce cardiac remodelling and negate some of the cardiovascular benefits of exercise. The implications of endurance exercise and athletic heart are becoming increasingly relevant as the popularity of endurance exercise has increased 20-fold within a generation. An apparent dose-response relationship exists between endurance exercise and left atrial dilatation. Repeated strenuous endurance exercise overloads atria, resulting in stretch-induced 'microtears', inflammation and endocardial scarring. Although these findings are observational in humans, similar mechanisms have recently been confirmed in animal models suggesting causation. Currently, it is not known whether a ceiling for endurance exercise exists, and, if so, what factors determine the threshold of harm. Although preliminary research is promising, much work remains if we are to understand the mechanisms underpinning atrial fibrillation in athletes.


Consecutive dose-finding trials adding lorazepam to the combination of metoclopramide plus dexamethasone: improved subjective effectiveness over the combination of diphenhydramine plus metoclopramide plus dexamethasone.
Four consecutive trials were undertaken to study lorazepam at each of three dosage levels and diphenhydramine when used in combination with iv metoclopramide and dexamethasone in patients receiving cisplatin at 120 mg/m2. The combination containing diphenhydramine had been the most effective treatment identified in prior trials. Earlier studies have found lorazepam to be a useful adjunct to other antiemetic agents. Sixty-five patients who had never received chemotherapy or antiemetics were directly observed in the hospital for 24 hours following cisplatin. Overall, 56% of the patients experienced no emesis and 78.5% of the patients had two or fewer vomiting episodes during the study period. No significant differences were noted in the number of patients who experienced no emesis or two or fewer episodes among the four trials. More sedation was seen with the lorazepam-containing regimens; other side effects were similar in type and severity. The trial using the highest dose of lorazepam (1.5 mg/m2) demonstrated significantly greater patient satisfaction (P = 0.039) and less anxiety during therapy (P = 0.02) when compared with the diphenhydramine combination. We conclude that combinations of iv metoclopramide plus dexamethasone with either diphenhydramine or lorazepam are well tolerated and effective in controlling cisplatin-induced emesis. The regimens containing lorazepam produced better subjective evaluations, and the combination using the highest lorazepam dose tested showed superior patient satisfaction and less anxiety during therapy.


Descriptive Analysis of Long- and Intermediate-Acting Insulin and Key Safety Outcomes in Adults with Type 2 Diabetes Mellitus.
As new biosimilar and follow-on insulins enter the market, more data are needed on safety, effectiveness, and patterns of use for these products to inform prescriber and patient decision-making regarding treatment. Additionally, data are needed regarding real-world patterns of use to inform future studies comparing the safety and effectiveness of bio-similars to already approved agents for diabetes treatment. To analyze the medication use patterns, adverse events, and availability of glycated hemoglobin (A1c) values for adult patients with type 2 diabetes mellitus (T2DM) who use long-acting insulin (LAI) or neutral protamine Hagedorn (NPH), an intermediate-acting insulin. We used the Biologics and Biosimilars Collective Intelligence Consortium's (BBCIC) distributed research network (DRN) for this descriptive analysis. The analysis time frame was January 1, 2011, to September 30, 2015, and included patients continuously insured for at least 183 days before the first date of a filled prescription for LAI or NPH insulin alone or with rapid- or short-acting insulin or sulfonylureas, whether newly starting insulin or switching to a different product. Insulin exposure episodes were the unit of analysis, and patients were classified in cohorts according to treatment. We followed patients until end of health plan enrollment or the end of the study period. We used occurrence of a study outcome, switch to another medication regimen, discontinuation of the current medication, or study end date to mark the end of an insulin episode. We describe demographics and availability of A1c values for analysis. Study outcomes included severe hypoglycemic events and major adverse cardiac events (MACE). We identified 103,951 patients with T2DM from a database of 39.1 million patients with commercial or Medicare Advantage pharmacy and medical benefits, who contributed 279,533 unique insulin exposure episodes. Most episodes (89%) included patients using LAI, and 52% of patients contributed data to 2 or more exposure cohorts. Insulin episodes lasted an average of 3.5 months, and patients had an average follow-up of 8.6 months. The unadjusted rate of severe hypoglycemic events requiring medical attention was 96.9 per 10,000 patient-years at risk (10kPYR). The unadjusted incident MACE rate was 676.9 events per 10kPYR. 38,330 T2DM patients in the BBCIC DRN had a baseline A1c available, and of those, less than 50% had a follow-up A1c result. Among patients with T2DM, our observed insulin patterns of use and rates of severe hypoglycemic outcomes and MACE are consistent with other studies. We noted a paucity of A1c results available, which implies that additional data sources may be needed to augment the BBCIC DRN. This study was coordinated and funded by the Biologics and Biosimilars Collective Intelligence Consortium (BBCIC) and represents the independent findings of the BBCIC Insulins Principal Investigator and the BBCIC Insulins Research Team. Lockhart is employed by the BBCIC and the Academy of Managed Care Pharmacy (AMCP). Eichelberger was employed by the BBCIC and AMCP at the time of this study. McMahill-Walraven is employed by Aetna, a CVS Health business. Panozzo, Marshall, and Brown are employed by Harvard Pilgrim Healthcare Institute. Aetna was reimbursed for data and analytic support from Harvard Pilgrim Healthcare Institute and the Reagan Udall Foundation for the U.S. Food and Drug Administration. Aetna receives external funding through research grants and subcontracts with Harvard Pilgrim Healthcare Institute, which are funded by the FDA, NIH, PCORI, BBCIC, Pfizer, and GSK; the Reagan-Udall Foundation for IMEDS; and PCORI for the ADAPTABLE Study. This work was previously presented as a poster at AMCP Nexus 2018; October 22-25, 2018; in Orlando, FL.


Effect of cyclic GMP-increasing agents nitric oxide and C-type natriuretic peptide on bovine chromaffin cell function: inhibitory role mediated by cyclic GMP-dependent protein kinase.
Both sodium nitroprusside (SNP), a nitric oxide (NO) generator, and C-type natriuretic peptide (CNP) have been found to raise cGMP levels in bovine chromaffin cells in a time- and concentration-dependent manner. The effect of these compounds on catecholamine secretion and calcium influx has also been studied, and both compounds were found to produce a slowly developing inhibitory effect on acetylcholine- or depolarization-stimulated catecholamine secretion and calcium increases without affecting the spontaneous release or the basal intracellular Ca2+ concentration. These inhibitory effects were observed only at high doses of acetylcholine or high levels of extracellular potassium and required concentrations of SNP or CNP very similar to those that increased cGMP levels. Preincubation with 100 microM zaprinast, a cGMP-phosphodiesterase inhibitor able to increase cGMP levels, mimicked the inhibitory effects of SNP and CNP. We investigated the effect of the soluble guanylate cyclase inhibitor methylene blue and the cGMP-dependent protein kinase (PKG) inhibitor 8-(4-chlorophenylthio)-guanosine 3',5'-cyclic monophosphorothioate, Rp isomer, on inhibition by SNP or CNP. Although methylene blue (10 microM) partially prevented the inhibitory effect of SNP, it did not do so for that produced by CNP, thus indicating that SNP acts through cGMP produced by the NO-activated guanylate cyclase. 8-(4-Chlorophenylthio)-guanosine 3',5'-cyclic monophosphorothioate, Rp isomer totally reversed both the SNP and CNP inhibitory effects. These results suggest that the activation of PKG mediates the inhibition induced by SNP and CNP. We successfully measured the PKG activity from cells preincubated with SNP or CNP, and our results show that this enzymatic activity increased with a time dependence very similar to the increase in the cGMP levels. Our results indicate that NO and CNP peptide inhibit secretagogue-stimulated catecholamine release via activation of soluble and particulate isoforms of the guanylate cyclase, respectively, presumably by inhibition of calcium entry through voltage-activated calcium channels. This inhibitory effects seems to be mediated by activation of the PKG.


'My brain couldn't move from planning a birth to planning a funeral': a qualitative study of parents' experiences of decisions after ending a pregnancy for fetal abnormality.
With increasing technology for screening and diagnostic testing for fetal abnormality in pregnancy, many more pregnant women and couples are faced with the decision to terminate a pregnancy often after receiving diagnostic test results in the second or third trimester of pregnancy. Whilst there is extensive research on people's experience of diagnostic testing and decision-making, there has been less research on people's experiences of decisions they face immediately following their termination. To describe the experiences of (often unanticipated) decisions that people face in the immediate aftermath of ending a pregnancy following diagnosis of serious fetal abnormality. Secondary analysis of narrative qualitative interview data. Participants were recruited throughout the United Kingdom (UK) and interviewed between 2004 and 2006. 38 individual mothers, 10 individual fathers and 10 couples who ended a pregnancy following diagnosis of fetal abnormality. Parents who had experienced the ending of a pregnancy following diagnosis of fetal abnormality described their experiences, and often distress, of facing painful decisions consequent upon their decision to terminate the pregnancy. What was striking from their accounts was their sense of being unprepared for these decisions. Often they suggested that they had received no information or forewarning of the 'choices' they would have to make, although information designed specifically for people in this situation is available in the UK. Many of the decisions that people face in the immediate aftermath of a termination for fetal abnormality are upsetting, and in some circumstances more so because they are not anticipated. Often parents report not receiving information which could be helpful until after these decisions have been made. Health professionals face difficult choices about what issues to raise with patients at this sensitive time, and the optimum time to inform patients of the extra decisions they will face. More research is needed on whether these decisions subsequent to a termination for fetal abnormality have long term implications for people's mental health and their ability to come to terms with their loss.


Sub-diffraction-limit localization imaging of a plasmonic nanoparticle pair with wavelength-resolved dark-field microscopy.
In this work, with wavelength-resolved dark-field microscopy, the center-of-mass localization information from nanoparticle pairs (i.e., spherical (45 nm in diameter) and rod (45 × 70 nm) shaped gold nanoparticle pairs with different gap distances and orientations) was explored and compared with the results determined by scanning electron microscopy (SEM) measurements. When the gap distance was less than 20 nm, the scattering spectrum of the nanoparticle pair was seriously modulated by the plasmonic coupling effect. The measured coordinate information determined by the optical method (Gaussian fitting) was not consistent with the true results determined by SEM measurement. A good correlation between the optical and SEM measurements was achieved when the gap distance was further increased (e.g., 20, 40 and 60 nm). Under these conditions, well-defined scattering peaks assigned to the corresponding individual nanoparticles could be distinguished from the obtained scattering spectrum. These results would afford valuable information for the studies on single plasmonic nanoparticle imaging applications with the optical microscopy method such as super-localization imaging, high precision single particle tracking in a crowding environment and so on.


Physical activity and fat mass gain in Mexican school-age children: a cohort study.
In México, the prevalence of unhealthy weight increased from 24% at 6 y to 33% at 12 y of age, opening a window of opportunity to better understand the pathogenesis of obesity. The objective of this study was to explore the association between time spent on medium, vigorous physical activity (MVPA) and concurrent gains in BMI, fat mass (FM) and fat-free mass (FFM), alternately, in a cohort of Mexican children followed from kindergarten (baseline) to 2nd grade elementary school (endline). The MVPA (5-d accelerometry), BMI, FM and FFM (air displacement plethysmography) were measured at baseline and endline. Associations between gains in BMI, FM and FFM and changes in MVPA were examined using lagged and dynamic regression models, controlling for energy intake and demographic variables. A total of 205 children were analyzed. Gender affected the effect of MVPA on FM gain. In girls, a high baseline MVPA predicted a lower FM gain (-0.96 kg, p=0.025) compared to low/medium MVPA. Increasing, decreasing or having a persistently high MVPA predicted a lower FM gain (range -1.6 to -1.03 kg, p<0.05) compared to persistently low MVPA. In boys, increases in MVPA were associated with higher gains in BMI (+0.76 kg/m2, p=0.04) and FFM (+1.1 kg, p=0.01) compared to persistently low MVPA. These results support a protective role of MVPA on FM gain in girls, suggesting that it may play a crucial role in the development of obesity. Further research on the gender effect of MVPA is warranted to better understand its role in the prevention and control of overweight and obesity.


Postnatal growth standards for preterm infants: the Preterm Postnatal Follow-up Study of the INTERGROWTH-21(st) Project.
Charts of size at birth are used to assess the postnatal growth of preterm babies on the assumption that extrauterine growth should mimic that in the uterus. The INTERGROWTH-21(st) Project assessed fetal, newborn, and postnatal growth in eight geographically defined populations, in which maternal health care and nutritional needs were met. From these populations, the Fetal Growth Longitudinal Study selected low-risk women starting antenatal care before 14 weeks' gestation and monitored fetal growth by ultrasonography. All preterm births from this cohort were eligible for the Preterm Postnatal Follow-up Study, which included standardised anthropometric measurements, feeding practices based on breastfeeding, and data on morbidity, treatments, and development. To construct the preterm postnatal growth standards, we selected all live singletons born between 26 and before 37 weeks' gestation without congenital malformations, fetal growth restriction, or severe postnatal morbidity. We did analyses with second-degree fractional polynomial regression models in a multilevel framework accounting for repeated measures. Fetal and neonatal data were pooled from study sites and stratified by postmenstrual age. For neonates, boys and girls were assessed separately. From 4607 women enrolled in the study, there were 224 preterm singleton births, of which 201 (90%) were enrolled in the Preterm Postnatal Follow-up Study. Variance component analysis showed that only 0·2% and 4·0% of the total variability in postnatal length and head circumference, respectively, could be attributed to between-site differences, justifying pooling the data from all study sites. Preterm growth patterns differed from those for babies in the INTERGROWTH-21(st) Newborn Size Standards. They overlapped with the WHO Child Growth Standards for term babies by 64 weeks' postmenstrual age. Our data have yielded standards for postnatal growth in preterm infants. These standards should be used for the assessment of preterm infants until 64 weeks' postmenstrual age, after which the WHO Child Growth Standards are appropriate. Size-at-birth charts should not be used to measure postnatal growth of preterm infants. Bill & Melinda Gates Foundation.


The HIV care cascade: a systematic review of data sources, methodology and comparability.
The cascade of HIV diagnosis, care and treatment (HIV care cascade) is increasingly used to direct and evaluate interventions to increase population antiretroviral therapy (ART) coverage, a key component of treatment as prevention. The ability to compare cascades over time, sub-population, jurisdiction or country is important. However, differences in data sources and methodology used to construct the HIV care cascade might limit its comparability and ultimately its utility. Our aim was to review systematically the different methods used to estimate and report the HIV care cascade and their comparability. A search of published and unpublished literature through March 2015 was conducted. Cascades that reported the continuum of care from diagnosis to virological suppression in a demographically definable population were included. Data sources and methods of measurement or estimation were extracted. We defined the most comparable cascade elements as those that directly measured diagnosis or care from a population-based data set. Thirteen reports were included after screening 1631 records. The undiagnosed HIV-infected population was reported in seven cascades, each of which used different data sets and methods and could not be considered to be comparable. All 13 used mandatory HIV diagnosis notification systems to measure the diagnosed population. Population-based data sets, derived from clinical data or mandatory reporting of CD4 cell counts and viral load tests from all individuals, were used in 6 of 12 cascades reporting linkage, 6 of 13 reporting retention, 3 of 11 reporting ART and 6 of 13 cascades reporting virological suppression. Cascades with access to population-based data sets were able to directly measure cascade elements and are therefore comparable over time, place and sub-population. Other data sources and methods are less comparable. To ensure comparability, countries wishing to accurately measure the cascade should utilize complete population-based data sets from clinical data from elements of a centralized healthcare setting, where available, or mandatory CD4 cell count and viral load test result reporting. Additionally, virological suppression should be presented both as percentage of diagnosed and percentage of estimated total HIV-infected population, until methods to calculate the latter have been standardized.


Evaluation of the neuroprotective effect of cannabinoids in a rat model of Parkinson's disease: importance of antioxidant and cannabinoid receptor-independent properties.
We have recently demonstrated that two plant-derived cannabinoids, Delta9-tetrahydrocannabinol and cannabidiol (CBD), are neuroprotective in an animal model of Parkinson's disease (PD), presumably because of their antioxidant properties. To further explore this issue, we examined the neuroprotective effects of a series of cannabinoid-based compounds, with more selectivity for different elements of the cannabinoid signalling system, in rats with unilateral lesions of nigrostriatal dopaminergic neurons caused by local application of 6-hydroxydopamine. We used the CB1 receptor agonist arachidonyl-2-chloroethylamide (ACEA), the CB2 receptor agonist HU-308, the non-selective agonist WIN55,212-2, and the inhibitors of the endocannabinoid inactivation AM404 and UCM707, all of them administered i.p. Daily administration of ACEA or WIN55,212-2 did not reverse 6-hydroxydopamine-induced dopamine (DA) depletion in the lesioned side, whereas HU-308 produced a small recovery that supports a possible involvement of CB2 but not CB1 receptors. AM404 produced a marked recovery of 6-hydroxydopamine-induced DA depletion and tyrosine hydroxylase deficit in the lesioned side. Possibly, this is caused by the antioxidant properties of AM404, which are derived from the presence of a phenolic group in its structure, rather than by the capability of AM404 to block the endocannabinoid transporter, because UCM707, another transporter inhibitor devoid of antioxidant properties, did not produce the same effect. None of these effects were observed in non-lesioned contralateral structures. We also examined the timing for the effect of CBD to provide neuroprotection in this rat model of PD. We found that CBD, as expected, was able to recover 6-hydroxydopamine-induced DA depletion when it was administered immediately after the lesion, but it failed to do that when the treatment started 1 week later. In addition, the effect of CBD implied an upregulation of mRNA levels for Cu,Zn-superoxide dismutase, a key enzyme in endogenous defenses against oxidative stress. In summary, our results indicate that those cannabinoids having antioxidant cannabinoid receptor-independent properties provide neuroprotection against the progressive degeneration of nigrostriatal dopaminergic neurons occurring in PD. In addition, the activation of CB2 (but not CB1) receptors, or other additional mechanisms, might also contribute to some extent to the potential of cannabinoids in this disease.


[Mechanisms of drug resistance and cancer stem cells].
Although the success of anticancer treatments has been increasing annually, drug resistance remains the dominant cause of death of cancer patients. Initial therapy often leaves residual dis-ease that leads to repeated tumor development or to loss of its sensitivity to available therapy. One reason of residual disease formation is the presence of cancer stem cells (CSCs). CSCs have been identified as a small population of cells that is capable of self renewal and differentiation. It is supposed that these cells are responsible for cancer initiation, progression, metastasis, recurrence and drug resistance. Over the past years, much attention has been paid to development of CSCs related therapies and to identification of key molecules involved in controlling the specific properties of CSCs populations. This article reviews the basic mechanisms of drug resistance in relation to cancer stem cells.


Pharmacokinetic characteristics, pharmacodynamic effect and in vivo antiviral efficacy of liver-targeted interferon alpha.
Interferon alpha (IFNα) is used for the treatment of hepatitis B virus infection, and whilst efficacious, it is associated with multiple adverse events caused by systemic exposure to interferon. We therefore hypothesise that targeting IFN directly to the intended site of action in the liver would reduce exposure in blood and peripheral tissue and hence improve the safety and tolerability of IFNα therapy. Furthermore we investigated whether directing IFN to the reservoir of infection in the liver may improve antiviral efficacy by increasing local concentration in target organs and tissues. Our previous results show that the mIFNα2 fused to an ASGPR specific liver targeting antibody, DOM26h-196-61, results in a fusion protein which retains the activity of both fusion partners when measured in vitro. In vivo targeting of the liver by mIFNα2-DOM26h-196-61, hereafter referred to as targeted mIFNα2, was observed in microSPECT imaging studies in mice. In this study we show by pharmacokinetic analysis that antibody mediated liver-targeting results in increased uptake and exposure of targeted mIFNα2 in target tissues, and correspondingly reduced uptake and exposure in systemic circulation, clearance organs and non-target tissues. We also show that cytokine activity and antiviral activity of liver-targeted IFN is observed in vivo, but that, contrary to expectations, liver-targeting of mIFNα2 using ASGPR specific dAbs actually leads to a reduced pharmacodynamic effect in target organs and lower antiviral activity in vivo when compared to non-targeted mIFNα2-dAb fusions.


Less is more: Burkholderia pseudomallei and chronic melioidosis.
The Gram-negative bacterium Burkholderia pseudomallei is the causative agent of melioidosis, a serious infectious disease of humans and animals. Once considered an esoteric tropical disease confined to Southeast Asia and northern Australia, research on B. pseudomallei has recently gained global prominence due to its classification as a potential bioterrorism agent by countries such as the United States and also by increasing numbers of case reports from regions where it is not endemic. An environmental bacterium typically found in soil and water, assessing the true global prevalence of melioidosis is challenged by the fact that clinical symptoms associated with B. pseudomallei infection are extremely varied and may be confused with diverse conditions such as lung cancer, tuberculosis, or Staphyloccocus aureus infection. These diagnostic challenges, coupled with lack of awareness among clinicians, have likely contributed to underdiagnosis and the high mortality rate of melioidosis, as initial treatment is often either inappropriate or delayed. Even after antibiotic treatment, relapses are frequent, and after resolution of acute symptoms, chronic melioidosis can also occur, and the symptoms can persist for months to years. In a recent article, Price et al. [mBio 4(4):e00388-13, 2013, doi:10.1128/mBio.00388-13] demonstrate how comparative genomic sequencing can reveal the repertoire of genetic changes incurred by B. pseudomallei during chronic human infection. Their results have significant clinical ramifications and highlight B. pseudomallei's ability to survive in a wide range of potential niches within hosts, through the acquisition of genetic adaptations that optimize fitness and resource utilization.


Defective modulation of angiotensin II-induced renal vasoconstriction in hypertensive rats.
In a previous study we observed that the ability of intravenous infusions of prostaglandin I2 to attenuate vasoconstriction caused by intravenous infusions of angiotensin II was reduced in the renal but not mesenteric vasculature of spontaneously hypertensive rats (SHR). One objective of the current study was to determine whether a renal defect in the angiotensin II/prostaglandin I2 interaction in SHR could be confirmed even when confounding hemodynamic changes induced by intravenous infusions of prostaglandin I2 were avoided. The second objective was to determine whether abnormal modulation of angiotensin II-induced renal vasoconstriction was present even in SHR that were maintained normotensive from an early age. Four-week-old SHR and normotensive Wistar-Kyoto rats were randomized to receive either normal drinking water or drinking water containing captopril (100 mg/kg per day). At 14 to 18 weeks of age, rats were pretreated with indomethacin to block the production of endogenous prostaglandin I2, and changes in mesenteric and renal vascular resistances induced by suprarenal/supramesenteric aortic infusions of angiotensin II (10, 30, and 100 ng/kg per minute) were elicited in the presence and absence of aortic infusions of prostaglandin I2 (0.1 and 0.3 micrograms/kg per minute). Data were analyzed globally using four- and three-factor ANOVAs. The ability of prostaglandin I2 to attenuate the renal vasoconstrictor response to angiotensin II was strain specific (P = .0138), and this strain-specific interaction was not influenced by chronic treatment with captopril (P = .3526).(ABSTRACT TRUNCATED AT 250 WORDS)


The View-Master Health Study Focus Groups.
Trichloroethylene (TCE), a common groundwater contaminant, was found at high levels at an Oregon work site in 1998. According to a recent report released by the National Research Council, "the evidence on carcinogenic risk and other health hazards from exposure to trichloroethylene has strengthened since 2001." A convenience sample of 13 former workers from the Oregon work site was recruited for a series of focus groups. Information obtained on plant processes, safety procedures, attitudes regarding medical record access, and opinions about proxy accuracy was subjected to qualitative content analysis. Workers recalled few safety policies and no training or support for control of safety. Most thought co-workers and family members would be the best source of proxy exposure information and favored granting access to medical records. Job-role mobility confirmed the importance of using a job or task exposure matrix. Information obtained will be used in development of an exposure assessment interview tool.


Tracking the rise of the "expert patient" in evolving paradigms of HIV care.
In the context of rolling out antiretroviral treatment programmes in resource-constrained settings, "expert patients" are hailed as an important step towards greater involvement of people living with HIV (PLHIV) in HIV care, and in addressing the human resources crisis. The rise of the "expert patient" in HIV care partly echoes the move towards greater patient agency and self-management that informs health services delivery in industrialised countries. However, the "expert patient" also represents a particular moment in the trajectory of PLHIVs' quest for greater access to treatment and health care. In this paper, we examine the unique evolution and 'instrumentalisation' of PLHIV in public health in relation to shifts in the epidemiology, interventions and socio-political response related to HIV since the 1980s. Synthesised broadly as "risk-based", "empowerment", "structural" and "activist" approaches, four key paradigms are identified that attribute different levels of citizenship for PLHIV, that is the motivation, agency and responsibility to act not only on their own interests, but also on behalf of collective interests in HIV prevention and care. We problematise the concept of the patient-citizen by drawing attention to the context-specificity of such notions as patient identity, capacity and rights as well as the structural realities of access to treatment, and broader engagement with the health system. We conclude by offering some thoughts on the applicability of 'expert patient' approaches across a broad range of settings.


Biophysical and structural characterization of the thioredoxin-binding domain of protein kinase ASK1 and its interaction with reduced thioredoxin.
Apoptosis signal-regulating kinase 1 (ASK1), a mitogen-activated protein kinase kinase kinase, plays a key role in the pathogenesis of multiple diseases. Its activity is regulated by thioredoxin (TRX1) but the precise mechanism of this regulation is unclear due to the lack of structural data. Here, we performed biophysical and structural characterization of the TRX1-binding domain of ASK1 (ASK1-TBD) and its complex with reduced TRX1. ASK1-TBD is a monomeric and rigid domain that forms a stable complex with reduced TRX1 with 1:1 molar stoichiometry. The binding interaction does not involve the formation of intermolecular disulfide bonds. Residues from the catalytic WCGPC motif of TRX1 are essential for complex stability with Trp(31) being directly involved in the binding interaction as suggested by time-resolved fluorescence. Small-angle x-ray scattering data reveal a compact and slightly asymmetric shape of ASK1-TBD and suggest reduced TRX1 interacts with this domain through the large binding interface without inducing any dramatic conformational change.


Intraspecific Niche Variation Drives Abundance-Occupancy Relationships in Freshwater Fish Communities.
A positive relationship between occupancy and average local abundance of species is found in a variety of taxa, yet the mechanisms driving this association between abundance and occupancy are still enigmatic. Here we show that freshwater fishes exhibit a positive abundance-occupancy relationship across 125 Swedish lakes. For a subset of 9 species from 11 lakes, we estimated species-specific diet breadth from stable isotopes, within-lake habitat breadth from catch data for littoral and pelagic nets, adaptive potential from genetic diversity, abiotic niche position, and dispersal capacity. Average local abundance was mainly positively associated with both within-lake habitat and diet breadth, that is, species with larger intraspecific variation in niche space had higher abundances. No measure was a good predictor of occupancy, indicating that occupancy may be more directly related to abundance or abiotic conditions than to niche breadth per se. This study suggests a link between intraspecific niche variation and a positive abundance-occupancy relationship and implies that management of freshwater fish communities, whether to conserve threatened or control invasive species, should initially be aimed at niche processes.


Simple Monte Carlo model for crowd dynamics.
In this paper, we introduce a simple Monte Carlo method for simulating the dynamics of a crowd. Within our model a collection of hard-disk agents is subjected to a series of two-stage steps, implying (i) the displacement of one specific agent followed by (ii) a rearrangement of the rest of the group through a Monte Carlo dynamics. The rules for the combined steps are determined by the specific setting of the granular flow, so that our scheme should be easily adapted to describe crowd dynamics issues of many sorts, from stampedes in panic scenarios to organized flow around obstacles or through bottlenecks. We validate our scheme by computing the serving times statistics of a group of agents crowding to be served around a desk. In the case of a size homogeneous crowd, we recover intuitive results prompted by physical sense. However, as a further illustration of our theoretical framework, we show that heterogeneous systems display a less obvious behavior, as smaller agents feature shorter serving times. Finally, we analyze our results in the light of known properties of nonequilibrium hard-disk fluids and discuss general implications of our model.


Vestibular Co-stimulation in Adults with a Cochlear Implant.
Vestibular co-stimulation is a side effect of cochlear implant stimulation. The electrical currents delivered by the cochlear implant can spread toward the vestibular system and thus stimulate it. The aim of the study is to evaluate whether it is feasible to functionally restore the balance by modifying the vestibular co-stimulation. Four adult patients, who had received a commercially available cochlear implant previously, were enrolled. Counterbalanced biphasic pulses were presented as bursts or as an amplitude-modulated biphasic pulse train (modulation frequencies ranging from 1 to 500 Hz) at the participant's upper comfortable level for electrical stimulation. Subjective sensations and vestibular-mediated eye movements were used for evaluating the possible effects of vestibular co-stimulation. One participant experienced a cyclic tilting of his head in response to an amplitude-modulated biphasic pulse train with a modulation frequency of 2 and 400 Hz. However, during a follow-up visit, the sensation could not be replicated. Subjective vestibular sensations or vestibular-mediated eye movements could not be electrically evoked with a commercially available cochlear implant in 4 adult patients with almost normal vestibular function. Therefore, customized design of the hard-, firm-, and/or software of the commercially available cochlear implant might be necessary in order to electrically restore vestibular performance.


Thymosin-β4: A key modifier of renal disease.
There is an urgent need for new treatments for chronic kidney disease (CKD). Thymosin-β4 is a peptide that reduces inflammation and fibrosis and has the potential to restore endothelial and epithelial cell injury, biological processes involved in the pathophysiology of CKD. Therefore, thymosin-β4 could be a novel therapeutic direction for CKD. Here, we review the current evidence on the actions of thymosin-β4 in the kidney in health and disease. Using transgenic mice, two recent studies have demonstrated that endogenous thymosin-β4 is dispensable for healthy kidneys. In contrast, lack of endogenous thymosin-β4 exacerbates mouse models of glomerular disease and angiotensin-II-induced renal injury. Administration of exogenous thymosin-β4, or its metabolite, Ac-SDKP, has shown therapeutic benefits in a range of experimental models of kidney disease. The studies conducted so far reveal a protective role for thymosin-β4 in the kidney and have shown promising results for the therapeutic potential of exogenous thymosin-β4 in CKD. Further studies should explore the mechanisms by which thymosin-β4 modulates kidney function in different types of CKD. Ac-SDKP treatment has beneficial effects in many experimental models of kidney disease, thus supporting its potential use as a new treatment strategy.


Formation of the bicoid morphogen gradient: an mRNA gradient dictates the protein gradient.
The Bicoid (Bcd) protein gradient is generally believed to be established in pre-blastoderm Drosophila embryos by the diffusion of Bcd protein after translation of maternal mRNA, which serves as a strictly localized source of Bcd at the anterior pole. However, we previously published evidence that the Bcd gradient is preceded by a bcd mRNA gradient. Here, we have revisited and extended this observation by showing that the bcd mRNA and Bcd protein gradient profiles are virtually identical at all times. This confirms our previous conclusion that the Bcd gradient is produced by a bcd mRNA gradient rather than by diffusion. Based on our observation that bcd mRNA colocalizes with Staufen (Stau), we propose that the bcd mRNA gradient forms by a novel mechanism involving quasi-random active transport of a Stau-bcd mRNA complex through a nonpolar microtubular network, which confines the bcd mRNA to the cortex of the embryo.


Human blood mononuclear cell in vitro cytokine response before and after two different strenuous exercise bouts in the presence of bloodroot and Echinacea extracts.
The purpose of this multidisciplinary investigation was to characterize cytokine production by human blood mononuclear cells after 2 contrasting exercise bouts (a maximal graded oxygen consumption [VO(2)max] test and 90 min of cycling at 85% of ventilatory threshold [VT]) when stimulated in vitro with extracts from bloodroot (Sanguinaria canadensis), coneflower (Echinacea tennesseensis), or solvent vehicle controls. Blood was sampled pre- and post-exercise. Production of TNF, IL-1beta, and IL-10 were measured at 24, 48, and 72 h, respectively. In the VO(2)max test there was a main effect of exercise such that exercise increased cytokine synthesis and a main effect of stimulant such that bloodroot extracts significantly increased cytokine production compared to other stimulants or controls. In the 90-min bout, there was a main effect of exercise for TNF and IL-1beta (but not IL-10) such that exercise decreased cytokine synthesis and a main effect of stimulant such that bloodroot extracts significantly increased cytokine production compared to other stimulants or controls, with exercisexstimulant interactions for both IL-1beta and IL-10. A similar though weaker effect was seen with Echinacea extracts; subsequent biochemical analyses suggested this was related to alkamide decay during 3 years undisturbed storage at ultralow (-80 degrees C) temperature. In this study, the VO(2)max test was associated with enhanced cytokine production whereas the 90-min cycling at 85% VT was associated with suppressed cytokine production. Bloodroot extracts were able to increase cytokine production in both contexts. Herbal extracts purported to offset exercise-associated effects on immune activity warrant continued investigation.


High rates of hepatitis C virus reinfection and spontaneous clearance of reinfection in people who inject drugs: a prospective cohort study.
Hepatitis C virus reinfection and spontaneous clearance of reinfection were examined in a highly characterised cohort of 188 people who inject drugs over a five-year period. Nine confirmed reinfections and 17 possible reinfections were identified (confirmed reinfections were those genetically distinct from the previous infection and possible reinfections were used to define instances where genetic differences between infections could not be assessed due to lack of availability of hepatitis C virus sequence data). The incidence of confirmed reinfection was 28.8 per 100 person-years (PY), 95%CI: 15.0-55.4; the combined incidence of confirmed and possible reinfection was 24.6 per 100 PY (95%CI: 16.8-36.1). The hazard of hepatitis C reinfection was approximately double that of primary hepatitis C infection; it did not reach statistical significance in confirmed reinfections alone (hazard ratio [HR]: 2.45, 95%CI: 0.87-6.86, p=0.089), but did in confirmed and possible hepatitis C reinfections combined (HR: 1.93, 95%CI: 1.01-3.69, p=0.047) and after adjustment for the number of recent injecting partners and duration of injecting. In multivariable analysis, shorter duration of injection (HR: 0.91; 95%CI: 0.83-0.98; p=0.019) and multiple recent injecting partners (HR: 3.12; 95%CI: 1.08-9.00, p=0.035) were independent predictors of possible and confirmed reinfection. Time to spontaneous clearance was shorter in confirmed reinfection (HR: 5.34, 95%CI: 1.67-17.03, p=0.005) and confirmed and possible reinfection (HR: 3.10, 95%CI: 1.10-8.76, p-value=0.033) than primary infection. Nonetheless, 50% of confirmed reinfections and 41% of confirmed or possible reinfections did not spontaneously clear. Hepatitis C reinfection and spontaneous clearance of hepatitis C reinfection were observed at high rates, suggesting partial acquired natural immunity to hepatitis C virus. Public health campaigns about the risks of hepatitis C reinfection are required.


HIV seroprevalence in a London same-day testing clinic.
To determine the prevalence of HIV infection among people attending a confidential, non-genitourinary medicine based testing clinic that provides HIV antibody test results on the day of consultation. Retrospective analysis of data collected on 2635 individuals attending the Same-Day HIV Testing Clinic at the Royal Free Hospital, London between March 1992 and February 1993. A total of 1612 men and 1023 women were tested for HIV antibody. The primary risk for HIV infection was heterosexual (71.7%; 1889 out of 2635) and homosexual contact (24.5%; 646 out of 2635). Fifty-four individuals were given positive HIV test results (46 men, median age 34 years; eight women, median age 27 years). Overall HIV seroprevalence was 2.0% (95% confidence interval, 1.5-2.5). HIV seroprevalence was highest among homosexual men (6.5%; 41 out of 635) and injecting drug users (5.7%; four out of 70). The rates for heterosexual men and women were 0.2% (two out of 915) and 0.7% (seven out of 974), respectively. Of the 54 individuals who were HIV-antibody-positive, 44 were Centers for Disease Control and Prevention stage II/III, eight stage IV and one was tested at the time of seroconversion (stage I; data were not available for one patient). Of the total numbers attending this clinic 27% (702 out of 2635) had previously been tested and received a negative result. Of a total of 54 HIV-antibody-positive individuals, 40% (21) had previously received a negative test result. The data suggest that seroprevalence amongst homosexual men attending a designated HIV testing clinic in London is lower than that reported by genitourinary clinic based testing sites. The large number of repeat testers who subsequently became infected with HIV suggests that there is a population requiring specific targetting for HIV risk reduction.


Effects of Ginkgo biloba extract EGb761 on neural differentiation of stem cells offer new hope for neurological disease treatment.
Stem cell transplantation has brought new hope for the treatment of neurological diseases. The key to stem cell therapy lies in inducing the specific differentiation of stem cells into nerve cells. Because the differentiation of stem cells in vitro and in vivo is affected by multiple factors, the final differentiation outcome is strongly associated with the microenvironment in which the stem cells are located. Accordingly, the optimal microenvironment for inducing stem cell differentiation is a hot topic. EGb761 is extracted from the leaves of the Ginkgo biloba tree. It is used worldwide and is becoming one of the focuses of stem cell research. Studies have shown that EGb761 can antagonize oxygen free radicals, stabilize cell membranes, promote neurogenesis and synaptogenesis, increase the level of brain-derived neurotrophic factors, and replicate the environment required during the differentiation of stem cells into nerve cells. This offers the possibility of using EGb761 to induce the differentiation of stem cells, facilitating stem cell transplantation. To provide a comprehensive reference for the future application of EGb761 in stem cell therapy, we reviewed studies investigating the influence of EGb761 on stem cells. These started with the composition and neuropharmacology of EGb761, and eventually led to the finding that EGb761 and some of its important components play important roles in the differentiation of stem cells and the protection of a beneficial microenvironment for stem cell transplantation.


Predictive factors for the placebo effect in clinical trials for dry eye: a pooled analysis of three clinical trials.
Placebo effect is one of the methodological difficulties in dry eye clinical trials. If we could elucidate the tendencies of the placebo response and find predictors, we could reduce the placebo response in clinical trials for dry eye. In this study, we investigated the predictive factors for the placebo effect in dry eye clinical trials. A total of 205 patients with dry eye assigned to the placebo arms of three placebo-controlled randomised clinical trials were analysed by simple and multivariable regression analysis. The corneal fluorescein (FL) staining score and dry eye symptoms were studied at week 4. The variables of interest included gender, age, complications of Sjögren's syndrome, Schirmer's test I value, tear break-up time and conjunctival hyperaemia score. We also conducted a stratified analysis according to the patients' age. Among all the studied endpoints, the baseline scores were significantly related to the corresponding placebo response. In addition, for the FL score and the dryness score, age was a significant predictor of the placebo response (p=0.04 and p<0.0001, respectively). Stratified analysis by age showed that patients more than 40 years of age are more likely to have a stronger placebo response in the FL and dryness scores. The baseline scores and age were predictive factors of the placebo response in frequently used endpoints, such as FL score or dryness symptoms. These patient characteristics can be controlled by study design, and our findings enable the design of more efficient placebo-controlled studies with good statistical power.


Relating the Past with the Present: Information Integration and Segregation during Ongoing Narrative Processing.
This study examined how the brain dynamically updates event representations by integrating new information over multiple minutes while segregating irrelevant input. A professional writer custom-designed a narrative with two independent storylines, interleaving across minute-long segments (ABAB). In the last (C) part, characters from the two storylines meet and their shared history is revealed. Part C is designed to induce the spontaneous recall of past events, upon the recurrence of narrative motifs from A/B, and to shed new light on them. Our fMRI results showed storyline-specific neural patterns, which were reinstated (i.e., became more active) during storyline transitions. This effect increased along the processing timescale hierarchy, peaking in the default mode network. Similarly, the neural reinstatement of motifs was found during Part C. Furthermore, participants showing stronger motif reinstatement performed better in integrating A/B and C events, demonstrating the role of memory reactivation in information integration over intervening irrelevant events.


Sequential analysis in a Bayesian model of diastolic blood pressure measurement.
A sequential method for diagnosing or excluding hypertension based on the Bayesian model of diastolic blood pressure presented in a companion article is presented. The likelihood ratio method of Wald is modified to include the effects of a prior probability distribution and to constrain the strategy to achieve specified positive and negative predictive values. The resulting formulas for upper and lower limits to diagnose and exclude diastolic hypertension can be evaluated using a hand calculator and a table of areas of the standard normal distribution. The strategy is illustrated for a population having a blood pressure distribution similar to that of the cohort screened for participation in the Hypertension Detection and Follow-up Program, with 90 mm Hg as the cutoff defining hypertension and required positive and negative predictive values of 95%. The performance of the strategy was simulated using Monte Carlo methods. The median number of readings required for diagnosis is three, and 80% of subjects are diagnosed in 11 or fewer readings. In contrast to the strategy's 95% predictive values, a fixed-number-of-measurements strategy requiring the same mean number of measurements has a positive predictive value of only 83% and a negative predictive value of 96%. When the parameters of the model have been properly measured or estimated, this method is practical, efficient, and accurate for diagnosing hypertension in a known population.


Requirement for the eIF4E binding proteins for the synergistic down-regulation of protein synthesis by hypertonic conditions and mTOR inhibition.
The protein kinase mammalian target of rapamycin (mTOR) regulates the phosphorylation and activity of several proteins that have the potential to control translation, including p70S6 kinase and the eIF4E binding proteins 4E-BP1 and 4E-BP2. In spite of this, in exponentially growing cells overall protein synthesis is often resistant to mTOR inhibitors. We report here that sensitivity of wild-type mouse embryonic fibroblasts (MEFs) to mTOR inhibitors can be greatly increased when the cells are subjected to the physiological stress imposed by hypertonic conditions. In contrast, protein synthesis in MEFs with a double knockout of 4E-BP1 and 4E-BP2 remains resistant to mTOR inhibitors under these conditions. Phosphorylation of p70S6 kinase and protein kinase B (Akt) is blocked by the mTOR inhibitor Ku0063794 equally well in both wild-type and 4E-BP knockout cells, under both normal and hypertonic conditions. The response of protein synthesis to hypertonic stress itself does not require the 4E-BPs. These data suggest that under certain stress conditions: (i) translation has a greater requirement for mTOR activity and (ii) there is an absolute requirement for the 4E-BPs for regulation by mTOR. Importantly, dephosphorylation of p70S6 kinase and Akt is not sufficient to affect protein synthesis acutely.


Reflectance confocal microscopy criteria of lichen planus-like keratosis.
Lichen planus-like keratosis (LPLK) may be difficult to differentiate from melanoma and other skin cancers on sun-damaged skin based on clinical and dermoscopic examination. Reflectance confocal microscopy (RCM) allows evaluation of skin lesions at high resolution. The aim of this study was to identify criteria for specific diagnosis of LPLK using in vivo RCM. Lesions included in the study were derived from patients presenting for skin examination at a private dermatology practice specializing in skin cancer. We retrospectively analysed RCM features of 28 biopsy-proven LPLK and compared them to RCM findings in skin cancers on sun-damaged skin, including five in situ squamous cell carcinomas, six actinic keratoses, seven superficial basal cell carcinomas and eight melanomas. The main RCM features of LPLK and their relative frequencies were: (i) typical honeycomb pattern of the spinous layer (78.6%); (ii) elongated cords and/or bulbous projections at the dermal-epidermal junction (75%); and (iii) numerous plump-bright cells and/or bright stellate spots in the superficial dermis (92.9%). These RCM features correlated with the following histopathological findings respectively: (i) spinous-granular layers without significant atypia of keratinocytes; (ii) elongated, bulbous rete ridges; and (iii) dense infiltration of melanophages and lymphocytes in superficial dermis. We propose diagnostic criteria that classify correctly 71.4% of LPLK, while avoiding misclassification of any of the skin cancers in the present series as LPLK. We identified RCM criteria for diagnosis of LPLK that correlate well with histopathological findings and that allow differentiation of LPLK from skin cancer.


Exploratory multivariate analysis of the effect of fatty fish consumption and medicinal use on heart rate and heart rate variability data.
The overall aim of the present study was to explore the relationship between medicinal use and fatty fish consumption on heart rate variability (HRV) and heart rate (HR) in a group of forensic inpatients on a variety of medications. A total of 49 forensic inpatients, randomly assigned to a fish group (n = 27) or a control group (n = 22) were included in the present study. Before and by the end of the food intervention period HR and HRV were measured during an experimental test procedure. An additional aim of this paper is to show how multivariate data analysis can highlight differences and similarities between the groups, thus being a valuable addition to traditional statistical hypothesis testing. The results indicate that fish consumption may have a positive effect on both HR and HRV regardless of medication, but that the influence of medication is strong enough to mask the true effect of fish consumption. Without correcting for medication, the fish group and control group become indistinguishable (p = 0.0794, Cohen's d = 0.60). The effect of medication is demonstrated by establishing a multivariate regression model that estimates HR and HRV in a recovery phase based on HR and HRV data recorded during psychological tests. The model performance is excellent for HR data, but yields poor results for HRV when employed on participants undergoing the more severe medical treatments. This indicates that the HRV behavior of this group is very different from that of the participants on no or lower level of medication. When focusing on the participants on a constant medication regime, a substantial improvement in HRV and HR for the fish group compared to the control group is indicated by a principal component analysis and t-tests (p = 0.00029, Cohen's d = 2.72). In a group of psychiatric inpatients characterized by severe mental health problems consuming different kinds of medication, the fish diet improved HR and HRV, indices of both emotional regulation and physical health.


Is accurate and reliable blood loss estimation the 'crucial step' in early detection of postpartum haemorrhage: an integrative review of the literature.
Postpartum haemorrhage (PPH) is the leading cause of maternal mortality in low-income countries and severe maternal morbidity in many high-income countries. Poor outcomes following PPH are often attributed to delays in the recognition and treatment of PPH. Experts have suggested that improving the accuracy and reliability of blood loss estimation is the crucial step in preventing death and morbidity from PPH. However, there is little guidance on how this can be achieved. The aim of this integrative review was to evaluate the various methods of assessing maternal blood loss during childbirth. A systematic, integrative review of published research studies was conducted. All types of studies were included if they developed, tested, or aimed to improve methods and skills in quantifying blood loss during childbirth, or explored experiences of those involved in the process. Thirty-six studies were included that evaluated the accuracy of visual estimation; tested methods to improve skills in measurement; examined their effect on PPH diagnosis and treatment, and / or explored additional factors associated with blood loss evaluation. The review found that health professionals were highly inaccurate at estimating blood loss as a volume. Training resulted in short term improvements in skills but these were not retained and did not improve clinical outcomes. Multi-faceted interventions changed some clinical practices but did not reduce the incidence of severe PPH or the timing of responses to excessive bleeding. Blood collection bags improved the accuracy of estimation but did not prevent delays or progression to severe PPH. Practitioners commonly used the nature and speed of blood flow, and the condition of the woman to indicate that the blood loss was abnormal. Early diagnosis of PPH should improve maternal outcomes, but there is little evidence that this can be achieved through improving the accuracy of blood loss volume measurements. The diagnosis may rely on factors other than volume, such as speed of blood flow and nature of loss. A change in direction of future research is required to explore these in more detail.


Liver-stage malaria parasites vulnerable to diverse chemical scaffolds.
Human malaria infection begins with a one-time asymptomatic liver stage followed by a cyclic symptomatic blood stage. All high-throughput malaria drug discovery efforts have focused on the cyclic blood stage, which has limited potential for the prophylaxis, transmission blocking, and eradication efforts that will be needed in the future. To address these unmet needs, a high-throughput phenotypic liver-stage Plasmodium parasite screen was developed to systematically identify molecules with liver-stage efficacy. The screen recapitulates liver-stage infection by isolating luciferase-expressing Plasmodium berghei parasites directly from the salivary glands of infected mosquitoes, adding them to confluent human liver cells in 384-well plates, and measuring luciferase activity after a suitable incubation period. Screening 5,375 known bioactive compounds identified 37 liver-stage malaria inhibitors with diverse modes of action, as shown by inhibition time course experiments. Further analysis of the hits in the Food and Drug Administration-approved drug subset revealed compounds that seem to act specifically on the liver stage of infection, suggesting that this phase of the parasite's life cycle presents a promising area for new drug discovery. Notably, many active compounds in this screen have molecular structures and putative targets distinctly different from those of known antimalarial agents.


Hardwood tree survival in heavy ground cover on reclaimed land in West Virginia: mowing and ripping effects.
Current West Virginia coal mining regulations emphasize reforestation as a preferred postmining land use on surface mined areas. Some mined sites reclaimed to pasture are being converted to forests. In the spring of 2001, we compared the establishment and growth of five hardwood tree species on a reclaimed West Virginaia surface mine with compacted soils and a heavy grass groundcover. We planted 1-yr-old seedlings of five species (black cherry [Prunus serotina Ehrh.], red oak [Quercus rubra L.], yellow poplar [Liriodendron tulipifera L.], black walnut [Juglans nigra L.], and white ash [Fraxinus americana L.]) into sites that were mowed and unmowed on north- and south-facing aspects. We applied a ripping treatment, which loosened the compacted soils and disturbed the heavy ground cover. First year results showed >80% survival for all species. After 7 yr black cherry survival averaged 36%, red oak 47%, yellow poplar 66%, black walnut 80%, and white ash 98% across all sites and treatments. Seedling survival was best on north, unmowed, and ripped areas. Average growth (height x diameter(2)) of trees after 7 yr was greatest with white ash (434 cm(3)), followed by yellow poplar (256 cm(3)) and black walnut (138 cm(3)), then by black cherry (31 cm(3)) and red oak (27 cm(3)). Browsing by wildlife had a negative impact on tree growth especially on south aspect sites. Overall, mowing reduced survival of black cherry, red oak, and yellow poplar, but not for black walnut and white ash. Ripping increased survival of black cherry, red oak, and yellow poplar. Growth of all species was improved with ripping. Using inverse linear-quadratic plateau models, the time required for tree survival to stabilize varied from 1 yr for white ash to 6 to 9 yr for the other species.


Symptomatic BK Virus Infection Is Associated With Kidney Function Decline and Poor Overall Survival in Allogeneic Hematopoietic Stem Cell Recipients.
Nephropathy due to BK virus (BKV) infection is an evolving challenge in patients undergoing hematopoietic stem cell transplantation (HSCT). We hypothesized that BKV infection was a marker of kidney function decline and a poor prognostic factor in HSCT recipients who experience this complication. In this retrospective study, we analyzed all patients who underwent their first allogeneic HSCT at our institution between 2004 and 2012. We evaluated the incidence of persistent kidney function decline, which was defined as a confirmed reduction in estimated glomerular filtration rate of at least 25% from baseline using the Chronic Kidney Disease Epidemiology equation. Cox proportional hazard regression was used to model the cause-specific hazard of kidney function decline, and the Fine-Gray method was used to account for the competing risks of death. Among 2477 recipients of a first allogeneic HSCT, BK viruria was detected in 25% (n = 629) and kidney function decline in 944 (38.1%). On multivariate analysis, after adjusting for age, sex, acute graft-versus-host disease (GVHD), chronic GVHD, preparative conditioning regimen, and graft source, BK viruria remained a significant risk factor for kidney function decline (p < 0.001). In addition, patients with BKV infection and kidney function decline experienced worse overall survival. After allogeneic HSCT, BKV infection was strongly and independently associated with subsequent kidney function decline and worse patient survival after HSCT.


[Quantitative changes in fibro-architectonics of the human cerebral cortex from birth to 12 years of age].
The peculiarities of fibroarchitectonics in topographically and functionally distinct cortical zones (including temporo-parieto-occipital subarea, occipital, pre-, postcentral, and frontal areas) were studied in the brain specimens of 74 children of different ages (from birth to 12 years) taken with yearly intervals, while those in the first year of life--with monthly intervals. Frontal sections stained with Nissl, Peters and Golgi methods were studied using computer analysis of optical images. The data were obtained on the increment rates of radial fiber fascicles' thickness, distances between fascicles and age dynamics of ratios of volume densities of neurons and fibers in areas 3, 4p, 6op, 17, 19, 37ac, 44 and 32/10. It was shown that age-related transformations of fibroarchitectonics in the areas of sensomotor, somatosensory, occipital, temporo-parieto-occipital and frontal cortex were heterochronic and took place with different intensity; most pronounced changes were found to occur during the first 2-3 years, while less intensive ones were observed up to the age of 9-12 years.


Deciphering the premature mortality in PIGA-CDG - An untold story.
Congenital disorder of glycosylation (CDG) due to a defective phosphatidylinositol glycan anchor biosynthesis class A protein (PIGA) is a severe X-linked developmental and epileptic encephalopathy. Seizures are often treatment refractory, and patients have intellectual disability and global developmental delay. Previous reports have suggested that patients with PIGA-CDG have a high risk of premature mortality. This study aimed to evaluate the observed high mortality and the causes of death in PIGA-CDG patients. We reviewed the literature and collected additional unpublished patients through an international network. In total, we reviewed the data of 88 patients of whom 30 patients born alive were deceased, and the overall mortality before the age of 20 years was 30 % (26/88). Age at death ranged from 15 days to 48 years of life. The median age at death was two years and more than half of the patients deceased in early childhood. The PIGA-specific mortality rate/1000 person-years was 44.9/1000 person-years (95 %, CI 31.4-64.3). There were no cases of definite or probable sudden unexpected death in epilepsy (SUDEP) and half of the patients died due to respiratory failure (15/30, 50 %) or possible SUDEP (3/30, 10 %). Three patients (10 %) died from severe cardiomyopathy, liver failure and gastrointestinal bleeding, respectively. The cause of death was unclassified in nine patients (30 %). Autopsies were rarely performed and the true cause of death remains unknown for the majority of patients. Our data indicate an increased risk of premature death in patients with PIGA-CDG when compared to most monogenic developmental and epileptic encephalopathies.


Differential expression of heat shock protein 70 gene between the cortex and caudate after transient focal cerebral ischaemia in rats.
In relation to changes of total protein synthesis, induction of 70-kDa heat shock protein (HSP70) mRNA was examined by Northern blot and in situ hybridization after 30 min of transient middle cerebral artery (MCA) occlusion of rats. HSP70 mRNA was not present in the control condition of brain. With reperfusion, the mRNA was greatly induced along with the recovery of total protein synthesis in the cerebral cortex and lateral caudate of the ipsilateral hemisphere. However, the level of the mRNA reached a maximum earlier in the lateral caudate (at 3 h) than in the cortex (at 8 h), and the maximum amount of the mRNA was much smaller in the caudate than in the cortex. Total protein synthesis in the lateral caudate did not completely recover until 7 days. Histological examination showed a severe damage in cells of lateral caudate, while cells in the cortex were almost normal at 7 days. No difference in the brain temperature was observed between the two regions. These results show that the induction of HSP70 mRNA correlates with the recovery of protein synthesis in brain cells after a transient ischaemia, and that the HSP70 gene expression is different at the transcriptional level between the cortical and caudate cells after the transient ischaemia.


Cortical 5-HT2A receptor signaling modulates anxiety-like behaviors in mice.
Serotonin [5-hydroxytryptamine (5-HT)] neurotransmission in the central nervous system modulates depression and anxiety-related behaviors in humans and rodents, but the responsible downstream receptors remain poorly understood. We demonstrate that global disruption of 5-HT2A receptor (5HT2AR) signaling in mice reduces inhibition in conflict anxiety paradigms without affecting fear-conditioned and depression-related behaviors. Selective restoration of 5HT2AR signaling to the cortex normalized conflict anxiety behaviors. These findings indicate a specific role for cortical 5HT2AR function in the modulation of conflict anxiety, consistent with models of cortical, "top-down" influences on risk assessment.


Deficits and variations in patients' experience with making 9 common medical decisions: the DECISIONS survey.
Although many researchers have examined patient involvement and patient-provider interactions within specific clinical environments, no nationally representative data exist to characterize patient perceptions of decision making and patient-provider communications across multiple common medical decisions. To identify deficits and variations in the patient experience of making common medical decisions about initiation of prescription medications for hypertension, hypercholesterolemia, or depression; screening tests for colorectal, breast, or prostate cancer; and surgeries for knee or hip replacement, cataracts, or lower back pain, as well as to identify factors associated with patient confidence in the decisions. National sample of US adults identified by random-digit dialing. Cross-sectional survey conducted from November 2006 to May 2007. Included 2473 English-speaking adults age 40 and older who reported undertaking 1 or more of the above 9 medical actions or discussing doing so with a health care provider within the past 2 years. Patients reported who initiated discussions and made the final decisions, how much discussion of pros and cons occurred, whether they were asked about their preferences, and their confidence that the decision "was the right one." The proportion of patient-driven decisions varied significantly across decisions (range: blood pressure: 16% to knee/hip replacement: 48%). Most patients (78%-85%) reported that providers made a recommendation, and such recommendations generally favored taking medical action. Fewer patients reported that providers asked them about their preferences (range: colon cancer screening: 34% to knee/hip replacement: 80%) or discussed reasons not to take action (range: breast cancer screening: 20% to lower back surgery: 80%). Decision confidence was higher among patients who reported primarily making the decision themselves (odds ratio [OR] = 14.6, P < 0.001) or having been asked for their preference (OR = 1.32, P < 0.01) and was lower among patients whose patient-provider discussions included cons (OR = 0.74, P = 0.008). Recall biases may affect patients' memories of their decision-making processes. DECISIONS participants reported wide variations in the proportion of discussions that included a conversation about reasons not to take action or a conversation about patients' preferences about what they would like to do. These factors appear directly related to patients' confidence that the decision was "right."


Immune cell-endothelial cell interactions in the bovine corpus luteum.
Early embryonic mortality accounts for a substantial portion of reproductive failure in agriculturally important livestock, including the dairy cow. The maintenance of early pregnancy requires a fully functional corpus luteum (CL) that is not susceptible to regression following fertilization, yet the cellular mechanisms of luteal regression are not clearly understood. Immune-cell accumulation within the CL at the time of regression is a well-documented phenomenon in a variety of species. In the dairy cow, immune-cell accumulation precedes luteal regression by several days and coincides with an increase in expression of the chemokine monocyte chemoattractant protein 1 (CCL2), suggesting that immune-mediated events promote tissue destruction. Recent studies indicate that endothelial cells comprising the CL are a primary source of CCL2 secretion. Moreover, although uterine-derived prostaglandin F(2α) (PGF) initiates luteal regression in the cow, PGF does not directly provoke CCL2 secretion by luteal endothelial cells. Instead, PGF-induced luteal regression is thought to require cooperative interaction among immune cells, endothelial cells, and steroidogenic cells of the CL to further promote CCL2 secretion, enhance immune-cell recruitment, and eliminate luteal tissue. This brief review focuses on putative interactions between immune cells and endothelial cells derived from the bovine CL that result in enhanced CCL2 expression and the elaboration of other inflammatory mediators (for example, cytokines), which perpetuate luteal regression. Fundamental knowledge of immune-endocrine interactions within the reproductive system of cows has relevance to other CL-bearing mammals, including humans and endangered animals, particularly in the development of methods to control and/or improve fertility. Thus, it is a timely topic for this symposium concerning ecological immunology and public health.


Algorithm for the Use of Biochemical Markers of Bone Turnover in the Diagnosis, Assessment and Follow-Up of Treatment for Osteoporosis.
Increased biochemical bone turnover markers (BTMs) measured in serum are associated with bone loss, increased fracture risk and poor treatment adherence, but their role in clinical practice is presently unclear. The aim of this consensus group report is to provide guidance to clinicians on how to use BTMs in patient evaluation in postmenopausal osteoporosis, in fracture risk prediction and in the monitoring of treatment efficacy and adherence to osteoporosis medication. A working group with clinical scientists and osteoporosis specialists was invited by the Scientific Advisory Board of European Society on Clinical and Economic Aspects of Osteoporosis, Osteoarthritis and Musculoskeletal Diseases (ESCEO). Serum bone formation marker PINP and resorption marker βCTX-I are the preferred markers for evaluating bone turnover in the clinical setting due to their specificity to bone, performance in clinical studies, wide use and relatively low analytical variability. BTMs cannot be used to diagnose osteoporosis because of low sensitivity and specificity, but can be of value in patient evaluation where high values may indicate the need to investigate some causes of secondary osteoporosis. Assessing serum levels of βCTX-I and PINP can improve fracture prediction slightly, with a gradient of risk of about 1.2 per SD increase in the bone marker in addition to clinical risk factors and bone mineral density. For an individual patient, BTMs are not useful in projecting bone loss or treatment efficacy, but it is recommended that serum PINP and βCTX-I be used to monitor adherence to oral bisphosphonate treatment. Suppression of the BTMs greater than the least significant change or to levels in the lower half of the reference interval in young and healthy premenopausal women is closely related to treatment adherence. In conclusion, the currently available evidence indicates that the principal clinical utility of BTMs is for monitoring oral bisphosphonate therapy.


Antibodies against OspA epitopes of Borrelia burgdorferi cross-react with neural tissue.
Neurological sequela of chronic Lyme disease include encephalopathy, myelopathy and peripheral neuropathy. These have generally been attributed to either persistent infection or pathogen-induced autoimmunity. In this study, we investigated the presence of cross-reactive human neural epitopes that share amino acid sequences with Borrelia burgdorferi OspA protein. Sequence similarity analysis was carried out by searching known cDNA sequences from brain tissue. The cDNA database search yielded three sequences that were identical to sequences in OspA. Corresponding peptides were synthesized and antibodies were generated against them in rabbits. Antibodies against two of the homologous OspA peptides were found to react with neurons in human brain, spinal cord and dorsal root ganglia by immunohistochemistry.


Salvage ablative therapy in prostate cancer: international multidisciplinary consensus on trial design.
Salvage ablative therapy (SAT) has been developed as a form of localized treatment for localized recurrence of prostate cancers following radiation therapy. To better address the utility of SAT, prospective clinical trials must address the aspects of accepted standards in the initial evaluation, treatment, follow-up, and outcomes in the oncology community. We undertook this study to achieve consensus on uniform standardized trial design for SAT trials. A literature search was performed and an international multidisciplinary group of experts was identified. A questionnaire was constructed and sent out to 71 participants in 3 consecutive rounds according to the Delphi method. The project was concluded with a face-to-face meeting in which the results were reviewed and conclusions were formulated. Patients with recurrent disease after radiation therapy were considered candidates for a SAT trial using any ablation scenario performed with cryotherapy or high-intensity focused ultrasound. It is feasible to compare different sources of energy or to compare with historical data on salvage radical prostatectomy outcomes. The primary objective should be to assess the efficacy of the treatment for negative biopsy rate at 12 months. Secondary objectives should include safety parameters and quality-of-life assessment. Exclusion criteria should include evidence of local or distant metastases. The optimal biopsy strategy is image-guided targeted biopsies. Follow-up includes multiparametric magnetic resonance imaging, prostate-specific antigen level, and quality of life for at least 5 years. A multidisciplinary board from international experts reached consensus on trial design for SAT in prostate cancer and provides a standard for designing a feasible SAT trial.