Levels and variation in overuse of fecal occult blood testing in the Veterans Health Administration.
Policy-makers have called for efforts to reduce overuse of cancer screening tests, including colorectal cancer screening (CRCS). Overuse of CRCS tests other than colonoscopy has not been well documented. To estimate levels and correlates of fecal occult blood test (FOBT) overuse in a national Veterans Health Administration (VHA) sample. Observational Participants included 1,844 CRCS-eligible patients who responded to a 2007 CRCS survey conducted in 24 VHA facilities and had one or more FOBTs between 2003 and 2009. We combined survey data on race, education, and income with administrative data on region, age, gender, CRCS procedures, and outpatient visits to estimate overuse levels and variation. We coded FOBTs as overused if they were conducted <10 months after prior FOBT, <9.5 years after prior colonoscopy, or <4.5 years after prior barium enema. We used multinomial logistic regression models to examine variation in overuse by reason (sooner than recommended after prior FOBT; sooner than recommended after colonoscopy, barium enema, or a combination of procedures), adjusting for clustering of procedures within patients, and patients within facilities. Of 4,236 FOBTs received by participants, 885 (21 %) met overuse criteria, with 323 (8 %) sooner than recommended after FOBT, and 562 (13 %) sooner than recommended after other procedures. FOBT overuse varied across facilities (9-32 %, p<0.0001) and region (12-23 %, p< .0012). FOBT overuse after prior FOBT declined between 2003 and 2009 (8 %-5 %, p= .0492), but overuse after other procedures increased (11-19 %, p= .0002). FOBT overuse of both types increased with number of outpatient visits (OR 1.15, p<0.001), but did not vary by patient demographics. More than 11 % of overused FOBTs were followed by colonoscopy within 12 months. Many FOBTs are performed sooner than recommended in the VHA. Variation in overuse by facility, region, and outpatient visits suggests addressing FOBT overuse will require system-level solutions.


A major cell surface antigen of Coccidioides immitis which elicits both humoral and cellular immune responses.
Multinucleate parasitic cells (spherules) of Coccidioides immitis isolates produce a membranous outer wall component (SOW) in vitro which has been reported to be reactive with antibody from patients with coccidioidal infection, elicits a potent proliferative response of murine immune T cells, and has immunoprotective capacity in a murine model of coccidioidomycosis. To identify the antigenic components of SOW, the crude wall material was first subjected to Triton X-114 extraction, and a water-soluble fraction derived from this treatment was examined for protein composition and reactivity in humoral and cellular immunoassays. Protein electrophoresis revealed that the aqueous fraction of three different isolates of C. immitis each contained one or two major glycoproteins (SOWgps), distinguished by their molecular sizes, which ranged from 58 to 82 kDa. The SOWgps, however, showed identical N-terminal amino acid sequences, and each was recognized by sera from patients with C. immitis infection. Antibody raised against the purified 58-kDa glycoprotein (SOWgp58) of the Silveira isolate was used for Western blot and immunolocalization analyses. Expression of SOWgp was shown to be parasitic phase specific, and the antigen was localized to the membranous SOW. The water-soluble fraction of SOW and the purified SOWgp58 were tested for the ability to stimulate proliferation of human peripheral monocytic cells (PBMC). The latter were obtained from healthy volunteers with positive skin test reaction to spherulin, a parasitic-phase antigen of C. immitis, and from volunteers who showed no skin test reaction to the same antigen. The SOW preparations stimulated proliferation of PBMC from skin test-positive but not skin test-negative donors, and the activated cells secreted gamma interferon, which is indicative of a T helper 1 pathway of immune response. Results of this study suggest that SOWgp is a major parasitic cell surface-expressed antigen that elicits both humoral and cellular immune responses in patients with coccidioidal infection.


Incidence of rabies virus complement-fixing antibodies in unvaccinated dogs, humans and livestock in Anambra State of Nigeria.
A total of 376 serum samples from dogs, humans and livestock were examined for complement-fixing (CF) antibodies against rabies virus. High CF antibody titres (up to 1:1024) were detected among unvaccinated dogs aged 3 months and above as in vaccinated ones, thus establishing endemicity of the virus in the area. An antibody titre of 1:128 was detected in the serum of a puppy aged below 3 months and is unlikely to be due to residual maternal antibodies. It rather provides evidence for seroconversion at that age and further establishes endemicity of the virus. Moderate to high CF antibody titres (greater than or equal to 1:64) were detected in 44.9% of the unvaccinated dogs, 6.3% of the cattle and 2.8% of the humans. These antibody titres indicate either exposure to rabies virus or inapparent infection by rabies-related viruses.


Analysis of tubulin proteins and peptides in neuronal and non-neuronal tissues using immobilized pH gradients.
The accurate identification of the individual protein products of multi-gene families is essential to the interpretation of data from a wide range of experimental approaches including molecular biology, protein chemistry, and cell biology. We have adapted immobilized pH gradient isoelectric focusing to provide high resolution of tubulin proteins. Here we use these techniques to investigate the heterogeneity of tubulin in several neuronal and non-neuronal tissues to provide an accurate evaluation of isotubulin composition. Of the ten sources examined, the greatest number of isotubulins was found in whole adult brain. Tubulin isolated from either neonate human or rat brain consists predominantly of the more basic alpha and beta isotubulins found in adult brain. Cerebrum, cerebellum, medulla and caudate nucleus all contain the same large number of isotubulins as in whole brain, but in varying proportions. Liver, kidney and spleen isotubulin populations are all similar to each other and consist of a simpler distribution than any neuronal tissue examined. The majority of the tubulin protein in these non-neuronal tissues is composed of only the most basic alpha tubulins and intermediately-charged beta tubulins. No isotubulins were identified that were unique to these three non-neuronal tissues. Tubulin from neuroblastoma cells has an isotubulin distribution grossly similar to non-neuronal sources but additionally contains two basic beta isotubulins found in adult brain that are absent from non-neuronal tissues.


Group vs individual approaches to understanding the clinical significance of differences or changes in quality of life.
This article focuses on the traversing of group and individual levels of quality-of-life data. A deductive approach is used to address the extent to which group data can be used to estimate clinical significance at the individual level. An inductive approach is used to evaluate the extent to which individual change data can be brought to the group level to define clinical significance. Both approaches have benefits and drawbacks. This article addresses how clinical significance can be defined for an individual when the threshold for meaningfulness is drawn from group data. It also addresses the condition under which one can use the same threshold difference for group vs individual differences or changes. A sample inductive approach explores the means to identify a clinically significant result or change, with use of insights from cognitive psychology. In most deductive approaches, the identification of a clinically significant difference or change requires identification of a criterion (or at least an interpretable anchor) against which the significance of a change in respondent score is compared.


The Development of Audio-Visual Integration for Temporal Judgements.
Adults combine information from different sensory modalities to estimate object properties such as size or location. This process is optimal in that (i) sensory information is weighted according to relative reliability: more reliable estimates have more influence on the combined estimate and (ii) the combined estimate is more reliable than the component uni-modal estimates. Previous studies suggest that optimal sensory integration does not emerge until around 10 years of age. Younger children rely on a single modality or combine information using inappropriate sensory weights. Children aged 4-11 and adults completed a simple audio-visual task in which they reported either the number of beeps or the number of flashes in uni-modal and bi-modal conditions. In bi-modal trials, beeps and flashes differed in number by 0, 1 or 2. Mutual interactions between the sensory signals were evident at all ages: the reported number of flashes was influenced by the number of simultaneously presented beeps and vice versa. Furthermore, for all ages, the relative strength of these interactions was predicted by the relative reliabilities of the two modalities, in other words, all observers weighted the signals appropriately. The degree of cross-modal interaction decreased with age: the youngest observers could not ignore the task-irrelevant modality-they fully combined vision and audition such that they perceived equal numbers of flashes and beeps for bi-modal stimuli. Older observers showed much smaller effects of the task-irrelevant modality. Do these interactions reflect optimal integration? Full or partial cross-modal integration predicts improved reliability in bi-modal conditions. In contrast, switching between modalities reduces reliability. Model comparison suggests that older observers employed partial integration, whereas younger observers (up to around 8 years) did not integrate, but followed a sub-optimal switching strategy, responding according to either visual or auditory information on each trial.


The neuroactive potential of the human gut microbiota in quality of life and depression.
The relationship between gut microbial metabolism and mental health is one of the most intriguing and controversial topics in microbiome research. Bidirectional microbiota-gut-brain communication has mostly been explored in animal models, with human research lagging behind. Large-scale metagenomics studies could facilitate the translational process, but their interpretation is hampered by a lack of dedicated reference databases and tools to study the microbial neuroactive potential. Surveying a large microbiome population cohort (Flemish Gut Flora Project, n = 1,054) with validation in independent data sets (ntotal = 1,070), we studied how microbiome features correlate with host quality of life and depression. Butyrate-producing Faecalibacterium and Coprococcus bacteria were consistently associated with higher quality of life indicators. Together with Dialister, Coprococcus spp. were also depleted in depression, even after correcting for the confounding effects of antidepressants. Using a module-based analytical framework, we assembled a catalogue of neuroactive potential of sequenced gut prokaryotes. Gut-brain module analysis of faecal metagenomes identified the microbial synthesis potential of the dopamine metabolite 3,4-dihydroxyphenylacetic acid as correlating positively with mental quality of life and indicated a potential role of microbial γ-aminobutyric acid production in depression. Our results provide population-scale evidence for microbiome links to mental health, while emphasizing confounder importance.


Engineering anastomosis between living capillary networks and endothelial cell-lined microfluidic channels.
This paper reports a method for generating an intact and perfusable microvascular network that connects to microfluidic channels without appreciable leakage. This platform incorporates different stages of vascular development including vasculogenesis, endothelial cell (EC) lining, sprouting angiogenesis, and anastomosis in sequential order. After formation of a capillary network inside the tissue chamber via vasculogenesis, the adjacent microfluidic channels are lined with a monolayer of ECs, which then serve as the high-pressure input ("artery") and low pressure output ("vein") conduits. To promote a tight interconnection between the artery/vein and the capillary network, sprouting angiogenesis is induced, which promotes anastomosis of the vasculature inside the tissue chamber with the EC lining along the microfluidic channels. Flow of fluorescent microparticles confirms the perfusability of the lumenized microvascular network, and minimal leakage of 70 kDa FITC-dextran confirms physiologic tightness of the EC junctions and completeness of the interconnections between artery/vein and the capillary network. This versatile device design and its robust construction methodology establish a physiological transport model of interconnected perfused vessels from artery to vascularized tissue to vein. The system has utility in a wide range of organ-on-a-chip applications as it enables the physiological vascular interconnection of multiple on-chip tissue constructs that can serve as disease models for drug screening.


Neuroprotective effect of Citrus kawachiensis (Kawachi Bankan) peels, a rich source of naringin, against global cerebral ischemia/reperfusion injury in mice.
Cerebral ischemia/reperfusion is known to induce the generation of reactive oxygen species and inflammatory responses. Numerous studies have demonstrated that naringin (NGIN) has anti-oxidant and anti-inflammatory properties. We previously reported that Citrus kawachiensis contains a large quantity of NGIN in its peel. In the present study, we orally (p.o.) administered dried peel powder of C. kawachiensis to mice of a transient global ischemia model and found in the hippocampus region that it 1) suppressed neuronal cell death, 2) reversed the reduction in the level of phosphorylated calcium-calmodulin-dependent protein kinase II, 3) had the tendency to reverse the reduction in the level of glutathione, and 4) blocked excessive activation of microglia and astrocytes. These results suggested that the dried peel powder of C. kawachiensis had a neuroprotective effect against ischemic brain via anti-oxidative and anti-inflammatory effects. We also showed that these effects of the dried peel powder were more powerful than those obtained with a comparable amount of NGIN alone.


Driving pressure and survival in the acute respiratory distress syndrome.
Mechanical-ventilation strategies that use lower end-inspiratory (plateau) airway pressures, lower tidal volumes (VT), and higher positive end-expiratory pressures (PEEPs) can improve survival in patients with the acute respiratory distress syndrome (ARDS), but the relative importance of each of these components is uncertain. Because respiratory-system compliance (CRS) is strongly related to the volume of aerated remaining functional lung during disease (termed functional lung size), we hypothesized that driving pressure (ΔP=VT/CRS), in which VT is intrinsically normalized to functional lung size (instead of predicted lung size in healthy persons), would be an index more strongly associated with survival than VT or PEEP in patients who are not actively breathing. Using a statistical tool known as multilevel mediation analysis to analyze individual data from 3562 patients with ARDS enrolled in nine previously reported randomized trials, we examined ΔP as an independent variable associated with survival. In the mediation analysis, we estimated the isolated effects of changes in ΔP resulting from randomized ventilator settings while minimizing confounding due to the baseline severity of lung disease. Among ventilation variables, ΔP was most strongly associated with survival. A 1-SD increment in ΔP (approximately 7 cm of water) was associated with increased mortality (relative risk, 1.41; 95% confidence interval [CI], 1.31 to 1.51; P<0.001), even in patients receiving "protective" plateau pressures and VT (relative risk, 1.36; 95% CI, 1.17 to 1.58; P<0.001). Individual changes in VT or PEEP after randomization were not independently associated with survival; they were associated only if they were among the changes that led to reductions in ΔP (mediation effects of ΔP, P=0.004 and P=0.001, respectively). We found that ΔP was the ventilation variable that best stratified risk. Decreases in ΔP owing to changes in ventilator settings were strongly associated with increased survival. (Funded by Fundação de Amparo e Pesquisa do Estado de São Paulo and others.).


Structural Analysis of Hippocampal Kinase Signal Transduction.
Kinases are a major clinical target for human diseases. Identifying the proteins that interact with kinases in vivo will provide information on unreported substrates and will potentially lead to more specific methods for therapeutic kinase regulation. Here, endogenous immunoprecipitations of evolutionally distinct kinases (i.e., Akt, ERK2, and CAMK2) from rodent hippocampi were analyzed by mass spectrometry to generate three highly confident kinase protein-protein interaction networks. Proteins of similar function were identified in the networks, suggesting a universal model for kinase signaling complexes. Protein interactions were observed between kinases with reported symbiotic relationships. The kinase networks were significantly enriched in genes associated with specific neurodevelopmental disorders providing novel structural connections between these disease-associated genes. To demonstrate a functional relationship between the kinases and the network, pharmacological manipulation of Akt in hippocampal slices was shown to regulate the activity of potassium/sodium hyperpolarization-activated cyclic nucleotide-gated channel(HCN1), which was identified in the Akt network. Overall, the kinase protein-protein interaction networks provide molecular insight of the spatial complexity of in vivo kinase signal transduction which is required to achieve the therapeutic potential of kinase manipulation in the brain.


How training affects Australian paediatricians' management of obesity.
Secondary care could be the optimal sector for managing child and adolescent obesity, given low primary care uptake and limited tertiary services. We aimed to determine Australian paediatricians' self-reported competence and training in managing obesity and, in a linked patient-level audit, whether these predict rates of measurement and obesity diagnosis. Australian Paediatric Research Network members completed an online survey, plus a prospective patient-level audit of up to 100 consecutive consultations over 2 weeks. self-reported competencies, training in and use of clinical skills in obesity and its comorbidities. Audit: paediatricians reported each child's height, weight, age, sex and diagnoses including overweight/obesity. Of 166 (44.7% response) paediatricians, most felt very/quite competent in assessing (89%) and managing (68%) obesity, but few in making a difference to obesity (20%) or managing hypertension (45%), insulin resistance (32%), fatty liver disease (22%) or dyslipidaemia (21%). The audit of 200 (66.2% response) paediatricians included 8345 patients. On average paediatricians recorded height and weight for 66.5% of patients (SD 30.0%, range 0-100%). Of the 296 (12.3%) patients obese by CDC cutpoints, 118 (39.9%) were diagnosed as obese; perceived competence increased the odds of recording this diagnosis but not measurement. Training levels were low, showed little association with measurement or obesity diagnosis, and skills learnt were not routinely used. There is a clear need for better paediatrician training in obesity management. However, care and outcomes for obese children are unlikely to improve unless effective management models can be operationalised systematically.


Immunohistochemical and in situ localization of Crimean-Congo hemorrhagic fever (CCHF) virus in human tissues and implications for CCHF pathogenesis.
Crimean-Congo hemorrhagic fever (CCHF) is a potentially fatal disease that occurs in parts of Africa, Asia, and eastern Europe, and that is caused by a recently emerged bunyavirus. Rapid laboratory diagnosis of CCHF infection is essential and is currently performed by virus isolation and serology. Histopathologic studies have been limited to a small number of cases, and little is known about the cellular tropism of CCHF virus and the pathogenesis of this disease. We conducted a retrospective case analysis of 12 patients with a diagnosis of CCHF infection, confirmed by virus isolation, who were evaluated at the Special Pathogens Unit, National Institute for Virology, South Africa. The clinicopathologic features of CCHF and the diagnostic role of virus isolation as compared with serology, immunohistochemistry, and in situ hybridization were evaluated. Additionally, the distribution of CCHF virus in human tissues was examined. The clinical and histopathologic features of CCHF resemble those of other viral hemorrhagic fevers. Of the 12 patients with virus isolation-confirmed CCHF infection, 5 were positive by serology, 10 by immunohistochemistry, and 5 by in situ hybridization. Immunohistochemistry and in situ hybridization analyses showed that the mononuclear phagocytes, endothelial cells, and hepatocytes are main targets of infection. Association of parenchymal necrosis in liver with viral infection suggests that cell damage may be mediated by a direct viral cytopathic effect. The diagnosis of CCHF, suspected by history and clinical features, can be supported histopathologically. However, since the pathologic features resemble those of other viral hemorrhagic fevers, an unequivocal diagnosis can be made only by laboratory tests. The utility of immunohistochemistry as a sensitive and rapid diagnostic modality was established by the high degree of concordance with virus isolation. Infection of mononuclear phagocytes, endothelial cells, and hepatocytes may play a critical role in the pathogenesis of CCHF.


Fibromyalgia and the facts. Sense or nonsense.
A recent flurry of important studies has provided critical new information that is relevant to the contemporary understanding of the fibromyalgia syndrome. The concept that these patients represent solely a form of masked depression or a distinctive syndrome of somatization is not supported by the current facts. Rather it would appear that a characteristic peripheral nociceptive component is modulated by an interplay of complex central factors. A disruption of the neuroendocrine axis controlling growth hormone production may be the link between disturbed sleep and muscle pain, as growth hormone is produced predominantly during stage four sleep. A paradigm to link some of these newer findings is presented.


Monitoring the effect of heparin by measurement of activated clotting time during and after percutaneous transluminal coronary angioplasty.
The anticoagulant effect of heparin during percutaneous transluminal coronary angioplasty was monitored by measurements of the activated clotting time in two studies that compared the effects of a single bolus of heparin with those of a bolus of heparin combined with a continuous infusion of the drug. In a preliminary study 40 patients received a single heparin bolus of 10,000 units (protocol I) and a further 40 patients received both a heparin bolus of 10,000 and a continuous infusion of heparin at a rate of 2000 units per hour (protocol II). During the first 45 minutes, nine patients (23%) in protocol I but only two patients (5%) in protocol II were found to be inadequately anticoagulated. For 24 hours after angioplasty both groups received an infusion of heparin at the rate of 2000 units per hour which led to consistent anticoagulation in 73 (91%) of patients. In a subsequent randomised study, 40 patients received heparin according to either protocol I or II. Protocol II was again found to lead to a higher rate of adequate anticoagulation. During the first 60 minutes 11 patients (55%) in protocol I but only three patients (15%) in protocol II were inadequately anticoagulated. In addition, the activated clotting time of arterial blood in the first 30 minutes was significantly higher than that of venous blood in 70% of the patients. A bolus of heparin (10,000 units) together with an infusion of 2000 units per hour should be routinely given during coronary angioplasty. The effects of heparin, which can vary considerably from patient to patient, should be monitored by the measurement of the activated clotting time of arterial blood.


How to get the most out of your trainees in robotic thoracic surgery-"the coachability languages".
We are honored to have been invited to write this piece entitled, "How to get the most out of your trainees in robotic thoracic surgery". Perhaps a better question is "How can we optimally coach and inspire each resident and/or fellow to maximize their value and potential as people, physicians and surgeons during the span of their career?". As surgeons, we must recognize some of the subtle differences in alignment between ourselves and our trainees, appreciate the value of the trainee within our profession, understand that there is variability to the coaching style that each trainee best responds to, and acknowledge that the success of the people we train-which may be our only true legacy-depends on how we engage and inspire them.


Controversies in the physiological basis of the 'anaerobic threshold' and their implications for clinical cardiopulmonary exercise testing.
This article reviews the notion of the 'anaerobic threshold' in the context of cardiopulmonary exercise testing. Primarily, this is a review of the proposed mechanisms underlying the ventilatory and lactate response to incremental exercise, which is important to the clinical interpretation of an exercise test. Since such tests are often conducted for risk stratification before major surgery, a failure to locate or justify the existence of an anaerobic threshold will have some implications for clinical practice. We also consider alternative endpoints within the exercise response that might be better used to indicate a patient's capacity to cope with the metabolic demands encountered both during and following major surgery.


Quantitative dynamic multicompartmental analysis of cholecystokinin receptor movement in a living cell using dual fluorophores and reconstruction of confocal images.
Receptor regulation is a key component of the phenomenon of desensitization in response to agonist stimulation which protects cells from overstimulation. Receptor internalization is one part of this response, often quantified by the portion of saturable ligand binding which becomes resistant to acidic washes. It is now clear that this can include receptor in multiple distinct cellular compartments. We have developed a morphological technique involving dual fluorescent probes to delineate the plasmalemma and the ligand-occupied receptor using confocal microscopy, with analysis involving three-dimensional reconstruction and quantitation of receptor movement through each compartment. When a radioiodinated cholecystokinin (CCK) analogue occupied its receptor on the CHO-CCKR cell line, it became progressively more resistant to dissociation with acidic medium. Quantitation of receptor internalization in these cells over time using this dynamic morphological technique correlated with the acid-resistant receptor fraction, and provided the additional information of the cellular compartments traversed. This technique will have multiple applications to explore the cell-specific handling of this and other ligand-occupied receptors.


Cross-species functional alignment reveals evolutionary hierarchy within the connectome.
Evolution provides an important window into how cortical organization shapes function and vice versa. The complex mosaic of changes in brain morphology and functional organization that have shaped the mammalian cortex during evolution, complicates attempts to chart cortical differences across species. It limits our ability to fully appreciate how evolution has shaped our brain, especially in systems associated with unique human cognitive capabilities that lack anatomical homologues in other species. Here, we develop a function-based method for cross-species alignment that enables the quantification of homologous regions between humans and rhesus macaques, even when their location is decoupled from anatomical landmarks. Critically, we find cross-species similarity in functional organization reflects a gradient of evolutionary change that decreases from unimodal systems and culminates with the most pronounced changes in posterior regions of the default mode network (angular gyrus, posterior cingulate and middle temporal cortices). Our findings suggest that the establishment of the default mode network, as the apex of a cognitive hierarchy, has changed in a complex manner during human evolution - even within subnetworks.


Progression of conventional cardiovascular risk factors and vascular disease risk in individuals: insights from the PROG-IMT consortium.
Averaged measurements, but not the progression based on multiple assessments of carotid intima-media thickness, (cIMT) are predictive of cardiovascular disease (CVD) events in individuals. Whether this is true for conventional risk factors is unclear. An individual participant meta-analysis was used to associate the annualised progression of systolic blood pressure, total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol with future cardiovascular disease risk in 13 prospective cohort studies of the PROG-IMT collaboration (n = 34,072). Follow-up data included information on a combined cardiovascular disease endpoint of myocardial infarction, stroke, or vascular death. In secondary analyses, annualised progression was replaced with average. Log hazard ratios per standard deviation difference were pooled across studies by a random effects meta-analysis. In primary analysis, the annualised progression of total cholesterol was marginally related to a higher cardiovascular disease risk (hazard ratio (HR) 1.04, 95% confidence interval (CI) 1.00 to 1.07). The annualised progression of systolic blood pressure, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol was not associated with future cardiovascular disease risk. In secondary analysis, average systolic blood pressure (HR 1.20 95% CI 1.11 to 1.29) and low-density lipoprotein cholesterol (HR 1.09, 95% CI 1.02 to 1.16) were related to a greater, while high-density lipoprotein cholesterol (HR 0.92, 95% CI 0.88 to 0.97) was related to a lower risk of future cardiovascular disease events. Averaged measurements of systolic blood pressure, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol displayed significant linear relationships with the risk of future cardiovascular disease events. However, there was no clear association between the annualised progression of these conventional risk factors in individuals with the risk of future clinical endpoints.


Validity conditions for the radiative transfer equation.
We compare the radiative transfer equation for media with constant refractive index with the radiative transfer equation for media with spatially varying refractive indices [J. Opt. A Pure App. Opt. 1, L1 (1999)] and obtain approximate conditions under which the former equation is accurate for modeling light propagation in scattering media with spatially varying refractive indices. These conditions impose restrictions on the variations of the refractive index, the gradient of the refractive index, the divergence of the rays, the frequency of modulation, and the widths of light pulses, which are related to the mean refractive index, the absorption coefficient, and the reduced scattering coefficient of the medium. Each condition is geometrically interpreted. Some implications of the results are discussed.


Being popular can be healthy or unhealthy: stress, social network diversity, and incidence of upper respiratory infection.
Diverse social contacts are generally associated with better health. However, diverse contacts can increase exposure to infectious agents. This should increase risk for disease, particularly among those whose host resistance is otherwise compromised (e.g., stressed individuals). In this prospective study, healthy college students who completed questionnaires assessing social network diversity and stressful life events were subsequently interviewed weekly for 12 weeks to track incidence of upper respiratory infections (URIs). URI episodes were defined by a symptom criterion and by clinically verified self-reported illness. Stress and diversity of social contacts interacted; diversity was associated with more illnesses among those with more stressful life events and slightly fewer illnesses among those with fewer stressful life events. Associations remained after controlling for neuroticism.


Self-aligning prosthetic device for older patients with vascular-related amputations: protocol for a randomised feasibility study (the STEPFORWARD study).
The majority of older patients with a transtibial amputation are prescribed a standard (more rigid, not self-aligning) prosthesis. These are mostly suitable for level walking, and cannot adjust to different sloped surfaces. This makes walking more difficult and less energy efficient, possibly leading to longer term disuse. A Cochrane Review concluded that there was insufficient evidence to recommend any individual type of prosthetic ankle-foot mechanism. This trial will establish the feasibility of conducting a large-scale trial to assess the effectiveness and cost-effectiveness of a self-aligning prosthesis for older patients with vascular-related amputations and other health issues compared with a standard prosthesis. This feasibility trial is a pragmatic, parallel group, randomised controlled trial (RCT) comparing standard treatment with a more rigid prosthesis versus a self-aligning prosthesis. The target sample size is 90 patients, who are aged 50 years and over, and have a transtibial amputation, where amputation aetiology is mostly vascular-related or non-traumatic. Feasibility will be measured by consent and retention rates, a plausible future sample size over a 24-month recruitment period and completeness of outcome measures. Qualitative interviews will be carried out with trial participants to explore issues around study processes and acceptability of the intervention. Focus groups with staff at prosthetics centres will explore barriers to successful delivery of the trial. Findings from the qualitative work will be integrated with the feasibility trial outcomes in order to inform the design of a full-scale RCT. Ethical approval was granted by Yorkshire and the Humber-Leeds West Research Ethics Committee on 4 May 2018. The findings will be disseminated via peer-reviewed research publications, articles in relevant newsletters, presentations at relevant conferences and the patient advisory group. ISRCTN15043643.


Cardiorespiratory fitness in young adulthood and the development of cardiovascular disease risk factors.
Low cardiorespiratory fitness is an established risk factor for cardiovascular and total mortality; however, mechanisms responsible for these associations are uncertain. To test whether low fitness, estimated by short duration on a maximal treadmill test, predicted the development of cardiovascular disease risk factors and whether improving fitness (increase in treadmill test duration between examinations) was associated with risk reduction. Population-based longitudinal cohort study of men and women 18 to 30 years of age in the Coronary Artery Risk Development in Young Adults (CARDIA) study. Participants who completed the treadmill examination according to the Balke protocol at baseline were followed up from 1985-1986 to 2000-2001. A subset of participants (n = 2478) repeated the exercise test in 1992-1993. Incident type 2 diabetes, hypertension, the metabolic syndrome (defined according to National Cholesterol Education Program Adult Treatment Panel III), and hypercholesterolemia (low-density lipoprotein cholesterol > or =160 mg/dL [4.14 mmol/L]). During the 15-year study period, the rates of incident diabetes, hypertension, the metabolic syndrome, and hypercholesterolemia were 2.8, 13.0, 10.2, and 11.7 per 1000 person-years, respectively. After adjustment for age, race, sex, smoking, and family history of diabetes, hypertension, or premature myocardial infarction, participants with low fitness (<20th percentile) were 3- to 6-fold more likely to develop diabetes, hypertension, and the metabolic syndrome than participants with high fitness (> or =60th percentile), all P<.001. Adjusting for baseline body mass index diminished the strength of these associations to 2-fold (all P<.001). In contrast, the association between low fitness and hypercholesterolemia was modest (hazard ratio [HR], 1.4; 95% confidence interval [CI], 1.1-1.7; P =.02) and attenuated to marginal significance after body mass index adjustment (P =.13). Improved fitness over 7 years was associated with a reduced risk of developing diabetes (HR, 0.4; 95% CI, 0.2-1.0; P =.04) and the metabolic syndrome (HR, 0.5; 95% CI, 0.3-0.7; P<.001), but the strength and significance of these associations was reduced after accounting for changes in weight. Poor fitness in young adults is associated with the development of cardiovascular disease risk factors. These associations involve obesity and may be modified by improving fitness.


A comparison of the effects of electrode implantation and targeting on pattern classification accuracy for prosthesis control.
The use of surface versus intramuscular electrodes as well as the effect of electrode targeting on pattern-recognition-based multifunctional prosthesis control was explored. Surface electrodes are touted for their ability to record activity from relatively large portions of muscle tissue. Intramuscular electromyograms (EMGs) can provide focal recordings from deep muscles of the forearm and independent signals relatively free of crosstalk. However, little work has been done to compare the two. Additionally, while previous investigations have either targeted electrodes to specific muscles or used untargeted (symmetric) electrode arrays, no work has compared these approaches to determine if one is superior. The classification accuracies of pattern-recognition-based classifiers utilizing surface and intramuscular as well as targeted and untargeted electrodes were compared across 11 subjects. A repeated-measures analysis of variance revealed that when only EMG amplitude information was used from all available EMG channels, the targeted surface, targeted intramuscular, and untargeted surface electrodes produced similar classification accuracies while the untargeted intramuscular electrodes produced significantly lower accuracies. However, no statistical differences were observed between any of the electrode conditions when additional features were extracted from the EMG signal. It was concluded that the choice of electrode should be driven by clinical factors, such as signal robustness/stability, cost, etc., instead of by classification accuracy.


Dating the genetic bottleneck of the African cheetah.
The cheetah is unusual among fields in exhibiting near genetic uniformity at a variety of loci previously screened to measure population genetic diversity. It has been hypothesized that a demographic crash or population bottleneck in the recent history of the species is causal to the observed monomorphic profiles for nuclear coding loci. The timing of a bottleneck is difficult to assess, but certain aspects of the cheetah's natural history suggest it may have occurred near the end of the last ice age (late Pleistocene, approximately 10,000 years ago), when a remarkable extinction of large vertebrates occurred on several continents. To further define the timing of such a bottleneck, the character of genetic diversity for two rapidly evolving DNA sequences, mitochondrial DNA and hypervariable minisatellite loci, was examined. Moderate levels of genetic diversity were observed for both of these indices in surveys of two cheetah subspecies, one from South Africa and one from East Africa. Back calculation from the extent of accumulation of DNA diversity based on observed mutation rates for VNTR (variable number of tandem repeats) loci and mitochondrial DNA supports a hypothesis of an ancient Pleistocene bottleneck that rendered the cheetah depauperate in genetic variation for nuclear coding loci but would allow sufficient time for partial reconstitution of more rapidly evolving genomic DNA segments.


What benefit could be derived from on-line adaptive prostate radiotherapy using rectal diameter as a predictor of motion?
This study investigated a relationship between rectum diameter and prostate motion during treatment with a view to reducing planning target volume (PTV) margins for an adaptive protocol. One hundred and ninety-four cone-beam computed tomography (CBCT) images of 10 patients were used to relate rectum diameter on CBCT to prostate intrafraction displacement. A threshold rectum diameter was used to model the impact of an adaptive PTV margin on rectum and bladder dose. Potential dose escalation with a 6 mm uniform margin adaptive protocol was compared to a PTV margin of 10 mm expansion of the clinical target volume (CTV) except 6 mm posterior. Of 194 fractions, 104 had a maximum rectal diameter of ≤3.5 cm. The prostate displaced ≤4 mm in 102 of those fractions. Changing from a standard to an adaptive PTV margin reduced the volume of rectum receiving 25, 50, 60, and 70 Gy by around 12, 9, 10, and 16%, respectively and bladder by approximately 21, 27, 29, and 35%, respectively. An average dose escalation of 4.2 Gy may be possible with an adaptive prostate radiotherapy protocol. In conclusion, a relationship between the prostate motion and the diameter of the rectum on CBCT potentially could enable daily adaptive radiotherapy which can be implemented from the first fraction.


The potential of target-specific oral anticoagulants for the acute and long-term treatment of venous thromboembolism.
Venous thromboembolism (VTE) comprises both deep vein thrombosis and pulmonary embolism. VTE is a leading cause of morbidity and mortality worldwide and its increasing incidence and prevalence are a major health concern. The primary medical objective during the acute phase of VTE treatment is to prevent thrombus extension and embolization. Extended treatment aims to prevent or minimize long-term complications, such as recurrent VTE, post-thrombotic syndrome and chronic thromboembolic pulmonary hypertension. Anticoagulant therapy has been the mainstay of treatment for VTE and traditionally involves initial therapy with heparin, overlapping with and followed by a vitamin K antagonist. Although effective, standard heparin/vitamin K antagonist therapy has several limitations that can be overcome by more recently developed target-specific oral anticoagulants (TSOACs). These agents have predictable pharmacokinetics, a rapid onset of action and few drug-drug or drug-food interactions. Furthermore, TSOACs offer convenient anticoagulation without the need for routine coagulation monitoring and dose adjustment. The efficacy and safety data from phase III clinical trials support the use of TSOACs for VTE treatment, including in special patient populations. Risk-stratification tools and strategies have been developed to assist physicians in managing anticoagulation treatment. Rivaroxaban is the first TSOAC to gain widespread approval for the treatment of acute deep vein thrombosis and pulmonary embolism and the long-term prevention of recurrent VTE as monotherapy. Dabigatran has also been approved for this indication recently. TSOACs, especially as monotherapy, represent a paradigm shift in clinical practice for the management of patients with VTE.


Monolithic integration of microlens arrays and fiber holder arrays in poly(methyl methacrylate) with fiber self-centering.
For many fiber applications the precise spacing from one fiber to another and the collimation of the outgoing light are important subjects. In the case of three-dimensional arrangements, such as fiber arrays, the complexity increases. The method of proton irradiation of poly(methyl methacrylate) permits the fabrication of slightly fan-shaped fiber holder arrays that can correct inhomogeneities of the fiber diameters in the micrometer range. Even a three-dimensional monolithic integration of fiber holders and corresponding microlenses can be achieved.


Effect of the trap crop, Solanum sisymbriifolium, on Globodera pallida, Globodera tabacum, and Globodera ellingtonae.
The effect of the nematode trap crop Solanum sisymbriifolium was assessed against three Globodera spp., the potato cyst nematode Globodera pallida (in Idaho), the recently described Globodera ellingtonae (in Oregon), and the tobacco cyst nematode Globodera tabacum (in Connecticut) in field trials. At all locations the ability of S. sisymbriifolium to reduce Globodera encysted second-stage juveniles (J2) in egg densities compared to fallow was considered. For G. ellingtonae, the impact of planting and termination dates of S. sisymbriifolium on final egg densities was also evaluated; and for G. pallida, the ability of the nematode to reproduce on potato (Solanum tuberosum) after exposure to S. sisymbriifolium was determined. Encysted J2 in egg densities of all three Globodera spp. declined from 25 to 68% after trap cropping with S. sisymbriifolium. For G. pallida, S. sisymbriifolium reduced final encysted J2 in egg density by 23 to 50% compared to the fallow treatment, and significantly decreased G. pallida reproduction on potato after exposure to S. sisymbriifolium by 99 to 100% compared to the fallow treatment (P < 0.0001). For G. ellingtonae, the planting date of S. sisymbriifolium in May or June did not impact final egg densities (P = 0.32). Rather, percentage reduction in G. ellingtonae encysted J2 in egg density was most influenced by the length of time to which nematodes were exposed to S. sisymbriifolium, with 30 and 81% reduction after 6 vs 12 wk of exposure, respectively (P < 0.0001). Similar levels of nematode reduction after S. sisymbriifolium were observed for G. tabacum after 12 to 14 wk of exposure to the trap crop; G. tabacum density changes consisted of a 114% increase after susceptible tobacco, a 65% decrease after resistant tobacco, and an 88% decrease after S. sisymbriifolium compared to bare soil. In conclusion, this research demonstrates the widespread applicability of S. sisymbriifolium in reducing a diversity of Globodera spp. present in the USA. The effect of the nematode trap crop Solanum sisymbriifolium was assessed against three Globodera spp., the potato cyst nematode Globodera pallida (in Idaho), the recently described Globodera ellingtonae (in Oregon), and the tobacco cyst nematode Globodera tabacum (in Connecticut) in field trials. At all locations the ability of S. sisymbriifolium to reduce Globodera encysted second-stage juveniles (J2) in egg densities compared to fallow was considered. For G. ellingtonae, the impact of planting and termination dates of S. sisymbriifolium on final egg densities was also evaluated; and for G. pallida, the ability of the nematode to reproduce on potato (Solanum tuberosum) after exposure to S. sisymbriifolium was determined. Encysted J2 in egg densities of all three Globodera spp. declined from 25 to 68% after trap cropping with S. sisymbriifolium. For G. pallida, S. sisymbriifolium reduced final encysted J2 in egg density by 23 to 50% compared to the fallow treatment, and significantly decreased G. pallida reproduction on potato after exposure to S. sisymbriifolium by 99 to 100% compared to the fallow treatment (P < 0.0001). For G. ellingtonae, the planting date of S. sisymbriifolium in May or June did not impact final egg densities (P = 0.32). Rather, percentage reduction in G. ellingtonae encysted J2 in egg density was most influenced by the length of time to which nematodes were exposed to S. sisymbriifolium, with 30 and 81% reduction after 6 vs 12 wk of exposure, respectively (P < 0.0001). Similar levels of nematode reduction after S. sisymbriifolium were observed for G. tabacum after 12 to 14 wk of exposure to the trap crop; G. tabacum density changes consisted of a 114% increase after susceptible tobacco, a 65% decrease after resistant tobacco, and an 88% decrease after S. sisymbriifolium compared to bare soil. In conclusion, this research demonstrates the widespread applicability of S. sisymbriifolium in reducing a diversity of Globodera spp. present in the USA.


Discovery and development of the Polo-like kinase inhibitor volasertib in cancer therapy.
Owing to their integral involvement in cell cycle regulation, the Polo-like kinase (Plk) family, particularly Plk1, has emerged as an attractive therapeutic target in oncology. In recent years, several Plk1 inhibitors have been developed, with some agents showing encouraging results in early-phase clinical trials. This review focuses on volasertib (BI 6727; an investigational agent), a potent and selective Plk inhibitor. Volasertib has shown promising activity in various cancer cell lines and xenograft models of human cancer. Trials performed to date suggest that volasertib has clinical efficacy in a range of malignancies, with the most promising results seen in patients with acute myeloid leukemia (AML). Encouragingly, recent phase II data have demonstrated that volasertib combined with low-dose cytarabine (LDAC) was associated with higher response rates and improved event-free survival than LDAC alone in patients with previously untreated AML. Based on these observations, and its presumably manageable safety profile, volasertib is currently in phase III development as a potential treatment for patients with AML who are ineligible for intensive remission induction therapy. Given that many patients with AML are of an older age and frail, this constitutes an area of major unmet need. In this review, we discuss the biologic rationale for Plk1 inhibitors in cancer, the clinical development of volasertib to date in solid tumors and AML, and the future identification of biomarkers that might predict response to volasertib and help determine the role of this agent in the clinic.


Persistent Wolbachia and cultivable bacteria infection in the reproductive and somatic tissues of the mosquito vector Aedes albopictus.
Commensal and symbiotic microbes have a considerable impact on the behavior of many arthropod hosts, including hematophagous species that transmit pathogens causing infectious diseases to human and animals. Little is known about the bacteria associated with mosquitoes other than the vectorized pathogens. This study investigated Wolbachia and cultivable bacteria that persist through generations in Ae. albopictus organs known to host transmitted arboviruses, such as dengue and chikungunya. We used culturing, diagnostic and quantitative PCR, as well as in situ hybridization, to detect and locate bacteria in whole individual mosquitoes and in dissected tissues. Wolbachia, cultivable bacteria of the genera Acinetobacter, Comamonas, Delftia and Pseudomonas co-occurred and persisted in the bodies of both males and females of Ae. albopictus initially collected in La Réunion during the chikungunya outbreak, and maintained as colonies in insectaries. In dissected tissues, Wolbachia and the cultivable Acinetobacter can be detected in the salivary glands. The other bacteria are commonly found in the gut. Quantitative PCR estimates suggest that Wolbachia densities are highest in ovaries, lower than those of Acinetobacter in the gut, and approximately equal to those of Acinetobacter in the salivary glands. Hybridization using specific fluorescent probes successfully localized Wolbachia in all germ cells, including the oocytes, and in the salivary glands, whereas the Acinetobacter hybridizing signal was mostly located in the foregut and in the anterior midgut. Our results show that Proteobacteria are distributed in the somatic and reproductive tissues of mosquito where transmissible pathogens reside and replicate. This location may portend the coexistence of symbionts and pathogens, and thus the possibility that competition or cooperation phenomena may occur in the mosquito vector Ae. albopictus. Improved understanding of the vectorial system, including the role of bacteria in the vector's biology and competence, could have major implications for understanding viral emergences and for disease control.


Microfluidic transwell inserts for generation of tissue culture-friendly gradients in well plates.
Gradients of biochemical molecules play a key role in many physiological processes such as axon growth, tissue morphogenesis, and trans-epithelium nutrient transport, as well as in pathophysiological phenomena such as wound healing, immune response, bacterial invasion, and cancer metastasis. In this paper, we report a microfluidic transwell insert for generating quantifiable concentration gradients in a user-friendly and modular format that is compatible with conventional cell cultures and with tissue explant cultures. The device is simply inserted into a standard 6-well plate, where it hangs self-supported at a distance of ~250 μm above the cell culture surface. The gradient is created by small microflows from the device, through an integrated track-etched porous membrane, into the cell culture well. The microfluidic transwell can deliver stable, quantifiable gradients over a large area with extremely low fluid shear stress to dissociated cells or tissue explants cultured independently on the surface of a 6-well plate. We used finite-element modeling to describe the porous membrane flow and molecular transport and to predict gradients generated by the device. Using the device, we applied a gradient of the chemotactic peptide N-formyl-met-leu-phe (fMLP) to a large population of HL-60 cells (a neutrophil cell line) and directly observed the migration with time-lapse microscopy. On quantification of the chemotactic response with an automated tracking algorithm, we found 74% of the cells moving towards the gradient. Additionally, the modular design and low fluid shear stress made it possible to apply gradients of growth factors and second messengers to mouse retinal explant cultures. With a simplified interface and well-defined gradients, the microfluidic transwell device has potential for broad applications to gradient-sensing biology.


Self-inhibition of steroid secretion by amphibian adrenocortical cells is not mediated through glucocorticoid receptors.
To investigate a possible direct action of glucocorticoids on adrenal steroidogenesis, the effect of corticosterone on the conversion of pregnenolone into various metabolites by frog adrenal tissue was examined. Frog interrenal slices were incubated with [3H]pregnenolone (1 mCi/ml) and the various labelled metabolites analysed by reverse-phase high-performance liquid chromatography. With the methanol gradient used, five identified steroids were resolved: progesterone, 11-deoxycorticosterone, corticosterone, 18-hydroxycorticosterone and aldosterone. Corticosterone (10 micrograms/ml) induced a 45-80% decrease in all steroids synthesized from [3H]pregnenolone. In contrast, the glucocorticoid agonist dexamethasone did not reduce the rate of conversion of pregnenolone into its metabolites. In addition, the inhibitory effect of corticosterone was not reversed by the specific glucocorticoid antagonist RU 43044. These results show that corticosterone exerts a direct inhibitory effect on adrenal steroid secretion. In addition, our data indicate that the ultra-short regulation induced by corticosterone is not mediated through glucocorticoid receptors.


In vitro heart valve tissue engineering.
Heart valve replacement represents the most common surgical therapy for end-stage valvular heart diseases. A major drawback all contemporary heart valve replacements have in common is the lack of growth, repair, and remodeling capabilities. To overcome these limitations, the emerging field of tissue engineering is focusing on the in vitro generation of functional, living heart valve replacements. The basic approach uses starter matrices of either decellularized xenogeneic or biopolymeric materials configured in the shape of the heart valve and subsequent cell seeding. Moreover, in vitro strategies using mechanical loading in bioreactor systems have been developed to improve tissue maturation. This chapter gives a short overview of the current concepts and provides detailed methods for in vitro heart valve tissue engineering.


Photoelectric artefact from optogenetics and imaging on microelectrodes and bioelectronics: New Challenges and Opportunities.
Bioelectronics, electronic technologies that interface with biological systems, are experiencing rapid growth in terms of technology development and applications, especially in neuroscience and neuroprosthetic research. The parallel growth with optogenetics and in vivo multi-photon microscopy has also begun to generate great enthusiasm for simultaneous applications with bioelectronic technologies. However, emerging research showing artefact contaminated data highlight the need for understanding the fundamental physical principles that critically impact experimental results and complicate their interpretation. This review covers four major topics: 1) material dependent properties of the photoelectric effect (conductor, semiconductor, organic, photoelectric work function (band gap)); 2) optic dependent properties of the photoelectric effect (single photon, multiphoton, entangled biphoton, intensity, wavelength, coherence); 3) strategies and limitations for avoiding/minimizing photoelectric effects; and 4) advantages of and applications for light-based bioelectronics (photo-bioelectronics).


Higher dietary flavonol intake is associated with lower incidence of type 2 diabetes.
Substantial experimental evidence suggests that several flavonoid classes are involved in glucose metabolism, but few clinical or epidemiologic studies exist that provide supporting human evidence for this relationship. The objective of this study was to determine if habitual intakes of specific flavonoid classes are related to incidence of type 2 diabetes (T2D). We followed 2915 members of the Framingham Offspring cohort who were free of T2D at baseline from 1991 to 2008. Diabetes was defined by either elevated fasting glucose (≥7.0 mmol/L) or initiation of hypoglycemic medication during follow-up. Dietary intakes of 6 flavonoid classes and total flavonoids were assessed using a validated, semiquantitative food frequency questionnaire. We observed 308 incident cases of T2D during a mean follow-up period of 11.9 y (range 2.5-16.8 y). After multivariable adjusted, time-dependent analyses, which accounted for long-term flavonoid intake during follow-up, each 2.5-fold increase in flavonol intake was associated with a 26% lower incidence of T2D [HR = 0.74 (95% CI: 0.61, 0.90); P-trend = 0.003] and each 2.5-fold increase in flavan-3-ol intake was marginally associated with an 11% lower incidence of T2D [HR = 0.89 (95% CI: 0.80, 1.00); P-trend = 0.06]. No other associations between flavonoid classes and risk of T2D were observed. Our observations support previous experimental evidence of a possible beneficial relationship between increased flavonol intake and risk of T2D.


A randomized parallel-group dietary study for stages II-IV ovarian cancer survivors.
Few studies have examined the dietary habits of ovarian cancer survivors. Therefore, we conducted a study to assess the feasibility and impact of two dietary interventions for ovarian cancer survivors. In this randomized, parallel-group study, 51 women (mean age, 53 years) diagnosed with stages II-IV ovarian cancer were recruited and randomly assigned to a low fat, high fiber (LFHF) diet or a modified National Cancer Institute diet supplemented with a soy-based beverage and encapsulated fruit and vegetable juice concentrates (FVJCs). Changes in clinical measures, serum carotenoid and tocopherol levels, dietary intake, anthropometry, and health-related quality of life (HRQOL) were assessed with paired t-tests. The recruitment rate was 25%, and the retention rate was 75% at 6 months. At baseline, 28% and 45% of women met guidelines for intake of fiber and of fruits and vegetables, respectively. After 6 months, total serum carotenoid levels and α- and β-carotene concentrations were significantly increased in both groups (P<0.01); however, β-carotene concentrations were increased more in the FVJC group. Serum β-cryptoxanthin levels, fiber intake (+5.2g/day), and daily servings of juice (+0.9 servings/day) and vegetables (+1.3 servings/day) were all significantly increased in the LFHF group (all P<0.05). Serum levels of albumin, lutein and zeaxanthin, retinol, and retinyl palmitate were significantly increased in the FVJC group (all P<0.05). No changes in cancer antigen-125, anthropometry, or HRQOL were observed. Overall, this study supports the feasibility of designing dietary interventions for stages II-IV ovarian cancer survivors and provides preliminary evidence that a low fat high fiber diet and a diet supplemented with encapsulated FVJC may increase phytonutrients in ovarian cancer survivors.


Endocytic vesicle rupture is a conserved mechanism of cellular invasion by amyloid proteins.
Numerous pathological amyloid proteins spread from cell to cell during neurodegenerative disease, facilitating the propagation of cellular pathology and disease progression. Understanding the mechanism by which disease-associated amyloid protein assemblies enter target cells and induce cellular dysfunction is, therefore, key to understanding the progressive nature of such neurodegenerative diseases. In this study, we utilized an imaging-based assay to monitor the ability of disease-associated amyloid assemblies to rupture intracellular vesicles following endocytosis. We observe that the ability to induce vesicle rupture is a common feature of α-synuclein (α-syn) assemblies, as assemblies derived from WT or familial disease-associated mutant α-syn all exhibited the ability to induce vesicle rupture. Similarly, different conformational strains of WT α-syn assemblies, but not monomeric or oligomeric forms, efficiently induced vesicle rupture following endocytosis. The ability to induce vesicle rupture was not specific to α-syn, as amyloid assemblies of tau and huntingtin Exon1 with pathologic polyglutamine repeats also exhibited the ability to induce vesicle rupture. We also observe that vesicles ruptured by α-syn are positive for the autophagic marker LC3 and can accumulate and fuse into large, intracellular structures resembling Lewy bodies in vitro. Finally, we show that the same markers of vesicle rupture surround Lewy bodies in brain sections from PD patients. These data underscore the importance of this conserved endocytic vesicle rupture event as a damaging mechanism of cellular invasion by amyloid assemblies of multiple neurodegenerative disease-associated proteins, and suggest that proteinaceous inclusions such as Lewy bodies form as a consequence of continued fusion of autophagic vesicles in cells unable to degrade ruptured vesicles and their amyloid contents.


No panacea for drainage after percutaneous nephrolithotomy.
The optimal postoperative drainage system for patients undergoing percutaneous nephrolithotomy (PCNL) has not yet been determined. Each of the different tubes available has theoretical advantages and disadvantages. The objective of this study was to determine if any one system of post-PCNL drainage has any significant advantage over the others. Sixty patients were randomized to either a 24F Re-entry tube (Group 1), an 8F pigtail catheter (Group 2), or a double-J stent (Group 3) (N = 20 for all groups). The double-J stent patients also had an 18F Councill catheter as a nephrostomy tube, which was removed on the morning of postoperative day 1. The three groups were well-matched for age, stone size, operative time, and mean number of percutaneous tracts (P > 0.05 for all). Data collected included intraoperative findings, postoperative analog pain scores and narcotic usage, presence of extravasation on postoperative nephrostogram, presence of perinephric fluid on postoperative ultrasound scans, length of stay, and whether leakage from the nephrostomy site persisted more than 48 hours after tube removal. There were no statistically significant differences among the three groups in terms of change in hematocrit or number of patients requiring blood transfusion; in the incidence of tube blockage, extravasation, or presence of perinephritic fluid; or in complication rates, length of hospital stay, or persistent leakage after tube removal. There was a strong trend to less postoperative leakage with the 8F pigtail than the Reentry tube (P = 0.05). Postoperative analog pain scores and narcotic usage were equivalent for all three groups. Patients Groups 1 and 2 were free of all tubes sooner than patients in Group 3 (P < 0.001). This study did not demonstrate an overwhelming advantage of any one drainage system over the others. All three systems were equally tolerated by patients. There may be less chance of prolonged nephrostomy-site leakage with an 8F pigtail catheter.


SIRT6 regulates osteogenic differentiation of rat bone marrow mesenchymal stem cells partially via suppressing the nuclear factor-κB signaling pathway.
Sirtuin 6 (SIRT6) is a NAD-dependent deacetylase involved in lifespan regulation. To evaluate the effect of SIRT6 on osteogenesis, rat bone marrow mesenchymal stem cells (rBMSCs) with enhanced or reduced SIRT6 function were developed. We observed that SIRT6 knockdown significantly reduced the mRNA levels of several key osteogenic markers in vitro, including alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), and osteocalcin, while overexpression of SIRT6 enhanced their expression. Additionally, SIRT6 knockdown activated nuclear factor-κB (NF-κB) transcriptional activity and upregulated the expression of acetyl-NF-κB p65 (Lys310). The decreased osteogenic differentiation ability of rBMSCs could be partially rescued by the addition of NF-κB inhibitor BAY 11-7082. Furthermore, SIRT6 overexpression in rBMSCs combined with the use of collagen/chitosan/hydroxyapatite scaffold could significantly boost new bone formation in rat cranial critical-sized defects, as determined by microcomputed tomography and histological examination. These data confirm that SIRT6 is mainly located in the nuclei of rBMSCs and plays an essential role in their normal osteogenic differentiation, partly by suppressing NF-κB signaling.


Novel division level bacterial diversity in a Yellowstone hot spring.
A culture-independent molecular phylogenetic survey was carried out for the bacterial community in Obsidian Pool (OP), a Yellowstone National Park hot spring previously shown to contain remarkable archaeal diversity (S. M. Barns, R. E. Fundyga, M. W. Jeffries, and N. R. Page, Proc. Natl. Acad. Sci. USA 91:1609-1613, 1994). Small-subunit rRNA genes (rDNA) were amplified directly from OP sediment DNA by PCR with universally conserved or Bacteria-specific rDNA primers and cloned. Unique rDNA types among > 300 clones were identified by restriction fragment length polymorphism, and 122 representative rDNA sequences were determined. These were found to represent 54 distinct bacterial sequence types or clusters (> or = 98% identity) of sequences. A majority (70%) of the sequence types were affiliated with 14 previously recognized bacterial divisions (main phyla; kingdoms); 30% were unaffiliated with recognized bacterial divisions. The unaffiliated sequence types (represented by 38 sequences) nominally comprise 12 novel, division level lineages termed candidate divisions. Several OP sequences were nearly identical to those of cultivated chemolithotrophic thermophiles, including the hydrogen-oxidizing Calderobacterium and the sulfate reducers Thermodesulfovibrio and Thermodesulfobacterium, or belonged to monophyletic assemblages recognized for a particular type of metabolism, such as the hydrogen-oxidizing Aquificales and the sulfate-reducing delta-Proteobacteria. The occurrence of such organisms is consistent with the chemical composition of OP (high in reduced iron and sulfur) and suggests a lithotrophic base for primary productivity in this hot spring, through hydrogen oxidation and sulfate reduction. Unexpectedly, no archaeal sequences were encountered in OP clone libraries made with universal primers. Hybridization analysis of amplified OP DNA with domain-specific probes confirmed that the analyzed community rDNA from OP sediment was predominantly bacterial. These results expand substantially our knowledge of the extent of bacterial diversity and call into question the commonly held notion that Archaea dominate hydrothermal environments. Finally, the currently known extent of division level bacterial phylogenetic diversity is collated and summarized.


Enhancing sensorimotor BCI performance with assistive afferent activity: An online evaluation.
An important goal in Brain-Computer Interfacing (BCI) is to find and enhance procedural strategies for users for whom BCI control is not sufficiently accurate. To address this challenge, we conducted offline analyses and online experiments to test whether the classification of different types of motor imagery could be improved when the training of the classifier was performed on the data obtained with the assistive muscular stimulation below the motor threshold. 10 healthy participants underwent three different types of experimental conditions: a) Motor imagery (MI) of hands and feet b) sensory threshold neuromuscular electrical stimulation (STM) of hands and feet while resting and c) sensory threshold neuromuscular electrical stimulation during performance of motor imagery (BOTH). Also, another group of 10 participants underwent conditions a) and c). Then, online experiments with 15 users were performed. These subjects received neurofeedback during MI using classifiers calibrated either on MI or BOTH data recorded in the same experiment. Offline analyses showed that decoding MI alone using a classifier based on BOTH resulted in a better BCI accuracy compared to using a classifier based on MI alone. Online experiments confirmed accuracy improvement of MI alone being decoded with the classifier trained on BOTH data. In addition, we observed that the performance in MI condition could be predicted on the basis of a more pronounced connectivity within sensorimotor areas in the frequency bands providing the best performance in BOTH. These finding might offer a new avenue for training SMR-based BCI systems particularly for users having difficulties to achieve efficient BCI control. It might also be an alternative strategy for users who cannot perform real movements but still have remaining afferent pathways (e.g., ALS and stroke patients).


Laterality influences schooling position in rainbowfish, Melanotaenia spp.
Cerebral lateralization is a widespread trait among animals, is often manifested as side biases in behaviour (laterality) and has been suggested to provide fitness benefits. Here we examined the influence of laterality on the organisation of fish schools using rainbowfish (Melanotaenia spp) as model species. The pattern and strength of laterality for each individual was determined by examining eye preferences whilst examining their reflection in a mirror. Schools of four fish of known laterality were then created and the preferred position for each fish within the school was repeatedly observed in a flume. Fish which showed right eye preferences in the mirror test preferentially adopted a position on the left side of the school. Conversely, fish that showed left eye preferences in the mirror test or where non-lateralised preferentially adopted a position slightly to the right side of the school. However, this general pattern varied depending on the species and sex of the school. Our results strongly implicate individual laterality in the geometry of school formation.


Epistasis, phenotypic disequilibrium and contrasting associations with climate in the land snail Theba pisana.
Hotter conditions favour effectively unbanded (EUB) shells in the snail Theba pisana. T. pisana is also polymorphic for colour of the shell's apex, determined by a pair of alleles at a locus linked to the banding locus. Apex colour is epistatic to shell banding, such that banded snails with a dark apex have darker bands. Annual censuses over 22 years across an ecotone between a sheltered Acacia thicket and open dune vegetation showed a persistent association of both EUB shells and pale apex with the Open habitat. The parallel variation was due partly to strong phenotypic disequilibrium, as the combination of EUB with dark apex was rare. Nevertheless, in fully banded shells the frequency of pale apex was also higher in the Open habitat, confirming independent, parallel associations of the two contributors to paleness. Within the Acacia habitat, temporal variation of the frequencies of banding morphs was much greater than for apex colour, and EUB shells were associated with hotter summers. Consistent with its primary effect only on the very small snails, apex colour did not vary with summer conditions, but instead, higher frequencies of pale apices were associated with sunnier winters. The intensity of selection was lower on apex colour than shell banding, due partly to the constraint of phenotypic disequilibrium. The shell traits in T. pisana are an example of complex responses to climatic variation, in which phenotypic disequilibrium constrains evolution of apex colour, but separate mechanisms of selection are evident.


New strategies to inhibit KEAP1 and the Cul3-based E3 ubiquitin ligases.
E3 ubiquitin ligases that direct substrate proteins to the ubiquitin-proteasome system are promising, though largely unexplored drug targets both because of their function and their remarkable specificity. CRLs [Cullin-RING (really interesting new gene) ligases] are the largest group of E3 ligases and function as modular multisubunit complexes constructed around a Cullin-family scaffold protein. The Cul3-based CRLs uniquely assemble with BTB (broad complex/tramtrack/bric-à-brac) proteins that also homodimerize and perform the role of both the Cullin adapter and the substrate-recognition component of the E3. The most prominent member is the BTB-BACK (BTB and C-terminal Kelch)-Kelch protein KEAP1 (Kelch-like ECH-associated protein 1), a master regulator of the oxidative stress response and a potential drug target for common conditions such as diabetes, Alzheimer's disease and Parkinson's disease. Structural characterization of BTB-Cul3 complexes has revealed a number of critical assembly mechanisms, including the binding of an N-terminal Cullin extension to a bihelical '3-box' at the C-terminus of the BTB domain. Improved understanding of the structure of these complexes should contribute significantly to the effort to develop novel therapeutics targeted to CRL3-regulated pathways.


Structural basis for enantiomer binding and separation of a common beta-blocker: crystal structure of cellobiohydrolase Cel7A with bound (S)-propranolol at 1.9 A resolution.
Cellobiohydrolase Cel7A (previously called CBH 1), the major cellulase produced by the mould fungus Trichoderma reesei, has been successfully exploited as a chiral selector for separation of stereo-isomers of some important pharmaceutical compounds, e.g. adrenergic beta-blockers. Previous investigations, including experiments with catalytically deficient mutants of Cel7A, point unanimously to the active site as being responsible for discrimination of enantiomers. In this work the structural basis for enantioselectivity of basic drugs by Cel7A has been studied by X-ray crystallography. The catalytic domain of Cel7A was co-crystallised with the (S)-enantiomer of a common beta-blocker, propranolol, at pH 7, and the structure of the complex was determined and refined at 1. 9 A resolution. Indeed, (S)-propranolol binds at the active site, in glucosyl-binding subsites -1/+1. The catalytic residues Glu212 and Glu217 make tight salt links with the secondary amino group of (S)-propranolol. The oxygen atom attached to the chiral centre of (S)-propranolol forms hydrogen bonds to the nucleophile Glu212 O(epsilon1) and to Gln175 N(epsilon2), whereas the aromatic naphthyl moiety stacks with the indole ring of Trp376 in site +1. The bidentate charge interaction with the catalytic glutamate residues is apparently crucial, since no enantioselectivity has been obtained with the catalytically deficient mutants E212Q and E217Q. Activity inhibition experiments with wild-type Cel7A were performed in conditions close to those used for crystallisation. Competitive inhibition constants for (R)- and (S)-propranolol were determined at 220 microM and 44 microM, respectively, corresponding to binding free energies of 20 kJ/mol and 24 kJ/mol, respectively. The K(i) value for (R)-propranolol was 57-fold lower than the highest concentration, 12.5 mM, used in co-crystallisation experiments. Still several attempts to obtain a complex with the (R)-enantiomer have failed. By using cellobiose as a selective competing ligand, the retention of the enantiomers of propranolol on the chiral stationary phase (CSP) based on Cel7A mutant D214N were resolved into enantioselective and non- selective binding. The enantioselective binding was weaker for both enantiomers on D214N-CSP than on wild-type-CSP.


What is the best dose of nature and green exercise for improving mental health? A multi-study analysis.
Green exercise is activity in the presence of nature. Evidence shows it leads to positive short and long-term health outcomes. This multistudy analysis assessed the best regime of dose(s) of acute exposure to green exercise required to improve self-esteem and mood (indicators of mental health). The research used meta-analysis methodology to analyze 10 UK studies involving 1252 participants. Outcomes were identified through a priori subgroup analyses, and dose-responses were assessed for exercise intensity and exposure duration. Other subgroup analyses included gender, age group, starting health status, and type of habitat. The overall effect size for improved self-esteem was d = 0.46 (CI 0.34-0.59, p < 0.00001) and for mood d = 0.54 (CI 0.38-0.69, p < 0.00001). Dose responses for both intensity and duration showed large benefits from short engagements in green exercise, and then diminishing but still positive returns. Every green environment improved both self-esteem and mood; the presence of water generated greater effects. Both men and women had similar improvements in self-esteem after green exercise, though men showed a difference for mood. Age groups: for self-esteem, the greatest change was in the youngest, with diminishing effects with age; for mood, the least change was in the young and old. The mentally ill had one of the greatest self-esteem improvements. This study confirms that the environment provides an important health service.


Radiofrequency ablation of primary and metastatic hepatic malignancies.
The majority of patients with primary of metastatic hepatic tumors are not candidates for resection, because of tumor size, location near major intrahepatic blood vessels precluding a margin-negative resection, multifocality, or inadequate hepatic function related to coexistent cirrhosis. Radiofrequency ablation (RFA) is an evolving technology being used to treat patients with unresectable primary and metastatic hepatic cancers. RFA produces coagulative necrosis of the tumor through local tissue heating. Liver tumors are treated percutaneously, laparoscopically, or during laparotomy, using ultrasonography to identify tumors and to guide placement of the RFA needle electrode. For tumors smaller than 2.0 cm in diameter, one or two deployments of the monopolar multiple-array needle electrode are sufficient to produce complete coagulative necrosis of the tumor. However, with increasing size of the tumor, there is a concomitant increase in the number of deployments of the needle electrode and the overall time necessary to produce complete coagulative necrosis of the tumor. In general, RFA is a safe, well-tolerated, effective treatment for unresectable hepatic malignancies less than 6.0 cm in diameter. Effective treatment of larger tumors awaits the development of more powerful, larger array monopolar and bipolar RFA technologies.


Cytotoxicity of selenium nanoparticles in rat dermal fibroblasts.
Ventilator-associated pneumonia is a deadly nosocomial infection caused by contaminated endotracheal tubes. It has been shown that polyvinyl chloride (PVC, the endotracheal tube substrate) coated with elemental selenium nanoparticles reduces bacterial adherence and proliferation on PVC by over 99%. However, it is not known if selenium nanoparticles elicit a cytotoxic effect in vitro. The purpose of this study was to investigate the cytotoxic effects of PVC coated with selenium nanoparticles on fibroblasts, which are mammalian cells central to endotracheal tube intubation. Different concentrations of selenium nanoparticles were precipitated onto the PVC surface by reduction of selenium salts using glutathione. Characterization of PVC coated with selenium nanoparticles was done by scanning electron microscopy, energy dispersive x-ray, and contact angle measurements. For the cytotoxicity experiments, fibroblasts were seeded at a density of 5000 cm(2) onto PVC coated with three different concentrations of selenium nanoparticles (high, medium, low) and incubated for 4 hours (adhesion) as well as for 24 hours and 72 hours (proliferation). The half-maximal inhibitory concentration (IC(50)) value was determined after 72 hours using an ultrahigh concentration. MTT assays were used to assess cell viability at the indicated time points. The three concentrations of selenium nanoparticles did not elicit a cytotoxic effect after 72 hours (P < 0.01, n = 3). It was found that the IC(50)value was at the ultrahigh concentration of selenium nanoparticles. The nanoparticulate elemental selenium concentration previously shown to decrease the function of bacteria was shown not to cause a cytotoxic effect on fibroblasts in vitro. These findings demonstrate great selectivity between bacteria and healthy cells, and are a viable option for coating endotracheal tubes in order to prevent ventilator-associated pneumonia.


Mindfulness-based stress reduction, mindfulness-based cognitive therapy, and Zen meditation for depression, anxiety, pain, and psychological distress.
Mindfulness has been described as a practice of learning to focus attention on moment-bymoment experience with an attitude of curiosity, openness, and acceptance. Mindfulness practices have become increasingly popular as complementary therapeutic strategies for a variety of medical and psychiatric conditions. This paper provides an overview of three mindfulness interventions that have demonstrated effectiveness for psychiatric symptoms and/or pain. The goal of this review is to provide a synopsis that practicing clinicians can use as a clinical reference concerning Zen meditation, mindfulness-based stress reduction (MBSR), and mindfulness-based cognitive therapy (MBCT). All three approaches originated from Buddhist spiritual practices, but only Zen is an actual Buddhist tradition. MBSR and MBCT are secular, clinically based methods that employ manuals and standardized techniques. Studies indicate that MBSR and MBCT have broad-spectrum antidepressant and antianxiety effects and decrease general psychological distress. MBCT is strongly recommended as an adjunctive treatment for unipolar depression. The evidence suggests that both MBSR and MBCT have efficacy as adjunctive interventions for anxiety symptoms. MBSR is beneficial for general psychological health and stress management in those with medical and psychiatric illness as well as in healthy individuals. Finally, MBSR and Zen meditation have a role in pain management.


Large-scale application of highly-diluted bacteria for Leptospirosis epidemic control.
Leptospirosis is a zoonotic disease of major importance in the tropics where the incidence peaks in rainy seasons. Natural disasters represent a big challenge to Leptospirosis prevention strategies especially in endemic regions. Vaccination is an effective option but of reduced effectiveness in emergency situations. Homeoprophylactic interventions might help to control epidemics by using highly-diluted pathogens to induce protection in a short time scale. We report the results of a very large-scale homeoprophylaxis (HP) intervention against Leptospirosis in a dangerous epidemic situation in three provinces of Cuba in 2007. Forecast models were used to estimate possible trends of disease incidence. A homeoprophylactic formulation was prepared from dilutions of four circulating strains of Leptospirosis. This formulation was administered orally to 2.3 million persons at high risk in an epidemic in a region affected by natural disasters. The data from surveillance were used to measure the impact of the intervention by comparing with historical trends and non-intervention regions. After the homeoprophylactic intervention a significant decrease of the disease incidence was observed in the intervention regions. No such modifications were observed in non-intervention regions. In the intervention region the incidence of Leptospirosis fell below the historic median. This observation was independent of rainfall. The homeoprophylactic approach was associated with a large reduction of disease incidence and control of the epidemic. The results suggest the use of HP as a feasible tool for epidemic control, further research is warranted.


Startle responses elicited by whiplash perturbations.
The human startle response produces muscle contractions throughout the body but the most brisk and synchronized contractions appear in the neck muscles. This response, which is greatest with the first exposure to a startling stimulus, could produce excessive and inappropriately directed muscle contractions that could explain the higher incidence of whiplash injuries in people who are unprepared for the collision. This study seeks neurophysiological evidence of startle responses in the neck muscles of 120 healthy subjects exposed to between 1 and 16 rear-end impacts or forward perturbations of different speeds. Startle responses were quantified by the synchronous electromyographic (EMG) activity between 10 and 20 Hz in bilaterally homologous sternocleidomastoid, scalene and cervical paraspinal neck muscles. Coherence analyses of EMGs from the left and right muscles were used to estimate synchrony for: (i) the first unexpected trial, (ii) subsequent habituated trials, and (iii) the superposition of habituated trials and a loud acoustic stimulus (40 ms, 124 dB sound). The peak in coherent EMG activity between contralateral muscle pairs in the 10-20 Hz bandwidth was related to startle. Synchrony in this bandwidth was observed between the left and right muscles during the first impact or whiplash-like perturbation. This synchrony decreased significantly in the habituated trials, but reappeared when the loud acoustic stimulus was introduced. Its presence in the first trial indicates that startle is part of the neuromuscular response to an unexpected rear-end impact. This startle component of the neuromuscular response could play a role in the aetiology of whiplash injuries.


Poly(ADP-ribose) glycohydrolase activity mediates post-traumatic inflammatory reaction after experimental spinal cord trauma.
The aim of the present study was to examine the role of poly-(ADP-ribose) glycohydrolase (PARG) on the modulation of the inflammatory response and tissue injury associated with neurotrauma. Spinal cord trauma was induced in wild-type (WT) mice by the application of vascular clips (force of 24 g) to the dura via a two-level T(6) to T(7) laminectomy. Spinal cord injury in WT mice resulted in severe trauma characterized by edema, neutrophil infiltration, and cytokine production followed by recruitment of other inflammatory cells, production of a range of inflammation mediators, tissue damage, apoptosis, and disease. The genetic disruption of the PARG gene in mice or the pharmacological inhibition of PARG with GPI 16552 [N-bis-(3-phenyl-propyl)9-oxo-fluorene-2,7-diamide] (40 mg/kg i.p. bolus), a novel and potent PARG inhibitor, significantly reduced the degree of spinal cord inflammation and tissue injury (histological score), neutrophil infiltration, cytokine production (tumor necrosis factor-alpha and interleukin-1beta), and apoptosis. In a separate experiment, we have clearly demonstrated that PARG inhibition significantly ameliorated the recovery of limb function. Taken together, our results indicate that PARG activity modulates the inflammatory response and tissue injury events associated with spinal cord trauma and participate in target organ damage under these conditions.


RECK-mediated inhibition of glioma migration and invasion.
RECK is an anti-tumoral gene whose activity has been associated with its inhibitory effects regulating MMP-2, MMP-9, and MT1-MMP. RECK level decreases as gliobastoma progresses, varying from less invasive grade II gliomas to very invasive human glioblastoma multiforme (GBM). Since RECK expression and glioma invasiveness show an inverse correlation, the aim of the present study is to investigate whether RECK expression would inhibit glioma invasive behavior. We conducted this study to explore forced RECK expression in the highly invasive T98G human GBM cell line. Expression levels as well as protein levels of RECK, MMP-2, MMP-9, and MT1-MMP were assessed by qPCR and immunoblotting in T98G/RECK+ cells. The invasion and migration capacity of RECK+ cells was inhibited in transwell and wound assays. Dramatic cytoskeleton modifications were observed in the T98G/RECK+ cells, when compared to control cells, such as the abundance of stress fibers (contractile actin-myosin II bundles) and alteration of lamellipodia. T98G/RECK+ cells also displayed phosphorylated focal adhesion kinase (P-FAK) in mature focal adhesions associated with stress fibers; whereas P-FAK in control cells was mostly associated with immature focal complexes. Interestingly, the RECK protein was predominantly localized at the leading edge of migrating cells, associated with membrane ruffles. Unexpectedly, introduced expression of RECK effectively inhibited the invasive process through rearrangement of actin filaments, promoting a decrease in migratory ability. This work has associated RECK tumor-suppressing activity with the inhibition of motility and invasion in this GBM model, which are two glioma characteristics responsible for the inefficiency of current available treatments.


Red and far-red light effects on a short-term behavioral response of a dinoflagellate.
Cessation of movement (stop response) is used as a criterion for light reception by the dinoflagellate Gyrodinium dorsum Kofoid. Brief irradiation (2 seconds at 470 nanometers) elicits a stop response in cells any time during the 6-minute interval after removal from growth lights. This stop response is inactivated by exposure for 4 minutes to 470-nanometer light prior to stimulation. Red light (620 nanometers) reactivates this stop response, and far-red light (700 nanometers) reverses this reactivation. This red-far-red photo reversibility is taken as evidence for phytochrome involvement.


GeneNotes--a novel information management software for biologists.
Collecting and managing information is a challenging task in a genome-wide profiling research project. Most databases and online computational tools require a direct human involvement. Information and computational results are presented in various multimedia formats (e.g., text, image, PDF, word files, etc.), many of which cannot be automatically processed by computers in biologically meaningful ways. In addition, the quality of computational results is far from perfect and requires nontrivial manual examination. The timely selection, integration and interpretation of heterogeneous biological information still heavily rely on the sensibility of biologists. Biologists often feel overwhelmed by the huge amount of and the great diversity of distributed heterogeneous biological information. We developed an information management application called GeneNotes. GeneNotes is the first application that allows users to collect and manage multimedia biological information about genes/ESTs. GeneNotes provides an integrated environment for users to surf the Internet, collect notes for genes/ESTs, and retrieve notes. GeneNotes is supported by a server that integrates gene annotations from many major databases (e.g., HGNC, MGI, etc.). GeneNotes uses the integrated gene annotations to (a) identify genes given various types of gene IDs (e.g., RefSeq ID, GenBank ID, etc.), and (b) provide quick views of genes. GeneNotes is free for academic usage. The program and the tutorials are available at: http://bayes.fas.harvard.edu/genenotes/. GeneNotes provides a novel human-computer interface to assist researchers to collect and manage biological information. It also provides a platform for studying how users behave when they manipulate biological information. The results of such study can lead to innovation of more intelligent human-computer interfaces that greatly shorten the cycle of biology research.


Predictors of outcome in patients with tuberculous meningitis.
To assess predictors of mortality and neurological sequelae in patients with tuberculous meningitis (TBM). Patients with TBM treated at 12 university hospitals in Turkey between 1985 and 1997 were evaluated using a standardised protocol applied retrospectively. Variables associated with hospital mortality as well as with the presence of neurological sequelae at 6 months were determined using logistic regression models. Four hundred and thirty-four patients between the ages of 13 and 83 years (mean 33 years) were evaluated. Sixty-eight per cent of these patients presented with Medical Research Council Stage II or III. One hundred and one patients (23.3%) died and 67 (27%) of evaluable survivors had neurological sequelae. In multi-variable analysis, convulsion (OR 3.3, 95%CI 1.2-9.0, P = 0.02), comatose mental status (OR 6.0, 95%CI 3.6-10.2, P = 0.01), and delayed or interrupted treatment (OR 5.1, 95%CI 2.4-11.2, P = 0.01) were shown to be predictors for mortality. The presence of extra-meningeal tuberculosis (OR 2.1, 95%CI 1.1-4.2, P = 0.035), cranial nerve palsy (OR 2.6, 95%CI 1.4-4.2, P = 0.01), hemiparesia/focal weakness (OR 9.3, 95%CI 3.8-22.6, P = 0.01), hemiplegia/multiple neurological deficit (OR 7.1, 95%CI 2.14-23.38, P = 0.01) and drowsiness (OR 4.2, 95%CI 2.04-8.82, P = 0.01) were independent predictors of neurological sequelae at 6 months following hospital discharge. The results of this study emphasise the importance of prompt and uninterrupted anti-tuberculosis therapy for tuberculous meningitis. The presence of seizures or coma on admission to hospital are important predictors for mortality, while the presence of focal neurological signs is a predictor for persistent neurological sequelae in survivors.


Sequential binding of agonists to the beta2 adrenoceptor. Kinetic evidence for intermediate conformational states.
The beta2 adrenoreceptor (beta2AR) is a prototypical G protein-coupled receptor (GPCR) activated by catecholamines. Agonist activation of GPCRs leads to sequential interactions with heterotrimeric G proteins, which activate cellular signaling cascades, and with GPCR kinases and arrestins, which attenuate GPCR-mediated signaling. We used fluorescence spectroscopy to monitor catecholamine-induced conformational changes in purified beta2AR. Here we show that upon catecholamine binding, beta2ARs undergo transitions to two kinetically distinguishable conformational states. Using a panel of chemically related catechol derivatives, we identified the specific chemical groups on the agonist responsible for the rapid and slow conformational changes in the receptor. The conformational changes observed in our biophysical assay were correlated with biologic responses in cellular assays. Dopamine, which induces only a rapid conformational change, is efficient at activating Gs but not receptor internalization. In contrast, norepinephrine and epinephrine, which induce both rapid and slow conformational changes, are efficient at activating Gs and receptor internalization. These results support a mechanistic model for GPCR activation where contacts between the receptor and structural determinants of the agonist stabilize a succession of conformational states with distinct cellular functions.


The face signature of fibrodysplasia ossificans progressiva.
Fibrodysplasia ossificans progressiva (FOP) causes extensive heterotopic bone formation due to heterozygous mutations in the glycine-serine activation domain of ACVR1 (ALK2), a bone morphogenetic protein type I receptor. Anecdotal observations of facial similarity have been made by clinicians and parents, but no objective quantitative analysis of the faces of FOP patients has ever been undertaken. We delineated the common facial characteristics of 55 individuals with molecularly confirmed FOP by analyzing their face signature (face shape difference normalized against age and sex matched controls) and associated face signature graphs (with face signatures as vertices and adjacency corresponding to greatest similarity). Our analysis identified 10 affected individuals whose face signature is more homogeneous than others with FOP. This distinct subgroup showed the previously identified reduced mandible as well as newly identified features: underdevelopment of the upper orbit/supra-orbital ridge; infra-orbital prominence; and, low-set ears. These findings strongly suggest that the canonical FOP mutation variably affects the postnatal morphogenesis of the normotopic cranial skeleton in the upper midface and mandible and may have important diagnostic and functional implications.