Quality of life impact of childhood skin conditions measured using the Children's Dermatology Life Quality Index (CDLQI): a meta-analysis.
The Children's Dermatology Life Quality Index (CDLQI) is the most widely used instrument for measuring the impact of skin disease on quality of life (QoL) in children. To provide a meta-analysis of all published QoL scores for a range of childhood skin conditions. Studies using the CDLQI questionnaire to measure QoL in skin conditions were identified by searching Medline and Embase from January 1995 (CDLQI creation) to December 2014. Studies were grouped according to condition and baseline scores were combined using meta-analysis. Sixty-seven studies using the CDLQI met the inclusion criteria. The overall estimated CDLQI scores for conditions reported more than once were [point estimate (95% confidence interval, CI), number of studies (n), score range]: atopic eczema [8·5 (7·1-9·8), n = 38, 0-29], acne [5·3 (1·9-8·5), n = 5, 0-30], alopecia [3·1 (0-7·7), n = 2, 0-6], molluscum contagiosum [3·5 (0·6-6·7), n = 5, 0-27], psoriasis [8·0 (3·9-12·1), n = 6, 0-29], scabies [9·2 (0·0-20·3), n = 2, 1-26], urticaria [7·1 (0-15·4), n = 2, 0-22], vitiligo [6·5 (0·7-12·2), n = 2, 0-20] and warts [2·9 (0-5·8), n = 4, 0-16]. Overall, the mean effect on QoL [weighted average CDLQI score 4·6 (95% CI 3·9-5·4)] for children with these conditions was small. However, many children were found to experience a very large impact on QoL (34% of children with atopic eczema, 10% with molluscum contagiosum and 1-5% with acne) in studies where the distributions of scores were provided. Most skin conditions in children have a 'small' mean effect on QoL. However, the range is large and a significant proportion of children with many common skin conditions will experience a very large effect on quality of life.


Clarity and diagnostic quality of digitized conventional intraoral radiographs.
To compare digital images of conventional radiographs with the original radiographs for perceived clarity of endodontic files, periapical lesions and carious lesions, and to establish the diagnostic value of the digital images. Four groups of ten radiographs were used: standardized bitewings demonstrating carious lesions, periapical radiographs of apical lesions, periapical radiographs showing endodontic files of various sizes at working length, and standardized periapical radiographs with size 08 files at working length. Radiographs were photographed using an Olympus C 2500-L digital camera and a Nikon D1X digital camera and were scanned using a Nikon Supercoolscan 4000 ED film/slide scanner. The digital images were then transferred to a Toshiba Satellite 2210 laptop. Three general dental practitioners compared each conventional radiograph with the three matching digital images. Images were ranked for clarity and were assessed for diagnostic quality. Data were analysed using General Estimating Equations. The clarity and diagnostic quality of the conventional radiographs were superior to the digital images produced by the three techniques (P < 0.001). No significant difference was found between the Nikon D1X and Camedia 2500-L cameras for clarity or diagnostic quality. The scanner was equivalent to the Camedia 2500-L camera for diagnostic quality, but was otherwise inferior to both cameras. Digitizing conventional radiographs using current high-grade digital cameras or scanners does not produce images of diagnostic quality. Improved resolution of viewing monitors is necessary to fully harness the potential of digital technology.


Highly multiparametric analysis by mass cytometry.
This review paper describes a new technology, mass cytometry, that addresses applications typically run by flow cytometer analyzers, but extends the capability to highly multiparametric analysis. The detection technology is based on atomic mass spectrometry. It offers quantitation, specificity and dynamic range of mass spectrometry in a format that is familiar to flow cytometry practitioners. The mass cytometer does not require compensation, allowing the application of statistical techniques; this has been impossible given the constraints of fluorescence noise with traditional cytometry instruments. Instead of "colors" the mass cytometer "reads" the stable isotope tags attached to antibodies using metal-chelating labeling reagents. Because there are many available stable isotopes, and the mass spectrometer provides exquisite resolution between detection channels, many parameters can be measured as easily as one. For example, in a single tube the technique allows for the ready detection and characterization of the major cell subsets in blood or bone marrow. Here we describe mass cytometric immunophenotyping of human leukemia cell lines and leukemia patient samples, differential cell analysis of normal peripheral and umbilical cord blood; intracellular protein identification and metal-encoded bead arrays.


Food consumption patterns in elementary school children.
Food consumption was investigated in children attending three elementary schools in urban Hamilton, Ontario. Dietary data were collected from 92 children in grades 2 to 4 through 24-hour dietary recalls (39% participation rate). Servings of four food groups were compared with recommended daily servings in Canada's Food Guide. The majority of students did not consume the recommended five daily servings of vegetables and fruit. On average, they consumed a high number of servings of "other foods," which were not included in the four food groups. More than 50% of the students did not consume the recommended daily servings of milk products, and only a small proportion (21%) drank milk during school lunch. We recommend that primary school educators promote the consumption of vegetables and fruits and milk products at school, either through healthy snack programs or educational programs.


Slimming World in Stop Smoking Services (SWISSS): study protocol for a randomized controlled trial.
Quitting smokers gain weight. This deters some from trying to stop smoking and may explain the increased incidence of type 2 diabetes after cessation. Dieting when stopping smoking may be counterproductive. Hunger increases cravings for smoking and tackling two behaviours together may undermine quitting success. A meta-analysis of randomized controlled trials (RCTs) showed individualized dietary support may prevent weight gain, although there is insufficient evidence whether it undermines smoking cessation. Commercial weight management providers (CWMPs), such as Slimming World, provide individualized dietary support for National Health Service (NHS) patients; however, there is no evidence that they can prevent cessation-related weight gain.Our objective is to determine whether attending Slimming World from quit date, through referral from NHS Stop Smoking Services, is more effective than usual care at preventing cessation-related weight gain. This RCT will examine the effectiveness of usual cessation support plus referral to Slimming World compared to usual cessation support alone. Healthy weight, overweight and obese adult smokers attending Stop Smoking Services will be included. The primary outcome is weight change in quitters 12 weeks post-randomization. Multivariable linear regression analysis will compare weight change between trial arms and adjust for known predictors of cessation-related weight gain.We will recruit 320 participants, with 160 participants in each arm. An alpha error rate of 5% and 90% power will detect a 2 kg (SD = 2.5) difference in weight gain at 12 weeks, assuming 20% remain abstinent by then. This trial will establish whether referral to the 12-week Slimming World programme plus usual care is an effective intervention to prevent cessation-related weight gain. If so, we will seek to establish whether weight control comes at the expense of a successful quit attempt in a further non-inferiority trial.Positive results from both these trials would provide a potential solution to cessation-related weight gain, which could be rolled out across England within Stop Smoking Services to better meet the needs of 0.75 million smokers stopping with NHS support every year. Current Controlled Trials ISRCTN65705512.


Does the ascending cholinergic projection inhibit or excite neurons in the rat thalamic reticular nucleus?
In rats anesthetized with urethan, neuronal activity was recorded in those portions of the thalamic reticular nucleus (TR) excitable by visual, somatosensory, or auditory input. Observations were made on changes in rate and pattern of discharge of these neurons during repetitive stimulation of the laterodorsal tegmental nucleus (LDT), which is composed of cholinergic neurons projecting to the thalamus. In general, TR neurons showed spontaneous activity consisting of sporadic bursts of several spikes and responded to sensory stimulation with bursts of spikes which repeated several times. Weak LDT stimulation depressed or eliminated the occurrence of both spontaneous and evoked burst discharges. When LDT stimulation was sufficiently strong, however, the majority of TR neurons resumed their tonic discharges. In some animals the cortical EEG was recorded simultaneously with unit recording in TR. Suppression of burst discharges in TR was obtained even with LDT stimulation weaker than the threshold for desynchronizing the EEG. The induction of tonic discharge, on the other hand, required stimulation strong enough to produce desynchronization. The effects of LDT stimulation, such as the suppression of bursts and the induction of tonic discharge, were mimicked by acetylcholine and were antagonized by scopolamine, both drugs being applied ionophoretically. Cooling of the visual cortex abolished LDT-induced tonic discharges of visual TR neurons. A recent report and our preliminary observation show that, when the resting potential is relatively hyperpolarized, TR neurons generated a burst of spikes superposed on a low-threshold broad spike, which is inactivated and replaced with tonic firing by depolarization. Supported by these facts, the present results suggest that cholinergic input depolarizes TR neurons directly and that further depolarization occurs indirectly via activated cortex when the LDT stimulation is sufficiently strong to desynchronize EEG.


Gridded global datasets for Gross Domestic Product and Human Development Index over 1990-2015.
An increasing amount of high-resolution global spatial data are available, and used for various assessments. However, key economic and human development indicators are still mainly provided only at national level, and downscaled by users for gridded spatial analyses. Instead, it would be beneficial to adopt data for sub-national administrative units where available, supplemented by national data where necessary. To this end, we present gap-filled multiannual datasets in gridded form for Gross Domestic Product (GDP) and Human Development Index (HDI). To provide a consistent product over time and space, the sub-national data were only used indirectly, scaling the reported national value and thus, remaining representative of the official statistics. This resulted in annual gridded datasets for GDP per capita (PPP), total GDP (PPP), and HDI, for the whole world at 5 arc-min resolution for the 25-year period of 1990-2015. Additionally, total GDP (PPP) is provided with 30 arc-sec resolution for three time steps (1990, 2000, 2015).


LEAD STUDIES : III. THE EFFECTS OF LEAD ON RED BLOOD CELLS. PART 1. CHANGES IN HEMOLYSIS.
It appears, from the investigations in other laboratories, that the anemia observed in cases of lead poisoning is due to destruction of blood rather than to diminished production of blood. The method of poisoning cells in vitro with lead was adopted in order to study this phenomenon, and distinct effects were thereby obtained, even when only 0.001 mg. of lead is added to approximately 5 billion washed red corpuscles. In order to obtain optimum results the usual dosage employed was ten times this or 0.01 mg. per 5 billion cells. The following changes were observed in cells so treated. 1. Such a marked increase in the resistance to hypotonic salt solution develops that complete hemolysis does not occur until the cells are exposed to a saline solution of 0.05 per cent. Untreated cells are completely hemolyzed in 0.25 or 0.225 per cent saline. 2. This reaction is quantitative and varies with the concentration of lead used. Under the conditions of our experiments this phenomenon seems to be unique. The effects of arsenic are very slight in comparison. 3. While from this reaction it may be concluded that lead increases cellular resistance, it also appears that it shortens the life of blood cells. This may be demonstrated by the much more rapid appearance of hemolysis than normal when the cells are merely allowed to stand in Ringer solution of any dilution. 4. In rabbits with acute lead poisoning these same phenomena may be noted in vivo. 5. Both phenomena may be changed in vitro by varying the time and temperature of the reaction and the concentration of lead, as Fici has already pointed out. 6. If normal cells stand in Ringer solution for 6 hours something diffuses into the solution which largely reduces the action of lead. After repeated washing these cells react with lead in the usual manner. 7. Small amounts of serum react with lead and eliminate its effects. Red blood cells, treated with a mixture of lead and blood serum, show normal hemolysis in hypotonic salt solution. 8. When lead is added to whole blood the only evidence of this neutralization is a marked decrease in the intensity of the reaction. This is probably explained by the fact that since both serum and cells are present together, the lead reacts with both simultaneously. This in turn explains how the reaction may occur in vivo. 9. The change in hemolysis does not appear in the blood of all species of animals. In the species in which it does occur, anemia and stippling of the red cells also tend to develop readily during lead intoxication. 10. In vitro, cells which have been exposed to lead are far more fragile than normal blood cells. Slight trauma causes them to hemolyze. In vivo this is probably an important causal factor in the increased destruction of blood and in the anemia of acute lead poisoning. 11. The serum of a rabbit which has been immunized to human blood, causes more rapid and complete hemolysis in cells treated with lead than in the control cells. No satisfactory explanation for this has been found.


Measurement of serum C-reactive protein concentration after renal transplantation.
A prospective study of serial serum C-reactive protein (CRP) concentrations was made on 21 patients who had received renal allografts. CRP was raised during 27 of 32 rejection episodes and in all of five episodes of rejection associated with infection. CRP values were persistently elevated in three irreversible rejection episodes. Significantly raised CRP concentrations were documented in 14 of 20 episodes of infection. In some cases CRP was a predictive indicator of rejection or infection. In all cases of infection or rejection associated with a rise in CRP, CRP values fell following successful treatment with appropriate agents. Serial CRP measurement used in conjunction with other clinical and biochemical parameters appears to be valuable clinically following renal transplantation.


Graphene mechanical oscillators with tunable frequency.
Oscillators, which produce continuous periodic signals from direct current power, are central to modern communications systems, with versatile applications including timing references and frequency modulators. However, conventional oscillators typically consist of macroscopic mechanical resonators such as quartz crystals, which require excessive off-chip space. Here, we report oscillators built on micrometre-size, atomically thin graphene nanomechanical resonators, whose frequencies can be electrostatically tuned by as much as 14%. Self-sustaining mechanical motion is generated and transduced at room temperature in these oscillators using simple electrical circuitry. The prototype graphene voltage-controlled oscillators exhibit frequency stability and a modulation bandwidth sufficient for the modulation of radiofrequency carrier signals. As a demonstration, we use a graphene oscillator as the active element for frequency-modulated signal generation and achieve efficient audio signal transmission.


Comparison of transverse and vertical skin incision for emergency cesarean delivery.
To compare incision-to-delivery intervals and related maternal and neonatal outcomes by skin incision in primary and repeat emergent cesarean deliveries. From 1999 to 2000, a prospective cohort study of all cesarean deliveries was conducted at 13 hospitals comprising the Eunice Kennedy Shriver National Institute of Child Health and Human Development's Maternal-Fetal Medicine Units Network. This secondary analysis was limited to emergent procedures, defined as those performed for cord prolapse, abruption, placenta previa with hemorrhage, nonreassuring fetal heart rate tracing, or uterine rupture. Incision-to-delivery intervals, incision-to-closure intervals, and maternal outcomes were compared by skin-incision type (transverse compared with vertical) after stratifying for primary compared with repeat singleton cesarean delivery. Neonatal outcomes were compared by skin-incision type. Of the 37,112 live singleton cesarean deliveries, 3,525 (9.5%) were performed for emergent indications of which 2,498 (70.9%) were performed by transverse and the remaining 1,027 (29.1%) by vertical incision. Vertical skin incision shortened median incision-to-delivery intervals by 1 minute (3 compared with 4 minutes, P<.001) in primary and 2 minutes (3 compared with 5 minutes, P<.001) in repeat cesarean deliveries. Total median operative time was longer after vertical skin incision by 3 minutes in primary (46 compared with 43 minutes, P<.001) and 4 minutes in repeat cesarean deliveries (56 compared with 52 minutes, P<.001). Neonates delivered through a vertical incision were more likely to have an umbilical artery pH of less than 7.0 (10% compared with 7%, P=.02), to be intubated in the delivery room (17% compared with 13%, P=.001), or to be diagnosed with hypoxic ischemic encephalopathy (3% compared with 1%, P<.001). In emergency cesarean deliveries, neonatal delivery occurred more quickly after a vertical skin incision, but this was not associated with improved neonatal outcomes. II.


Voltage- and time-dependent block by perhexiline of K+ currents in human atrium and in cells expressing a Kv1.5-type cloned channel.
Perhexiline maleate is an antianginal drug that has been shown to have antiarrhythmic effects in humans. To examine whether some of these clinical observations could be caused by block of cardiac K+ channels, we examined the effects of perhexiline on a rapidly activating delayed rectifier K+ channel (Kv1.5) cloned from human heart and stably expressed in human embryonic kidney cells as well as a corresponding K+ current (the ultra-rapid delayed rectifier, IKur) in human atrial myocytes. With the use of inside-out macropatches, we found that perhexiline inhibited Kv1.5 current in a time- and voltage-dependent manner with an IC50 value of 1.5 x 10(-6) M at +50 mV. Perhexiline reduced Kv1.5 tail current amplitude and slowed its decay relative to control. These data are consistent with blockade of open channels, probably from the intracellular surface. Perhexiline (3 microM) also blocked IKur in human atrial myocytes. The block that was observed was both time- and voltage-dependent in qualitatively similar ways to block of Kv1.5 channels. However, the time-dependent block of IKur by perhexiline was somewhat slower and its voltage-dependence steeper relative to its effects on Kv1.5. These data indicate that perhexiline blocks both cloned and native human cardiac K+ channels. Blockade of one or more types of voltage-dependent K+ channels may explain some of the electrophysiological effects of perhexiline observed in humans.


Evaluation of Severe Acute Respiratory Syndrome Coronavirus 2 Transmission Mitigation Strategies on a University Campus Using an Agent-Based Network Model.
Universities are faced with decisions on how to resume campus activities while mitigating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) risk. To provide guidance for these decisions, we developed an agent-based network model of SARS-CoV-2 transmission to assess the potential impact of strategies to reduce outbreaks. The model incorporates important features related to risk at the University of California San Diego. We found that structural interventions for housing (singles only) and instructional changes (from in-person to hybrid with class size caps) can substantially reduce the basic reproduction number, but masking and social distancing are required to reduce this to at or below 1. Within a risk mitigation scenario, increased frequency of asymptomatic testing from monthly to twice weekly has minimal impact on average outbreak size (1.1-1.9), but substantially reduces the maximum outbreak size and cumulative number of cases. We conclude that an interdependent approach incorporating risk mitigation, viral detection, and public health intervention is required to mitigate risk.


Reinterpreting the magnetic resonance signs of hemodynamic impairment in the brains of multiple sclerosis patients from the perspective of a recent discovery of outflow block in the extracranial veins.
Multiple sclerosis patients examined with perfusion magnetic resonance (MR) imaging techniques have been found to have patterns of abnormal blood flow. These include prolonged mean transit time, a trend toward decreased cerebral blood flow in the area of plaques, and decreased cerebral blood flow and prolonged mean transit time within normal-appearing white matter. Increased cerebral blood flow and volume and decreased mean transit time (compared with the baseline values before the relapse) were found to precede the development of plaques. In addition, susceptibility-weighted imaging utilizing deoxyhemoglobin as the contrast has revealed that venous blood in cerebral veins of multiple sclerosis patients is less deoxygenated compared with healthy controls. All these findings were traditionally interpreted as a sign of local flow disturbances mediated by inflammatory and neurodegenerative processes. However, recent findings of significant stenoses in the extracranial veins that drain the brain and spinal cord shed new light on these MR results. With the assumption that a majority, if not all, of multiple sclerosis patients exhibit such extracranial venous obstacles, the perfusion MR images of multiple sclerosis patients should be reinterpreted. Perhaps ongoing MR studies with respect to extracranial venous hemodynamics may decipher some of the unsolved puzzles related to this neurologic disease.


Thresholding technique for accurate analysis of density and geometry in QCT, pQCT and microCT images.
Computed tomography (CT) is widely used in the assessment of bone parameters in live patients and animals as well as bone samples. Quantitative analysis requires the segmentation of the bone from the surrounding tissue, and most segmentation methods rely on some type of thresholding technique. The aim of this communication is to highlight the influence of threshold selection on various bone parameters and recommend appropriate thresholds. Two types of information are of interest in bone analysis from images: geometric parameters and density parameters. We know from imaging theory that blurring is an inherent byproduct of all imaging methods. Depending on the threshold used for segmentation, the object boundary moves in space due to the sloping edge. It is, thus, critical to select the threshold that creates an object boundary that reflects the actual object size. Similarly, due to blurring, the imaged density shows erroneous values at the object boundaries. Such values must not be included for an accurate representation of the object density. Using a pQCT scanner and a bone phantom with known density and geometry, we show that the thresholds for geometry and density are different. The threshold for accurate geometric segmentation was 49% of the difference of the density between the adjacent tissues. The threshold for accurate density assessment was 95% of the maximum density value of the bone. These specific thresholds are valid only for the scanner tested; however, the principle for selecting the thresholds is valid across scanner platforms and scale of imaging.


Media created violence: a social determinant of mental health.
In today's high technological world, scientific discoveries contribute remarkable development to human life, but it could also have an adverse impact on mankind. Among all these advancements, media is one of the inventions which aims at capturing a countless group of viewers and transmit information via various mediums. Media violence is considered one of the hampering determinants which harms an individual psychologically. The primary goal of a health professional is to work for the maintenance of mental health. Therefore, it is imperative to create an understanding about the impact of media violence on mental health, particularly in the Pakistani context. Violence has become a major public health problem in Pakistan. The main cause of violence seems to be anger and frustration due to poverty, political conflicts, lack of education, and the overall governance approach in the country. Therefore, there is a prime need to think and work on this neglected area like conducting research and increasing public awareness, and to curb media violence.


Therapeutic options in treatment of advanced carcinoma of the prostate.
Considerable controversy continues to surround the therapy of metastatic carcinoma of the prostate. Until recently orchiectomy and diethylstilbestrol (DES) were the only treatment options available. The development of megestrol acetate is of interest because of its broad spectrum of activity and excellent patient acceptability. Interim results of a study comparing megestrol acetate 120 mg/d plus mini-dose DES 0.1 mg/d with DES 3 mg/d are reported. Megestrol acetate had minimal side effects, with 2% of patients withdrawing from the megestrol acetate arm because of toxicity, compared with 37% from the DES arm. Significant cardiovascular toxicity occurred in 33% of patients taking DES and in 7% taking megestrol acetate. Both therapies achieved permanent suppression of serum testosterone to castrate levels. Time to progression and overall survival were longer with DES treatment, 17 versus 23 months and 24 versus 44 months, respectively, but this was not significant (P = .34 and P = .16, respectively). A review of the literature on the treatment of metastatic carcinoma of the prostate is presented to determine what should be recommended as standard therapy. Total androgen blockade is analyzed critically and results of therapy are compared with other modalities. Based on efficacy, cost, toxicity, and patient acceptability, orchiectomy still should be considered standard therapy and total androgen blockade should be considered experimental.


Effect of Honey and Green Tea Solutions on Streptococcus mutans.
The aim of this cross-sectional in vivo study was to assess the effect of green tea and honey solutions on the level of salivary Streptococcus mutans. A convenient sample of 30 Saudi boys aged 7-10 years were randomly assigned into 2 groups of 15 each. Saliva sample was collected for analysis of level of S. mutans before rinsing. Commercial honey and green tea were prepared for use and each child was asked to rinse for two minutes using 10 mL of the prepared honey or green tea solutions according to their group. Saliva samples were collected again after rinsing. The collected saliva samples were prepared and colony forming unit (CFU) of S. mutans per mL of saliva was calculated. The mean number of S. mutans before and after rinsing with honey and green tea solutions were 2.28* 10(8)(2.622*10(8)), 5.64 *10(7)(1.03*10(8)), 1.17*10(9)(2.012*10(9)) and 2.59*10(8) (3.668*10(8)) respectively. A statistically significant reduction in the average number of S. mutans at baseline and post intervention in the children who were assigned to the honey (P=0.001) and green tea (P=0.001) groups was found. A single time mouth rinsing with honey and green tea solutions for two minutes effectively reduced the number of salivary S. mutans of 7-10 years old boys.


Haemodynamic responses to dopamine and dobutamine infusions as a function of duration of infusion.
In normal animals, dopamine and dobutamine produce similar haemodynamic effects. However, clinical observations on patients in heart failure suggest that dobutamine produces a more favourable and stable haemodynamic effect. One explanation is that the inotropic action of dopamine is largely an indirect one mediated through noradrenaline release, while that of dobutamine is direct. If this were true, one would predict that the haemodynamic effects of dopamine and dobutamine would be similar over short-term infusions, but that the response to dopamine would be attenuated with time as a result of noradrenaline depletion. Experiments are reported which are consistent with this explanation. 15-min infusions of graded doses of dopamine and dobutamine produced indistinguishable haemodynamic effects in a dog model in which the haemodynamic characteristics of heart failure were produced by a chronic aorta-left atrial shunt: dopamine also produced the expected increase in mesenteric and renal flows with this short-term exposure. With long-term infusions of dopamine (5 h), the specific mesenteric/renal vasodilatation was lost with time, and indices of myocardial contractility also were attenuated with time. In contrast, the haemodynamic actions of dobutamine were well maintained. Both amines lowered total peripheral resistance and ventricular filling pressures over the entire period. These results suggest that the haemodynamic responses to dopamine change with time, and that some of the discrepancies between animal observations (short term) and those in patients (invariably long term) can be explained on this basis rather than on the basis of species differences or underlying pathology. They also suggest that dopamine is a less suitable 'inotropic' drug than dobutamine.


Prototyping of poly(dimethylsiloxane) interfaces for flow gating, reagent mixing, and tubing connection in capillary electrophoresis.
Integrated microfluidic systems coupled with electrophoretic separations have broad application in biologic and chemical analysis. Interfaces for the connection of various functional parts play a major role in the performance of a system. Here, we developed a rapid prototyping method to fabricate monolithic poly(dimethylsiloxane) (PDMS) interfaces for flow-gated injection, online reagent mixing, and tube-to-tube connection in an integrated capillary electrophoresis (CE) system. The basic idea was based on the properties of PDMS: elasticity, transparency, and suitability for prototyping. The molds for these interfaces were prepared by using commercially available stainless steel wires and nylon lines or silica capillaries. A steel wire was inserted through the diameter of a nylon line and a cross format was obtained as the mold for PDMS casting of flow gates and 4-way mixers. These interfaces accommodated tubing connection through PDMS elasticity and provided easy visual trouble shooting. The flow gate used smaller channel diameters, thus reducing flow rate by 25-fold for effective gating compared with mechanically machined counterparts. Both PDMS mixers and the tube-to-tube connectors could minimize the sample dead volume by using an appropriate capillary configuration. As a whole, the prototyped PDMS interfaces are reusable, inexpensive, convenient for connection, and robust when integrated with the CE detection system. Therefore, these interfaces could see potential applications in CE and CE-coupled systems.


Management of human immunodeficiency virus infection in advanced age.
Human immunodeficiency virus (HIV)-positive patients treated with antiretroviral therapy now have increased life expectancy and develop chronic illnesses that are often seen in older HIV-negative patients. To address emerging issues related to aging with HIV. Screening older adults for HIV, diagnosis of concomitant diseases, management of multiple comorbid medical illnesses, social isolation, polypharmacy, and factors associated with end-of-life care are reviewed. Published guidelines and consensus statements were reviewed. PubMed and PsycINFO were searched between January 2000 and February 2013. Articles not appearing in the search that were referenced by reviewed articles were also evaluated. The population of older HIV-positive patients is rapidly expanding. It is estimated that by 2015 one-half of the individuals in the United States with HIV will be older than age 50. Older HIV-infected patients are prone to having similar chronic diseases as their HIV-negative counterparts, as well as illnesses associated with co-infections. Medical treatments associated with these conditions, when added to an antiretroviral regimen, increase risk for polypharmacy. Care of aging HIV-infected patients involves a need to balance a number of concurrent comorbid medical conditions. HIV is no longer a fatal disease. Management of multiple comorbid diseases is a common feature associated with longer life expectancy in HIV-positive patients. There is a need to better understand how to optimize the care of these patients.


Primary isoniazid prophylaxis against tuberculosis in HIV-exposed children.
The dual epidemic of human immunodeficiency virus (HIV) and tuberculosis is a major cause of sickness and death in sub-Saharan Africa. We conducted a double-blind, randomized, placebo-controlled trial of preexposure isoniazid prophylaxis against tuberculosis in HIV-infected children and uninfected children exposed to HIV during the perinatal period. We randomly assigned 548 HIV-infected and 804 HIV-uninfected infants (91 to 120 days of age) to isoniazid (10 to 20 mg per kilogram of body weight per day) or matching placebo for 96 weeks. All patients received bacille Calmette-Guérin (BCG) vaccination against tuberculosis within 30 days after birth. HIV-infected children had access to antiretroviral therapy. The primary outcome measures were tuberculosis disease and death in HIV-infected children and latent tuberculosis infection, tuberculosis disease, and death in HIV-uninfected children within 96 to 108 weeks after randomization. Antiretroviral therapy was initiated in 98.9% of HIV-infected children during the study. Among HIV-infected children, protocol-defined tuberculosis or death occurred in 52 children (19.0%) in the isoniazid group and 53 (19.3%) in the placebo group (P=0.93). Among HIV-uninfected children, there was no significant difference in the combined incidence of tuberculosis infection, tuberculosis disease, or death between the isoniazid group (39 children, 10%) and the placebo group (45 children, 11%; P=0.44). The rate of tuberculosis was 121 cases per 1000 child-years (95% confidence interval [CI], 95 to 153) among HIV-infected children as compared with 41 per 1000 child-years (95% CI, 31 to 52) among HIV-uninfected children. There were no significant differences in clinical or severe laboratory toxic effects between treatment groups. Primary isoniazid prophylaxis did not improve tuberculosis-disease-free survival among HIV-infected children or tuberculosis-infection-free survival among HIV-uninfected children immunized with BCG vaccine. Despite access to antiretroviral therapy, the burden of tuberculosis remained high among HIV-infected children. (Funded by the National Institutes of Health and Secure the Future; ClinicalTrials.gov number, NCT00080119.).


Manipulation and imaging of Kryptolebias marmoratus embryos.
The self-fertilizing mangrove killifish, Kryptolebias marmoratus, is an upcoming model species for a range of biological disciplines. To further establish this model in the field of developmental biology, we examined several techniques for embryonic manipulation and for imaging that can be used in an array of experimental designs. These methodological approaches can be divided into two categories: handling of embryos with and without their chorionic membrane. Embryos still enclosed in their chorion can be manipulated using an agarose bed or a methyl cellulose system, holding them in place and allowing their rotation to more specific angles and positions. Using these methods, we demonstrate microinjection of embryos and monitoring of fluorescent yolk syncytial nuclei (YSN) using both stereo and compound microscopes. For higher magnification imaging using compound microscopes as well as time-lapse analyses, embryos were dechorionated and embedded in low-melting-point agarose. To demonstrate this embedding technique, we further examined fluorescent YSN and also analyzed the yolk surface of K. marmoratus embryos. The latter was observed to provide an excellent imaging platform for study of the behavior and morphology of cells during embryonic development, for various types of cells. Our data demonstrate that K. marmoratus is an excellent model species for research in developmental biology, as methodological approaches for the manipulation and imaging of embryos are efficient and readily available.


Mathematical analysis of a model for the growth of the bovine corpus luteum.
The corpus luteum (CL) is an ovarian tissue that grows in the wound space created by follicular rupture. It produces the progesterone needed in the uterus to maintain pregnancy. Rapid growth of the CL and progesterone transport to the uterus require angiogenesis, the creation of new blood vessels from pre-existing ones, a process which is regulated by proteins that include fibroblast growth factor 2 (FGF2). In this paper we develop a system of time-dependent ordinary differential equations to model CL growth. The dependent variables represent FGF2, endothelial cells (ECs), luteal cells, and stromal cells (like pericytes), by assuming that the CL volume is a continuum of the three cell types. We assume that if the CL volume exceeds that of the ovulated follicle, then growth is inhibited. This threshold volume partitions the system dynamics into two regimes, so that the model may be classified as a Filippov (piecewise smooth) system. We show that normal CL growth requires an appropriate balance between the growth rates of luteal and stromal cells. We investigate how angiogenesis influences CL growth by considering how the system dynamics depend on the dimensionless EC proliferation rate, ρ₅. We find that weak (low ρ₅) or strong (high ρ₅) angiogenesis leads to 'pathological' CL growth, since the loss of CL constituents compromises progesterone production or delivery. However, for intermediate values of ρ₅, normal CL growth is predicted. The implications of these results for cow fertility are also discussed. For example, inadequate angiogenesis has been linked to infertility in dairy cows.


Point mutations throughout the GLI3 gene cause Greig cephalopolysyndactyly syndrome.
Greig cephalopolysyndactyly syndrome, characterized by craniofacial and limb anomalies (GCPS; MIM 175700), previously has been demonstrated to be associated with translocations as well as point mutations affecting one allele of the zinc finger gene GLI3. In addition to GCPS, Pallister-Hall syndrome (PHS; MIM 146510) and post-axial polydactyly type A (PAP-A; MIM 174200), two other disorders of human development, are caused by GLI3 mutations. In order to gain more insight into the mutational spectrum associated with a single phenotype, we report here the extension of the GLI3 mutation analysis to 24 new GCPS cases. We report the identification of 15 novel mutations present in one of the patient's GLI3 alleles. The mutations map throughout the coding gene regions. The majority are truncating mutations (nine of 15) that engender prematurely terminated protein products mostly but not exclusively N-terminally to or within the central region encoding the DNA-binding domain. Two missense and two splicing mutations mapping within the zinc finger motifs presumably also interfere with DNA binding. The five mutations identified within the protein regions C-terminal to the zinc fingers putatively affect additional functional properties of GLI3. In cell transfection experiments using fusions of the DNA-binding domain of yeast GAL4 to different segments of GLI3, transactivating capacity was assigned to two adjacent independent domains (TA(1)and TA(2)) in the C-terminal third of GLI3. Since these are the only functional domains affected by three C-terminally truncating mutations, we postulate that GCPS may be due either to haploinsufficiency resulting from the complete loss of one gene copy or to functional haploinsufficiency related to compromised properties of this transcription factor such as DNA binding and transactivation.


Is it beneficial to selectively boost high-risk tumor subvolumes? A comparison of selectively boosting high-risk tumor subvolumes versus homogeneous dose escalation of the entire tumor based on equivalent EUD plans.
To quantify and compare expected local tumor control and expected normal tissue toxicities between selective boosting IMRT and homogeneous dose escalation IMRT for the case of prostate cancer. Four different selective boosting scenarios and three different high-risk tumor subvolume geometries were designed to compare selective boosting and homogeneous dose escalation IMRT plans delivering the same equivalent uniform dose (EUD) to the entire PTV. For each scenario, differences in tumor control probability between both boosting strategies were calculated for the high-risk tumor subvolume and remaining low-risk PTV, and were visualized using voxel based iso-TCP maps. Differences in expected rectal and bladder complications were quantified using radiobiological indices (generalized EUD (gEUD) and normal tissue complication probability (NTCP)) as well as %-volumes. For all investigated scenarios and high-risk tumor subvolume geometries, selective boosting IMRT improves expected TCP compared to homogeneous dose escalation IMRT, especially when lack of control of the high-risk tumor subvolume could be the cause for tumor recurrence. Employing, selective boosting IMRT significant increases in expected TCP can be achieved for the high-risk tumor subvolumes. The three conventional selective boosting IMRT strategies, employing physical dose objectives, did not show significant improvement in rectal and bladder sparing as compared to their counterpart homogeneous dose escalation plans. However, risk-adaptive optimization, utilizing radiobiological objective functions, resulted in reduction in NTCP for the rectum when compared to its corresponding homogeneous dose escalation plan. Selective boosting is a more effective method than homogeneous dose escalation for achieving optimal treatment outcomes. Furthermore, risk-adaptive optimization increases the therapeutic ratio as compared to conventional selective boosting IMRT.


The Failure of All Mothers or the Mother of All Failures? Juror Perceptions of Failure to Protect Laws.
Failure to protect laws have been an area of interest among legal scholars, child and domestic violence advocates, and the social work and psychology fields with varying focuses on the societal underpinnings of the laws and their application. Despite the pervasive legal literature on "mother blaming" and the revictimization of battered women that these laws engender, few studies have empirically examined if "mother blaming" occurs in these cases, or if battered women are treated more harshly. The aim of the present study was to investigate the impact of defendant sex and presence of domestic violence on mock juror decision making in a failure to protect case. The influences of juror sex on decision making were also examined. Jury eligible community members read a summary of a case in which the defendant was charged with failing to protect their child from a third-party abuser. The sex of the defendant and the presence of domestic violence were manipulated. Participants then rendered a verdict, provided sentencing recommendations, and responded to attitudinal questions about the defendant and perpetrator. Mock jurors were more likely to find the defendant guilty and view the defendant more negatively when the defendant had been the victim of domestic violence. Defendant sex did not affect outcome measures; however, female jurors were more punitive than male jurors. Results are discussed in terms of "victim blaming" and labeling theory of intimate partner violence.


Removing or truncating connexin 43 in murine osteocytes alters cortical geometry, nanoscale morphology, and tissue mechanics in the tibia.
Gap junctions are formed from ubiquitously expressed proteins called connexins that allow the transfer of small signaling molecules between adjacent cells. Gap junctions are especially important for signaling between osteocytes and other bone cell types. The most abundant type of connexin in bone is connexin 43 (Cx43). The C-terminal domain of Cx43 is thought to be an important modulator of gap junction function but the role that this domain plays in regulating tissue-level mechanics is largely unknown. We hypothesized that the lack of the C-terminal domain of Cx43 would cause morphological and compositional changes as well as differences in how bone responds to reference point indentation (RPI) and fracture toughness testing. The effects of the C-terminal domain of Cx43 in osteocytes and other cell types were assessed in a murine model (C57BL/6 background). Mice with endogenous Cx43 in their osteocytes removed via a Cre-loxP system were crossed with knock-in mice which expressed Cx43 that lacked the C-terminal domain in all cell types due to the insertion of a truncated allele to produce the four groups used in the study. The main effect of removing the C-terminal domain from osteocytic Cx43 increased cortical mineral crystallinity (p=0.036) and decreased fracture toughness (p=0.017). The main effect of the presence of the C-terminal domain in other cell types increased trabecular thickness (p<0.001), cortical thickness (p=0.008), and average RPI unloading slope (p=0.004). Collagen morphology was altered when either osteocytes lacked Cx43 (p=0.008) or some truncated Cx43 was expressed in all cell types (p<0.001) compared to controls but not when only the truncated form of Cx43 was expressed in osteocytes (p=0.641). In conclusion, the presence of the C-terminal domain of Cx43 in osteocytes and other cell types is important to maintain normal structure and mechanical integrity of bone.


Prostatic carcinoma: a Pakistani perspective.
As in many countries of the world, prostatic adenocarcinoma is extremely common in Pakistan. In a study carried out in our section, it was the third commonest malignancy in males, comprising almost 7% of all malignant neoplasms. Furthermore, another investigation revealed it to be the commonest malignant neoplasm in males in the seventh and eight decades, comprising 15% and 25% of respectively of all malignant neoplasms in these age groups. Recent data extracted from radical prostatectomy specimens in our department have revealed that the majority of carcinomas are advanced (like most other cancers in our country) when they are first diagnosed or treated. The majority of cases in which radical prostatectomy was performed showed not only advanced stage (pT3a or pT3b) but also exhibited extra-prostatic extension and positive surgical margins. For a cancer which is so common in our population, the diagnostic and therapeutic modalities available are extremely limited.


From Undernutrition to Overnutrition: The Evolution of Overweight and Obesity among Young Men in Switzerland since the 19th Century.
The global obesity epidemic continues, new approaches are needed to understand the causes. We analyzed data from an evolutionary perspective, stressing developmental plasticity. We present diachronical height, weight, and BMI data for 702,902 Swiss male conscripts aged 18-20 years, a representative, standardized and unchanged data source. From 1875 to 1879, the height distribution was slightly left-skewed; 12.1% of the conscripts were underweight, overweight and obesity were rare. The BMI-to-height relationship was positive but not linear, and very short conscripts were particularly slim. Since the 1870s, Swiss conscripts became taller, a trend that markedly slowed in the 1990s. In contrast, weight increased in two distinct steps at the end of the 1980s and again after 2002. Since 2010, BMI did not increase but stabilized at a high level. The body of young men adapted differently to varying living conditions over time: First, less investment in height and weight under conditions of undernutrition and food uncertainty; second, more investment in height under more stable nutritional conditions; third, development of obesity during conditions of plateaued height growth, overnutrition, and decreasing physical activity. This example contributes to the evaluation of hypotheses on human developmental plasticity.


A randomized, double-blind comparison of clozapine and high-dose olanzapine in treatment-resistant patients with schizophrenia.
Clozapine, despite its side-effect burden, has been considered to be the drug of choice for patients with schizophrenia whose psychotic symptoms fail to respond adequately to other anti-psychotic drugs. There are conflicting data concerning the potential utility of olanzapine in treatment-resistant schizophrenia at doses beyond the 10- to 20-mg/day range that has proven to be effective for most nonrefractory patients with schizophrenia. The main objective of this study was to compare the efficacy and tolerability of high-dose olanzapine (target dose, 25-45 mg/day) and clozapine (300-900 mg/day) in patients with schizophrenia or schizoaffective disorder who had failed to respond adequately to prior treatment with other antipsychotic drugs. This 6-month, randomized, double-blind, parallel-group study compared the efficacy and tolerability of olanzapine (mean dose, 34 mg/day; N = 19) or clozapine (mean dose, 564 mg/day; N = 21) in patients with treatment-resistant schizophrenia or schizoaffective disorder, diagnosed according to DSM-IV criteria. Outcome measures included psychopathology, cognitive performance (as assessed with a comprehensive neuropsychological test battery), and tolerability. The study was conducted between May 2000 and December 2003. Robust and significant (mostly p < .001) improvement in multiple measures of psychopathology, mainly between 6 weeks and 6 months of treatment, was found in both treatment groups, with no significant difference between the 2 treatments except for the Global Assessment of Functioning score, which favored clozapine (p = .01). Improvement in some domains of cognition was significant-and equivalent for both drugs, as well. Nonsignificantly different improvement in Verbal List Learning-Immediate Recall (p < .05), Controlled Word Association Test (p < .05), and Digit Symbol Substitution Test (p < .001) was found. There were no significant differences in extrapyramidal symptoms. Weight gain was significantly (p = .01) greater with olanzapine. Olanzapine, at higher than customary doses, demonstrated similar efficacy to clozapine in treatment-resistant schizophrenia and schizoaffective disorder in this study. However, the small sample size precludes definitively concluding that the 2 treatments are equivalent, at these doses, in treatment-resistant schizophrenia. The metabolic side effects of olanzapine are a limitation in its use. ClinicalTrials.gov identifier NCT00179231.


The impact of acutely elevated intraocular pressure on the porcine optic nerve head.
To investigate the effects of acute elevations in intraocular pressure (IOP) on the cup, prelaminar, and lamina cribrosa regions of the porcine optic nerve head (ONH). Ex vivo imaging of 10 porcine ONHs was performed using spectral-domain optical coherence tomography (OCT). The IOP was manipulated with a pressure head and measured with a pressure transducer. Reference scans were taken at 0 mm Hg, before further scanning was performed at 7-mm Hg steps, up to 49 mm Hg. Morphometric parameters were measured across centrally located OCT B-scans at different IOPs, and the relationship between IOP and changes in these parameters was analyzed. As IOP increased from 0 to 49 mm Hg, mean cross-sectional cup area increased (28% ± 3%, P < 0.001), lamina cribrosa area decreased (18% ± 2%, P < 0.001), and prelaminar tissue area decreased (5.5% ± 0.5%, P < 0.001). Multivariate regression demonstrated that most of the change in cup area is associated with changes in both lamina cribrosa position and thickness (r = 0.89, P < 0.001). Acute elevations in IOP were shown to result in posterior displacement of ONH, as well as lamina cribrosa and prelaminar tissue deformation in the porcine ONH.


Progress and problems in the biology, diagnostics, and therapeutics of prion diseases.
The term "prion" was introduced by Stanley Prusiner in 1982 to describe the atypical infectious agent that causes transmissible spongiform encephalopathies, a group of infectious neurodegenerative diseases that include scrapie in sheep, Creutzfeldt-Jakob disease in humans, chronic wasting disease in cervids, and bovine spongiform encephalopathy in cattle. Over the past twenty years, the word "prion" has been taken to signify various subtly different concepts. In this article, we refer to the prion as the transmissible principle underlying prion diseases, without necessarily implying any specific biochemical or structural identity. When Prusiner started his seminal work, the study of transmissible spongiform encephalopathies was undertaken by only a handful of scientists. Since that time, the "mad cow" crisis has put prion diseases on the agenda of both politicians and the media. Significant progress has been made in prion disease research, and many aspects of prion pathogenesis are now understood. And yet the diagnostic procedures available for prion diseases are not nearly as sensitive as they ought to be, and no therapeutic intervention has been shown to reliably affect the course of the diseases. This article reviews recent progress in the areas of pathogenesis of, diagnostics of, and therapy for prion diseases and highlights some conspicuous problems that remain to be addressed in each of these fields.


Breeding systems in Echinococcus granulosus (Cestoda; Taeniidae): selfing or outcrossing?
We used the PCR-SSCP method followed by sequencing in order to assess the genetic variability of coding and noncoding parts of the genome of Echinococcus granulosus (Cestoda; Taeniidae) and to test whether or not the parasite populations are mainly self-fertilizing. For this, we analysed a sample of 110 E. granulosus metacestode isolates collected from different geographical regions (Southern Brazil, Europe and Australia) and from different intermediate hosts (ovine, bovine, human, macropod, swine and equine). Using appropriate controls, we were able to identify 4 strains in that sample (sheep, cattle, pig and horse strains). The high degree of genetic differentiation between strains, but not within, and the monomorphism found in most loci (EgAg4, EgActII, EgHbx2 and EgAg6-non-coding-EgAgB/1 and EgND1-coding) indicated that they are largely selfed. On the other hand, outcrossing was also shown to occur, since 5 potential hybrid genotypes between cattle and sheep strains were found in populations of Southern Brazil, but absent in other geographical areas. We suggest that both processes are adaptive. The article also reports, for the first time, the occurrence of the E. granulosus cattle strain in South America.


Alternative oxidase (AOX) over-expression improves cell expansion and elongation in cotton seedling exposed to cool temperatures.
Evidence that supports a relation between AOX expression and improvement in plant height, internode length, and total leaf area under cool temperature is shown. Cell expansion and elongation appear to be enhanced when AOX expression was increased. Cotton growth is sensitive to cool temperature during germination and early seedling development. Delayed emergence, seedling damage, and increased risk to disease are common. Late seasonal cool weather is a major factor limiting the consistent production of high-quality cotton lint in West Texas. Alternative oxidase functions in the inner membrane of the mitochondria via an alternative respiration pathway and serves as a multifunctional system for amelioration of abiotic and biotic stresses. Cotton seedling emergence and growth exposed to cool temperatures was examined in plants with enhanced AOX expression. Thirteen T1 seed lines showed 3 to 1 segregation for the T-DNA containing the tobacco AOX1 gene. Two over-expressing, single-copy, homozygous AOX lines (94-20T and 66-6T) and Null line (94-3N) were selected for examination. The transcript levels were ≈ 2 to 6 fold higher in the AOX lines compared to those of the Null line and wild-type in stem, leaf, root and boll tissues. The research examined the hypothesis that transgenic cotton with enhanced AOX expression will have enhanced growth traits under suboptimal cool temperatures. Improved plant height, internode length, plant height and internode length from second node, and total leaf area under cool temperatures were observed in AOX over-expression lines. This may be attributed to improved cell expansion and elongation characteristics in the AOX line.


Evidence for agonist-induced interaction of angiotensin receptor with a guanine nucleotide-binding protein in bovine adrenal zona glomerulosa.
Previous studies have documented that high affinity binding of [125I]angiotensin II to adrenal cortex receptors was modulated by guanine nucleotides. Since in other receptor systems, similar properties of hormone-receptor interactions were shown to be specific for agonists, we studied the differential binding characteristics of agonists and antagonists to this receptor using a new radiolabeled antagonist [125I] [Sar1,Ile8] angiotensin II. Receptor saturation studies indicate that the antagonist is binding to a homogeneous population of sites with a Kd of 0.6-2.0 nM and with a receptor density around 1 pmol/mg of protein. Competition curves using unlabeled antagonists are characterized by a slope factor of 1 and a single Kd of 1-3 nM. Addition of guanylylimidodiphosphate to the assay is absolutely without effect on radiolabeled antagonist binding. In contrast, competition curves using the full agonists angiotensin II, [Sar1]angiotensin II, angiotensin III, and [des-Arg]angiotensin III display slope factors of 0.79, 0.87, 0.70, and 0.84, respectively. These curves can be explained by two apparent forms of the receptor having high and low affinity for the agonist. The higher affinity form associated with these four agonists is characterized by a Kd of 1.2 nM, 0.25 nM, 0.8 nM, and 3 microM, and corresponds to 60, 56, 42, and 25% of angiotensin II-binding sites, respectively. The other form displays 13- to 33-fold lower affinity. Addition of guanine nucleotide to the assay results in a 2-4-fold shift to the right and a steepening (slope factor 0.9-1.0) of agonist competition curves. Angiotensin II receptors, occupied by the full agonist [131I] [Sar1] angiotensin II or by the antagonist [125I] [Sar1, Ile8]angiotensin II, were then solubilized with the nonionic detergent octylglucoside. Dissociation of the agonist [131I] [Sar1] angiotensin II from solubilized receptors is enhanced by guanylylimidodiphosphate or sodium acetate, while dissociation of the antagonist [125I] [Sar1, Ile8]angiotensin II displays little sensitivity towards guanine nucleotides or increased ionic strength. Inclusion of bile salts in the solubilization medium preferentially destabilizes receptor-bound agonist, presumably by interfering with protein-protein interactions required for high affinity agonist binding. Separation of radiolabeled agonist and antagonist-occupied solubilized receptor complexes by steric exclusion high performance liquid chromatography reveals that the agonist-occupied receptor complex behaves as a larger protein than the antagonist-occupied receptor complex.(ABSTRACT TRUNCATED AT 400 WORDS)


Statistical properties of autonomous flows in 2D active nematics.
We study the dynamics of a tunable 2D active nematic liquid crystal composed of microtubules and kinesin motors confined to an oil-water interface. Kinesin motors continuously inject mechanical energy into the system through ATP hydrolysis, powering the relative microscopic sliding of adjacent microtubules, which in turn generates macroscale autonomous flows and chaotic dynamics. We use particle image velocimetry to quantify two-dimensional flows of active nematics and extract their statistical properties. In agreement with the hydrodynamic theory, we find that the vortex areas comprising the chaotic flows are exponentially distributed, which allows us to extract the characteristic system length scale. We probe the dependence of this length scale on the ATP concentration, which is the experimental knob that tunes the magnitude of the active stress. Our data suggest a possible mapping between the ATP concentration and the active stress that is based on the Michaelis-Menten kinetics that governs the motion of individual kinesin motors.


What are the costs of marital conflict and dissolution to children's physical health?
Do parental marital conflict and dissolution influence the risk trajectory of children's physical health risk? This paper reviews evidence addressing this question in the context of understanding how early environmental adversities may trigger a succession of risks that lead to poor health in childhood and greater risk for chronic health problems in adulthood. We first review existing evidence linking marital conflict and dissolution to offspring's physical health outcomes. Next, we provide evidence supporting biopsychosocial pathways that may link marital conflict and dissolution with accelerated health risk trajectories across the lifespan. Specifically, we posit that consequential to the stresses associated with marital conflict and disruption, parenting practices are compromised, leading to offspring deficits in affective, behavioral, and cognitive domains. These deficits, in turn, are hypothesized to increase health risk through poor health behaviors and by altering physiological stress-response systems, including neuroendocrine, cardiovascular, and neurotransmitter functioning. On the basis of the available direct evidence and theoretically plausible pathways, it appears that there is a cost of marital conflict and disruption to children's health; however, more comprehensive investigations are needed to further elucidate this relationship. In the final section, we address limitations in the current literature and identify research that is needed to better evaluate the association between marital conflict and dissolution and children's physical health.


Significant Serum Progesterone Variations on the Day of Final Oocyte Maturation in Stimulated IVF Cycles.
Objective: To evaluate intraday serum progesterone levels on the day of final oocyte maturation in women undergoing ovarian stimulation in a GnRH-antagonist protocol. Study design, size, and duration: The study was done as a prospective observational study at a Private IVF centre in Muscat, Oman. 30 patients were recruited from May 2018 to March 2019. Patients: Thirty patients with primary/secondary infertility and an indication for ovarian stimulation for IVF/ICSI treatment. The study was registered at the clinicaltrials.gov under the number: NCT03519776. Main outcome measures: Progesterone levels at 4 time points (8 a.m., 11 a.m., 2 p.m. and 5 p.m.) on the day of final oocyte maturation. Results: A total of 120 samples from 30 patients were included in this prospective study. Progesterone levels on the day of final oocyte maturation showed a significant decline over the day with the mean values at 8 a.m.:1.0 ng/ml, at 11 a.m.:0.8 ng/ml, at 2 a.m.: 0.7 ng/ml and at 5 p.m.:0.6 ng/ml. The difference between the first and the last progesterone level was 0.4 ng/ml, reflecting a 37.8% decline of the progesterone level within 9 h and there was a highly significant decrease in the progesterone levels recorded between 8 a.m. and 11 a.m., between 8 a.m. and 2 p.m., between 8 a.m. and 5 p.m. and 11 a.m. and 5 p.m. (p < 0.001). Conclusion: The study findings have two clinically important conclusions: Firstly, progesterone levels on the day of final oocyte maturation decline significantly from the morning to the afternoon in patients, questioning the reliability of one arbitrarily taken progesterone level regarding the decision to perform a fresh embryo transfer or to cryopreserve the embryos. Secondly, declining progesterone levels 12 h after the last administration of gonadotropins support the theory that enhanced ovarian stimulation at the end of the follicular phase leads to an overload of the capacity of the enzymes metabolizing progesterone further on, therefore resulting in elevated progesterone levels in circulation.


Prevalence of blood parasites in European passeriform birds.
Variation in the prevalence of blood parasites among species of birds has been used to test hypotheses about the effects of sexual selection and parental investment on disease resistance, and how vector abundance influences infection. However, the factors causing this variation are still poorly understood. We assessed the statistical effects of biogeographic, plumage-related and life-history traits on the prevalence of the blood parasites Plasmodium, Haemoproteus, Leucocytozoon and Trypanosoma in European passerine birds. Most of the variation in parasite prevalence occurred at low taxonomic levels. Brighter male plumage and greater host body mass were associated with higher prevalence, explaining 32% of the total variation. Male plumage brightness remained a significant factor when we controlled for phylogenetic effects. These relationships were driven primarily by simuliid-transmitted parasites (Leucocytozoon, Trypanosoma), which were more frequent in species with northern distributions. Host species with greater maximum longevity and shorter nestling periods had higher prevalences of Plasmodium; however, the effect was not stable after controlling for phylogeny using pairwise contrasts. Coevolution between hosts and parasites appears to create temporal and spatial variation that disconnects haematozoan prevalence from evolutionarily conservative life-history traits while creating some positive associations with traits that are phylogenetically labile. Clearly, ecologists should be cautious in relating patterns of variation in haematozoan prevalence to particular host traits.


Sources of total, non-milk extrinsic, and intrinsic and milk sugars in the diets of older adults living in sheltered accommodation.
The WHO recommends limiting non-milk extrinsic sugars (NMES) consumption to < or = 10 % energy to reduce the risk of unhealthy weight gain and dental caries, and to restrict frequency of intake to < or = 4 times/d to reduce risk of dental caries. Older adults, especially those from low-income backgrounds, are at increased risk of dental caries, yet there is little information on sugars intake (frequency of intake and food sources) in this age group. The aim of this report is to present baseline data from a community-based dietary intervention study of older adults from socially deprived areas of North East England, on the quantity and sources of total sugars, NMES, and intrinsic and milk sugars, and on frequency of NMES intake. Dietary intake was assessed using two 3-d estimated food diaries, completed by 201 participants (170 female, thirty-one male) aged 65-85 years (mean 76.7 (sd 5.5) years) recruited from sheltered housing schemes. Total sugars represented 19.6 %, NMES 9.3 %, and intrinsic and milk sugars 10.3 % of daily energy intake. Eighty-one (40.3 %) exceeded the NMES intake recommendation. Mean frequency of NMES intake was 3.4 times/d. The fifty-three participants (26.4 %) who exceeded the frequency recommendation ( < or = 4 times/d) obtained a significantly greater percentage of energy from NMES compared with those participants who met the recommendation. The food groups 'biscuits and cakes' (18.9 %), 'soft drinks' (13.1 %) and 'table sugar' (11.1 %) made the greatest contributions to intakes of NMES. Interventions to reduce NMES intake should focus on limiting quantity and frequency of intake of these food groups.


A coalescent framework for comparing alternative models of population structure with genetic data: evolution of Celebes toads.
Isolation of populations eventually leads to divergence by genetic drift, but if connectivity varies over time, its impact on diversification may be difficult to discern. Even when the habitat patches of multiple species overlap, differences in their demographic parameters, molecular evolution and stochastic events contribute to differences in the magnitude and distribution of their genetic variation. The Indonesian island of Sulawesi, for example, harbours a suite of endemic species whose intraspecific differentiation or interspecific divergence may have been catalysed by habitat fragmentation. To further test this hypothesis, we have performed phylogenetic and coalescent-based analyses on molecular variation in mitochondrial and nuclear DNA of the Celebes toad (Bufo celebensis). Results support a role for habitat fragmentation that led to a population structure in these toads that closely matches distributions of Sulawesi macaque monkeys. Habitat fragmentation, therefore, may also have affected other groups on this island.


SU-E-T-114: Characterization of the Spatial Response Functions of Ionization Chambers for Photon Beam Dosimetry.
The finite extension of an ionization chamber gives rise to a spatial averaging effect, known as the "volume effect". In order to provide the appropriate corrections, the response functions along its lateral and longitudinal directions are characterized using Gaussian distributions, whose standard deviations slat and slong have been determined for a large set of clinical dosimeters. Nine cylindrical ionization chambers, two parallel-plate chambers and two 2D ionization chamber arrays have been examined by scanning rectangular photon fields along their short axes. The true profiles D(x) were known from scans with a small Si diode. The ionization chambers were aligned with their symmetry axes either perpendicular or parallel to the scan direction in order to obtain slat and slong separately. In a search process, D(x) was numerically convolved with normalized one-dimensional Gaussian kernels K(x) of varying s. The best fit between the convolution product D(x) * K(x) and the measured profile M(x) of the ionization chamber was used to determine parameters slat and slong of the Gaussian kernels. For both the lateral and longitudinal directions, very good agreement was found between M(x) and the convolution products of D(x) with Gaussian kernels K(x). For all chambers, their 2s values are similar to the cavity dimensions, which means that the "tails" of the Gaussian response functions reach into the exterior of the chambers, - an effect of the ranges of the secondary electrons. At higher photon energies response functions K(x) are slightly wider, but no detectable depth dependence has been observed. We have shown that the response functions of ionization chambers can be described by Gaussian distributions, confirming earlier observations, and we determined their standard deviations in both the lateral and longitudinal directions. Using these response functions, appropriate correction methods determined to eliminate the volume effect can be applied.


Influence of hypertension on aortic atherosclerosis in the Watanabe rabbit.
The effects of one-kidney, one clip Goldblatt hypertension on aortic atherosclerosis have been studied in the Watanabe heritable hyperlipidemic (WHHL) rabbit. Renovascular surgery was performed on WHHL rabbits at 3 months of age, and the rabbits were followed for periods of 3-6 months. Aortic atherosclerosis was assessed by measurement of intimal surface involvement with atherosclerotic lesions, determination of aortic free and ester cholesterol content, and microscopic examination. Systolic blood pressure increased by approximately 40-60 mm Hg in the renovascular surgical group as compared with the sham-operated group, but body weight, heart rate, serum cholesterol, and serum triglyceride were unaffected. Aortic atherosclerosis was increased in the hypertensive rabbits, even after 2-3 months of hypertension. At 3 months after renovascular surgery, the aortic surface area covered by atherosclerotic disease averaged 77 +/- 4.4% in hypertensive as compared with 16 +/- 3.3 in control rabbits. At 6 months after surgery, the values were 62 +/- 8.2% and 30 +/- 5.3% in the hypertensive and control rabbits, respectively. The differences in surface involvement and cholesterol content as a result of hypertension were particularly prominent in the descending thoracic aorta. Atherosclerotic lesions in the descending thoracic and abdominal aortic regions of normotensive WHHL rabbits were localized primarily to the ostia of branch vessels, but in the hypertensive rabbits, the involvement was typically very diffuse. No major differences in the nature of atherosclerotic lesions of comparable size were apparent by light microscopy. The results indicate that hypertension accelerates atherogenesis in the WHHL rabbit and suggest that this model may be valuable for studying the mechanisms by which such acceleration is induced.


Activation of innate immune-response genes in little brown bats (Myotis lucifugus) infected with the fungus Pseudogymnoascus destructans.
Recently bats have been associated with the emergence of diseases, both as reservoirs for several new viral diseases in humans and other animals and, in the northern Americas, as hosts for a devastating fungal disease that threatens to drive several bat species to regional extinction. However, despite these catastrophic events little Information is available on bat defences or how they interact with their pathogens. Even less is known about the response of bats to infection during torpor or long-term hibernation. Using tissue samples collected at the termination of an experiment to explore the pathogenesis of White Nose Syndrome in Little Brown Bats, we determined if hibernating bats infected with the fungus Pseudogymnoascus destructans could respond to infection by activating genes responsible for innate immune and stress responses. Lesions due to fungal infection and, in some cases, secondary bacterial infections, were restricted to the skin. However, we were unable to obtain sufficient amounts of RNA from these sites. We therefore examined lungs for response at an epithelial surface not linked to the primary site of infection. We found that bats responded to infection with a significant increase in lungs of transcripts for Cathelicidin (an anti-microbial peptide) as well as the immune modulators tumor necrosis factor alpha and interleukins 10 and 23. In conclusion, hibernating bats can respond to experimental P. destructans infection by activating expression of innate immune response genes.


From Targets to Action: Rolling up our Sleeves after Paris.
At the United Nations Climate Change Conference in Paris in 2015 ambitious targets for responding to the threat of climate change have been set: limiting global temperature increase to "well below 2 °C […] and to pursue efforts to limit the temperature increase to 1.5 °C". However, calculating the CO2 budget for 1.5 °C, it becomes clear that there is nearly no room left for future emissions. Scenarios suggest that negative emission technologies will play an even more important role for 1.5 °C than they already play for 2 °C. Especially against this background the feasibility of the target(s) is hotly debated, but this debate does not initiate the next steps that are urgently needed. Already the negotiations have featured the move from targets to implementation which is needed in the coming decade. Most importantly, there is an urgent need to develop and implement instruments that incentivize the rapid decarbonization. Moreover, it needs to be worked out how to link the climate and development agenda and prevent a buildup of coal power causing lock-in effects. Short term entry points into climate policy should now be in the focus instead of the fruitless debate on the feasibility of targets.


A New Application Using a Chromogenic Assay in a Plant Pathogen DNA Macroarray Detection System.
A DNA macroarray was previously developed to detect major fungal and oomycete pathogens of solanaceous crops. To provide a convenient alternative for researchers with no access to X-ray film-developing facilities, specific CCD cameras or Chemidoc XRS systems, a chromogenic detection method with sensitivity comparable with chemiluminescent detection, has been developed. A fungal (Stemphylium solani) and an oomycete (Phytophthora capsici) pathogen were used to develop the protocol using digoxigenin (DIG)-labeled targets. The internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (rDNA), including ITS1, 5.8S rDNA, and ITS2, was used as the target gene and polymerase chain reaction amplified as in the previous protocol. Various amounts of species-specific oligonucleotides on the array, quantities of DIG-labeled ITS amplicon, and hybridization temperatures were tested. The optimal conditions for hybridization were 55°C for 2 h using at least 10 pmol of each species-specific oligonucleotide and labeled target at 10 ng/ml of hybridization buffer. Incubation of the hybridized array with anti-DIG conjugated alkaline phosphatase substrates, NBT/BCIP, produced visible target signals between 1 and 3 h compared with 1 h in chemiluminescent detection. Samples from pure cultures, soil, and artificially inoculated plants were also used to compare the detection using chemiluminescent and chromogenic methods. Chromogenic detection was shown to yield similar results compared with chemiluminescent detection in regard to signal specificity, duration of hybridization between the array and targets, and cost, though it takes 1 to 2 h longer for the visualization process, thus providing a convenient alternative for researchers who lack darkroom facilities. To our knowledge, this is the first report of DNA macroarray detection of plant pathogens using a chromogenic method.


Commodity risk assessment of oak logs with bark from the US for the oak wilt pathogen Bretziella fagacearum under an integrated systems approach.
The European Commission submitted to the EFSA Panel on Plant Health a Dossier by USDA proposing a systems approach to mitigate the risk of entry of Bretziella fagacearum to the EU when trading oak logs with bark from the USA. Due to the forthcoming ban of methyl bromide (MB), the Dossier indicates sulfuryl fluoride (SF) as the substitute fumigant for this commodity. After collecting additional evidence from USDA,EU NPPOs, external experts and the published literature, the Panel performed a quantitative assessment on the likelihood of pest freedom for B. fagacearum at the point of entry in the EU, comparing the proposed systems approach with those already implemented by Commission Decision 2005/359/EC. The Panel provided also a non-quantitative assessment for all risk reduction options (RROs) proposed to be undertaken in the EU, from the point of entry to processing at the sawmill. The quantitative assessment until the EU point of entry, based on experts' judgement, indicated that: i) the most effective import option remains the current one with MB (95% certainty that between 9,573 and 10,000 containers per 10,000 would be free of B. fagacearum), followed by that with SF (95% certainty that between 8,639 and 10,000 containers per 10,000 would be free of B. fagacearum) and, last, by the other existing option based on delivering white oak logs in certain periods of the year to certain regions of the EU without fumigation (95% certainty, between 7,803 and 10,000 containers per 10,000). RROs proposed to be undertaken in the EU are expected to further reduce the risk of establishment of B. fagacearum, should these RROs be regulated, correctly implemented and checked by NPPOs. A wood pathway analysis is needed to quantitatively assess the importance of each measure and to optimise regulatory actions and risk management efforts.


White matter hyperintensity progression and late-life depression outcomes.
White matter hyperintensities (WMHs) are bright foci seen in the parenchyma of the brain on T2-weighted cranial magnetic resonance imaging (MRI) scans and are associated with geriatric depression. Because they are associated with age, they should increase in number and size over time. To our knowledge, this is the first longitudinal, volumetric MRI study of WMHs in depression. To determine if WMH progression over 2 years influences depression outcomes. Over 2 years, depressed subjects received antidepressant treatment according to a naturalistic somatic treatment algorithm designed to offer the best possible treatment to the individual. After the treatment period, depressed subjects were dichotomized based on whether they had reached and sustained remission during this period. One hundred thirty-three subjects aged 60 years or older meeting DSM-IV criteria for major depressive disorder. Cranial MRI was obtained at baseline and approximately 2 years later. White matter hyperintensity volume was measured in each hemisphere using a semiautomated segmentation process. Subjects were dichotomized based on achieving or not achieving remission of depressive symptoms, defined as a Montgomery-Asberg Depression Rating Scale score of 8 or less. The depressed subgroup that achieved and sustained remission had significantly less increases in WMH volume (11.5%) than did the group that did not achieve or sustain remission (31.6%) (P =.01). In a regression model, greater change in WMH volume was significantly associated with failure to sustain remission (P =.004) even when controlling for baseline depression severity, medical illness severity, age, sex, and race. Education was associated with achieving and sustaining remission (P =.02). Greater progression of WMH volume is associated with poor outcomes in geriatric depression. Future work is needed to develop means of slowing the rate of WMH progression and to determine whether this will lead to improved depression outcomes in elderly persons.


An emerging new concept for the management of type 2 diabetes with a paradigm shift from the glucose-centric to beta cell-centric concept of diabetes - an Asian perspective.
Recent advances in anti-diabetic medications and glucose monitoring have led to a paradigm shift in diabetes care. Newer anti-diabetic medications such as DPP-4 inhibitors, GLP-1 receptor agonists (GLP-1RAs), and SGLT2 inhibitors have enabled optimal glycemic control to be achieved without increasing the risk of hypoglycemia and weight gain. Treatment with GLP-1RAs and SGLT2 inhibitors has been demonstrated to improve cardiorenal outcomes, positioning these agents as the mainstay of treatment for patients with type 2 diabetes (T2DM). The development of these newer agents has also prompted a paradigm shift in the concept of T2DM, highlighting the importance of beta cell dysfunction in the pathophysiology of T2DM. Recent advances in pharmacotherapy for diabetes are summarized with a focus on the role of incretin-based drugs and SGLT2 inhibitors. The importance of a paradigm shift from a glucose-centric to a beta cell-centric concept of T2DM is also discussed, given from an Asian perspective. Management of T2DM including lifestyle modification as well as pharmacotherapy should be focused on reducing beta cell workload, to preserve functional beta cell mass. A paradigm shift from a glucose-centric to a beta cell-centric concept of T2DM enhances the implementation of person-centered diabetes care.


Presentation of cytosolic glycosylated peptides by human class I major histocompatibility complex molecules in vivo.
Antigens presented by class I major histocompatibility complex (MHC) molecules for recognition by cytotoxic T lymphocytes consist of 8-10-amino-acid-long cytosolic peptides. It is not known whether posttranslationally modified peptides are also presented by class I MHC molecules in vivo. Many different posttranslational modifications occur on cytoplasmic proteins, including a cytosolic O-beta-linked glycosylation of serine and threonine residues with N-acetylglucosamine (GlcNAc). Using synthetic glycopeptides carrying the monosaccharide O-beta-GlcNAc substitution on serine residues, we have shown that glycopeptides bind efficiently to class I MHC molecules and elicit a glycopeptide-specific cytotoxic T lymphocyte response in mice. In this study, we provide evidence that peptides presented by human class I MHC molecules in vivo encompass a small, significant amount of glycopeptides, constituting up to 0.1% of total peptide. Furthermore, we find that carbohydrate structures present on glycopeptides isolated from class I MHC molecules are dominated by the cytosolic O-beta-GlcNAc substitution, and synthetic peptides carrying this substitution are efficiently transported by TAP (transporter associated with antigen presentation) into the endoplasmic reticulum. Thus, in addition to unmodified peptides, posttranslationally modified cytosolic peptides carrying O-beta-linked GlcNAc can be presented by class I MHC molecules to the immune system.


"We don't wear it on our sleeve": Sickle cell disease and the (in)visible body in parts.
This paper approaches the lived experiences of patients with a genetically inherited chronic disease, sickle cell disease (SCD), through the lens of (in)visibility. SCD has been referred to as an "invisible" disease for a variety of interrelated reasons, including the difficulty of objectively measuring its characteristic symptoms, the lack of popular or specialist attention, and its characterization as a "black" disease. By mobilizing "invisibility" as a way of probing the day-to-day reinforcements of marginality, this article delves into how structural forces are experienced, interpreted, and negotiated by individual actors. To this end, we present ethnographic data collected from November 2009 until November 2013 with SCD patients and healthcare workers in Chicago. These data emphasize that rendering (in)visible is not a totalizing act, but rather meaningfully breaks the body into differentially visible and ideology-laden parts. More broadly, this indicates the need to rigorously question sources and effects of authority in biomedicine.


T cell source of type 1 cytokines determines illness patterns in respiratory syncytial virus-infected mice.
Manipulation of the cytokine microenvironment at the time of vaccination can influence immune responses to remote challenge, providing a strategy to study the molecular pathogenesis of respiratory syncytial virus (RSV) vaccine-enhanced disease in the mouse model. Although treatment with antibody against IL-4 or recombinant IL-12 (rIL-12) at the time of formalin-inactivated RSV vaccination induced a similar shift in the pattern of cytokine mRNA expression upon live virus challenge, anti-IL-4 treated mice had increased CD8+ cytotoxic T lymphocyte activity and reduced illness compared with rIL-12-treated mice. To define effector mechanisms responsible for these patterns, CD4+ and/or CD8+ T lymphocytes were selectively depleted in vivo at the time of RSV challenge. In rIL-12-treated mice, CD4+ lymphocytes made the largest contribution to IFN-gamma mRNA, RSV clearance, and illness, while in anti-IL-4 treated mice, CD8+ lymphocytes were the major effector. The effector responsible for virus clearance also mediated illness, suggesting that efficiency of virus clearance determined disease expression. These results demonstrate that the phenotype of effector cells involved in the immune response to virus challenge may be a more important determinant of disease than patterns of cytokine expression classically assigned to Th1 and Th2 lymphocytes.


Pretransplantation assessment of the risk of lymphoproliferative disorder.
Posttransplantation lymphoproliferative disorder (PTLD) is an uncommon but often fatal complication of solid organ transplantation that occurs in approximately 3% of patients. To determine the relative importance and relationship of potential risk factors for PTLD before transplantation (i.e., Epstein-Barr virus [EBV] serostatus of the recipient and the cytomegalovirus [CMV] sero-status of the recipient and the potential donor) and the principal risk factor after transplantation (immunosuppression with antilymphocyte antibody), we analyzed the findings for the first 381 consecutive adult nonrenal transplant recipients seen at Mayo Clinic. In the absence of the other risk factors, the incidence rate of PTLD for EBV-seronegative recipients was 24 times higher (95% confidence interval [CI]: 6.2, 89) than that for EBV-seropositive recipients. The additional risk factors of therapy with OKT3 for rejection and CMV seromismatch (i.e., a negative recipient and a positive donor) each further amplified this risk four- to sixfold. Together, all three risk factors acted synergistically to increase the incidence rate of fatal and/or CNS PTLD by a factor of 654 (CI: 368, 1,162) compared with the low incidence rate (.458 cases per 100 person years) when none of these risk factors were present. Pretransplantation determination of recipient EBV and CMV serostatus can identify a subgroup of patients whose risk for severe PTLD may preclude transplantation.


Cisplatin-induced apoptosis in p53-deficient renal cells via the intrinsic mitochondrial pathway.
Nephrotoxicity is the major limiting factor for the use of cisplatin in cancer therapy. Recent studies have demonstrated an important role for p53 in cisplatin-induced renal injury. Nevertheless, pharmacological and genetic blockade of p53 only provides partial renoprotective effects, suggesting the presence of p53-independent injury mechanisms. To understand the p53-independent mechanisms, we have now examined cisplatin-induced apoptosis in p53-deficient kidney cells. We show that cisplatin could induce Bax activation, cytochrome c release, and apoptosis in primary cultures of p53-deficient renal tubular cells, albeit at a level that was lower than in the wild-type cells. Cisplatin could also induce typical apoptosis in p53-deficient baby mouse kidney (BMK) cells. The apoptosis was caspase dependent and could be completely blocked by general caspase inhibitors. Bax and Bak, two key molecules in the mitochondrial pathway of apoptosis, were interdependently activated by cisplatin, with Bax translocation to and Bax/Bak oligomerization in mitochondria, leading to cytochrome c release. Importantly, cytochrome c release and apoptosis were diminished in Bax/Bak single or double-knockout BMK cells. Furthermore, overexpression of Bcl-2 could ameliorate cisplatin-induced cytochrome c release and apoptosis. Together, the results have demonstrated a p53-independent mechanism of cisplatin nephrotoxicity that involves the mitochondrial pathway of apoptosis.


Coordination design of cadmium ions at the 4-fold axis channel of the apo-ferritin cage.
Spherical protein cages with highly symmetrical structures provide unique environments for the conjugation of metal ions and metal nanoparticles. Ferritin has been widely studied as a template for the coordination of metal ions and metal nanoparticles in fundamental research and applications. However, it remains difficult to design metal coordination sites precisely. In this work, we describe the design and construction of new metal coordination sites by introducing Cys residues at the 4-fold symmetrical hydrophobic channel of apo-ferritin. X-ray crystal structure analyses of the mutants containing Cd(ii) ions show that the four or eight binding sites for Cd(ii) ions are located at the 4-fold symmetrical axis channel of apo-ferritin. It was found that the coordination number and configuration of Cd(ii) ions can be varied by adjusting the positions of the Cys residues at the symmetrical channels of the apo-ferritin cage.


Privatization of Biofilm Matrix in Structurally Heterogeneous Biofilms.
The self-produced biofilm provides beneficial protection for the enclosed cells, but the costly production of matrix components makes producer cells susceptible to cheating by nonproducing individuals. Despite detrimental effects of nonproducers, biofilms can be heterogeneous, with isogenic nonproducers being a natural consequence of phenotypic differentiation processes. For instance, in Bacillus subtilis biofilm cells differ in production of the two major matrix components, the amyloid fiber protein TasA and exopolysaccharides (EPS), demonstrating different expression levels of corresponding matrix genes. This raises questions regarding matrix gene expression dynamics during biofilm development and the impact of phenotypic nonproducers on biofilm robustness. Here, we show that biofilms are structurally heterogeneous and can be separated into strongly and weakly associated clusters. We reveal that spatiotemporal changes in structural heterogeneity correlate with matrix gene expression, with TasA playing a key role in biofilm integrity and timing of development. We show that the matrix remains partially privatized by the producer subpopulation, where cells tightly stick together even when exposed to shear stress. Our results support previous findings on the existence of "weak points" in seemingly robust biofilms as well as on the key role of linkage proteins in biofilm formation. Furthermore, we provide a starting point for investigating the privatization of common goods within isogenic populations.IMPORTANCE Biofilms are communities of bacteria protected by a self-produced extracellular matrix. The detrimental effects of nonproducing individuals on biofilm development raise questions about the dynamics between community members, especially when isogenic nonproducers exist within wild-type populations. We asked ourselves whether phenotypic nonproducers impact biofilm robustness, and where and when this heterogeneity of matrix gene expression occurs. Based on our results, we propose that the matrix remains partly privatized by the producing subpopulation, since producing cells stick together when exposed to shear stress. The important role of linkage proteins in robustness and development of the structurally heterogeneous biofilm provides an entry into studying the privatization of common goods within isogenic populations.


The fMRI BOLD signal tracks electrophysiological spectral perturbations, not event-related potentials.
Functional magnetic resonance imaging (fMRI) and electroencephalography (EEG) are primary tools of the psychological neurosciences. It is therefore important to understand the relationship between hemodynamic and electrophysiological responses. An early study by Huettel and colleagues found that the coupling of fMRI blood-oxygen-level-dependent signal (BOLD) and subdurally-recorded signal-averaged event-related potentials (ERPs) was not consistent across brain regions. Instead, a growing body of evidence now indicates that hemodynamic changes measured by fMRI reflect non-phase-locked changes in high frequency power rather than the phase-locked ERP. Here, we revisit the data from Huettel and colleagues and measure event-related spectral perturbations (ERSPs) to examine the time course of frequency changes. We found that, unlike the ERP, γ-ERSP power was consistently coupled with the hemodynamic response across three visual cortical regions. Stimulus duration modulated the BOLD signal and the γ-ERSP in the peri-calcarine and fusiform cortices, whereas there was no such modulation of either physiological signal in the lateral temporal-occipital cortex. This finding reconciles the original report with the more recent literature and demonstrates that the ERP and ERSP reflect dissociable aspects of neural activity.


X chromosome duplications affect a region of the chromosome they do not duplicate in Caenorhabditis elegans.
X chromosome duplications have been used previously to vary the dose of specific regions of the X chromosome to study dosage compensation and sex determination in Caenorhabditis elegans. We show here that duplications suppress and X-linked hypomorphic mutation and elevate the level of activity of an X-linked enzyme, although these two genes are located in a region of the X chromosome that is not duplicated. The effects do not depend on the region of the X chromosome duplicated and is stronger in strains with two doses of a duplication than in strains with one dose. This is evidence for a general elevation of X-linked gene expression in strains carrying X-chromosome duplications, consistent with the hypothesis that the duplications titrate a repressor acting on many X-linked genes.


Downregulation of the type 1 insulin-like growth factor receptor in mouse melanoma cells is associated with enhanced radiosensitivity and impaired activation of Atm kinase.
The type 1 insulin-like growth factor receptor (IGF1R) is required for growth, tumorigenicity and protection from apoptosis. IGF1R overexpression is associated with radioresistance in breast cancer. We used antisense (AS) RNA to downregulate IGF1R expression in mouse melanoma cells. Cells expressing AS-IGF1R transcripts were more radiosensitive in vitro and in vivo than controls. Also they showed reduced radiation-induced p53 accumulation and p53 serine 18 phosphorylation, and radioresistant DNA synthesis. These changes were reminiscent of the cellular phenotype of the human genetic disorder ataxia-telangiectasia (A-T), caused by mutations in the ATM gene. Cellular Atm protein levels were lower in AS-IGF1R-transfected cells than in control cells, although there was no difference in Atm expression at the transcriptional level. AS-IGF1R cells had detectable basal Atm kinase activity, but failed to induce kinase activity after irradiation. This suggests that IGF1R signalling can modulate the function of Atm, and supports the concept of targeted IGF1R downregulation as a potential treatment for malignant melanoma and other radioresistant tumours.