Bovine herpes virus gD protein produced in plants using a recombinant tobacco mosaic virus (TMV) vector possesses authentic antigenicity.
A tobacco mosaic virus (TMV)-based vector was utilized for expression of a cytosolic form of the bovine herpesvirus type 1 (BHV-1) protein glycoprotein D (gDc). Nicotiana benthamiana plants were harvested 7 days after inoculation with RNA transcripts derived from the TMV-gDc recombinant virus. Recombinant gDc protein of expected electrophoretic mobility accumulated in inoculated leaves to a concentration of about 20 micrograms/g of fresh leaf tissue. Oil-based vaccines were formulated with crude foliar extracts to immunize mice parentally. After a single injection, animals developed a sustained and specific response to both the isolated gD and native virus particles. Cattle vaccinated with the same gDc containing extracts developed specific humoral and cellular immune responses directed against both the viral gD and BHV-1 particles. Most importantly, animals vaccinated with the plant-produced gDc showed good levels of protection after challenge with the virulent BHV-1. Virus excretion was drastically reduced in these animals, reaching levels comparable to animals vaccinated with a commercial BHV-1 vaccine. The positive immunological characterization obtained for the gDc, indicated that an important part of the natural conformation was retained in the plant recombinant protein.


Feasibility and Effectiveness of a Peer Referral Incentive Intervention to Promote Male Circumcision Uptake in Zambia.
Medical male circumcision is a promising HIV prevention tool in countries with generalized HIV epidemics, but demand creation interventions are needed to support scale-up. We piloted a peer referral intervention in which circumcision clients were offered incentives for referring their peers for circumcision. The intervention was implemented between June 2014 and February 2015 in 6 randomly selected health facilities in Southern Province, Zambia. For the first 5 months, circumcision clients ≥18 years of age were given referral vouchers that allowed them to refer up to 5 peers for circumcision within a 3-month period. An incentive of US$2 was offered for each referral. The primary outcome was the number of circumcisions performed per month in each facility. To assess the effect of the intervention, a difference-in-difference analysis was performed using longitudinal data from the intervention facilities and 22 nonintervention facilities. A questionnaire was also implemented to understand men's perceptions of the intervention. During the 8-month intervention period, 1222 men over 18 years of age were circumcised in intervention facilities. In the first 5 months, 699 circumcision clients were enrolled and 385 clients brought a referral voucher given to them by an enrolled client. Difference-in-difference analyses did not show a significant increase in circumcisions performed in intervention facilities. However, circumcision clients reported that the referral incentive motivated them to encourage their friends to seek male circumcision. Peer referrals were also reported to be an important factor in men's decisions because 78% of clients who were referred reported that talking with a circumcised friend was important for their decision to get circumcised. The peer referral incentive intervention for male circumcision was feasible and acceptable. However, the intervention did not have a significant effect on demand for male circumcision. Barriers to circumcision and features of the intervention may have limited the effect of the intervention. Further efforts regarding encouraging male-to-male communication and evaluations with larger sample sizes are needed.


Nickel Oxide Interstratified agr-Zirconium Phosphate, a Composite Exhibiting Ferromagnetic Behavior.
As part of an ongoing research program to synthesize novel pillared layered materials, nickel and cobalt hydroxyacetates were inserted between the layers of amine intercalates of alpha-zirconium phosphate. The structure of the resultant nickel composite, derived from x-ray powder data, was found to consist of a three-tiered layer of nickel atoms bridged by hydroxo and acetato groups. Heating to 420 degrees C converted the hydroxyacetate layers to oxide and imparted ordered magnetic domains to the composite. The phosphate layers appear to act as a template directing the growth of the inserted layers in this class of composite materials.


Role of social media and the Internet in pathways to care for adolescents and young adults with psychotic disorders and non-psychotic mood disorders.
Although psychosis often occurs during adolescence, there has been little research on how the ubiquitously used Internet and social media could impact pathways to care. We examined how youth with psychotic spectrum disorders (PSD) versus non-psychotic mood disorders (NPMD) use online resources in the early illness stages. Social media use and pathways to care data were collected using a semi-structured interview from 80 youth (PSD = 40 and NPMD = 40) aged 12-21 years within 2 years of symptom onset. A total of 97.5% of participants (mean age = 18.3 years) regularly used social media, spending approximately 2.6 ± 2.5 h per day online. There were 22.4% of our sample (PSD = 19.4%, NPMD = 25.0%, P = 0.56) who reported waiting to reach out for help believing that symptoms would disappear. A total of 76.5% (PSD = 67.5%, NPMD = 85.0%, P = 0.06) noticed social media habit changes during symptom emergence. Thirty per cent reported discussing their symptoms on social media (PSD = 22.5%, NPMD = 37.5%, P = 0.14). NPMD patients sought information most on how to stop symptoms (40.0% vs. 13.5%, P = 0.01), while PSD youth were more commonly interested in what caused their symptoms (21.6% vs. 15.0%, P = 0.45). More PSD patients (42.9% vs. 25.0%, P = 0.10) would prefer to receive mental health information via the Internet. Altogether, 63.6% (PSD = 64.9%, NPMD = 62.5%, P = 0.83) were amenable to clinicians proactively approaching them via social media during symptom emergence. A total of 74.3% (PSD = 78.4%, NPMD = 70.0%, P = 0.40) liked the idea of obtaining help/advice from professionals via social media. The Internet and social media provide an unparalleled opportunity to supplement and potentially transform early intervention services, and acceptance of this approach appears to be high.


LoyalTracker: Visualizing Loyalty Dynamics in Search Engines.
The huge amount of user log data collected by search engine providers creates new opportunities to understand user loyalty and defection behavior at an unprecedented scale. However, this also poses a great challenge to analyze the behavior and glean insights into the complex, large data. In this paper, we introduce LoyalTracker, a visual analytics system to track user loyalty and switching behavior towards multiple search engines from the vast amount of user log data. We propose a new interactive visualization technique (flow view) based on a flow metaphor, which conveys a proper visual summary of the dynamics of user loyalty of thousands of users over time. Two other visualization techniques, a density map and a word cloud, are integrated to enable analysts to gain further insights into the patterns identified by the flow view. Case studies and the interview with domain experts are conducted to demonstrate the usefulness of our technique in understanding user loyalty and switching behavior in search engines.


Fully automatic recognition of the temporal phases of facial actions.
Past work on automatic analysis of facial expressions has focused mostly on detecting prototypic expressions of basic emotions like happiness and anger. The method proposed here enables the detection of a much larger range of facial behavior by recognizing facial muscle actions [action units (AUs)] that compound expressions. AUs are agnostic, leaving the inference about conveyed intent to higher order decision making (e.g., emotion recognition). The proposed fully automatic method not only allows the recognition of 22 AUs but also explicitly models their temporal characteristics (i.e., sequences of temporal segments: neutral, onset, apex, and offset). To do so, it uses a facial point detector based on Gabor-feature-based boosted classifiers to automatically localize 20 facial fiducial points. These points are tracked through a sequence of images using a method called particle filtering with factorized likelihoods. To encode AUs and their temporal activation models based on the tracking data, it applies a combination of GentleBoost, support vector machines, and hidden Markov models. We attain an average AU recognition rate of 95.3% when tested on a benchmark set of deliberately displayed facial expressions and 72% when tested on spontaneous expressions.


Sequencing at the syllabic and supra-syllabic levels during speech perception: an fMRI study.
The processing of fluent speech involves complex computational steps that begin with the segmentation of the continuous flow of speech sounds into syllables and words. One question that naturally arises pertains to the type of syllabic information that speech processes act upon. Here, we used functional magnetic resonance imaging to profile regions, using a combination of whole-brain and exploratory anatomical region-of-interest (ROI) approaches, that were sensitive to syllabic information during speech perception by parametrically manipulating syllabic complexity along two dimensions: (1) individual syllable complexity, and (2) sequence complexity (supra-syllabic). We manipulated the complexity of the syllable by using the simplest syllable template-a consonant and vowel (CV)-and inserting an additional consonant to create a complex onset (CCV). The supra-syllabic complexity was manipulated by creating sequences composed of the same syllable repeated six times (e.g., /pa-pa-pa-pa-pa-pa/) and sequences of three different syllables each repeated twice (e.g., /pa-ta-ka-pa-ta-ka/). This parametrical design allowed us to identify brain regions sensitive to (1) syllabic complexity independent of supra-syllabic complexity, (2) supra-syllabic complexity independent of syllabic complexity and, (3) both syllabic and supra-syllabic complexity. High-resolution scans were acquired for 15 healthy adults. An exploratory anatomical ROI analysis of the supratemporal plane (STP) identified bilateral regions within the anterior two-third of the planum temporale, the primary auditory cortices as well as the anterior two-third of the superior temporal gyrus that showed different patterns of sensitivity to syllabic and supra-syllabic information. These findings demonstrate that during passive listening of syllable sequences, sublexical information is processed automatically, and sensitivity to syllabic and supra-syllabic information is localized almost exclusively within the STP.


A rare combination of amniotic constriction band with osteogenesis imperfecta.
Amniotic constriction bands and osteogenesis imperfecta are disorders arising from a collagen defect. We report a rare association of amniotic bands with osteogenesis imperfecta in a child. The child was born with multiple amniotic bands involving the right leg, both hands and both feet. Multiple fractures of long bones of lower limbs occurred in childhood due to trivial trauma. Deformities of the femur and tibia due to malunion with osteopenia and blue sclerae were present. The patient was treated with z plasty of constriction band of the right tibia and bisphosphonate for osteogenesis imperfecta. This rare association of both collagen diseases may provide further insight for the pathogenesis of these diseases.


Diagnostic value of maximal-outer-diameter and maximal-mural-thickness in use of ultrasound for acute appendicitis in children.
To evaluate the maximal-outer-diameter (MOD) and the maximal-mural-thickness (MMT) of the appendix in children with acute appendicitis and to determine their optimal cut-off values to diagnose acute appendicitis. In total, 164 appendixes from 160 children between 1 and 17 years old (84 males, 76 females; mean age, 7.38 years) were examined by high-resolution abdominal ultrasound for acute abdominal pain and the suspicion of acute appendicitis. We measured the MOD and the MMT at the thickest point of the appendix. Patients were categorized into two groups according to their medical records: patients who had surgery (surgical appendix group) and patients who did not have surgery (non-surgical appendix group). Data were analyzed by MedCalc v.9.3. The rank sum test (Mann-Whitney test) was used to evaluate the difference in the MOD and the MMT between the two groups. ROC curve analysis was used to determine the optimal cut-off value of the MOD and the MMT on diagnosis of acute appendicitis. There were 121 appendixes (73.8%) in the non-surgical appendix group and 43 appendixes (26.2%) in the surgical appendix group. The median MOD differed significantly between the two groups (0.37 cm vs 0.76 cm, P < 0.0001), and the median MMT also differed (0.15 cm vs 0.33 cm, P < 0.0001). The optimal cut-off value of the MOD and the MMT for diagnosis of acute appendicitis in children was > 0.57 cm (sensitivity 95.4%, specificity 93.4%) and > 0.22 cm (sensitivity 90.7%, specificity 79.3%), respectively. The MOD and the MMT are reliable criteria to diagnose acute appendicitis in children. An MOD > 0.57 cm and an MMT > 0.22 cm are the optimal criteria.


Spotlight on the fetus: how physical activity during pregnancy influences fetal health: a narrative review.
Before and during pregnancy, women often aim to improve their lifestyle so as to provide a healthier environment for their developing child. It remains unresolved, however, as to whether physical activity (PA) during pregnancy poses a possible risk or whether it might even have beneficial effects on the developing child. There is increasing evidence that PA during pregnancy is indeed beneficial to maternal physiological and psychological health and that it is generally not detrimental to the fetal cardiovascular system and neuronal function in the developing child. This also led to international recommendations for PAs during pregnancy. In the current review, we aimed to comprehensively assess the evidence of beneficial and harmful effects of maternal PA, including high-performance sports, on fetal development. The different mental and body-based relaxation techniques presented here are frequently performed during pregnancy. We found a considerable number of studies addressing these issues. In general, neither low key, moderate maternal PA nor relaxation techniques were observed to have a harmful effect on the developing child. However, we identified some forms of PA which could have at least a transient unfavourable effect. Notably, the literature currently available does not provide enough evidence to enable us to make a general conclusive statement on this subject. This is due to the lack of longitudinal studies on the metabolic and cognitive effects of regular PA during pregnancy and the wide diversity of methods used. In particular, the kind of PA investigated in each study differed from study to study.


Mass drug administration of azithromycin for trachoma reduces the prevalence of genital Chlamydia trachomatis infection in the Solomon Islands.
Chlamydia trachomatis is the most common bacterial sexually transmitted infection and is frequently asymptomatic; ocular C. trachomatis strains cause trachoma. Mass drug administration (MDA) of azithromycin for trachoma might also reduce the prevalence of genital C. trachomatis. In a survey conducted in the Solomon Islands in 2014, prior to MDA, the prevalence of genital C. trachomatis was 20.3% (95% CI 15.9% to 25.4%). We conducted a survey to establish the impact of MDA with azithromycin on genital C. trachomatis. Women attending three community outpatient clinics, predominantly for antenatal care, 10 months after MDA with azithromycin given for trachoma elimination, were enrolled in this survey. Self-taken high vaginal swabs were for C. trachomatis and Neisseria gonorrhoeae using the BD Probetec strand displacement assay. 298 women were enrolled. C. trachomatis infection was diagnosed in 43 women (14.4%, 95% CI 10.6% to 18.9%) and N. gonorrhoeae in 9 (3%, 95% CI 1.4% to 5.7%). The age-adjusted OR for C. trachomatis infection was consistent with a significant decrease in the prevalence of C. trachomatis following MDA (OR 0.58, 95% CI 0.37 to 0.94, p=0.027). There was no change in the prevalence of N. gonorrhoeae between following MDA (OR 0.51, 95% CI 0.22 to 1.22, p=0.13). This study demonstrated a 40% reduction in the age-adjusted prevalence of genital C. trachomatis infection following azithromycin MDA for trachoma elimination.


Cortically coupled computer vision for rapid image search.
We describe a real-time electroencephalography (EEG)-based brain-computer interface system for triaging imagery presented using rapid serial visual presentation. A target image in a sequence of nontarget distractor images elicits in the EEG a stereotypical spatiotemporal response, which can be detected. A pattern classifier uses this response to reprioritize the image sequence, placing detected targets in the front of an image stack. We use single-trial analysis based on linear discrimination to recover spatial components that reflect differences in EEG activity evoked by target versus nontarget images. We find an optimal set of spatial weights for 59 EEG sensors within a sliding 50-ms time window. Using this simple classifier allows us to process EEG in real time. The detection accuracy across five subjects is on average 92%, i.e., in a sequence of 2500 images, resorting images based on detector output results in 92% of target images being moved from a random position in the sequence to one of the first 250 images (first 10% of the sequence). The approach leverages the highly robust and invariant object recognition capabilities of the human visual system, using single-trial EEG analysis to efficiently detect neural signatures correlated with the recognition event.


The desired moral attitude of the physician: (I) empathy.
In professional medical ethics, the physician traditionally is obliged to fulfil specific duties as well as to embody a responsible and trustworthy personality. In the public discussion, different concepts are suggested to describe the desired underlying attitude of physicians. In this article, one of them--empathy--is presented in an interpretation that is meant to depicture (together with the two additional concepts compassion and care) this attitude. Therefore empathy in the clinical context is defined as the adequate understanding of the inner processes of the patient concerning his health-related problems. Adequacy is scrutinized on behalf of the emotional and subjective involvement of he physician, and on the necessary dependence on medical-moral-goals. In the present interpretation, empathy alone is no guarantee of the right moral attitude, but a necessary instrumental skill in order to perceive and treat a patient as an individual person. The concepts of compassion and care that will be discussed in two forthcoming articles are necessary parts to describe the desired moral attitude of the physician more completely.


Oriental herbs as a source of novel anti-androgen and prostate cancer chemopreventive agents.
Androgen and androgen receptor (AR) signaling are crucial for the genesis of prostate cancer (PCa), which can often develop into androgen-ligand-independent diseases that are lethal to the patients. Recent studies show that even these hormone-refractory PCa require ligand-independent AR signaling for survival. As current chemotherapy is largely ineffective for PCa and has serious toxic sideeffects, we have initiated a collaborative effort to identify and develop novel, safe and naturally occurring agents that target AR signaling from Oriental medicinal herbs for the chemoprevention and treatment of PCa. We highlight our discovery of decursin from an Oriental formula containing Korean Angelica gigas Nakai (Dang Gui) root as a novel anti-androgen/AR agent. We have identified the following mechanisms to account for the specific anti-AR actions: rapid block of AR nuclear translocation, inhibition of binding of 5alpha-dihydrotestesterone to AR and increased proteasomal degradation of AR protein. Furthermore, decursin lacks the agonist activity of the "pure" anti-androgen bicalutamide and is more potent than bicalutamide in inducing PCa apoptosis. Structure-activity analyses reveal a critical requirement of the side-chain on decursin or its structural isomer decursinol angelate for anti-AR, cell cycle arrest and proapoptotic activities. This work demonstrates the feasibility of using activity-guided fractionation in cell culture assays combined with mechanistic studies to identify novel anti-androgen/ AR agents from complex herbal mixtures.


Who presents past the gestational age limit for first trimester abortion in the public sector in Mexico City?
To identify socio-demographic factors associated with presenting for abortion services past the gestational age (GA) limit (12 weeks), and thus not receiving services, in Mexico City's public sector first trimester abortion program. We used clinical data from four high volume sites in the Interrupción Legal de Embarazo (ILE) program, 2007-2015. We used descriptive statistics to quantify the proportion of women who did not receive an abortion due to presenting past the gestational age limit. We used multivariable logistic regression to identify associations between women's characteristics and presenting past the GA limit and calculated predicted probabilities of late presentation for key characteristics. Our sample included 52,391 women, 8.10% (n = 4,246) of whom did not receive abortion services due to presenting past the GA limit. Adolescents (12-17) made up 8.69% of the total sample and 13.40% of those presenting past the GA limit (p< 0.05). In multivariable analyses, all age groups of adult women had significantly lower odds than adolescents of presenting past the limit (aOR = 0.77, aOR = 0.63, aOR = 0.58 and aOR = 0.37 for 19-24, 25-29, 30-39, and > = 40 years' old respectively). Women living in Mexico City and with higher levels of education had lower odds of presenting past the GA limit, and there was an educational gradient across all age groups. In the multivariable predicted probability models, adolescents at every level of education have significantly higher probabilities of not receiving an abortion due to presenting past the gestational age limit compared with adults (among women with a primary education: 11.75% adolescents vs. 9.02-4.26% across adult age groups). Our results suggest that continued efforts are needed to educate women, especially younger and less educated women, about early pregnancy recognition. In addition, all women need information about the availability of first trimester legal abortion to ensure timely access to abortion services.


Does Living Closer to a University Increase Educational Attainment? A Longitudinal Study of Aspirations, University Entry, and Elite University Enrolment of Australian Youth.
Geography remains a critical factor that shapes the development of aspirations, attainment, and choice in young people. We focus on the role of geography on university entry and aspirations due to the increasing requirement in society for a higher education qualification for access to prestigious positions in society. Using a large representative longitudinal database (N = 11,999; 50 % male; 27 % provincial or rural; 2 % Indigenous) of Australia youth we explore the association between distance to a university campus and the critical attainment outcomes of university entry and enrolment in an elite university as well as critical predictors of these outcomes in access to information resources (i.e., university outreach programs) and university aspirations. In doing so, we provide new insight into distance effects, and the extent that these are due to selection, cost, and community influence. Our findings suggest that distance is significantly associated with both university expectations and entrance, with an especially large impact upon young people from low socioeconomic backgrounds. However, we also find little evidence that distance is related to attending a university led information session. Our conclusion is that distance effects cannot be fully explained by selection in terms of academic achievement and socioeconomic status, and that anticipatory decisions and costs are the most likely drivers of the distance effect.


Beneficial influence of postmenopausal estrogen therapy on serum adhesion molecules is independent of the route and dose of administration.
In current literature, the immune-inflammatory theory of atherosclerosis is widely discussed. The role of how adhesion molecules contribute to the development of atheromatic plaques is especially underlined. 120 females in menopausal age were included in the study between 2004 and 2009. All the women were of menopausal age (51±3 years), from southern Poland, with FSH levels above 30 mIU/ml, and complaining of menopausal symptoms that disturbed normal daily activity. The study was conducted over a 6 month period. Three groups of 40 randomized patients were selected. The control group consisted of 40 volunteers, who were also from southern Poland, in good health, without menopausal symptoms, or menstrual periods in the last 6 months. Control subjects were match according to age and weight, with FSH levels above 25 mIU/ml and normal TSH and prolactin values. All patients, in the treatment and control groups were seronegative for Chlamydia pneumonia throughout the duration of the study. After 6 months, hormonal therapy was found to significantly reduce levels of sICAM-1 and sVCAM-1 in all treated groups compared to the control group and the results were statistically significant. Alternatively, in the latter group, we observed increased levels of the investigated adhesion molecules (group I: 37.5 µg/24h transdermal estradiol + dydrogesteron; group II 50 µg/24h transdermal estradiol + medroxyprogesteron; group III 1mg of oral estradiol + noretisteron sICAM-1 and control group; using paired Wilcoxon test). All of the investigated estrogen therapy schemas have a favorable impact on the blood levels of sICAM-1 and sVCAM-1 in postmenopausal women without cardiovascular risk factors, reducing their concentration.


Evolution of cartesian genetic programs for development of learning neural architecture.
Although artificial neural networks have taken their inspiration from natural neurological systems, they have largely ignored the genetic basis of neural functions. Indeed, evolutionary approaches have mainly assumed that neural learning is associated with the adjustment of synaptic weights. The goal of this paper is to use evolutionary approaches to find suitable computational functions that are analogous to natural sub-components of biological neurons and demonstrate that intelligent behavior can be produced as a result of this additional biological plausibility. Our model allows neurons, dendrites, and axon branches to grow or die so that synaptic morphology can change and affect information processing while solving a computational problem. The compartmental model of a neuron consists of a collection of seven chromosomes encoding distinct computational functions inside the neuron. Since the equivalent computational functions of neural components are very complex and in some cases unknown, we have used a form of genetic programming known as Cartesian genetic programming (CGP) to obtain these functions. We start with a small random network of soma, dendrites, and neurites that develops during problem solving by repeatedly executing the seven chromosomal programs that have been found by evolution. We have evaluated the learning potential of this system in the context of a well-known single agent learning problem, known as Wumpus World. We also examined the harder problem of learning in a competitive environment for two antagonistic agents, in which both agents are controlled by independent CGP computational networks (CGPCN). Our results show that the agents exhibit interesting learning capabilities.


Comparing different definitions of prediabetes with subsequent risk of diabetes: an individual participant data meta-analysis involving 76 513 individuals and 8208 cases of incident diabetes.
There are currently five widely used definition of prediabetes. We compared the ability of these to predict 5-year conversion to diabetes and investigated whether there were other cut-points identifying risk of progression to diabetes that may be more useful. We conducted an individual participant meta-analysis using longitudinal data included in the Obesity, Diabetes and Cardiovascular Disease Collaboration. Cox regression models were used to obtain study-specific HRs for incident diabetes associated with each prediabetes definition. Harrell's C-statistics were used to estimate how well each prediabetes definition discriminated 5-year risk of diabetes. Spline and receiver operating characteristic curve (ROC) analyses were used to identify alternative cut-points. Sixteen studies, with 76 513 participants and 8208 incident diabetes cases, were available. Compared with normoglycemia, current prediabetes definitions were associated with four to eight times higher diabetes risk (HRs (95% CIs): 3.78 (3.11 to 4.60) to 8.36 (4.88 to 14.33)) and all definitions discriminated 5-year diabetes risk with good accuracy (C-statistics 0.79-0.81). Cut-points identified through spline analysis were fasting plasma glucose (FPG) 5.1 mmol/L and glycated hemoglobin (HbA1c) 5.0% (31 mmol/mol) and cut-points identified through ROC analysis were FPG 5.6 mmol/L, 2-hour postload glucose 7.0 mmol/L and HbA1c 5.6% (38 mmol/mol). In terms of identifying individuals at greatest risk of developing diabetes within 5 years, using prediabetes definitions that have lower values produced non-significant gain. Therefore, deciding which definition to use will ultimately depend on the goal for identifying individuals at risk of diabetes.


Unravelling the coupling of surface plasmons in carbon nanotubes by near-field nanoscopy.
We experimentally and theoretically investigated the propagation and subwavelength coupling of surface plasmons in carbon nanotube pairs (CNTPs). We found two plasmon modes in the Fourier transformed near-field images of CNTPs, while only one propagating plasmon mode in the single carbon nanotube (CNT). Using the finite element method (FEM), we calculated plasmon coupling between two carbon nanotubes. We attributed the observed two plasmon modes to the symmetric and asymmetric coupling of the plasmons between the two CNTs. Using numerical simulation, we further investigated the plasmon coupling strength and found that it is sensitive to the separating distance of CNTs. When the separation is larger than 5 nm, the plasmon coupling attenuates. We show that near-field optical imaging is a feasible way to characterize the plasmon coupling of CNTs, even if the separation of the CNTs is only a few nanometers.


Binge-like ethanol consumption increases corticosterone levels and neurodegneration whereas occupancy of type II glucocorticoid receptors with mifepristone is neuroprotective.
Excessive ethanol (EtOH) use leads to impaired memory and cognition. Using a rat model of binge-like intoxication, we tested whether elevated corticosterone (Cort) levels contribute to the neurotoxic consequences of EtOH exposure. Rats were adrenalectomized (Adx) and implanted with cholesterol pellets, or cholesterol pellets containing Cort in order to achieve basal, medium, or high blood concentrations of Cort. Intragastric EtOH or an isocaloric control solution was given three times daily for 4 days to achieve blood alcohol levels ranging between 200 and 350 mg/dl. Mean 24-hour plasma levels of Cort were ∼110 and ∼40 ng/ml in intact EtOH-treated and intact control animals, respectively. Basal Cort replacement concentrations in EtOH-treated Adx animals did not exacerbate alcohol-induced neurodegeneration in the hippocampal dentate gyrus (DG) or the entorhinal cortex (EC) as observed by amino-cupric silver staining. In contrast, Cort replacement pellets resulting in plasma Cort levels twofold higher (medium) than normal, or greater than twofold higher (high) in Adx-Cort-EtOH animals increased neurodegeneration. In separate experiments, pharmacological blockade of the Type II glucocorticoid (GC) receptor was initiated with mifepristone (RU38486; 0, 5, 15 mg/kg/day, i.p.). At the higher dose, mifepristone decreased the number of degenerating hippocampal DG cells in binge-EtOH-treated intact animals, whereas, only a trend for reduction was observed in 15 mg/kg/day mifepristone-treated animals in the EC, as determined by fluoro-jade B staining. These results suggest that elevated circulating Cort in part mediates EtOH-induced neurotoxicity in the brain through activation of Type II GC receptors.


Interdisciplinary structural follow-up of surgical newborns: a prospective evaluation.
Information on physical and developmental outcomes of children with anatomical congenital anomalies (CAs) may indicate the need for early intervention and reduce impact on the child's life and parental burden. From 1999 to 2003, 101 children with CA (76.5% of initial survivors) were seen 6-monthly in a tertiary children's hospital. Growth, neurologic outcome, mental and psychomotor development as determined with the Bayley Scales of Infant Development, and categorization of predictive sociodemographic and medical variables of the children were evaluated prospectively and longitudinally. Congenital diaphragmatic hernia (CDH) and esophageal atresia patients showed impaired growth, that is, both height for age (-1.5 standard deviation score [SDS]) and weight for height (-1.0 SDS). Overall neurologic outcome was normal, however, suspect or abnormal for 40% of CDH patients. Overall mental development was normal, but psychomotor scores were significantly lower than the norm (95% confidence interval, 83.8-92.2 at 6 months and 87.9-98.5 at 24 months). Sex, maternal age, socioeconomic status, CA, severity-of-disease covariables, and need of medical appliances at home could predict negative outcome significantly (P < .05). The CA survivors show impaired growth and psychomotor developmental delay up to age 2 years. This warrants specific follow-up programs and infrastructure for these patients.


Breastfeeding and hospitalization for diarrheal and respiratory infection in the United Kingdom Millennium Cohort Study.
The objective of this study was to measure the effect of breastfeeding on hospitalization for diarrheal and lower respiratory tract infections in the first 8 months after birth in contemporary United Kingdom. The study was a population-based survey (sweep 1 of the United Kingdom Millennium Cohort Study). Data on infant feeding, infant health, and a range of confounding factors were available for 15,890 healthy, singleton, term infants who were born in 2000-2002. The main outcome measures were parental report of hospitalization for diarrhea and lower respiratory tract infection in the first 8 months after birth. Seventy percent of infants were breastfed (ever), 34% received breast milk for at least 4 months, and 1.2% were exclusively breastfed for at least 6 months. By 8 months of age, 12% of infants had been hospitalized (1.1% for diarrhea and 3.2% for lower respiratory tract infection). Data analyzed by month of age, with adjustment for confounders, show that exclusive breastfeeding, compared with not breastfeeding, protects against hospitalization for diarrhea and lower respiratory tract infection. The effect of partial breastfeeding is weaker. Population-attributable fractions suggest that an estimated 53% of diarrhea hospitalizations could have been prevented each month by exclusive breastfeeding and 31% by partial breastfeeding. Similarly, 27% of lower respiratory tract infection hospitalizations could have been prevented each month by exclusive breastfeeding and 25% by partial breastfeeding. The protective effect of breastfeeding for these outcomes wears off soon after breastfeeding cessation. Breastfeeding, particularly when exclusive and prolonged, protects against severe morbidity in contemporary United Kingdom. A population-level increase in exclusive, prolonged breastfeeding would be of considerable potential benefit for public health.


Anti-TNFalpha therapy of rheumatoid arthritis: what can we learn about chronic disease?
The importance of tumour necrosis factor (TNF)alpha in rheumatoid arthritis (RA) was initially proposed on the basis of analysis of cytokine gene regulation at the local site of the disease, the synovium. This was then verified in animal models and established in an extensive series of clinical trials, culminating in now 250000 treated patients with either of two approved TNF inhibitors, antibody or fusion protein. The degree and magnitude of clinical benefit has enabled analyses of the mechanism by which anti-TNF benefits, and hence insights into important steps in the disease process. It was found that essentially all aspects of RA were ameliorated, and important mechanisms of benefit involved diminution of multiple pro-inflammatory cytokines, adhesion molecules and chemokines, leading to reduced cell trafficking, reduced angiogenesis and most importantly halting of joint destruction. What of the problems? Safety is better than prior drugs, but there is a small increase in severe infections, smaller than might have been anticipated. Cost is the major drawback limiting greater use. In view of the central pathological processes down-regulated, and their role in many diseases, the early clinical success of anti-TNF in RA led to subsequent successful trials and registration in Crohn's disease and juvenile rheumatoid arthritis, and successful trials in ankylosing spondylitis, psoriasis and psoriatic arthritis. The era of anti-cytokine therapeutics is just dawning.


Introducing an online community into a clinical education setting: a pilot study of student and staff engagement and outcomes using blended learning.
There are growing reasons to use both information and communication functions of learning technologies as part of clinical education, but the literature offers few accounts of such implementations or evaluations of their impact. This paper details the process of implementing a blend of online and face-to-face learning and teaching in a clinical education setting and it reports on the educational impact of this innovation. This study designed an online community to complement a series of on-site workshops and monitored its use over a semester. Quantitative and qualitative data recording 43 final-year medical students' and 13 clinical educators' experiences with this blended approach to learning and teaching were analysed using access, adoption and quality criteria as measures of impact. The introduction of the online community produced high student ratings of the quality of learning and teaching and it produced student academic results that were equivalent to those from face-to-face-only learning and teaching. Staff had mixed views about using blended learning. Projects such as this take skilled effort and time. Strong incentives are required to encourage clinical staff and students to use a new mode of communication. A more synchronous or multi-channel communication feedback system might stimulate increased adoption. Cultural change in clinical teaching is also required before clinical education can benefit more widely from initiatives such as this.


Are patient-held vaccination records associated with improved vaccination coverage rates?
The goal was to determine whether patient-held vaccination records improve vaccination rates. The public-use files of the 2004-2006 National Immunization Survey, a national, validated survey of households with children 19 to 35 months of age, were used. The main outcome was up-to-date (UTD) vaccination status (4 diphtheria-tetanus-acellular pertussis/diphtheria-tetanus vaccine, 3 poliovirus vaccine, 1 measles vaccine, 3 Haemophilus influenza type B vaccine, and 3 hepatitis B vaccine doses), and the main predictor was the use of a vaccination record. Control variables were race/ethnicity, maternal education, poverty status, language, number of children in the home, state of residence, and number of health care providers. Overall, 80.8% of children were UTD, and 40.8% of children had vaccination records. Children with vaccination records were more likely to be UTD (83.9% vs 78.6%; P < .0001). The largest effects associated with vaccination records were seen for children with multiple providers, comparing with and without a vaccination record (82.8% vs 71.9%; P < .0001), those with low maternal education, (81.6% vs 72.9%; P < .0001), and those with > or =4 children in the household, (76% vs 69.6%; P < .004). Logistic regression predicting UTD status and controlling for race/ethnicity, maternal education, poverty level, language, number of children in the home, and number of vaccine providers revealed the vaccination record to be associated with a 62% increase in the odds of UTD status (odds ratio: 1.62 [95% confidence interval: 1.49-1.77]). Use of patient-held vaccination records is an easily implemented strategy that is associated with increased immunization rates. A greater effect was seen in groups at risk for underimmunization. Methods to incorporate and to ensure effective use of these records should be implemented.


Quantitation of substitutions at amino acid 70 in hepatitis C virus genotype 1b.
Substitutions of amino acid (aa) 70 in the core region of hepatitis C virus genotype 1b (HCV 1b) are a predictor of the non-virological response to pegylated interferon plus ribavirin (PEG-IFN/RBV) therapy. The aim of our study was to develop quantitative real-time reverse transcription polymerase chain reaction (qPCR) assays to quantify wild-type (70 W) and mutant (70 M) strains of HCV 1b. We used the TaqMan system to quantify strains 70 W and 70 M. Codon 70 in the HCV 1b core region can be either CGN or CAN, therefore degenerate TaqMan minor groove binder (MGB) probes with inosine were used. We determined detection limits, sensitivity and specificity of the methods developed. Direct sequencing and cloning of the HCV core region were used to confirm the reliability of our new system. Serum samples from 138 Chinese patients infected with HCV 1b were examined with the system we developed and compared with results obtained from the Roche TaqMan RT-PCR HCV RNA quantitation system. Degenerate MGB probes were able to clearly distinguish 70 W from 70 M. The detection limit was 10³ copies/mL. Cross-reactivity tests confirmed the specificity of our method. Our system can effectively quantify 70 W and 70 M for 99.6% of patients with HCV 1b. Further tests involving cloning and sequencing confirmed the reliability of our system. We developed an assay system using degenerate TaqMan MGB probes with inosine to quantify wild-type and mutant viral RNAs of the HCV 1b core region at aa 70. Our developed assay system had high levels of sensitivity and accuracy, and could prove useful in investigating dynamic changes during PEG-IFN/RBV therapy to assess virological responses.


Strain vs. charge mediated magnetoelectric coupling across the magnetic oxide/ferroelectric interfaces.
We utilize polarized neutron reflectometry (PNR) in consort with ab initio based density functional theory (DFT) calculations to study magnetoelectric coupling at the interface of a ferroelectric PbZr0.2Ti0.8O3 (PZT) and magnetic La0.67Sr0.33MnO3 (LSMO) heterostructure grown on a Nb-doped SrTiO3 (001) substrate. Functional device working conditions are mimicked by gating the heterostructure with a Pt top electrode to apply an external electric field, which alters the magnitude and switches the direction of the ferroelectric (FE) polarization, across the PZT layer. PNR results show that the gated PZT/LSMO exhibits interfacial magnetic phase modulation attributed to ferromagnetic (FM) to A-antiferromagnetic (A-AF) phase transitions resulting from hole accumulation. When the net FE polarization points towards the interface (positive), the interface doesn't undergo a magnetic phase transition and retains its global FM ordered state. In addition to changes in the interfacial magnetic ordering, the global magnetization of LSMO increases while switching the polarization from positive to negative and decreases vice versa. DFT calculations indicate that this enhanced magnetization also correlates with an out of plane tensile strain, whereas the suppressed magnetization for positive polarization is attributed to out of plane compressive strain. These calculations also show the coexistence of FM and A-AF phases at zero out of plane strain. Charge modulations throughout the LSMO layer appear to be unaffected by strain, suggesting that these charge mediated effects do not significantly change the global magnetization. Our PNR results and DFT calculations are in consort to verify that the interfacial magnetic modulations are due to co-action of strain and charge mediated effects with the strain and charge effects dominant at different length scale.


Radical Reactions of Alkyl 2-Bromo-2,2-difluoroacetates with Vinyl Ethers: "Omitted" Examples and Application for the Synthesis of 3,3-Difluoro-GABA.
Addition reactions of perfluoroalkyl radicals to ordinary or polyfluorinated alkenes have been frequently used to synthesize perfluoroalkylated organic compounds. Here ethyl/methyl 2-bromo-2,2-difluoroacetate, diethyl (bromodifluoromethyl)phosphonate, [(bromodifluoromethyl)sulfonyl]benzene, and ethyl 2-bromo-2-fluoroacetate were involved in Na2S2O4-mediated radical additions to vinyl ethers in the presence of alcohols to give difluoro or monofluoroacetyl-substituted acetals or corresponding difluoromethylphosphonate- and (difluoromethylphenyl)sulfonyl-substituted alkyl acetals. This methodology has also been applied as a key step in the synthesis of hitherto unknown 3,3-difluoro-GABA, completing the series of isomeric difluoro GABAs. Comparison of the pKa values of 3-fluoro- and 3,3-difluoro-GABA with that of the fluorine free parent compound showed that introduction of each fluorine lead to acidification of both the amino and the carboxyl functions by approximately one unit.


Reduced expression of proteolipid protein 2 increases ER stress-induced apoptosis and autophagy in glioblastoma.
Proteolipid protein 2 (PLP2) is an integral ion channel membrane protein of the endoplasmic reticulum. The protein has been shown to be highly expressed in many cancer types, but its importance in glioma progression is poorly understood. Using publicly available datasets (Rembrandt, TCGA and CGGA), we found that the expression of PLP2 was significantly higher in high-grade gliomas than in low-grade gliomas. We confirmed these results at the protein level through IHC staining of high-grade (n = 56) and low-grade glioma biopsies (n = 16). Kaplan-Meier analysis demonstrated that increased PLP2 expression was associated with poorer patient survival. In functional experiments, siRNA and shRNA PLP2 knockdown induced ER stress and increased apoptosis and autophagy in U87 and U251 glioma cell lines. Inhibition of autophagy with chloroquine augmented apoptotic cell death in U87- and U251-siPLP2 cells. Finally, intracranial xenografts derived from U87- and U251-shPLP2 cells revealed that loss of PLP2 reduced glioma growth in vivo. Our results therefore indicate that increased PLP2 expression promotes GBM growth and that PLP2 represents a potential future therapeutic target.


Replacing corn with glycerol in diets for transition dairy cows.
Expansion of the biofuels industry has increased the availability of glycerol as an alternative feed for dairy cows. The objective of this study was to determine the effects of glycerol on feed intake, milk production, rumen volatile fatty acids, and metabolic parameters in transition dairy cows. Multiparous Holstein cows were fed diets containing either high-moisture corn (n=11) or glycerol (n=12) from -28 to +56 d relative to calving. Glycerol was included at 11.5 and 10.8% of the ration dry matter for the pre- and postpartum diets, respectively. Prepartum feed intake was not changed by glycerol feeding (14.9 vs. 14.6 kg/d, control vs. glycerol) nor did postpartum feed intake differ (19.8 vs. 20.7 kg/d, control vs. glycerol). Overall milk yield did not differ (35.8 vs. 37 kg/d, control vs. glycerol) and milk composition, milk urea nitrogen, somatic cells, and energy balance were not different with glycerol feeding. Blood glucose content was decreased in cows fed glycerol during the prepartum period (59.1 vs. 53.4 mg/dL), and β-hydroxybutyrate concentration was increased (0.58 vs. 0.82 mmol/L, control vs. glycerol). Concentrations of blood nonesterified fatty acids did not differ between the treatment groups, and no response to glycerol for blood metabolites during the postpartum period was observed. Total rumen volatile fatty acid concentrations (mmol/L) did not differ between treatments, but proportions of rumen propionate and butyrate were greater for cows fed glycerol (22.7 vs. 28.6% of propionate, control vs. glycerol; and 11.5 vs. 15.3% of butyrate, control vs. glycerol) at the expense of acetate (61.4 vs. 51.5%, control vs. glycerol). These data indicate that glycerol is a suitable replacement for corn grain in diets for transition dairy cows.


Postnatal depressive symptoms and child psychological development at 10 years: a prospective study of longitudinal data from the South African Birth to Twenty cohort.
In high-income countries, maternal postnatal depression is associated with adverse outcomes in the child. However, few studies have investigated this relation in countries of low and middle income. Furthermore, to our knowledge, no studies have followed up cohorts into later childhood. We aimed to investigate whether maternal depression 6 months after birth is associated with psychological difficulties in a socioeconomically disadvantaged South African cohort of children at age 10 years. Birth to Twenty is a prospective, longitudinal, birth-cohort study based in the Soweto area of Johannesburg, South Africa. Mothers and children in this cohort have been followed up at timepoints ranging from before birth to age 10 years. Maternal mood was measured at 6 months with the Pitt depression inventory and at 10 years with the Centre for Epidemiologic Studies depression scale (CES-D). Child psychological functioning was assessed at 10 years with the South African child assessment schedule (SACAS). Our primary outcome was psychological development of children at age 10 years, measured by total score on the SACAS. Secondary outcomes were scores on externalising and internalising subscales of the SACAS. We used t tests to compare psychological outcomes between children whose mother had postnatal depression at 6 months and those whose mother did not have postnatal depression. We examined associations between maternal postnatal depression and child psychological outcomes by multivariate linear-regression analysis, adjusting for socioeconomic status and maternal depression at 10 years, and we used logistic regression to provide odds ratios for associations identified by linear regression. 1866 mothers completed the Pitt depression inventory 6 months after the birth of their child; of these, 453 (24%) had symptoms of postnatal depression. At the 10-year assessment, 1012 mothers completed the CES-D questionnaire, of whom 747 (74%) were judged to have depression. Sociodemographic characteristics did not differ between mothers with and without depression at both 6 months and 10 years. After adjusting for socioeconomic status and maternal depression at 10 years, children whose mothers had postnatal depression at 6 months were more than twice as likely to have significant psychological difficulties 10 years later compared with children whose mothers did not have postnatal depression at 6 months (adjusted odds ratio 2·26, 95% CI 1·23-4·16). Maternal postnatal depression is associated with adverse psychological outcomes in children up to 10 years later in countries of low and middle income. In view of the increased prevalence of postnatal depression in these settings, this finding has important implications for policy and interventions for children and their mothers. Wellcome Trust (UK), Medical Research Council of South Africa, Human Science Research Council (South Africa), University of the Witwatersrand.


Intrapulmonary metastasis of non-small cell lung cancer: a prognostic assessment.
According to the revised TNM classification in 1997, intrapulmonary metastasis within the same lobe of the primary tumor is designated as T4 and intrapulmonary metastasis in a different lobe is M1. However, their prognostic implications remain unclear. To assess their prognoses, we retrospectively analyzed the postoperative survival of patients with and without intrapulmonary metastasis. From January 1982 to December 1996, 2340 patients with non-small cell lung cancer underwent surgical resection. The survival of patients having complete resection (n = 1534) was analyzed according to their intrapulmonary metastasis status: patients without intrapulmonary metastasis (n = 1393), those with metastasis in the same lobe (n = 105), and those with metastasis in a different lobe (n = 18). For comparison, patients with T4 disease without intrapulmonary metastasis in the same lobe (n = 54) and those with M1 disease without metastasis in a different lobe (distant M1, n = 18) were also analyzed. The overall 5-year survivals were as follows: no intrapulmonary metastasis, 60%; stage T4 disease with no intrapulmonary metastasis, 34%; pulmonary metastasis in the same lobe, 34%; pulmonary metastasis in a different lobe, 11%; and distant M1, 6%. The differences in survival between patients with no pulmonary metastasis and those with metastasis in the same lobe (P <.001, log-rank test) and between patients with metastasis in the same lobe and those with distant M1 (P <.001) were significant. In contrast, there was no significant difference between patients with metastasis in the same lobe and those with T4 disease and no intrapulmonary metastasis or between patients with metastasis to a different lobe and those with distant M1. Prognostically, intrapulmonary metastasis within the same lobe of the primary tumor was comparable with T4 and that in a different lobe was comparable with M1. In terms of postoperative prognosis, the revised TNM classification for intrapulmonary metastasis seems to be appropriate.


Renal Deletion of 12 kDa FK506-Binding Protein Attenuates Tacrolimus-Induced Hypertension.
Tacrolimus is a widely used immunosuppressive drug that inhibits the phosphatase calcineurin when bound to the 12 kDa FK506-binding protein (FKBP12). When this binding occurs in T cells, it leads to immunosuppression. Tacrolimus also causes side effects, however, such as hypertension and hyperkalemia. Previously, we reported that tacrolimus stimulates the renal thiazide-sensitive sodium chloride cotransporter (NCC), which is necessary for the development of hypertension. However, it was unclear if tacrolimus-induced hypertension resulted from tacrolimus effects in renal epithelial cells directly or in extrarenal tissues, and whether inhibition of calcineurin was required. To address these questions, we developed a mouse model in which FKBP12 could be deleted along the nephron. FKBP12 disruption alone did not cause phenotypic effects. When treated with tacrolimus, however, BP and the renal abundance of phosphorylated NCC were lower in mice lacking FKBP12 along the nephron than in control mice. Mice lacking FKBP12 along the nephron also maintained a normal relationship between plasma potassium levels and the abundance of phosphorylated NCC with tacrolimus treatment. In cultured cells, tacrolimus inhibited dephosphorylation of NCC. Together, these results suggest that tacrolimus causes hypertension predominantly by inhibiting calcineurin directly in cells expressing NCC, indicating thiazide diuretics may be particularly effective for lowering BP in tacrolimus-treated patients with hypertension.


Integrated acousto-optic heterodyning device modules in LiNbO(3) substrate.
This paper reports the realization of integrated acousto-optic (AO) device modules that combine a wideband AO Bragg cell, an ion-milled Bragg diffraction grating, and a titanium-indiffused protonexchanged waveguide lens in a Y-cut LiNbO(3) substrate, 1 x 8 x 16 mm(3) in size, to perform optical heterodyning, and their application to rf signal processing. These integrated AO heterodyne modules have demonstrated the capabilities for channelized detection of the amplitude, the frequency, and the phase of wideband rf signals and thus the capability to perform interferometric rf spectral analysis with significantly improved performances over the conventional AO Bragg cells. The single-unit (basic) modules have provided single-tone simultaneous and two-tone third-order spurious-free dynamic ranges of 51 and 40 dB, respectively, and a bandwidth of 205 MHz centered at 350 MHz at the optical wavelength of 0.6328 microm, the optical power of 1.0 mW, and the drive power of 50 mW/rf signal input. Furthermore the dual-unit modu es that consist of a pair of identical basic heterodyne devices in the same LiNbO(3) waveguide substrate, also 1 x 8 x 16 mm(3) in size, have been constructed and used to measure the angle of arrival of the rf signals.


SERCA2a tyrosine nitration coincides with impairments in maximal SERCA activity in left ventricles from tafazzin-deficient mice.
The sarco/endoplasmic reticulum Ca2+ -ATPase (SERCA) is imperative for normal cardiac function regulating both muscle relaxation and contractility. SERCA2a is the predominant isoform in cardiac muscles and is inhibited by phospholamban (PLN). Under conditions of oxidative stress, SERCA2a may also be impaired by tyrosine nitration. Tafazzin (Taz) is a mitochondrial-specific transacylase that regulates mature cardiolipin (CL) formation, and its absence leads to mitochondrial dysfunction and excessive production of reactive oxygen/nitrogen species (ROS/RNS). In the present study, we examined SERCA function, SERCA2a tyrosine nitration, and PLN expression/phosphorylation in left ventricles (LV) obtained from young (3-5 months) and old (10-12 months) wild-type (WT) and Taz knockdown (TazKD ) male mice. These mice are a mouse model for Barth syndrome, which is characterized by mitochondrial dysfunction, excessive ROS/RNS production, and dilated cardiomyopathy (DCM). Here, we show that maximal SERCA activity was impaired in both young and old TazKD LV, a result that correlated with elevated SERCA2a tyrosine nitration. In addition PLN protein was decreased, and its phosphorylation was increased in TazKD LV compared with control, which suggests that PLN may not contribute to the impairments in SERCA function. These changes in expression and phosphorylation of PLN may be an adaptive response aimed to improve SERCA function in TazKD mice. Nonetheless, we demonstrate for the first time that SERCA function is impaired in LVs obtained from young and old TazKD mice likely due to elevated ROS/RNS production. Future studies should determine whether improving SERCA function can improve cardiac contractility and pathology in TazKD mice.


CD28 expressed on malignant plasma cells induces a prosurvival and immunosuppressive microenvironment.
Interactions between the malignant plasma cells of multiple myeloma and stromal cells within the bone marrow microenvironment are essential for myeloma cell survival, mirroring the same dependence of normal bone marrow-resident long-lived plasma cells on specific marrow niches. These interactions directly transduce prosurvival signals to the myeloma cells and also induce niche production of supportive soluble factors. However, despite their central importance, the specific molecular and cellular components involved remain poorly characterized. We now report that the prototypic T cell costimulatory receptor CD28 is overexpressed on myeloma cells during disease progression and in the poor-prognosis subgroups and plays a previously unrecognized role as a two-way molecular bridge to support myeloid stromal cells in the microenvironment. Engagement by CD28 to its ligand CD80/CD86 on stromal dendritic cell directly transduces a prosurvival signal to myeloma cell, protecting it against chemotherapy and growth factor withdrawal-induced death. Simultaneously, CD28-mediated ligation of CD80/CD86 induces the stromal dendritic cell to produce the prosurvival cytokine IL-6 (involving novel cross-talk with the Notch pathway) and the immunosuppressive enzyme IDO. These findings identify CD28 and CD80/CD86 as important molecular components of the interaction between myeloma cells and the bone marrow microenvironment, point to similar interaction for normal plasma cells, and suggest novel therapeutic strategies to target malignant and pathogenic (e.g., in allergy and autoimmunity) plasma cells.


A six-member SNP assay on the iPlex MassARRAY platform provides a rapid and affordable alternative for typing major African Staphylococcus aureus types.
Data on the clonal distribution of Staphylococcus aureus in Africa are scanty, partly due to the high costs and long turnaround times imposed by conventional genotyping methods such as spa and multilocus sequence typing (MLST), which means there is a need for alternative typing approaches. This study evaluated the discriminatory power, cost of and time required for genotyping Kenyan staphylococcal isolates using iPlex MassARRAY compared to conventional methods. Fifty-four clinical S. aureus isolates from three counties were characterized using iPlex MassARRAY, spa and MLST typing methods. Ten single-nucleotide polymorphisms (SNPs) from the S. aureus MLST loci were assessed by MassARRAY. The MassARRAY assay identified 14 unique SNP genotypes, while spa typing and MLST revealed 22 spa types and 21 sequence types (STs) that displayed unique regional distribution. spa type t355 (ST152) was the dominant type overall while t037/t2029 (ST 241) dominated among the methicillin-resistant S. aureus (MRSA) isolates. MassARRAY showed 83 % and 82 % accuracy against spa typing and MLST, respectively, in isolate classification. Moreover, MassARRAY identified all MRSA strains and a novel spa type. MassARRAY had a reduced turnaround time (<12 h) compared to spa typing (4 days) and MLST (20 days). The MassARRAY reagent and consumable costs per isolate were approximately $18 USD compared to spa typing ($30 USD) and MLST ($126 USD). This study demonstrated that iPlex MassARRAY can be adapted as a useful surveillance tool to provide a faster, more affordable and fairly accurate method for genotyping African S. aureus isolates to identify clinically significant genotypes, MRSA strains and emerging strain types.


A simple visual system without neurons in jellyfish larvae.
Earlier detailed studies of cnidarian planula larvae have revealed a simple nervous system but no eyes or identifiable light sensing structures. Here, we describe the planula of a box jellyfish, Tripedalia cystophora, and report that these larvae have an extremely simple organization with no nervous system at all. Their only advanced feature is the presence of 10-15 pigment-cup ocelli, evenly spaced across the posterior half of the larval ectoderm. The ocelli are single cell structures containing a cup of screening pigment filled with presumably photosensory microvilli. These rhabdomeric photoreceptors have no neural connections to any other cells, but each has a well-developed motor-cilium, appearing to be the only means by which light can control the behaviour of the larva. The ocelli are thus self-contained sensory-motor entities, making a nervous system superfluous.


Dimethylglycine Deficiency and the Development of Diabetes.
Experimental studies have suggested possible protective effects of dimethylglycine (DMG) on glucose metabolism. DMG is degraded to glycine through a DMG-dehydrogenase (DMGDH)-catalyzed reaction, and this is the only known pathway for the breakdown of DMG in mammals. In this study, we aimed to identify the strongest genetic determinant of circulating DMG concentration and to investigate its associations with metabolic traits and incident diabetes. In the cohort with full metabolomics data (n = 709), low plasma levels of DMG were significantly associated with higher blood glucose levels (P = 3.9E(-4)). In the genome-wide association study (GWAS) of the discovery cohort (n = 5,205), the strongest genetic signal of plasma DMG was conferred by rs2431332 at the DMGDH locus, where the major allele was associated with lower DMG levels (P = 2.5E(-15)). The same genetic variant (major allele of rs2431332) was also significantly associated with higher plasma insulin (P = 0.019), increased HOMA insulin resistance (P = 0.019), and an increased risk of incident diabetes (P = 0.001) in the pooled analysis of the discovery cohort together with the two replication cohorts (n = 20,698 and n = 7,995). These data are consistent with a possible causal role of DMG deficiency in diabetes development and encourage future studies examining if inhibition of DMGDH, or alternatively, supplementation of DMG, might prove useful for the treatment/prevention of diabetes.


Elusive copy number variation in the mouse genome.
Array comparative genomic hybridization (aCGH) to detect copy number variants (CNVs) in mammalian genomes has led to a growing awareness of the potential importance of this category of sequence variation as a cause of phenotypic variation. Yet there are large discrepancies between studies, so that the extent of the genome affected by CNVs is unknown. We combined molecular and aCGH analyses of CNVs in inbred mouse strains to investigate this question. Using a 2.1 million probe array we identified 1,477 deletions and 499 gains in 7 inbred mouse strains. Molecular characterization indicated that approximately one third of the CNVs detected by the array were false positives and we estimate the false negative rate to be more than 50%. We show that low concordance between studies is largely due to the molecular nature of CNVs, many of which consist of a series of smaller deletions and gains interspersed by regions where the DNA copy number is normal. Our results indicate that CNVs detected by arrays may be the coincidental co-localization of smaller CNVs, whose presence is more likely to perturb an aCGH hybridization profile than the effect of an isolated, small, copy number alteration. Our findings help explain the hitherto unexplored discrepancies between array-based studies of copy number variation in the mouse genome.


Strong effects of coral species on the diversity and structure of reef fish communities: A multi-scale analysis.
While there is increasing evidence for habitat specialization in coral reef fishes, the extent to which different corals support different fish communities is not well understood. Here we quantitatively assess the relative importance of different coral species in structuring fish communities and evaluate whether sampling scale and coral colony size affect the perceived strength of fish-habitat relationships. Fish communities present on colonies of eight coral species (Porites cylindrica, Echinopora horrida, Hydnophora rigida, Stylophora pistillata, Seriatopora hystrix, Acropora formosa, A. tenuis and A. millepora) were examined in the Lizard Island lagoon, Great Barrier Reef, Australia. Additionally, the differences in fish communities supported by three coral species (P. cylindrica, E. horrida, H. rigida) were investigated at three spatial scales of sampling (2x2 m, 1x1 m, 0.5x0.5 m). Substantial differences in fish communities were observed across the different coral species, with E. horrida and H. rigida supporting the most fish species and individuals. Coral species explained more of the variability in fish species richness (20.9-53.6%), than in fish abundance (0-15%). Most coral species supported distinctive fish communities, with dissimilarities ranging from 50 to 90%. For three focal coral species, a greater amount of total variation in fish species richness and fish abundance was evident at a larger scale of sampling. Together, these results indicate that the structure of reef fish communities is finely tuned to coral species. Loss of preferred coral species could have profound effects on reef fish biodiversity, potentially more so than would be predicted on the basis of declining coral cover alone.


Alanine aminotransferase decreases with age: the Rancho Bernardo Study.
Serum alanine aminotransferase (ALT) is a marker of liver injury. The 2005 American Gastroenterology Association Future Trends Committee report states that serum ALT levels remain constant with age. This study examines the association between serum ALT and age in a community-dwelling cohort in the United States. A cross-sectional study of 2,364 (54% female) participants aged 30-93 years from the Rancho Bernardo Study cohort who attended a research clinic visit in 1984-87. Demographic, metabolic co-variates, ALT, bilirubin, gamma glutamyl transferase (GGT), albumin, and adiposity signaling biomarkers (leptin, IL-6, adiponectin, ghrelin) were measured. Participants were divided into four-groups based upon age quartile, and multivariable-adjusted least squares of means (LSM) were examined (p for trend <0.05). ALT decreased with increasing age, with mean ALT levels (IU/L) of 23, 21, 20, and 17 for those between quartile ages 30-62, 63-71, 72-77, and 78-93 years (p<0.0001). Trends of decreasing LSM ALT with age and the decreasing prevalence of categorically defined elevated serum ALT with age remained robust after adjusting for sex, alcohol use, metabolic syndrome components, and biomarkers of adiposity (p-value <0.0001), and was not materially changed after adjusting for bilirubin, GGT, and albumin. ALT levels decrease with age in both men and women independent of metabolic syndrome components, adiposity signaling biomarkers, and other commonly used liver function tests. Further studies are needed to understand the mechanisms responsible for a decline in ALT with age, and to establish the optimal cut-point of normal ALT in the elderly.


Phencyclidine analogs and precursors: rotarod and lethal dose studies in the mouse.
A series of phencyclidine (PCP) related analogs, carbonitrile synthetic precursors and two monohydroxylated metabolites were compared pharmacologically in mice for their ability to produce ataxia using the rotarod method and toxicologically for their acute 4-hr lethality. The slope of the PCP dose-ataxic response curve was steeper than those of diazepam, pentobarbital, morphine and ketocyclazocine but not the slope of the sigma agonist, N-allylnormetazocine curve. Responses for all analogs, metabolites and precursors produced curves parallel to that of PCP. Ataxia potencies of all PCP-related compounds ranged from 0.05 to 2.15 X PCP and durations of action ranged from 18 to 65 min. N-ethyl-1-phenylcyclohexylamine, 1-[1-(2-thienyl)-cyclohexyl]-piperidine and 1-[1-(2-thienyl)-cyclohexyl]-pyrrolidine were most potent and least potent were 1-(1-phenyl-cyclohexyl)-4-methylpiperidine, the phenyl and thienyl morpholines and 4-phenyl-4-piperidinocyclohexanol. Among the PCP analogs, modifying the piperidine or aromatic ring effected changes only in potency. Seizures and respiratory depression characterized the lethal effects of PCP, its analogs, metabolites and precursors. However, the precursors failed to elicit the stereotyped movements and hyperactivity that preceded seizures produced by the other compounds. Overall potencies for lethality relative to PCP covered a narrow range (0.16-1.83) with the carbonitrile precursors being most potent. Therapeutic indices indicated relatively large margins of safety for 1-[1-(2-thienyl)-cyclohexyl]-piperidine, 1-[1-(2-thienyl)-cyclohexyl]-piperidine, N-ethyl-1-phenylcyclohexylamine and ketamine and the smallest were for 1-(1-phenylcyclohexyl)-4-methylpiperidine, the metabolite 4-phenyl-4-piperidinocyclohexanol and the three precursors.(ABSTRACT TRUNCATED AT 250 WORDS)


On the assessment of preferences for health and duration: maximal endurable time and better than dead preferences.
Several studies revealed difficulties with the valuation and analysis of health states deemed worse than dead. These problems may be linked to maximal endurable time (MET) preferences, the phenomenon that for severe states better than dead (BTD), shorter durations are often preferred to longer durations. To test the association between the duration of health states and their valuation. A representative sample of 123 Dutch respondents (age range, 18-45 years) valued 5 EQ-5D health states. With a straightforward method using BTD preferences, respondents indicated whether a state of a certain duration is better, equal to, or worse than dead. To validate these BTD preferences, MET preferences (whether a longer duration of a health state is better, equal, or worse than a shorter duration) were collected. BTD and MET preferences were strongly related (P < 0.001). For severe health states, although still judged as better than dead, BTD preferences curved downwards with increasing duration. Such curved BTD patterns occurred in 28% of the respondents, especially for more severe states (P < 0.001). BTD preferences revealed that the value of moderate and severe states declines with increasing duration, suggesting that health and duration interact. For states worse than dead versus states better than dead, traditional valuation techniques have the drawback that different preference questions are used. Using BTD preferences, however, a single simple preference question can assess states better than dead, as well as states worse than dead.


Revealing the spatial shifting pattern of COVID-19 pandemic in the United States.
We describe the use of network modeling to capture the shifting spatiotemporal nature of the COVID-19 pandemic. The most common approach to tracking COVID-19 cases over time and space is to examine a series of maps that provide snapshots of the pandemic. A series of snapshots can convey the spatial nature of cases but often rely on subjective interpretation to assess how the pandemic is shifting in severity through time and space. We present a novel application of network optimization to a standard series of snapshots to better reveal how the spatial centres of the pandemic shifted spatially over time in the mainland United States under a mix of interventions. We find a global spatial shifting pattern with stable pandemic centres and both local and long-range interactions. Metrics derived from the daily nature of spatial shifts are introduced to help evaluate the pandemic situation at regional scales. We also highlight the value of reviewing pandemics through local spatial shifts to uncover dynamic relationships among and within regions, such as spillover and concentration among states. This new way of examining the COVID-19 pandemic in terms of network-based spatial shifts offers new story lines in understanding how the pandemic spread in geography.


Persecutory delusions: a cognitive perspective on understanding and treatment.
A spectrum of severity of paranoia (unfounded thoughts that others are deliberately intending to cause harm) exists within the general population. This is unsurprising: deciding whether to trust or mistrust is a vital aspect of human cognition, but accurate judgment of others' intentions is challenging. The severest form of paranoia is persecutory delusions, when the ideas are held with strong conviction. This paper presents a distillation of a cognitive approach that is being translated into treatment for this major psychiatric problem. Persecutory delusions are viewed as threat beliefs, developed in the context of genetic and environmental risk, and maintained by several psychological processes including excessive worry, low self-confidence, intolerance of anxious affect and other internal anomalous experiences, reasoning biases, and the use of safety-seeking strategies. The clinical implication is that safety has to be relearned, by entering feared situations after reduction of the influence of the maintenance factors. An exciting area of development will be a clinical intervention science of how best to enhance learning of safety to counteract paranoia.


Angiotensin II up-regulates the leukemia-associated Rho guanine nucleotide exchange factor (RhoGEF), a regulator of G protein signaling domain-containing RhoGEF, in vascular smooth muscle cells.
In vascular smooth muscle, stimulation of heterotrimeric G protein-coupled receptors (GPCRs) by various contractile agonists activates intracellular signaling molecules to result in an increase in cytosolic Ca2+ and the subsequent phosphorylation of myosin light chain (MLC) by Ca2+/calmodulin-dependent MLC kinase. In addition, a portion of agonist-induced contraction is partially mediated by the Ca2+-independent activation of the small G protein RhoA and a downstream target, Rho-kinase. The activation of RhoA is controlled by several regulatory proteins, including guanine nucleotide exchange factors (GEFs). GEFs activate RhoA by promoting the release of GDP and then facilitating the binding of GTP. There are three Rho-specific GEFs (RhoGEFs) in vascular smooth muscle that contain a binding domain [regulator of G protein signaling (RGS) domain] capable of linking GPCRs to RhoA activation: PDZ-RhoGEF, leukemia-associated RhoGEF (LARG), and p115RhoGEF. We hypothesized that RGS domain-containing RhoGEFs, especially LARG, participate in linking GPCR to RhoA activation in vascular smooth muscle. We observed that angiotensin II up-regulates LARG via the AT1 receptor, and this up-regulation is signaled via the phosphatidylinositol 3-kinase pathway. Furthermore, angiotensin II treatment caused a small, but significant, increase in the component of contractile responses sensitive to Rho-kinase antagonism. These observations support the hypothesis that RhoGEFs, particularly LARG, participate in linking GPCR to RhoA activation in vascular smooth muscle.


Simplified Bayesian sensitivity analysis for mismeasured and unobserved confounders.
We examine situations where interest lies in the conditional association between outcome and exposure variables, given potential confounding variables. Concern arises that some potential confounders may not be measured accurately, whereas others may not be measured at all. Some form of sensitivity analysis might be employed, to assess how this limitation in available data impacts inference. A Bayesian approach to sensitivity analysis is straightforward in concept: a prior distribution is formed to encapsulate plausible relationships between unobserved and observed variables, and posterior inference about the conditional exposure-disease relationship then follows. In practice, though, it can be challenging to form such a prior distribution in both a realistic and simple manner. Moreover, it can be difficult to develop an attendant Markov chain Monte Carlo (MCMC) algorithm that will work effectively on a posterior distribution arising from a highly nonidentified model. In this article, a simple prior distribution for acknowledging both poorly measured and unmeasured confounding variables is developed. It requires that only a small number of hyperparameters be set by the user. Moreover, a particular computational approach for posterior inference is developed, because application of MCMC in a standard manner is seen to be ineffective in this problem.


Advancing Leadership Skills: A Multiyear Examination of LEND Trainee Self-Efficacy Growth.
Purpose The current healthcare system requires Maternal and Child Health (MCH) professionals with strong interdisciplinary leadership competence. MCH training programs utilize a conceptual framework for leadership and 12 validated MCH Leadership Competencies. Examining Trainee Perceived Leadership Competence (TPLC) through the competencies has the potential to inform our understanding of leadership development. Description Five cohorts of NH-ME leadership education in neurodevelopmental disabilities trainees (n = 102) completed the MCH Leadership Competencies Self-Assessment at three time points. Paired-sample t tests examined TPLC scores. A one-way analysis of variance tested for statistically significant differences in mean difference scores. A General Linear Model was used to examine the extent to which TPLC scores changed when controlling for specific variables. Assessment Statistically significant differences in mean scores between Time 1 and Time 3 were found. Cohen's d effect sizes fell in the moderate range. A one-way ANOVA demonstrated significant differences between groups in the spheres of self and others. TPLC mean scores between Time 1 and Time 3 in the sphere of wider community had the highest increases in four out of five cohorts. Age, discipline, experience, and relationship to disability did not contribute to the model. Conclusion On average, cohorts began the year with very different evaluations of their leadership competence but finished the year with similar scores. This suggests participation in the NH-ME LEND Program consistently supported the development of leadership self-identity. Small sample sizes limit the ability to draw definitive conclusions from these results. Further study with a larger sample may reveal relationships between cohort characteristics and change scores.


Identification of hierarchical chromatin domains.
The three-dimensional structure of the genome is an important regulator of many cellular processes including differentiation and gene regulation. Recently, technologies such as Hi-C that combine proximity ligation with high-throughput sequencing have revealed domains of self-interacting chromatin, called topologically associating domains (TADs), in many organisms. Current methods for identifying TADs using Hi-C data assume that TADs are non-overlapping, despite evidence for a nested structure in which TADs and sub-TADs form a complex hierarchy. We introduce a model for decomposition of contact frequencies into a hierarchy of nested TADs. This model is based on empirical distributions of contact frequencies within TADs, where positions that are far apart have a greater enrichment of contacts than positions that are close together. We find that the increase in contact enrichment with distance is stronger for the inner TAD than for the outer TAD in a TAD/sub-TAD pair. Using this model, we develop the TADtree algorithm for detecting hierarchies of nested TADs. TADtree compares favorably with previous methods, finding TADs with a greater enrichment of chromatin marks such as CTCF at their boundaries. A python implementation of TADtree is available at http://compbio.cs.brown.edu/software/ email@example.com Supplementary data are available at Bioinformatics online.


Chemical-gene relation extraction using recursive neural network.
In this article, we describe our system for the CHEMPROT task of the BioCreative VI challenge. Although considerable research on the named entity recognition of genes and drugs has been conducted, there is limited research on extracting relationships between them. Extracting relations between chemical compounds and genes from the literature is an important element in pharmacological and clinical research. The CHEMPROT task of BioCreative VI aims to promote the development of text mining systems that can be used to automatically extract relationships between chemical compounds and genes. We tested three recursive neural network approaches to improve the performance of relation extraction. In the BioCreative VI challenge, we developed a tree-Long Short-Term Memory networks (tree-LSTM) model with several additional features including a position feature and a subtree containment feature, and we also applied an ensemble method. After the challenge, we applied additional pre-processing steps to the tree-LSTM model, and we tested the performance of another recursive neural network model called Stack-augmented Parser Interpreter Neural Network (SPINN). Our tree-LSTM model achieved an F-score of 58.53% in the BioCreative VI challenge. Our tree-LSTM model with additional pre-processing and the SPINN model obtained F-scores of 63.7 and 64.1%, respectively.Database URL: https://github.com/arwhirang/recursive_chemprot.


A new minimally-invasive method for microinjection into the mouse spinal dorsal horn.
Noninvasive gene delivery to the spinal dorsal horn (SDH) remains challenging because existing methods to directly microinject vectors require laminectomy, which leads to tissue damage and inflammation. Such responses might hamper accurate readouts of cellular and behavioural effects of an introduced gene. Here we develop a new minimally-invasive SDH microinjection technique without the need of laminectomy in which a microcapillary is inserted into the SDH parenchyma through an intervertebral space. Using this method, we microinjected adeno-associated virus with an astrocytic promoter into the SDH and achieved efficient gene expression in an astrocyte-specific manner without gliosis, neuronal loss or inflammation. Furthermore, astrocytic loss- and gain-of-function of the transcription factor STAT3 by expressing a dominant-negative form and a constitutive-active form of STAT3, respectively, demonstrated the necessity and sufficiency of astrocytic STAT3 in the maintenance of neuropathic pain following peripheral nerve injury, a debilitating chronic pain state in which currently available treatments are frequently ineffective. Thus, our technique enables manipulation of gene expression in cell type- and spatial-specific manners without adverse effects, and may be useful for research in SDH physiology and pathology.


Genetic analyses of the human eye colours using a novel objective method for eye colour classification.
In this study, we present a new objective method for measuring the eye colour on a continuous scale that allows researchers to associate genetic markers with different shades of eye colour. With the use of the custom designed software Digital Iris Analysis Tool (DIAT), the iris was automatically identified and extracted from high resolution digital images. DIAT was made user friendly with a graphical user interface. The software counted the number of blue and brown pixels in the iris image and calculated a Pixel Index of the Eye (PIE-score) that described the eye colour quantitatively. The PIE-score ranged from -1 to 1 (brown to blue). The software eliminated the need for user based interpretation and qualitative eye colour categories. In 94% (570) of 605 analyzed eye images, the iris region was successfully extracted and a PIE-score was calculated. A very high correlation between the PIE-score and the human perception of eye colour was observed. The correlations between the PIE-scores and the six IrisPlex SNPs (HERC2 rs12913832, OCA2 rs1800407, SLC24A4 rs12896399, TYR rs1393350, SLC45A2 rs16891982 and IRF4 rs12203592) were analyzed in 570 individuals. Significant differences (p<10(-6)) in the PIE-scores of the individuals typed as HERC2 rs12913832 G (PIE=0.99) and rs12913832 GA (PIE=-0.71) or A (PIE=-0.87) were observed. We adjusted for the effect of HERC2 rs12913832 and showed that the quantitative PIE-scores were significantly associated with SNPs with minor effects (OCA2 rs1800407, SLC24A4 rs12896399 and TYR rs1393350) on the eye colour. We evaluated the two published prediction models for eye colour (IrisPlex [1] and Snipper[2]) and compared the predictions with the PIE-scores. We found good concordance with the prediction from individuals typed as HERC2 rs12913832 G. However, both methods had difficulties in categorizing individuals typed as HERC2 rs12913832 GA because of the large variation in eye colour in HERC2 rs12913832 GA individuals. With the use of the DIAT software and the PIE-score, it will be possible to automatically compare the iris colour of large numbers of iris images obtained by different studies and to perform large meta-studies that may reveal loci with small effects on the eye colour.


Impact of economic, regulatory, and patent policies on innovation in cancer chemoprevention.
Chemoprevention agents are an emerging new scientific area that holds out the promise of delaying or avoiding a number of common cancers. These new agents face significant scientific, regulatory, and economic barriers, however, which have limited investment in their research and development (R&D). These barriers include above-average clinical trial scales, lengthy time frames between discovery and Food and Drug Administration approval, liability risks (because they are given to healthy individuals), and a growing funding gap for early-stage candidates. The longer time frames and risks associated with chemoprevention also cause exclusivity time on core patents to be limited or subject to significant uncertainties. We conclude that chemoprevention uniquely challenges the structure of incentives embodied in the economic, regulatory, and patent policies for the biopharmaceutical industry. Many of these policy issues are illustrated by the recently Food and Drug Administration-approved preventive agents Gardasil and raloxifene. Our recommendations to increase R&D investment in chemoprevention agents include (a) increased data exclusivity times on new biological and chemical drugs to compensate for longer gestation periods and increasing R&D costs; chemoprevention is at the far end of the distribution in this regard; (b) policies such as early-stage research grants and clinical development tax credits targeted specifically to chemoprevention agents (these are policies that have been very successful in increasing R&D investment for orphan drugs); and (c) a no-fault liability insurance program like that currently in place for children's vaccines.


Clonal analysis of a human antibody response. Quantitation of precursors of antibody-producing cells and generation and characterization of monoclonal IgM, IgG, and IgA to rabies virus.
We quantitated and characterized the changes in the human B cell repertoire, at the clonal level, before and after immunization with rabies virus. Moreover, we generated 10 monoclonal cell lines producing IgM, IgG, and IgA antibodies to the virus. We found that in healthy subjects, not previously exposed to the virus, nearly 2% of the circulating B lymphocytes were committed to the production of antibodies that bound the virus. These B cells expressed the surface CD5 molecule. The antibodies they produced were polyreactive IgM that displayed a relatively low affinity for the virus components (Kd, 1.0-2.4 x 10(-6) g/microliters). After immunization, different anti-virus (IgG and IgA) antibody-producing cells consistently appeared in the circulation and increased from less than 0.005% to greater than 10% of the total B cells committed to the production of IgG and IgA, respectively. Most of such B cells do not express CD5 and produce monoreactive antibodies of high affinity for rabies virus (Kd, 6.5 x 10(-9) to 1.2 x 10(-10) g/microliters). One of these IgG mAbs efficiently neutralized rabies virus in vitro and in vivo, as detailed elsewhere (Dietzschold, B., P. Casali, Y. Ueki, M. Gore, C. E. Rupprecht, A. L. Notkins, and H. Koprowski, manuscript submitted for publication). Hybridization experiments using probes specific for the different human V gene segment families revealed that cell precursors producing low affinity IgM binding to rabies virus utilized a restricted number of VH gene segments (i.e., only members of the VHIIIb subfamily), whereas cell precursors producing high affinity IgG and IgA to rabies virus utilized an assortment of different VH gene segments (i.e., members of the VHI, VHIII, VHIV, and VHVI families and VHIIIb subfamily). In conclusion, our studies show that EBV transformation in conjunction with limiting dilution technology and somatic cell hybridization techniques are useful methods for quantitating, at the B cell clonal level, the human antibody response to foreign Ags and for generating human mAbs of predetermined specificity and high affinity.


Thermal mapping of a scanning thermal microscopy tip.
Scanning thermal microscopy (SThM) is a very promising technique for local investigation of temperature and thermal properties of nanostructures with great application potential in contemporary nanoelectronics and nanotechnology. In order to increase the localization of SThM measurements, the size of probes has recently substantially decreased, which results in novel types of SThM probes manufactured with the use of modern silicon microfabrication technology. Quantitative SThM measurements with these probes need methods, which enable to assess the quality of thermal contact between the probe and the investigated surface. In this paper we propose a tip thermal mapping (TThM) procedure, which is used to estimate experimentally the distribution of power dissipated by the tip of an SThM probe. We also show that the proposed power dissipation model explains the results of active-mode SThM measurements and that the TThM procedure is reversible for a given probe and sample.


Fluorinated alkyl compounds including long chain carboxylic acids in wild bird livers from Japan.
A wide range of fluorinated alkyl compounds (FACs) has been reported in wildlife in various locations in the world. However, such information regarding Japanese wildlife is rarely found. In the present study, we investigated the occurrence of 21 FACs, including perfluorinated alkyl sulfonates (PFASs), perfluorinated carboxylates (PFCAs), and fluorotelomer acids, in the livers of 10 wild bird species from two regions in northern Japan. To avoid interferences, FACs were quantified by a recently developed method using acetonitrile and solid-phase extraction followed by an ion exchange HPLC column separation. Apart from perfluorooctane sulfonate (PFOS), which was found at the highest levels of all the compounds detected, several long chain perfluorinated carboxylates (PFCAs) from C8 to C16, particularly perfluorotetradecanoic acid (PFTeDA) and perfluorohexadecanoic acid (PFHxDA), were detected for the first time. Additionally, 7:3 FTCA, a fluorotelomer acid, was also detected in most swan livers from Miyagi prefecture and all the birds from Tochigi prefecture. However, none of the sulfonamides and unsaturated telomer acids were detected in any species. Swans seem to be the least exposed wild birds to FACs among the investigated birds, signifying that feeding habits may reflect FAC accumulation in wild birds. The highest total concentration of detected FACs was 405ngg(-1)wet wt., which was found in a Japanese sparrowhawk, indicating that the top predatory wild birds can accumulate several long chain carboxylic acids. However, the current FAC concentrations found in livers may suggest that these compounds alone would not cause a severe toxic effect in these species.


Biochemical and mechanical extracellular matrix properties dictate mammary epithelial cell motility and assembly.
Biochemical and mechanical cues of the extracellular matrix have been shown to play important roles in cell-matrix and cell-cell interactions. We have experimentally tested the combined influence of these cues to better understand cell motility, force generation, cell-cell interaction, and assembly in an in vitro breast cancer model. MCF-10A non-tumorigenic mammary epithelial cells were observed on surfaces with varying fibronectin ligand concentration and polyacrylamide gel rigidity. Our data show that cell velocity is biphasic in both matrix rigidity and adhesiveness. The maximum cell migration velocity occurs only at specific combination of substrate stiffness and ligand density. We found cell-cell interactions reduce migration velocity. However, the traction forces cells exert onto the substrate increase linearly with both cues, with cells in pairs exerting higher maximum tractions observed over single cells. A relationship between force and motility shows a maximum in single cell velocity not observed in cell pairs. Cell-cell adhesion becomes strongly favored on softer gels with elasticity ≤ 1250 Pascals (Pa), implying the existence of a compliance threshold that promotes cell-cell over cell-matrix adhesion. Finally on gels with stiffness similar to pre-malignant breast tissue, 400 Pa, cells undergo multicellular assembly and division into 3D spherical aggregates on a 2D surface.