New insights into establishment and maintenance of DNA methylation imprints in mammals.
Fundamental to genomic imprinting in mammals is the acquisition of epigenetic marks that differ in male and female gametes at 'imprinting control regions' (ICRs). These marks mediate the allelic expression of imprinted genes in the offspring. Much has been learnt about the nature of imprint marks, the times during gametogenesis at which they are laid down and some of the factors responsible especially for DNA methylation. Recent work has revealed that transcription and histone modifications are critically involved in DNA methylation acquisition, and these findings allow us to propose rational models for methylation establishment. A completely novel perspective on gametic DNA methylation has emerged from epigenomic profiling. Far more differentially methylated loci have been identified in gametes than known imprinted genes, which leads us to revise the notion that methylation of ICRs is a specifically targeted process. Instead, it seems to obey default processes in germ cells, giving rise to distinct patterns of DNA methylation in sperm and oocytes. This new insight, together with the identification of proteins that preserve DNA methylation after fertilization, emphasizes the key role played by mechanisms that selectively retain differential methylation at imprinted loci during early development. Addressing these mechanisms will be essential to understanding the specificity and evolution of genomic imprinting.


Adolescent computer use and alcohol use: what are the role of quantity and content of computer use?
The purpose of this study was to examine the relationship between computer use and alcohol use among adolescents. In particular, the goal of the research was to determine the role of lifetime drinking and past month drinking on quantity as measured by amount of time on the computer (for school work and excluding school work) and on content as measured by the frequency of a variety of activities on the internet (e.g., e-mail, searching for information, social networking, listen to/download music). Participants (aged 13-17 years and residing in the United States) were recruited via the internet to complete an anonymous survey online using a popular survey tool (N=270). Their average age was 16 and the sample was predominantly female (63% girls). A series of analyses was conducted with the computer use measures as dependent variables (hours on the computer per week for school work and excluding school work; various internet activities including e-mail, searching for information, social networking, listen to/download music) controlling for gender, age, academic performance and age of first computer use. Based on the results, past month drinkers used the computer more hours per week excluding school work than those who did not. As expected, there were no differences in hours based on alcohol use for computer use for school work. Drinking also had relationships with more frequent social networking and listening to/downloading music. These findings suggest that both quantity and content of computer use were related to adolescent drinking.


Role of dopamine D2 receptors in plasticity of stress-induced addictive behaviours.
Dopaminergic systems are implicated in stress-related behaviour. Here we investigate behavioural responses to chronic stress in dopamine D2 receptor knockout mice and find that anxiety-like behaviours are increased compared with wild-type mice. Repeated stress exposure suppresses cocaine-induced behavioural sensitization, cocaine-seeking and relapse behaviours in dopamine D2 receptor knockout mice. Cocaine challenge after drug withdrawal in cocaine-experienced wild-type or dopamine D2 receptor knockout mice is associated with inhibition of long-term depression in the nucleus accumbens, and chronic stress during withdrawal prevents inhibition after cocaine challenge in cocaine-experienced dopamine D2 receptor knockout mice, but not in wild-type mice. Lentiviral-induced knockdown of dopamine D2 receptors in the nucleus accumbens of wild-type mice does not affect basal locomotor activity, but confers stress-induced inhibition of the expression of cocaine-induced behavioural sensitization. Stressed mice depleted of dopamine D2 receptors do not manifest long-term depression inhibition. Our results suggest that dopamine D2 receptors have roles in regulating synaptic modification triggered by stress and drug addiction.


Networking Across Borders for Individuals with Organic Acidurias and Urea Cycle Disorders: The E-IMD Consortium.
Patients with organic acidurias (OAD) and urea cycle disorders (UCD) are at increased risk of disability, impaired quality of life and reduced life expectancy. Clinical care in any one centre is constrained by small patient numbers; and furthermore diagnostic and treatment strategies vary between metabolic centres and countries, resulting in significant inequalities and disparity in patient outcome. The overall objective of the EU-funded activity 'European registry and network for intoxication type metabolic diseases' (E-IMD) is to collect systematic data to improve the knowledge of these diseases, to develop consensus care guidelines and to provide detailed information materials for families and professionals. Within three years E-IMD has (1) established a network of 87 partners in 25 countries (2) set up a patient registry of more than 1,000 individuals with OAD and UCD, (3) launched a website ( www.e-imd.org ) including detailed information materials in 11 languages, (4) developed guidelines for OAD and UCD, (5) organised two teaching courses and various scientific meetings, (6) extended the IT platform clustering with other inherited metabolic diseases (IMD) and (7) strengthened the collaboration with other international scientific consortia. E-IMD has made important steps towards improving and sharing knowledge on OAD and UCD and harmonisation of diagnostic and therapeutic strategies. Through the establishment of a modular patient registry, clustering with other IMD and stepwise extension of the network, E-IMD has implemented the core components of a European Reference Network for rare diseases.


Impaired striatal Akt signaling disrupts dopamine homeostasis and increases feeding.
The prevalence of obesity has increased dramatically worldwide. The obesity epidemic begs for novel concepts and therapeutic targets that cohesively address "food-abuse" disorders. We demonstrate a molecular link between impairment of a central kinase (Akt) involved in insulin signaling induced by exposure to a high-fat (HF) diet and dysregulation of higher order circuitry involved in feeding. Dopamine (DA) rich brain structures, such as striatum, provide motivation stimuli for feeding. In these central circuitries, DA dysfunction is posited to contribute to obesity pathogenesis. We identified a mechanistic link between metabolic dysregulation and the maladaptive behaviors that potentiate weight gain. Insulin, a hormone in the periphery, also acts centrally to regulate both homeostatic and reward-based HF feeding. It regulates DA homeostasis, in part, by controlling a key element in DA clearance, the DA transporter (DAT). Upon HF feeding, nigro-striatal neurons rapidly develop insulin signaling deficiencies, causing increased HF calorie intake. We show that consumption of fat-rich food impairs striatal activation of the insulin-activated signaling kinase, Akt. HF-induced Akt impairment, in turn, reduces DAT cell surface expression and function, thereby decreasing DA homeostasis and amphetamine (AMPH)-induced DA efflux. In addition, HF-mediated dysregulation of Akt signaling impairs DA-related behaviors such as (AMPH)-induced locomotion and increased caloric intake. We restored nigro-striatal Akt phosphorylation using recombinant viral vector expression technology. We observed a rescue of DAT expression in HF fed rats, which was associated with a return of locomotor responses to AMPH and normalization of HF diet-induced hyperphagia. Acquired disruption of brain insulin action may confer risk for and/or underlie "food-abuse" disorders and the recalcitrance of obesity. This molecular model, thus, explains how even short-term exposure to "the fast food lifestyle" creates a cycle of disordered eating that cements pathological changes in DA signaling leading to weight gain and obesity.


Arabidopsis annexins AnnAt1 and AnnAt4 interact with each other and regulate drought and salt stress responses.
Annexins are Ca2+--and phospholipid-binding proteins that form an evolutionarily conserved multigene family throughout the animal and plant kingdoms. Two annexins, AnnAt1 and AnnAt4, have been identified as components in osmotic stress and abscisic acid signaling in Arabidopsis. Here, we report that AnnAt1 and AnnAt4 regulate plant stress responses in a light-dependent manner. The single-mutant annAt1 and annAt4 plants showed tolerance to drought and salt stress, which was greatly enhanced in double-mutant annAt1annAt4 plants, but AnnAt4-overexpressing transgenic plants (35S:AnnAt4) were more sensitive to stress treatments under long day conditions. Furthermore, expression of stress-related genes was altered in these mutant and transgenic plants. Upon dehydration and salt treatment, AtNCED3, encoding 9-cis-epoxycarotenoid dioxygenase, and P5CS1, encoding Δ-1-pyrroline-5-carboxylate synthase, which are key enzymes in ABA and proline synthesis, respectively, were highly induced in annAt1annAt4 plants and to a lesser extent in annAt1 and annAt4 plants, but not in 35S:AnnAt4 plants. While annAt1 plants were more drought sensitive, annAt4 plants were more tolerant in short days than in long days. In vitro and in vivo binding assays revealed that AnnAt1 and AnnAt4 bind to each other in a Ca2+-dependent manner. Our results suggest that AnnAt1 and AnnAt4 function cooperatively in response to drought and salt stress and their functions are affected by photoperiod.


Does Family Caregiver Burden Differ Between Elderly and Younger Caregivers in Supporting Dying Patients With Cancer? An Italian Study.
The aging of the world's population increasingly calls on older people to care for their cancer relatives. This scenario confronts clinicians involved with end-of-life care with an imposing challenge: elderly family caregivers could have a different perception of the burdens associated with assistance compared to their younger counterparts. Palliativists need to know what limits and resources of these new age categories of caregivers could be for a global management of dying patients with cancer and their family. To evaluate the caregiver burden in family caregivers supporting dying patients with cancer in order to compare the differences between 2 different caregivers age groups (younger vs elderly population). This is a cross-sectional study. A total of 174 family caregivers of hospice patients were interviewed through the Caregiver Burden Inventory (CBI). The sample group was divided into 2 subgroups aged <65 (younger group) and ≥65 years old (elderly group). Compared with younger caregivers, the elderly group reported significantly higher scores in the CBI-developmental subscale (P = .009) confirmed by the generalized linear model (multivariate) evaluation that included possible predictors in the model. No further differences were found between the 2 age groups in the other CBI scores (time-dependent, physical, social, emotional, and overall score). Elderly caregivers are at high risk for experiencing developmental burden. This finding could prompt mental health professionals to pay greater attention to the value that assistance to the family member can have on their personal story and on that of the family or couple.


Molecular Mechanism of J-Domain-Triggered ATP Hydrolysis by Hsp70 Chaperones.
Efficient targeting of Hsp70 chaperones to substrate proteins depends on J-domain cochaperones, which in synergism with substrates trigger ATP hydrolysis in Hsp70s and concomitant substrate trapping. We present the crystal structure of the J-domain of Escherichia coli DnaJ in complex with the E. coli Hsp70 DnaK. The J-domain interacts not only with DnaK's nucleotide-binding domain (NBD) but also with its substrate-binding domain (SBD) and packs against the highly conserved interdomain linker. Mutational replacement of contacts between J-domain and SBD strongly reduces the ability of substrates to stimulate ATP hydrolysis in the presence of DnaJ and compromises viability at heat shock temperatures. Our data demonstrate that the J-domain and the substrate do not deliver completely independent signals for ATP hydrolysis, but the J-domain, in addition to its direct influence on Hsp70s catalytic center, makes Hsp70 more responsive for the hydrolysis-inducing signal of the substrate, resulting in efficient substrate trapping.


Biophysical characterization of anticoagulant hemextin AB complex from the venom of snake Hemachatus haemachatus.
Hemextin AB complex from the venom of Hemachatus haemachatus is the first known natural anticoagulant that specifically inhibits the enzymatic activity of blood coagulation factor VIIa in the absence of factor Xa. It is also the only known heterotetrameric complex of two three-finger toxins. Individually only hemextin A has mild anticoagulant activity, whereas hemextin B is inactive. However, hemextin B synergistically enhances the anticoagulant activity of hemextin A and their complex exhibits potent anticoagulant activity. In this study we characterized the nature of molecular interactions leading to the complex formation. Circular dichroism studies indicate the stabilization of beta-sheet in the complex. Hemextin AB complex has an increased apparent molecular diameter in both gas and liquid phase techniques. The complex formation is enthalpically favorable and entropically unfavorable with a negative change in the heat capacity. Thus, the anticoagulant complex shows less structural flexibility than individual subunits. Both electrostatic and hydrophobic interactions are important for the complexation; the former driving the process and the latter helping in the stabilization of the tetramer. The tetramer dissociates into dimers and monomers with the increase in the ionic strength of the solution and also with increase in the glycerol concentration in the buffer. The two dimers formed under each of these conditions display distinct differences in their apparent molecular diameters and anticoagulant properties. Based on these results, we have proposed a model for this unique anticoagulant complex.


Short term response of patients with bronchiectasis to treatment with amoxycillin given in standard or high doses orally or by inhalation.
The effect of three amoxycillin treatment regimens on purulent secretions of patients with bronchiectasis has been studied. On the basis of information recorded on a diary card the patients were divided into three groups, according to the usual nature of their secretions: seven who produced mucoid sputum, which occasionally became purulent; seven whose secretions were usually mucopurulent but occasionally purulent; and 19 whose secretions were persistently purulent. Treatment with capsules of amoxycillin in a dosage of 250 mg three times a day resulted in clearance of purulent secretions in patients of the mucoid group when they were treated for a clinical exacerbation. The sputum remained clear in these patients for long periods before a further exacerbation (median 6 1/2, range 1-11 months). The mucopurulent-purulent group also responded to this dosage but sputum purulence returned more rapidly (median 9, range 4-31 days). Only three of the 19 (17%) patients with persistently purulent secretions showed a macroscopic response to this dosage, whereas seven (60%) of 12 patients who received the higher dosage (3 g sachets twice a day) responded. Among the failures, some responded to nebulised amoxycillin, suggesting that higher levels of amoxycillin in secretions are required in these patients. Macroscopic clearance of purulent secretions was finally achieved in most of the patients studied. The response was not always predictable from the results of sputum culture. Clearance of secretions by antibiotics was also identified by the patients, using a diary card score. Improvements in well being and in symptoms were noticed even in the group who usually produced mucopurulent and purulent secretions even though they appeared to be clinically stable before treatment.


Fruit and vegetable intake in African Americans income and store characteristics.
The purpose of this study was to examine whether the characteristics of retail food stores where African-American women shopped mediated the association between their income and intake of fruits and vegetables. Food store characteristics included store type (supermarket, specialty store, limited assortment store, independent grocer), store location (suburbs, city of Detroit), and perceptions of the selection/quality and affordability of fresh produce for sale. The analysis drew upon data from a probability sample of 266 African-American women living in 2001 in eastside Detroit, which had no supermarkets. Structural equation modeling was used to calculate a path model of direct and indirect effects. Women shopping at supermarkets and specialty stores consumed fruit and vegetables more often, on average, than those shopping at independent grocers. More positive perceptions of the selection/quality, but not affordability, of fresh produce at the retail outlet where they shopped was positively associated with intake, independent of store type and location as well as age, per capita income, and years of education. The results suggested an indirect association between income and fruit and vegetable intake; women with higher per capita incomes were more likely to shop at supermarkets than at other grocers, which in turn was associated with intake. Previous studies have shown that few supermarkets are located in the city of Detroit, a symptom of economic divestment over the past several decades. Results of this study suggest this may have negative implications for dietary quality, particularly among lower-income women.


A Method to Estimate Cadaveric Femur Cortical Strains During Fracture Testing Using Digital Image Correlation.
This protocol describes the method using digital image correlation to estimate cortical strain from high speed video images of the cadaveric femoral surface obtained from mechanical testing. This optical method requires a texture of many contrasting fiduciary marks on a solid white background for accurate tracking of surface deformation as loading is applied to the specimen. Immediately prior to testing, the surface of interest in the camera view is painted with a water-based white primer and allowed to dry for several minutes. Then, a black paint is speckled carefully over the white background with special consideration for the even size and shape of the droplets. Illumination is carefully designed and set such that there is optimal contrast of these marks while minimizing reflections through the use of filters. Images were obtained through high speed video capture at up to 12,000 frames/s. The key images prior to and including the fracture event are extracted and deformations are estimated between successive frames in carefully sized interrogation windows over a specified region of interest. These deformations are then used to compute surface strain temporally during the fracture test. The strain data is very useful for identifying fracture initiation within the femur, and for eventual validation of proximal femur fracture strength models derived from Quantitative Computed Tomography-based Finite Element Analysis (QCT/FEA).


Rift Valley fever virus epidemic in Kenya, 2006/2007: the entomologic investigations.
In December 2006, Rift Valley fever (RVF) was diagnosed in humans in Garissa Hospital, Kenya and an outbreak reported affecting 11 districts. Entomologic surveillance was performed in four districts to determine the epidemic/epizootic vectors of RVF virus (RVFV). Approximately 297,000 mosquitoes were collected, 164,626 identified to species, 72,058 sorted into 3,003 pools and tested for RVFV by reverse transcription-polymerase chain reaction. Seventy-seven pools representing 10 species tested positive for RVFV, including Aedes mcintoshi/circumluteolus (26 pools), Aedes ochraceus (23 pools), Mansonia uniformis (15 pools); Culex poicilipes, Culex bitaeniorhynchus (3 pools each); Anopheles squamosus, Mansonia africana (2 pools each); Culex quinquefasciatus, Culex univittatus, Aedes pembaensis (1 pool each). Positive Ae. pembaensis, Cx. univittatus, and Cx. bitaeniorhynchus was a first time observation. Species composition, densities, and infection varied among districts supporting hypothesis that different mosquito species serve as epizootic/epidemic vectors of RVFV in diverse ecologies, creating a complex epidemiologic pattern in East Africa.


Racial and Ethnic Differences in Health Care Utilization Following Severe Acute Brain Injury in the United States.
To examine racial and ethnic differences in the utilization of 3 interventions (tracheostomy placement, gastrostomy tube placement, and hospice utilization) among patients with severe acute brain injury (SABI). Retrospective cohort study. Data from the National Inpatient Sample, from 2002 to 2012. Adult patients with SABI defined as a primary diagnosis of stroke, traumatic brain injury, or post-cardiac arrest who received greater than 96 hours of mechanical ventilation. Race/ethnicity, stratified into 5 categories (white, black, Hispanic, Asian, and other). Data from 86 246 patients were included in the cohort, with a mean (standard deviation) age of 60 (18) years. In multivariable analysis, compared to white patients, black patients had an 20% increased risk of tracheostomy utilization (relative risk [RR]: 1.20, 95% CI: 1.16-1.24, P < .001), Hispanic patients had a 10% increased risk (RR: 1.10, 95% CI: 1.06-1.14, P < .001), Asian patients had an 8% increased risk (RR: 1.08, 95% CI: 1.01-1.16, P = .02), and other race patients had an 10% increased risk (RR: 1.10, 95% CI: 1.04-1.16, P < .001). A similar relationship was observed for gastrostomy utilization. In multivariable analysis, compared to white patients, black patients had a 25% decreased risk of hospice discharge (RR: 0.75, 95% CI: 0.67-0.85, P < .001), Hispanic patients had a 20% decreased risk (RR: 0.80, 95% CI: 0.69-0.94, P < .01), and Asian patients had a 47% decreased risk (RR: 0.53, 95% CI: 0.39-0.73, P < .001). There was no observed relationship between race/ethnicity and in-hospital mortality. Minority race was associated with increased utilization of tracheostomy and gastrostomy, as well as decreased hospice utilization among patients with SABI. Further research is needed to better understand the mechanisms underlying these race-based differences in critical care.


Association Between Calcium or Vitamin D Supplementation and Fracture Incidence in Community-Dwelling Older Adults: A Systematic Review and Meta-analysis.
The increased social and economic burdens for osteoporosis-related fractures worldwide make the prevention of such injuries a major public health goal. Previous studies have reached mixed conclusions regarding the association between calcium, vitamin D, or combined calcium and vitamin D supplements and fracture incidence in older adults. To investigate whether calcium, vitamin D, or combined calcium and vitamin D supplements are associated with a lower fracture incidence in community-dwelling older adults. The PubMed, Cochrane library, and EMBASE databases were systematically searched from the inception dates to December 24, 2016, using the keywords calcium, vitamin D, and fracture to identify systematic reviews or meta-analyses. The primary randomized clinical trials included in systematic reviews or meta-analyses were identified, and an additional search for recently published randomized trials was performed from July 16, 2012, to July 16, 2017. Randomized clinical trials comparing calcium, vitamin D, or combined calcium and vitamin D supplements with a placebo or no treatment for fracture incidence in community-dwelling adults older than 50 years. Two independent reviewers performed the data extraction and assessed study quality. A meta-analysis was performed to calculate risk ratios (RRs), absolute risk differences (ARDs), and 95% CIs using random-effects models. Hip fracture was defined as the primary outcome. Secondary outcomes were nonvertebral fracture, vertebral fracture, and total fracture. A total of 33 randomized trials involving 51 145 participants fulfilled the inclusion criteria. There was no significant association of calcium or vitamin D with risk of hip fracture compared with placebo or no treatment (calcium: RR, 1.53 [95% CI, 0.97 to 2.42]; ARD, 0.01 [95% CI, 0.00 to 0.01]; vitamin D: RR, 1.21 [95% CI, 0.99 to 1.47]; ARD, 0.00 [95% CI, -0.00 to 0.01]. There was no significant association of combined calcium and vitamin D with hip fracture compared with placebo or no treatment (RR, 1.09 [95% CI, 0.85 to 1.39]; ARD, 0.00 [95% CI, -0.00 to 0.00]). No significant associations were found between calcium, vitamin D, or combined calcium and vitamin D supplements and the incidence of nonvertebral, vertebral, or total fractures. Subgroup analyses showed that these results were generally consistent regardless of the calcium or vitamin D dose, sex, fracture history, dietary calcium intake, and baseline serum 25-hydroxyvitamin D concentration. In this meta-analysis of randomized clinical trials, the use of supplements that included calcium, vitamin D, or both compared with placebo or no treatment was not associated with a lower risk of fractures among community-dwelling older adults. These findings do not support the routine use of these supplements in community-dwelling older people.


Near-road modeling and measurement of cerium-containing particles generated by nanoparticle diesel fuel additive use.
Cerium oxide nanoparticles (nCe) are used as a fuel-borne catalyst in diesel engines to reduce particulate emissions, yet the environmental and human health impacts of the exhaust particles are not well understood. To bridge the gap between emission measurements and ambient impacts, size-resolved measurements of particle composition and mass concentration have been performed in Newcastle-upon-Tyne, United Kingdom, where buses have used an nCe additive since 2005. These observations show that the noncrustal cerium fraction thought to be associated with the use of nCe has a mass concentration ∼ 0.3 ng m(-3) with a size distribution peaking at 100-320 nm in aerodynamic diameter. Simulations with a near-roadway multicomponent sectional aerosol dynamic model predict that the use of nCe additives increases the number concentration of nuclei mode particles (<50 nm in diameter) while decreasing the total mass concentration. The near-road model predicts a downwind mass size distribution of cerium-containing particles peaking at 150 nm in aerodynamic diameter, a value similar to that measured for noncrustal cerium in Newcastle. This work shows that both the emission and atmospheric transformation of cerium-containing particles needs to be taken into account by regional modelers, exposure scientists, and policymakers when determining potential environmental and human health impacts.


Cortical representation of tympanic membrane movements due to pressure variation: an fMRI study.
Middle ear sensory information has never been localized in the homunculus of the somatosensory cortex (S1). We investigated the somatosensory representation of the middle ear in 15 normal hearing subjects. We applied small air pressure variations to the tympanic membrane while performing a 3T-fMRI study. Unilateral stimulations of the right ear triggered bilateral activations in the caudal part of the postcentral gyrus in Brodmann area 43 (BA 43) and in the auditory associative areas 42 (BA 42) and 22 (BA 22). BA 43 has been found to be involved in activities accompanying oral intake and could be more largely involved in pressure activities in the oropharynx area. The tympanic membrane is indirectly related to the pharynx area through the action of tensor tympani, which is a Eustachian tube muscle. The Eustachian tube muscles have a role in pressure equalization in the middle ear and also have a role in the pharyngeal phase of swallowing. Activation of BA 42 and BA 22 could reflect activations associated with the bilateral acoustic reflex triggered prior to self-vocalization to adjust air pressure in the oropharynx during speech. We propose that BA 43, 42, and 22 are the cortical areas associated with middle ear function. We did not find representation of tympanic membrane movements due to pressure in S1, but its representation in the postcentral gyrus in BA 43 seems to suggest that at least part of this area conveys pure somatosensory information.


Administration of general anaesthesia to a paediatric patient with osteopetrosis.
Osteopetrosis is a rare clinical syndrome characterised by the failure of bone resorption and remodelling, which causes multiple anatomical and physiological impairments. Pathological fractures can occur, in addition to, haemathological and metabolic impairments. Our patient was a 9-year-old girl diagnosed with osteopetrosis in the neonatal period. She had severe anaemia, thrombocytopaenia, hypocalcaemia, as well as growth and development delays. In this case report, the administration of general anaesthesia to the patient for a biopsy of the scalp and skull and a partial maxillectomy is presented.


The relevance of Heidegger's philosophy of technology for biomedical ethics.
Heidegger's thoughts on modern technology have received much attention in many disciplines and fields, but, with a few exceptions, the influence has been sparse in biomedical ethics. The reason for this might be that Heidegger's position has been misinterpreted as being generally hostile towards modern science and technology, and the fact that Heidegger himself never subjected medical technologies to scrutiny but was concerned rather with industrial technology and information technology. In this paper, Heidegger's philosophy of modern technology is introduced and then brought to bear on medical technology. Its main relevance for biomedical ethics is found to be that the field needs to focus upon epistemological and ontological questions in the philosophy of medicine related to the structure and goal of medical practice. Heidegger's philosophy can help us to see how the scientific attitude in medicine must always be balanced by and integrated into a phenomenological way of understanding the life-world concerns of patients. The difference between the scientific and the phenomenological method in medicine is articulated by Heidegger as two different ways of studying the human body: as biological organism and as lived body. Medicine needs to acknowledge the priority of the lived body in addressing health as a way of being-in-the-world and not as the absence of disease only. A critical development of Heidegger's position can provide us with a criterion for distinguishing the uses of medical technologies that are compatible with such an endeavor from the technological projects that are not.


Generating true minima in constrained variational formulations via modified Lagrange multipliers.
Variational principles are important in the investigation of large classes of physical systems. They can be used both as analytical methods as well as starting points for the formulation of powerful computational techniques such as dynamical optimization methods. Systems with charged objects in dielectric media and systems with magnetically active particles are important examples. In these examples and other important cases, the variational principles describing the system are required to obey a number of constraints. These constraints are implemented within the variational formulation by means of Lagrange multipliers. Such constrained variational formulations are in general not unique. For the application of efficient simulation methods, one must find specific formulations that satisfy a number of important conditions. An often required condition is that the functional be positive-definite, in other words, its extrema be actual minima. In this article, we present a general approach to attack the problem of finding, among equivalent variational functionals, those that generate true minima. The method is based on the modification of the Lagrange multiplier which allows us to generate large families of effective variational formulations associated with a single original constrained variational principle. We demonstrate its application to different examples and, in particular, to the important cases of Poisson and Poisson-Boltzmann equations. We show how to obtain variational formulations for these systems with extrema that are always minima.


Weight-related teasing and non-normative eating behaviors as predictors of weight loss maintenance. A longitudinal mediation analysis.
Weight loss maintenance is essential for the reduction of obesity-related health impairments. However, only a minority of individuals successfully maintain reduced weight in the long term. Research has provided initial evidence for associations between weight-related teasing (WRT) and greater non-normative eating behaviors. Further, first evidence was found for associations between non-normative eating behaviors and weight loss maintenance. Hence, the present study aimed to examine the predictive value of WRT for weight loss maintenance and the role of non-normative eating behaviors as possible mediators of this relationship. The study was part of the German Weight Control Registry that prospectively followed individuals who had intentionally lost at least 10% of their maximum weight and had maintained this reduced weight for at least one year. In N = 381 participants, retrospective WRT during childhood and adolescence, current non-normative eating behaviors (i.e., restrained, external, emotional eating), and change in body mass index (BMI, kg/m(2)) over two years were examined using self-report assessments. Structural equation modeling was used to analyze the assumed mediational relationship. As a result, a greater effect of retrospective WRT during childhood and adolescence predicted less successful adult weight loss maintenance over two years. Current emotional eating fully mediated this relationship while current restrained and external eating yielded no mediational effects. Hence, a greater effect of WRT predicted greater current emotional eating, which in turn predicted a smaller decrease or a greater increase in BMI. Our findings suggest that suffering from WRT during childhood and adolescence might lead to emotional eating which in turn impairs long-term weight loss maintenance. Thus, our results highlight the need for interventions aiming at reducing weight stigmatization and targeting emotional eating for successful long-term weight loss maintenance.


A Phase II Trial of Cabozantinib in Hormone Receptor-Positive Breast Cancer with Bone Metastases.
We assessed the antitumor activity of cabozantinib, a potent multireceptor oral tyrosine kinase inhibitor, in patients with hormone receptor-positive breast cancer with bone metastases. In this single-arm multicenter phase II study, patients received an initial starting dose of 100 mg, later reduced to 60 mg, per day. The primary endpoint was the bone scan response rate. Secondary endpoints included objective response rate by RECIST, progression-free survival (PFS), and overall survival (OS). Of 52 women enrolled, 20 (38%) experienced a partial response on bone scan and 6 (12%) had stable disease. Prior to the first repeat bone scan at 12 weeks, 19 (35%) patients discontinued study treatment because of early clinical progression or unacceptable toxicity. RECIST evaluation based on best overall response by computed tomography revealed stable disease in extraosseous tissues in 26 patients (50%) but no complete or partial responses. In 25 patients with disease control on bone scan at 12 weeks, only 3 (12%) patients developed extraosseous progression. The median PFS was 4.3 months, and median OS was 19.6 months. The most common grade 3 or 4 toxicities were hypertension (10%), anorexia (6%), diarrhea (6%), fatigue (4%), and hypophosphatemia (4%). Bone scans improved in 38% of patients with metastatic hormone receptor-positive breast cancer and remained stable in an additional 12% for a minimum duration of 12 weeks on cabozantinib. Further investigations should assess the activity of cabozantinib in combination with other hormonal and other breast cancer therapies and determine whether bone scan responses correlate with meaningful antitumor effects. ClinicalTrials.gov identifier. NCT01441947 IMPLICATIONS FOR PRACTICE: Most patients with metastatic hormone receptor-positive (HR+) breast cancer have bone involvement, and many have bone-only disease, which is difficult to evaluate for response. This phase II single-arm study evaluated the clinical activity of the small molecule MET/RET/VEGFR2 inhibitor cabozantinib in patients with metastatic HR+ breast cancer with bone metastases. This study met its primary endpoint, and cabozantinib treatment resulted in a significant bone scan response rate correlating with improved survival. This is the first study to use bone scan response as a primary endpoint in breast cancer. The results support further study of cabozantinib in HR+ breast cancer.


Association of Herpes Simplex Virus Testing with Hospital Length of Stay for Infants ≤60 Days of Age Undergoing Evaluation for Meningitis.
Although neonatal herpes simplex virus (HSV) causes significant morbidity, utilization of the cerebrospinal fluid (CSF) HSV polymerase chain reaction (PCR) test remains variable. Our objective was to examine the association of CSF HSV PCR testing with length of stay (LOS) in a 20-center retrospective cohort of hospitalized infants aged ≤60 days undergoing evaluation for meningitis after adjustment for patient-level factors and clustering by center. Of 20,496 eligible infants, 7,399 (36.1%) had a CSF HSV PCR test performed, and 46 (0.6% of those tested) had a positive test. Infants who had a CSF HSV PCR test performed had a 23% longer hospital LOS (incident rate ratio 1.23; 95% CI: 1.14-1.33). Targeted CSF HSV PCR testing may mitigate the impact on LOS for low-risk infants.


Perceptual and neural response to affective tactile texture stimulation in adults with autism spectrum disorders.
Autism spectrum disorders (ASD) are associated with differences in sensory sensitivity and affective response to sensory stimuli, the neural basis of which is still largely unknown. We used psychophysics and functional magnetic resonance imaging (fMRI) to investigate responses to somatosensory stimulation with three textured surfaces that spanned a range of roughness and pleasantness in a sample of adults with ASD and a control group. While psychophysical ratings of roughness and pleasantness were largely similar across the two groups, the ASD group gave pleasant and unpleasant textures more extreme average ratings than did controls. In addition, their ratings for a neutral texture were more variable than controls, indicating they are less consistent in evaluating a stimulus that is affectively ambiguous. Changes in brain blood oxygenation level-dependent (BOLD) signal in response to stimulation with these textures differed substantially between the groups, with the ASD group exhibiting diminished responses compared to the control group, particularly for pleasant and neutral textures. For the most unpleasant texture, the ASD group exhibited greater BOLD response than controls in affective somatosensory processing areas such as the posterior cingulate cortex and the insula. The amplitude of response in the insula in response to the unpleasant texture was positively correlated with social impairment as measured by the Autism Diagnostic Interview-Revised (ADI-R). These results suggest that people with ASD tend to show diminished response to pleasant and neutral stimuli, and exaggerated limbic responses to unpleasant stimuli, which may contribute to diminished social reward associated with touch, perpetuating social withdrawal, and aberrant social development.


The Rubber Hand Illusion: feeling of ownership and proprioceptive drift do not go hand in hand.
In the Rubber Hand Illusion, the feeling of ownership of a rubber hand displaced from a participant's real occluded hand is evoked by synchronously stroking both hands with paintbrushes. A change of perceived finger location towards the rubber hand (proprioceptive drift) has been reported to correlate with this illusion. To measure the time course of proprioceptive drift during the Rubber Hand Illusion, we regularly interrupted stroking (performed by robot arms) to measure perceived finger location. Measurements were made by projecting a probe dot into the field of view (using a semi-transparent mirror) and asking participants if the dot is to the left or to the right of their invisible hand (Experiment 1) or to adjust the position of the dot to that of their invisible hand (Experiment 2). We varied both the measurement frequency (every 10 s, 40 s, 120 s) and the mode of stroking (synchronous, asynchronous, just vision). Surprisingly, with frequent measurements, proprioceptive drift occurs not only in the synchronous stroking condition but also in the two control conditions (asynchronous stroking, just vision). Proprioceptive drift in the synchronous stroking condition is never higher than in the just vision condition. Only continuous exposure to asynchronous stroking prevents proprioceptive drift and thus replicates the differences in drift reported in the literature. By contrast, complementary subjective ratings (questionnaire) show that the feeling of ownership requires synchronous stroking and is not present in the asynchronous stroking condition. Thus, subjective ratings and drift are dissociated. We conclude that different mechanisms of multisensory integration are responsible for proprioceptive drift and the feeling of ownership. Proprioceptive drift relies on visuoproprioceptive integration alone, a process that is inhibited by asynchronous stroking, the most common control condition in Rubber Hand Illusion experiments. This dissociation implies that conclusions about feelings of ownership cannot be drawn from measuring proprioceptive drift alone.


High Prevalence of Asymptomatic Neurocysticercosis in an Endemic Rural Community in Peru.
Neurocysticercosis is a common helminthic infection of the central nervous system and an important cause of adult-onset epilepsy in endemic countries. However, few studies have examined associations between neurologic symptoms, serology and radiographic findings on a community-level. We conducted a population-based study of resident's ≥2 years old in a highly endemic village in Peru (pop. 454). We applied a 14 -question neurologic screening tool and evaluated serum for antibodies against Taenia solium cysticercosis using enzyme-linked immunoelectrotransfer blot (LLGP-EITB). We invited all residents ≥18 years old to have non-contrast computerized tomography (CT) of the head. Of the 385 residents who provided serum samples, 142 (36.9%) were seropositive. Of the 256 residents who underwent CT scan, 48 (18.8%) had brain calcifications consistent with NCC; 8/48 (17.0%) reported a history of headache and/or seizures. Exposure to T. solium is very common in this endemic community where 1 out of 5 residents had brain calcifications. However, the vast majority of people with calcifications were asymptomatic. This study reports a high prevalence of NCC infection in an endemic community in Peru and confirms that a large proportion of apparently asymptomatic residents have brain calcifications that could provoke seizures in the future.


Bladder tumours among rodent operatives handling ANTU.
The rodenticide ANTU (alpha-naphthylthiourea) was used in the United Kingdom mainly in the late 1940s and early 1950s. The product then contained up to 0.2% of beta-naphthylamine as an impurity, and it was finally withdrawn in 1967 as a suspected carcinogen. Fourteen cases of urothelial tumours among rodent operatives exposed to ANTU are reported: in one district four out of 27 staff were affected, and in another area two out of 10. These cases strongly suggest that the early ANTU manufactured in the United Kingdom posed a cancer hazard to users. ANTU is still made or used in various countries, though the current product may be relatively pure and no longer contaminated by beta-naphthylamine. Recent laboratory evidence shows that even pure ANTU is mutagenic in the Ames test, and the safety of this rodenticide may need review.


Epstein-Barr virus nuclear protein 2 transactivates a cis-acting CD23 DNA element.
Epstein-Barr virus (EBV) nuclear protein 2 (EBNA-2) is essential for B-lymphocyte growth transformation. EBNA-2 upregulates mRNAs encoding CD23, a B-lymphocyte surface protein closely associated with EBV-induced growth transformation. To further investigate this EBNA-2 effect, we searched in the genomic DNA spanning the type a and type b CD23 mRNA start sites for a cis-acting fragment that would render a promoter transactivatable by EBNA-2. An 800-bp CD23 DNA fragment (-335 to +465 relative to the type a CD23 mRNA start site) conferred EBNA-2 responsiveness to the herpes simplex virus thymidine kinase (TK) promoter when transfected into EBV-negative B-lymphoma cells. Deletional analysis identified a -275/-89 subfragment that was EBNA-2 responsive when cloned in either orientation and at variable distances upstream of the heterologous promoter. EBNA-2 and the cis-acting CD23 element increased TK-promoted mRNA and did not alter the herpes simplex virus TK promoter transcription start site. As expected, a type a CD23 promoter (-335/+80) which contained the EBNA-2-responsive element was transactivated by EBNA-2. As in EBV infection and stable EBNA-2 transfection, the CD23 DNA element in cis with heterologous or homologous promoters was less responsive to type 2 than to type 1 EBNA-2, whereas the EBNA-2-responsive DNA fragment from the EBV latent membrane protein 1 promoter was more responsive to the type 2 EBNA-2. These experiments delineate a 186-bp, EBNA-2-responsive cell DNA fragment and provide firm evidence that EBNA-2 transactivates transcription of cell genes. The greater type 1 versus type 2 EBNA-2 responsiveness of the CD23 promoter and the lack of a similar effect on the latent membrane protein 1 promoter is consistent with the hypothesis that greater cell gene transactivation by type 1 EBNA-2 is the basis for the more efficient growth-transforming properties of type 1 EBV.


Neurotoxicity of traffic-related air pollution.
The central nervous system is emerging as an important target for adverse health effects of air pollution, where it may contribute to neurodevelopmental and neurodegenerative disorders. Air pollution comprises several components, including particulate matter (PM) and ultrafine particulate matter (UFPM), gases, organic compounds, and metals. An important source of ambient PM and UFPM is represented by traffic-related air pollution, primarily diesel exhaust (DE). Human epidemiological studies and controlled animal studies have shown that exposure to air pollution, and to traffic-related air pollution or DE in particular, may lead to neurotoxicity. In particular, air pollution is emerging as a possible etiological factor in neurodevelopmental (e.g. autism spectrum disorders) and neurodegenerative (e.g. Alzheimer's disease) disorders. The most prominent effects caused by air pollution in both humans and animals are oxidative stress and neuro-inflammation. Studies in mice acutely exposed to DE (250-300μg/m3 for 6h) have shown microglia activation, increased lipid peroxidation, and neuro-inflammation in various brain regions, particularly the hippocampus and the olfactory bulb. An impairment of adult neurogenesis was also found. In most cases, the effects of DE were more pronounced in male mice, possibly because of lower antioxidant abilities due to lower expression of paraoxonase 2.


Total incidence and distribution of inflammatory joint diseases in a defined population: results from the Kuopio 2000 arthritis survey.
To study the incidence of inflammatory joint diseases in a defined population in Finland. We collected data for the year 2000 on a population of 87,000 inhabitants of Kuopio, Finland, of whom 20% were < 16 years of age. Information about the study was given through a local newspaper, and subjects attended one health center and 2 local hospitals for study. Inclusion criteria were that subjects have at least one peripheral joint with synovitis or signs of inflammation in sacroiliac, glenohumeral, or hip joints on the first visit. Incidence rates were calculated according to the diagnosis on the first visit, except for children, for whom diagnoses were established after 3 months' followup. A total of 188 adult incident cases (138 women, 50 men) and 11 children (8 girls, 3 boys) satisfied the inclusion criteria. The incidence of all arthritides was 230/100,000 (95% confidence interval 198.9-263.9) for the whole population; 271/100,000 (95% CI 233.7-312.7) for adults and 64/100,000 (95% CI 31.7-113.8) for children. Among adults the annual incidence of rheumatoid arthritis (RA), ankylosing spondylitis (AS), psoriatic arthritis (PsA), reactive arthritis (ReA), other spondyloarthropathies (SpA), connective tissue disease (CTD), crystalline arthritis, viral arthritis, and undifferentiated arthritis were 36, 7, 23, 10, 13, 9, 19, 7, and 149/100,000, respectively. The mean age at diagnosis was 49.4 +/- 16.3 years for all cases of arthritis among adults, about the same for both women and men. The mean age at diagnosis was 59.7 years in RA, 31.5 years in AS, 48.7 years in PsA, 38.0 years in ReA, 36.5 years in other SpA, 36.1 years in CTD, 65.0 years in crystalline arthritis, 53.3 years in viral arthritis, and 48.3 years in undifferentiated arthritis. Four of 11 children had juvenile idiopathic arthritis (JIA). The incidence of JIA was 23/100,000 in the population < 16 years of age. Of the remaining cases, 3 children had antibodies against Sindbis (Pogosta) virus and 4 had a transient monoarthritis. The overall incidence of arthritides among adults was slightly higher than previously reported from Finland. The incidence rates in the child population are in agreement with previous figures. These data are useful in planning the provision of health care.


Legal briefing: coerced treatment and involuntary confinement for contagious disease.
This issue's "Legal Briefing" column covers recent legal developments involving coerced treatment and involuntary confinement for contagious disease. Recent high profile court cases involving measles, tuberculosis, human immunodeficiency virus, and especially Ebola, have thrust this topic back into the bioethics and public spotlights. This has reignited debates over how best to balance individual liberty and public health. For example, the Presidential Commission for the Study of Bioethical Issues has officially requested public comments, held open hearings, and published a 90-page report on "ethical considerations and implications" raised by "U.S. public policies that restrict association or movement (such as quarantine)." Broadly related articles have been published in previous issues of The Journal of Clinical Ethics. We categorize recent legal developments on coerced treatment and involuntary confinement into the following six categories: 1. Most Public Health Confinement Is Voluntary 2. Legal Requirements for Involuntary Confinement 3. New State Laws Authorizing Involuntary Confinement 4. Quarantine Must Be as Least Restrictive as Necessary 5. Isolation Is Justified Only as a Last Resort 6. Coerced Treatment after Persistent Noncompliance.


Rapid experimental evolution of reproductive isolation from a single natural population.
Ecological speciation occurs when local adaptation generates reproductive isolation as a by-product of natural selection. Although ecological speciation is a fundamental source of diversification, the mechanistic link between natural selection and reproductive isolation remains poorly understood, especially in natural populations. Here, we show that experimental evolution of parasite body size over 4 y (approximately 60 generations) leads to reproductive isolation in natural populations of feather lice on birds. When lice are transferred to pigeons of different sizes, they rapidly evolve differences in body size that are correlated with host size. These differences in size trigger mechanical mating isolation between lice that are locally adapted to the different sized hosts. Size differences among lice also influence the outcome of competition between males for access to females. Thus, body size directly mediates reproductive isolation through its influence on both intersexual compatibility and intrasexual competition. Our results confirm that divergent natural selection acting on a single phenotypic trait can cause reproductive isolation to emerge from a single natural population in real time.


Prevalence of traditional cardiovascular risk factors among Nigerians with stroke.
Stroke is an important cause of morbidity and mortality worldwide. The case fatality rates from stroke are two- to three-fold higher in sub-Saharan Africa, including Nigeria, than in the developed world, mainly because of limited healthcare facilities and untreated risk factors. The aim was to determine the prevalence of traditional cardiovascular risk factors among Nigerians with stroke and compare the prevalence of risk factors between young and older adults with stroke. The study was cross-sectional in design, and was carried out on stroke patients who were 15 years of age or older, in the medical wards and neurology clinic of Aminu Kano Teaching Hospital, Nigeria. Data was collated consecutively over six months. A total of 81 patients were studied. Sixteen of them (19.8%) were under 45 years old (group 1) while the remaining 65 patients (80.2%) were 45 years or older (group 2). All patients had at least one risk factor. One-third of group 1 patients (37.5%) and 81.5% of group 2 patients had three or more cardiovascular risk factors (p = 0.0004). The most widespread risk factor in all patients, particularly in group 2 patients was systemic hypertension, while dyslipidaemia was most common among group 1 patients. Recurrent stroke was significantly more common among group 2 than group 1 patients (30.8 and 6.3% respectively) (p = 0.045). Cardiovascular risk factors, particularly hypertension and dyslipidaemia were prevalent in the studied patients with stroke. The older patients in group 2 had more multiple-risk factors than the younger ones in group 1. Secondary prevention strategies including detection and treatment of risk factors may curtail the burden of the disease.


[The use nicergoline in the treatment of diabetes mellitus].
Presented herein is a literature review aimed at investigating the appropriateness and possibility of using nicergoline (sermion) for treatment of patients suffering from diabetes mellitus. The analysis includes the most clinically significant results of scientific studies. The material to be reviewed was retrieved using the following key words: 'nicergoline', 'sermion', and 'diabetes mellitus' (with their respective Russian equivalents) in such databases as Medline, PubMed, ScienceDirect, PMC, Cochrane, as well as archives of both Russian and foreign journals, guidelines (clinical guidelines on rendering medical care for patients with diabetes mellitus, selected lectures on endocrinology). A broad spectrum of action and no significant side effects have made it possible to use this drug in various pathological conditions. At the same time, because of limited experience of using nicergoline for vascular diseases and an insufficient number of the carried out studies the precise role of this therapeutic agent in clinical practice has not yet been conclusively defined. Special attention is given to the analysis of efficacy of nicergoline in atherosclerosis and diabetes mellitus.


Changes in Fatty Acid Composition of Human Milk in Response to Cold-Like Symptoms in the Lactating Mother and Infant.
Infants rely on their innate immune systems to protect them from infection. Human milk (HM) contains fatty acids (FAs) and monoacylglycerols that are known to exhibit antiviral and antibacterial properties in vitro. The specific fat content of HM may potentially affect the efficacy of this antimicrobial activity. This preliminary study investigates whether the proportions of FA in HM change in response to infections, leading to cold-like symptoms in the mother or infant. Milk samples were obtained from mothers (n = 26) when they and their infants were healthy, and when mother, infant, or both suffered cold-like symptoms. The milk was hydrolysed and FA proportions were measured using gas chromatography. Fifteen FAs were recorded, of which eight were detected in sufficient quantities for statistical analysis. The proportions of capric (C10:0) and lauric acids (C12:0) in HM were significantly lower, and palmitic acid (C16:0) was higher when mothers and infants were ill compared to healthy samples. Palmitoleic (C16:1, n-7) and stearic acid (C18:0) proportions were higher in HM when the infant was unwell, but were not related to maternal health. Whilst the differences detected were small (less than 0.5%), the effects may be additive and potentially have a protective function. The value of further studies is certainly indicated.


A comparison of the growth-promoting, lipolytic, diabetogenic and immunological properties of pituitary and recombinant-DNA-derived bovine growth hormone (somatotropin).
The physiological mechanisms by which growth hormone (somatotropin) exerts its several metabolic activities remain poorly understood. In particular, there is disagreement as to whether the diabetogenic and lipolytic activities of the hormone are intrinsic properties of the molecule or are the result of contamination with other pituitary components. The availability of recombinant-DNA-derived bovine growth hormone (rebGH) presented an opportunity to compare the biological activities of rebGH and pituitary bGH in the absence of pituitary contaminants. Pituitary bGH and rebGH were immunologically identical in the radioimmunoassay for bGH, and good agreement was obtained for the potency of the latter measured by radioimmunoassay (1.6 units/mg) and the dwarf-mouse bioassay (1.4 units/mg). The lipolytic activity in vitro was examined by measuring the release of glycerol from rat epididymal fat maintained in the presence of dexamethasone (0.2 microgram/ml) and the material to be tested (0.1 and 0.2 mg/ml). Although two preparations of pituitary bGH stimulated a significant (P less than 0.01) increase in glycerol production, neither rebGH nor recombinant-DNA-derived chicken GH was lipolytic. However, when rebGH was intravenously injected into three sheep (0.15 mg/kg), the increase in plasma nonesterified fatty acids was similar to that measured with the same dose of pituitary bGH. Insulin-tolerance tests were conducted in sheep before and after treatment with rebGH and pituitary bGH (four subcutaneous injections of 0.15 mg/kg). Although the effect of rebGH was less than that of the pituitary hormone, both significantly impaired the ability of insulin to lower the concentration of plasma glucose. These data suggest that the lipolytic and diabetogenic activities of bGH are intrinsic properties of the molecule. However, the lipolytic activity may only become apparent after either modification of the molecule in vivo or activation of a lipolytic intermediate.


Podcasting as a Learning Tool in Medical Education: Prior to and During the Pandemic Period
Podcasting as a Learning Tool in Medical Education: Before and During the Pandemic Period Podcasts have seen significant growth as a medium for medical education over the last 15 years. The COVID-19 pandemic altered the way in which medical education is delivered to learners, including medical students, resident physicians, fellows, and practicing clinicians in the form of continuing medical education. A literature search using Google Scholar, PubMed, and NCBI was conducted to analyze and discuss how podcasts are utilized in medical education-both before and during the pandemic-and how this form of asynchronous education may influence clinical decision-making and patient outcomes. Finally, this review discusses how learners' habits while using podcasts may affect the way in which the information is internalized and the future of using podcasts to supplement medical education.


Alignments of endocrine, anthropometric, and metabolic parameters in type 2 diabetes after intervention with an Okinawa-based Nordic diet.
An Okinawa-based Nordic diet with moderately low carbohydrate content and high fat and protein content has been shown to improve anthropometry and metabolism in type 2 diabetes. The objectives of this study were to measure plasma or serum levels of hormones regulating energy metabolism and metabolic control, that is, cholecystokinin (CCK), Cortisol, C-peptide, ghrelin, glucagon, glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP), insulin, leptin, plasminogen activator inhibitor-1 (PAI-1), polypeptide YY (PYY), resistin, and visfatin after this diet intervention, and to determine partial correlations between hormonal levels and anthropometric and metabolic responses. A total of 30 patients (17 women) with type 2 diabetes, mean age 57.5 ± 8.2 years, and body mass index (BMI) 29.9 ± 4.1 kg/m2 were served the diet for 12 weeks. Fasting hormones were measured by Luminex and enzyme-linked immunosorbent assay (ELISA) before study start and after 12 and 28 weeks, along with anthropometric and metabolic parameters. The levels of CCK (P = 0.005), cortisol (P = 0.015), C-peptide (P = 0.022), glucagon (P = 0.003), GLP-1 (P = 0.013), GIP (P < 0.001), insulin (P = 0.004), leptin (P < 0.001), and PYY (P < 0.001) were lowered after dietary intervention. These reduced levels only remained for PYY at week 28 (P = 0.002), when also ghrelin (P = 0.012) and visfatin (P = 0.021) levels were reduced. Changes of glucose values correlated with changed levels of C-peptide and PYY (P < 0.001), insulin (P = 0.002), and PAI-1 (P = 0.009); changes of triglyceride values with changed levels of C-peptide, insulin, and PYY (P < 0.001) and PAI-1 (P = 0.005); changes of insulin resistance with changes of leptin levels (P = 0.003); and changes of BMI values with changed levels of C-peptide, insulin, and leptin (P < 0.001). Okinawa-based Nordic diet in type 2 diabetes has significant impact on the endocrine profile, which correlates with anthropometric and metabolic improvements.


C-Terminal Alpha-1 Antitrypsin Peptide: A New Sepsis Biomarker with Immunomodulatory Function.
Systemic inflammatory response syndrome (SIRS) is a life threatening condition and the leading cause of death in intensive care units. Although single aspects of pathophysiology have been described in detail, numerous unknown mediators contribute to the progression of this complex disease. The aim of this study was to elucidate the pathophysiological role of CAAP48, a C-terminal alpha-1 antitrypsin fragment, that we found to be elevated in septic patients and to apply this peptide as diagnostic marker for infectious and noninfectious etiologies of SIRS. Incubation of human polymorphonuclear neutrophils with synthetic CAAP48, the SNP-variant CAAP47, and several control peptides revealed intense neutrophil activation, induction of neutrophil chemotaxis, reduction of neutrophil viability, and release of cytokines. We determined the abundance of CAAP48 in patients with severe sepsis, severe SIRS of noninfectious origin, and viral infection. CAAP48 levels were 3-4-fold higher in patients with sepsis compared to SIRS of noninfectious origin and allowed discrimination of those patients with high sensitivity and specificity. Our results suggest that CAAP48 is a promising discriminatory sepsis biomarker with immunomodulatory functions, particularly on human neutrophils, supporting its important role in the host response and pathophysiology of sepsis.


Efficacy of Farabloc as an analgesic in primary fibromyalgia.
The goal of our study was to determine the efficacy of Farabloc, an electromagnetic shielding fabric compared to placebo fabric when worn as a nightgown, as an analgesic in patients hospitalized with fibromyalgia. In a rheumatologic and rehabilitation hospital, we performed a phase 1, single-blind study of patients using Farabloc (F) or placebo (P) gowns for 8 h per night during the 20-day hospitalization and a phase 2, single-blind crossover study of patients using both F and P gowns randomly and alternatively switching after 10 of 21 days hospitalization (phase 1: 42 F, mean age 49.02 years, 35 female, 7 male; 84 P, mean age 48.08 years, 72 female, 12 males; phase 2: 25 F/P, P/F, or P/P, mean age 44.0 years, 24 female, 1 male). The study involved randomly selected and blinded use of hospital gown 8 h per night of either F or P fabric. The main outcome measures were changes from admission or midpoint to discharge in quantity of pain (QN), quality of pain (QL), and paracetamol use (PU). In phase 1, all three variables significantly favored F over P when using paired t test, two sample t test, Mann-Whitney, and analysis of covariance tests. QN was reduced (F = -2.03 -/+ 0.99*, P = 0.59 -/+ 0.71). QL was reduced (F = -10.64 -/+ 5.69*, P = -2.54 -/+ 3.40). PU was reduced (F = 10.69 -/+ 6.68*, P = 26.12 -/+ 9.37). In phase 2, comparing midpoint to discharge levels in the three variables again favored P/F over F/P and P/P (>0.001): QN (P/F +16.00 -/+ 8.35* F/P -13.27 -/+ 11.40), QL (P/F +8.71 -/+ 4.75* F/P -6.55 -/+ 5.59), and PU (F -9.29 -/+ 4.39* P -18.00 -/+ 5.27) (*p = <0.001). Patients with fibromyalgia had less pain after sleeping in a gown made of Farabloc than with a placebo fabric. This suggests that Farabloc, an electromagnetic shielding fabric, has analgesic properties in fibromyalgia. Reduced pain observation is consistent with previous studies in phantom limb pain and delayed onset muscle pain. Limitations of this study include single blind design, small sample size, and in phase 2, a lack of washout period and a F/F group.


Deficits in behavioral inhibition predict treatment engagement in prison inmates.
Many inmates do not respond favorably to standard treatments routinely offered in prison. Executive cognitive functioning and emotional regulation may play a key role in treatment responsivity. During intake into treatment, inmates (N = 224) were evaluated for executive functioning, emotional perception, stress reactivity (salivary cortisol), IQ, psychological and behavioral traits, prior drug use, child and family background, and criminal histories and institutional behavior. Outcome measures included program completion, treatment readiness, responsivity and gain, and the Novaco Reaction to Provocation Questionnaire. Relative deficits in behavioral inhibition significantly predicted treatment outcomes, more so than background, psychological, or behavioral variables, and other neurocognitive and emotional regulatory measures. Future replications of these results have potential to improve assessment and treatment of offenders who are otherwise intractable.


Political regimes, political ideology, and self-rated health in Europe: a multilevel analysis.
Studies on political ideology and health have found associations between individual ideology and health as well as between ecological measures of political ideology and health. Individual ideology and aggregate measures such as political regimes, however, were never examined simultaneously. Using adjusted logistic multilevel models to analyze data on individuals from 29 European countries and Israel, we found that individual ideology and political regime are independently associated with self-rated health. Individuals with rightwing ideologies report better health than leftwing individuals. Respondents from Eastern Europe and former Soviet republics report poorer health than individuals from social democratic, liberal, Christian conservative, and former Mediterranean dictatorship countries. In contrast to individual ideology and political regimes, country level aggregations of individual ideology are not related to reporting poor health. This study shows that although both individual political ideology and contextual political regime are independently associated with individuals' self-rated health, individual political ideology appears to be more strongly associated with self-rated health than political regime.


Abundance and community structure of ammonia-oxidizing bacteria in activated sludge from different geographic regions in China.
Detailed ecological information on ammonia-oxidizing bacteria (AOB) in activated sludge of wastewater treatment plants (WWTPs) is very important to improve the efficiency of wastewater treatment. In this study, activated sludge samples were collected from seven municipal WWTPs located in seven cities in China, and real-time quantitative polymerase chain reaction (qPCR), as well as construction of clone libraries combined with correlation-based data analysis was performed. Further, the effect of geographic distribution and some water quality parameters on the ecological distribution of AOB in activated sludge from WWTPs were investigated. The geographic distribution, the influent concentration of total nitrogen (TN) and ammonia nitrogen (NH4+-N) had significant effects on the abundance of AOB (P < 0.05). However, the community structure of AOB were not significantly affected by geographic distribution, but by water quality parameters including the concentrations of TN and NH4+-N. N. oligotropha lineage was the dominant AOB group in the wastewater treatment systems. The results obtained in this study provide useful information to understand some aspects of the ecological information and influencing factors of AOB in geographically distributed WWTPs.


[The IC3D classification of the corneal dystrophies].
The recent availability of genetic analyses has demonstrated the shortcomings of the current phenotypic method of corneal dystrophy classification. Abnormalities in different genes can cause a single phenotype, whereas different defects in a single gene can cause different phenotypes. Some disorders termed corneal dystrophies do not appear to have a genetic basis. The purpose of this study was to develop a new classification system for corneal dystrophies, integrating up-to-date information on phenotypic description, pathologic examination, and genetic analysis. The International Committee for Classification of Corneal Dystrophies (IC3D) was created to devise a current and accurate nomenclature. This anatomic classification continues to organize dystrophies according to the level chiefly affected. Each dystrophy has a template summarizing genetic, clinical, and pathologic information. A category number from 1 through 4 is assigned, reflecting the level of evidence supporting the existence of a given dystrophy. The most defined dystrophies belong to category 1 (a well-defined corneal dystrophy in which a gene has been mapped and identified and specific mutations are known) and the least defined belong to category 4 (a suspected dystrophy where the clinical and genetic evidence is not yet convincing). The nomenclature may be updated over time as new information regarding the dystrophies becomes available. The IC3D Classification of Corneal Dystrophies is a new classification system that incorporates many aspects of the traditional definitions of corneal dystrophies with new genetic, clinical, and pathologic information. Standardized templates provide key information that includes a level of evidence for there being a corneal dystrophy. The system is user-friendly and upgradeable and can be retrieved on the website www.corneasociety.org/ic3d .


Electrophysiological correlates of stimulus-driven reorienting deficits after interference with right parietal cortex during a spatial attention task: a TMS-EEG study.
TMS interference over right intraparietal sulcus (IPS) causally disrupts behaviorally and EEG rhythmic correlates of endogenous spatial orienting before visual target presentation [Capotosto, P., Babiloni, C., Romani, G. L., & Corbetta, M. Differential contribution of right and left parietal cortex to the control of spatial attention: A simultaneous EEG-rTMS study. Cerebral Cortex, 22, 446-454, 2012; Capotosto, P., Babiloni, C., Romani, G. L., & Corbetta, M. Fronto-parietal cortex controls spatial attention through modulation of anticipatory alpha rhythms. Journal of Neuroscience, 29, 5863-5872, 2009]. Here we combine data from our previous studies to examine whether right parietal TMS during spatial orienting also impairs stimulus-driven reorienting or the ability to efficiently process unattended stimuli, that is, stimuli outside the current focus of attention. Healthy volunteers (n = 24) performed a Posner spatial cueing task while their EEG activity was being monitored. Repetitive TMS (rTMS) was applied for 150 msec simultaneously to the presentation of a central arrow directing spatial attention to the location of an upcoming visual target. Right IPS-rTMS impaired target detection, especially for stimuli presented at unattended locations; it also caused a modulation of the amplitude of parieto-occipital positive ERPs peaking at about 480 msec (P3) post-target. The P3 significantly decreased for unattended targets and significantly increased for attended targets after right IPS-rTMS as compared with sham stimulation. Similar effects were obtained for left IPS stimulation albeit in a smaller group of volunteers. We conclude that disruption of anticipatory processes in right IPS has prolonged effects that persist during target processing. The P3 decrement may reflect interference with postdecision processes that are part of stimulus-driven reorienting. Right IPS is a node of functional interaction between endogenous spatial orienting and stimulus-driven reorienting processes in human vision.


Variability in pediatric utilization of trauma facilities in California: 1999 to 2005.
We identify geographic, system, and socioeconomic differences between injured children cared for within and outside of state-designated trauma centers. This was a nonconcurrent observational study of a population-based sample from the California Office of Statewide Health Planning and Development Public Patient Discharge Database 1999 to 2005. Patients were 1 to 14 years of age, with International Classification of Diseases, Ninth Revision (ICD-9) diagnostic codes indicative of trauma. Injury Severity Scores were calculated from ICD-9 codes and categorized into severity categories. Outcomes were hospitalization in a trauma or nontrauma center. Children with severe injury who resided 0 to 10, 11 to 25, 26 to 50, and more than 50 miles from a trauma center were hospitalized in these centers at rates of 80.0%, 71.2%, 51.4%, and 28.5%, respectively. Children with severe injury who were living in a county with a trauma center were hospitalized in these centers at rates of 78.8%, whereas children living in a county without a trauma center were hospitalized in trauma centers at rates of 39.0%. Children with severe injury and public, private non-health maintenance organization (HMO), and HMO insurance were hospitalized in trauma centers at rates of 77.7%, 68.0%, and 55.4%, respectively. Age, injury severity, insurance type, residence in a county with a trauma center, and proximity to a trauma center were identified as predictors of trauma center utilization by logistic regression. We demonstrate considerable variation in the utilization pattern of trauma specialty care for children with moderate and severe injuries. Children with HMO and private insurance are cared for less often in trauma centers than those with public insurance, even after adjustment for other variables. Increased distance to a trauma center, as well as lack of trauma center within a county, also decreases trauma center utilization. These results suggest that assessing trauma center utilization patterns in total populations of children may identify opportunities for improved referral policies and practices as part of a larger effort to ensure high-quality trauma care for all children in need.


Squamous cell carcinoma of suprapubic cystostomy tract in a male with locally advanced primary urethral malignancy.
A 65-year-old man with stricture urethra underwent drainage of periurethral abscess and suprapubic cystostomy (SPC) placement. He presented to us 3 months later with a fungating ulcer at the site of perineal incision, the biopsy of which revealed squamous cell carcinoma (SCC). He underwent a total penile amputation, wide local excision scrotum, radical urethrocystoprostatectomy, ileal conduit with the en-bloc excision of the SPC tract. Histopathological examination of the suprapubic tract also revealed SCC. This is the first documented case of SCC of a suprapubic tract in the presence of primary urethral SCC.


Long-term effects of a multifaceted intervention to encourage the choice of the oral route for proton pump inhibitors: an interrupted time-series analysis.
Drugs are often given intravenously even when the patient is able to swallow and when an oral form would be more cost-effective. Evaluation of the impact of a multifaceted intervention on the early switch from intravenous to oral administration of proton pump inhibitors (PPI) in a hospital setting. The interrupted time series of intravenous PPI consumption was analysed. At a French University Hospital, the Drug Committee, composed of multidisciplinary pharmacy and medical staff, addressed the issue of increasing consumption of intravenous PPI drugs (May 2003). Letters to department heads, academic analyses from members of the Drug Committee, paper reminders at the point of care and audit-feedbacks by pharmacists. Monitoring of consumption and repeated reminder letters were planned. EFFECT OF CHANGE: The consumption of PPI was stable before the first intervention (mean level: 954 units/month). An immediate decrease occurred after the first Drug Committee letter (30% relative reduction, 95% CI -16% to -46%; p<0.001) with a significant trend change during the first multifaceted intervention (-24 units/month, 95% CI -42 to -7; p = 0.007). After the end of the outreach visits (July 2004), the consumptions increased (+32 units/month, 95% CI: 14 to 50, p<0.001). The second intervention had no significant impact. A complex intervention (audit, feedbacks, outreach visits) had an effect on practice. It was not sustained even after a less resource-intensive intervention. Other types of interventions are needed that could be continuously implemented to improve ordering practices long term.


PTCDA molecules on an InSb(001) surface studied with atomic force microscopy.
PTCDA (3,4,9,10-perylene-tetracarboxylic-dianhydride) molecular structures assembled on an InSb(001) c(8 × 2) reconstructed surface have been studied using frequency modulated atomic force microscopy. The high-resolution imaging of the structures is possible through repulsive interactions, using the constant height scanning mode. During initial stages of growth the [110] diffusion channel dominates as indicated by formation of long PTCDA molecular chains parallel to the [110] crystallographic direction on the InSb surface. For a single monolayer coverage a wetting layer of PTCDA is formed. Finally it is shown that the PTCDA/InSb is a promising system for building molecular nanostructures by manipulation of single molecules with the AFM tip.


Precursors of cognitive impairments in psychotic disorders: a population-based study.
Cognitive deficits have been found to be more prevalent in psychotic than in other disorders. Longitudinal research has shown that these deficits were generally already existent before onset of illness and are therefore not necessarily attributable to neurodegenerative processes. This study investigated whether both low IQ and markers of premorbid cognitive dysfunction independently contribute to an increased risk for psychoses. In a cross-sectional study about 50,000 young Swiss males completed a survey of intellectual problems in childhood/adolescence and other vulnerability factors during military call-up in 2005/2006. Subsequently, military IQ assessments were carried out on the entire sample. Diagnostic assessments were conducted according to International Classification of Diseases-10th edition (ICD-10). Low, especially performance, IQ was highly associated with an increased risk for psychotic disorders after adjusting for preexisting cognitive deficits and covariates, while in other disorders this association was less marked. Furthermore, preexisting intellectual problems emerged as important risk factors for psychoses. Our results confirm the importance of low IQ as characteristic of psychoses. Although premorbid intellectual deficits are common in people who go on to develop psychosis, neurodegenerative disease processes may also precipitate further declines in fluid cognitive functions. Assessment of cognitive functioning should be taken into account in early detection of psychoses.


Curcumin attenuates oxidative damage in animals treated with a renal carcinogen, ferric nitrilotriacetate (Fe-NTA): implications for cancer prevention.
Curcumin (diferuloylmethane), a biologically active ingredient derived from rhizome of the plant Curcuma longa, has potent anticancer properties as demonstrated in a plethora of human cancer cell lines/animal carcinogenesis model and also acts as a biological response modifier in various disorders. We have reported previously that dietary supplementation of curcumin suppresses renal ornithine decarboxylase (Okazaki et al. Biochim Biophys Acta 1740:357-366, 2005) and enhances activities of antioxidant and phase II metabolizing enzymes in mice (Iqbal et al. Pharmacol Toxicol 92:33-38, 2003) and also inhibits Fe-NTA-induced oxidative injury of lipids and DNA in vitro (Iqbal et al. Teratog Carcinog Mutagen 1:151-160, 2003). This study was designed to examine whether curcumin possess the potential to suppress the oxidative damage caused by kidney-specific carcinogen, Fe-NTA, in animals. In accord with previous report, at 1 h after Fe-NTA treatment (9.0 mg Fe/kg body weight intraperitoneally), a substantial increased formation of 4-hydroxy-2-nonenal (HNE)-modified protein adducts in renal proximal tubules of animals was observed. Likewise, the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and protein reactive carbonyl, an indicator of protein oxidation, were also increased at 1 h after Fe-NTA treatment in the kidneys of animals. The prophylactic feeding of animals with 1.0% curcumin in diet for 4 weeks completely abolished the formation of (i) HNE-modified protein adducts, (ii) 8-OHdG, and (iii) protein reactive carbonyl in the kidneys of Fe-NTA-treated animals. Taken together, our results suggest that curcumin may afford substantial protection against oxidative damage caused by Fe-NTA, and these protective effects may be mediated via its antioxidant properties. These properties of curcumin strongly suggest that it could be used as a cancer chemopreventive agent.


Structures of neuroligin-1 and the neuroligin-1/neurexin-1 beta complex reveal specific protein-protein and protein-Ca2+ interactions.
Neurexins and neuroligins provide trans-synaptic connectivity by the Ca2+-dependent interaction of their alternatively spliced extracellular domains. Neuroligins specify synapses in an activity-dependent manner, presumably by binding to neurexins. Here, we present the crystal structures of neuroligin-1 in isolation and in complex with neurexin-1 beta. Neuroligin-1 forms a constitutive dimer, and two neurexin-1 beta monomers bind to two identical surfaces on the opposite faces of the neuroligin-1 dimer to form a heterotetramer. The neuroligin-1/neurexin-1 beta complex exhibits a nanomolar affinity and includes a large binding interface that contains bound Ca2+. Alternatively spliced sites in neurexin-1 beta and in neuroligin-1 are positioned nearby the binding interface, explaining how they regulate the interaction. Structure-based mutations of neuroligin-1 at the interface disrupt binding to neurexin-1 beta, but not the folding of neuroligin-1 and confirm the validity of the binding interface of the neuroligin-1/neurexin-1 beta complex. Our results provide molecular insights for understanding the role of cell-adhesion proteins in synapse function.


Breast Milk Consumption in Preterm Neonates and Cardiac Shape in Adulthood.
Preterm birth relates to long-term alterations in cardiac morphology and function. Understanding whether preterm postnatal life is a tractable period of cardiovascular development that can be positively altered by nutrition is relevant to long-term outcomes. We hypothesized that being fed human breast milk during early postnatal life is beneficial to long-term cardiac structure and function in preterm-born individuals compared with infant formulas. A total of 926 preterm-born infants originally took part in a randomized controlled trial of postnatal milk-feeding regimens between 1982 and 1985 across 5 different UK centers. Preterm-born individuals were randomly assigned to either breast milk donated by unrelated lactating women or nutrient-enriched formulas. We followed 102 individuals from this cohort: 30 of whom had been randomized to being fed exclusively human milk and 16 to being fed exclusively formula. As a comparison group, we recruited an additional 102 individuals born term to uncomplicated pregnancies. Cardiac morphology and function were assessed by MRI. Preterm-born individuals fed exclusively human milk as infants had increased left and right ventricular end-diastolic volume index (+9.73%, P = .04 and +18.2%, P < .001) and stroke volume index (+9.79%, P = .05 and +22.1%, P = .01) compared with preterm-born individuals who were exclusively formula fed as infants. This study provides the first evidence of a beneficial association between breast milk and cardiac morphology and function in adult life in those born preterm and supports promotion of human milk for the care of preterm infants to reduce long-term cardiovascular risk.


Acute respiratory distress syndrome subphenotypes and differential response to simvastatin: secondary analysis of a randomised controlled trial.
Precision medicine approaches that target patients on the basis of disease subtype have transformed treatment approaches to cancer, asthma, and other heterogeneous syndromes. Two distinct subphenotypes of acute respiratory distress syndrome (ARDS) have been identified in three US-based clinical trials, and these subphenotypes respond differently to positive end-expiratory pressure and fluid management. We aimed to investigate whether these subphenotypes exist in non-US patient populations and respond differently to pharmacotherapies. HARP-2 was a multicentre, randomised controlled trial of simvastatin (80 mg) versus placebo done in general intensive care units (ICUs) at 40 hospitals in the UK and Ireland within 48 h of onset of ARDS. The primary outcome was ventilator-free days, and secondary outcomes included non-pulmonary organ failure-free days and mortality. In a secondary analysis of HARP-2, we applied latent class analysis to baseline data without consideration of outcomes to identify subphenotypes, and we compared clinical outcomes across subphenotypes and treatment groups. 540 patients were recruited to HARP-2. One patient withdrew consent for the use of their data, so data from 539 patients were analysed. In our secondary analysis, a two-class (two subphenotype) model was an improvement over a one-class model (p<0·0001), with 353 (65%) patients in the hypoinflammatory subphenotype group and 186 (35%) in the hyperinflammatory subphenotype group. Additional classes did not improve model fit. Clinical and biological characteristics of the two subphenotypes were similar to previous studies. Patients with the hyperinflammatory subphenotype had fewer ventilator-free days (median 2 days [IQR 0-17] vs 18 [IQR 0-23]; p<0·0001), fewer non-pulmonary organ failure-free days (15 [0-25] vs 27 [21-28]; p<0·0001), and higher 28-day mortality (73 [39%] vs 59 [17%]; p<0·0001) than did those with the hypoinflammatory subphenotype. Although HARP-2 found no difference in 28-day survival between placebo and simvastatin, significantly different survival was identified across patients stratified by treatment and subphenotype (p<0·0001). Specifically, within the hyperinflammatory subphenotype, patients treated with simvastatin had significantly higher 28-day survival than did those given placebo (p=0·008). A similar pattern was observed for 90-day survival. Two subphenotypes of ARDS were identified in the HARP-2 cohort, with distinct clinical and biological features and disparate clinical outcomes. The hyperinflammatory subphenotype had improved survival with simvastatin compared with placebo. These findings support further pursuit of predictive enrichment strategies in critical care clinical trials. UK Efficacy and Mechanism Evaluation Programme and National Institutes of Health.


Probing the contacts of a low-affinity substrate with a membrane-embedded transport protein using 1H-13C cross-polarisation magic-angle spinning solid-state NMR.
Solid-state NMR combined with sample deuteration was used to probe the proximity of the low-affinity substrate D-glucose to its binding site within the Escherichia coli sugar transport protein GalP. Samples of E. coli inner membranes with amplified expression of GalP were incubated in D(2)O with D-[(13)C(6)]glucose and (13)C NMR signals from the substrate were assigned in two-dimensional dipolar-assisted rotational resonance (DARR) spectra. The signals were confirmed as representing D-glucose bound to GalP as the peaks were abolished after the substrate was displaced from the specific site with the inhibitor forskolin. The (13)C chemical shift values for D-[(13)C(6)]glucose in solution revealed some differences compared to those for ligand bound to GalP, the differences being most pronounced for positions C1 and C2, and especially for C1 in the α-anomer. (13)C cross-polarization build-up was measured for C1 and C2 of D-[(13)C(6)]glucose and D-[(2)H(7), (13)C(6)]glucose in GalP membranes suspended in D(2)O. The build-up curves for the deuterated substrate reflect intermolecular (1)H-(13)C interactions between the protein and the fully deuterated substrate; the signal build-up suggests that the α-anomer is situated closer to the protein binding site than is the β-anomer, consistent with its relatively high signal intensities and more pronounced chemical shift changes in the 2D-correlation spectra. These results demonstrate the utility of solid-state NMR combined with sample deuteration for mapping the binding interface of low affinity ligands with membrane proteins.


Amplitude variations on a MEMS-based extreme adaptive optics coronagraph testbed.
High-contrast imaging techniques such as coronagraphy are expected to play an important role in the imaging of extrasolar planets. Instruments like the Gemini Planet Imager (GPI) or the Spectro-Polar-Imetric High-Contrast Exoplanet Research (SPHERE) require high-dynamic range, achieved using coronagraphs to block light coming from the parent star. An extremely good adaptive optics (AO) system is required to reduce dynamic atmospheric wavefront errors to 50-100 nm rms. Systematic wavefront errors must also be controlled at the nanometer-equivalent level to remove persistent speckle artifacts. While precision AO systems can control wavefront phase errors at this level, systematic amplitude or intensity errors can also produce speckle artifacts and are uncontrolled by traditional AO phase conjugation. On the Laboratory for Adaptive Optics (LAO) extreme AO testbed, we observed a discrepancy between the coronagraphic image profile and the profile predicted by simple simulations using the measured optical phase, which could potentially be explained by amplitude variations. Measurements showed up to 7% rms intensity changes across the microelectrical mechanical (MEM) plane of the system. We identified potential sources of amplitude variation and compared them to a Fresnel model of the system. One potential concern was the surface structure of the MEM system's (MEMS) deformable mirror, but analysis shows that it induces at most 2% rms variation. The bulk of the observed intensity variation is due to nonuniform illumination of the system by the input single-mode fiber and phase errors mixing into amplitude at the nonpupil-plane due to the Talbot effect, coupled with residual astigmatism in the pupil imager.


Protein crystallization promotes type 2 immunity and is reversible by antibody treatment.
Although spontaneous protein crystallization is a rare event in vivo, Charcot-Leyden crystals (CLCs) consisting of galectin-10 (Gal10) protein are frequently observed in eosinophilic diseases, such as asthma. We found that CLCs derived from patients showed crystal packing and Gal10 structure identical to those of Gal10 crystals grown in vitro. When administered to the airways, crystalline Gal10 stimulated innate and adaptive immunity and acted as a type 2 adjuvant. By contrast, a soluble Gal10 mutein was inert. Antibodies directed against key epitopes of the CLC crystallization interface dissolved preexisting CLCs in patient-derived mucus within hours and reversed crystal-driven inflammation, goblet-cell metaplasia, immunoglobulin E (IgE) synthesis, and bronchial hyperreactivity (BHR) in a humanized mouse model of asthma. Thus, protein crystals may promote hallmark features of asthma and are targetable by crystal-dissolving antibodies.


Leukocyte Subset Changes in Response to a 164-km Road Cycle Ride in a Hot Environment.
The purpose of this observational study was to determine the circulating leukocyte subset response to completing the 2013 Hotter'N Hell Hundred recreational 164-km road cycle event in a hot and humid environmental condition. Twenty-eight men and four women were included in this study. Whole blood samples were obtained 1-2 hours before (PRE) and immediately after (POST) the event. Electronic sizing/sorting and cytometry were used to determine complete blood counts (CBC) including neutrophil, monocyte, and lymphocyte subsets. The concentration of circulating total leukocytes (103·μL-1) increased 134% from PRE to POST with the greatest increase in neutrophils (319%, p<0.0001). Circulating monocytes (including macrophages) increased 24% (p=0.004) and circulating lymphocytes including B and T cells increased 53% (p<0.0001). No association was observed between rolling time or relative intensity and leukocyte subset. Completing the Hotter n' Hell Hundred (HHH), a 100 mile recreational cycling race in extreme (hot and humid) environmental conditions, induces a substantial increase in total leukocytes in circulation. The contribution of increases in specific immune cell subsets is not equal, with neutrophils increasing to greater than 4-fold starting values from PRE to POST race. It is likely that exercise in stressful environmental conditions affects the complement of circulating immune cells, although activational state and characterization of specific leukocyte subsets remains unclear. The observed increase in circulating cell sub-populations suggests that the circulating immune surveillance system may be acutely affected by exercise in hot and humid conditions.


Large-scale genome-wide association studies and meta-analyses of longitudinal change in adult lung function.
Genome-wide association studies (GWAS) have identified numerous loci influencing cross-sectional lung function, but less is known about genes influencing longitudinal change in lung function. We performed GWAS of the rate of change in forced expiratory volume in the first second (FEV1) in 14 longitudinal, population-based cohort studies comprising 27,249 adults of European ancestry using linear mixed effects model and combined cohort-specific results using fixed effect meta-analysis to identify novel genetic loci associated with longitudinal change in lung function. Gene expression analyses were subsequently performed for identified genetic loci. As a secondary aim, we estimated the mean rate of decline in FEV1 by smoking pattern, irrespective of genotypes, across these 14 studies using meta-analysis. The overall meta-analysis produced suggestive evidence for association at the novel IL16/STARD5/TMC3 locus on chromosome 15 (P = 5.71 × 10(-7)). In addition, meta-analysis using the five cohorts with ≥3 FEV1 measurements per participant identified the novel ME3 locus on chromosome 11 (P = 2.18 × 10(-8)) at genome-wide significance. Neither locus was associated with FEV1 decline in two additional cohort studies. We confirmed gene expression of IL16, STARD5, and ME3 in multiple lung tissues. Publicly available microarray data confirmed differential expression of all three genes in lung samples from COPD patients compared with controls. Irrespective of genotypes, the combined estimate for FEV1 decline was 26.9, 29.2 and 35.7 mL/year in never, former, and persistent smokers, respectively. In this large-scale GWAS, we identified two novel genetic loci in association with the rate of change in FEV1 that harbor candidate genes with biologically plausible functional links to lung function.