Menstrual Suppression in Gender Minority Youth
The purpose of this case series was to evaluate menstrual suppression in sex assigned at birth female adolescents identifying as male or gender non-conforming. A retrospective chart review of four gender minority youth (GMY), age 14-17, was performed for gender identity history, type and success of menstrual suppression, method satisfaction, side effects and improvement in menstrual distress. Menstrual suppression was successful in three patients, one patient discontinued use due to side effects that caused an increase in gender dysphoria. Menstrual distress and bleeding pattern improved in the majority of GMY in this series but side effects, as well as contraindications, may limit their use. In conclusion, menstrual dysphoria can be life-threatening for GMY and it is important that clinicians consider menstrual suppression in GMY with menstrual dysphoria. This series emphasizes the importance of individualized treatment plans.


Corner excitations in the 2D triangle-shaped topological insulator with chiral superconductivity on the triangular lattice.
The 2D triangle-shaped C 3-symmetric topological insulator with the chiral superconducting coupling on the triangular lattice is investigated. While such a system cannot provide the topologically protected corner excitations, we report the presence of the nontopological corner excitations with energy value to lie in the first-order edge spectrum gap. Though these excitations are not topologically protected, they appear for a rather wide range of the parameters values and are robust against the boundary defects and weak disorder. We reveal the presence of the Majorana corner states, which appear along the line in the parameter space.


Appropriateness of antibiotic prophylaxis for major surgery in Korea.
To determine the appropriateness of antibiotic prophylaxis regimens for major surgery in Korea. Retrospective study using a written survey for each patient who underwent arthroplasty, colon surgery, or hysterectomy. Six tertiary hospitals in Seoul and Gyeonggi Province. From each hospital, a maximum of 150 patients who underwent each type of surgery were randomly chosen for the study. Of 2,644 eligible patients, 1,914 patients were included in the analysis; 677 of these patients underwent arthroplasty, 578 underwent colon surgery, and 659 underwent hysterectomy. Nineteen patients were excluded from the analyses of the class and number of antibiotics used for prophylaxis because they underwent multiple surgeries at different sites. For each of the 1,895 remaining patients, antibiotic prophylaxis involved a mean (+/-SD) of 2.8 +/- 0.9 classes of antibiotics. The most commonly prescribed agents were cephalosporins (prescribed for 1,875 [98.9%] of the patients) and aminoglycosides (1,404 [74.1%]). A total of 1,574 (83.1%) of patients received at least 2 classes of antibiotics simultaneously. Only 15 (0.8%) of 1,895 patients received antibiotic prophylaxis in accordance with published guidelines. Of 506 patients for whom the initial dose of antibiotics was evaluated, 374 (73.9%) received an appropriate initial dose. Of the 1,676 patients whose medical records included information about antibiotic administration relative to the time of surgery, only 188 (11.2%) received antibiotic prophylaxis an hour or less before the surgical incision was made. Of the 1,748 patients whose medical records included information about duration of surgery, antibiotic prophylaxis was discontinued 24 hours or less after surgery for only 3 (0.2%) of the patients. Most patients who had major surgery in Korea received inappropriate antibiotic prophylaxis. Measures to improve the appropriateness of antibiotic prophylaxis are urgently required.


STOCK MARKET CRASH AND EXPECTATIONS OF AMERICAN HOUSEHOLDS.
This paper utilizes data on subjective probabilities to study the impact of the stock market crash of 2008 on households' expectations about the returns on the stock market index. We use data from the Health and Retirement Study that was fielded in February 2008 through February 2009. The effect of the crash is identified from the date of the interview, which is shown to be exogenous to previous stock market expectations. We estimate the effect of the crash on the population average of expected returns, the population average of the uncertainty about returns (subjective standard deviation), and the cross-sectional heterogeneity in expected returns (disagreement). We show estimates from simple reduced-form regressions on probability answers as well as from a more structural model that focuses on the parameters of interest and separates survey noise from relevant heterogeneity. We find a temporary increase in the population average of expectations and uncertainty right after the crash. The effect on cross-sectional heterogeneity is more significant and longer lasting, which implies substantial long-term increase in disagreement. The increase in disagreement is larger among the stockholders, the more informed, and those with higher cognitive capacity, and disagreement co-moves with trading volume and volatility in the market.


An Epidemiological Analysis of Head Injuries in Taiwan.
Traumatic head injuries occur frequently in Taiwan, having catastrophic consequences for the victims, their families, and society as a whole. However, little is known about the risk factors at the population level in Taiwan. The primary aim of this study was to obtain more information on these variables and their relationships. Another aim was to analyze the effects of independent variables such as sex, age, residency, pre-existing conditions, mechanisms of injury, associated injuries, and severity on the probability of in-hospital death. Using the 2007⁻2008 total admissions claim dataset from Taiwan's National Health Insurance system, total admissions due to acute head injury were selected for further analysis. The obtained data included patient demographics and trauma hospitalization rate. A total of 99,391 patients were admitted with head injury, 48,792 of which had moderate-to-severe head injury. There were 4935 cases recorded as in-hospital mortality and the standardized in-hospital mortality rate was 10.7 deaths per 100,000 person-years. The mortality rate increased with age. After adjustments, male sex, age older than 54 years, living in a rural area, lower monthly income, a Charlson comorbidity index greater than one, being a pedestrian hit by a motor vehicle, fall from a height, and having significant chest, abdominal, or lower extremity injury increased the risk of death during admission. This population-based analysis provides information about the incidence rate and death rate for admissions in Taiwan due to acute head injury and the factors that affect in-hospital mortality. Our results that highlight the risk factors for adverse outcome can help us prevent or improve rural area trauma care of head injury patients in the future.


Sphingomyelinase activity of Trichomonas vaginalis extract and subfractions.
Trichomoniasis is one of the most common acute sexually transmitted curable diseases, and it is disseminated worldwide generating more than 170 million cases annually. Trichomonas vaginalis is the parasite that causes trichomoniasis and has the ability to destroy cell monolayers of the vaginal mucosa in vitro. Sphingomyelinases (SMase) are enzymes that catalyze the hydrolysis of sphingomyelin into ceramide and phosphorylcholine. Ceramide appears to be a second messenger lipid in programmed apoptosis, cell differentiation, and cell proliferation. Sphingomyelinase is probably a major source of ceramide in cells. Signal transduction mediated by ceramide leads cells to produce cytokine induced apoptosis during several inflammatory responses. SMase are also relevant toxins in several microorganisms. The main objective of this research is to identify SMase activity of T. vaginalis in the total extract (TE), P30, and S30 subfractions from brooked trophozoites. It was found that these fractions of T. vaginalis have SMase activity, which comes principally from P30 subfraction and was mainly type C. Enzymatic activity of SMase increased linearly with time and is pH dependent with two peaks by pH 5.5 and pH 7.5. The addition of manganese to the reaction mixture increased the SMase activity by 1.97.


Reference bias in reports of drug trials.
Articles published before 1985 describing double blind trials of two or more non-steroidal anti-inflammatory drugs in rheumatoid arthritis were examined to see whether there was any bias in the references they cited. Althogether 244 articles meeting the criteria were found through a Medline search and through examining the reference lists of the articles retrieved. The drugs compared in the studies were classified as new or as control drugs and the outcome of the trial as positive or not positive. The reference lists of all papers with references to other trials on the new drug were then examined for reference bias. Positive bias was judged to have occurred if the reference list contained a higher proportion of references with a positive outcome for that drug than among all the articles assumed to have been available to the authors (those published more than two years earlier than the index article). Altogether 133 of the 244 articles were excluded for various reasons--for example, 44 because of multiple publication and 19 because they had no references. Among the 111 articles analysed bias was not possible in the references of 35 (because all the references gave the same outcome); 10 had a neutral selection of references, 22 a negative selection, and 44 a positive selection--a significant positive bias. This bias was not caused by better scientific standing of the cited articles over the uncited ones. Thus retrieving literature by scanning reference lists may produce a biased sample of articles, and reference bias may also render the conclusions of an article less reliable.


Adenovirus-mediated gene therapy of hepatocellular carcinoma using cancer-specific gene expression.
Most patients with hepatocellular carcinoma have an elevated alpha-feto-protein (AFP) level. This high level of AFP expression is transcriptionally controlled by the 5'-flanking sequence of the AFP gene. Using the 5'-flanking sequence as a promoter for the herpes simplex virus thymidine kinase (HSV-TK) gene in an adenoviral vector (Av1AFPTK1), the therapeutic efficacy of adenovirus-mediated HSV-TK gene transduction, followed by ganciclovir (GCV) administration, was studied in tumors in athymic nude mice. Av1AFPTK1 transduction of two cell lines demonstrated HSV-TK enzyme activity only in the AFP-producing cells (HuH7) and not in the AFP nonproducing cells (SK-Hep-1). As expected, only transduced HuH7 cells were killed by GCV treatment. Transduction by an adenoviral vector harboring a Rous sarcoma virus promoter and HSV-TK gene (Av1TK1) showed enzymatic activity and GCV killing in both cell lines. All HuH7 tumors that were transduced with either Av1AFPTK1 or Av1TK1 completely regressed after GCV treatment. On the other hand, there was complete regression of SK-Hep-1 tumors only when treated with Av1TK1 and GCV and not when treated with Av1AFPTK1 and GCV. Thus, cell-specific killing was achieved by adenoviral vector containing AFP promoter for the HSV-TK gene and GCV treatment.


Haplotype and phenotype analysis of six recurrent BRCA1 mutations in 61 families: results of an international study.
Several BRCA1 mutations have now been found to occur in geographically diverse breast and ovarian cancer families. To investigate mutation origin and mutation-specific phenotypes due to BRCA1, we constructed a haplotype of nine polymorphic markers within or immediately flanking the BRCA1 locus in a set of 61 breast/ovarian cancer families selected for having one of six recurrent BRCA1 mutations. Tests of both mutations and family-specific differences in age at diagnosis were not significant. A comparison of the six mutations in the relative proportions of cases of breast and ovarian cancer was suggestive of an effect (P = .069), with 57% of women presumed affected because of the 1294 del 40 BRCA1 mutation having ovarian cancer, compared with 14% of affected women with the splice-site mutation in intron 5 of BRCA1. For the BRCA1 mutations studied here, the individual mutations are estimated to have arisen 9-170 generations ago. In general, a high degree of haplotype conservation across the region was observed, with haplotype differences most often due to mutations in the short-tandem-repeat markers, although some likely instances of recombination also were observed. For several of the instances, there was evidence for multiple, independent, BRCA1 mutational events.


Nutrition and physical activity in NAFLD: an overview of the epidemiological evidence.
Nonalcoholic fatty liver disease (NAFLD) has been recognized as a major health burden. The high prevalence of NAFLD is probably due to the contemporary epidemics of obesity, unhealthy dietary pattern, and sedentary lifestyle. The efficacy and safety profile of pharmacotherapy in the treatment of NAFLD remains uncertain and obesity is strongly associated with hepatic steatosis; therefore, the first line of treatment is lifestyle modification. The usual management of NAFLD includes gradual weight reduction and increased physical activity (PA) leading to an improvement in serum liver enzymes, reduced hepatic fatty infiltration, and, in some cases, a reduced degree of hepatic inflammation and fibrosis. Nutrition has been demonstrated to be associated with NAFLD and Non-alcoholic steatohepatitis (NASH) in both animals and humans, and thus serves as a major route of prevention and treatment. However, most human studies are observational and retrospective, allowing limited inference about causal associations. Large prospective studies and clinical trials are now needed to establish a causal relationship. Based on available data, patients should optimally achieve a 5%-10% weight reduction. Setting realistic goals is essential for long-term successful lifestyle modification and more effort must be devoted to informing NAFLD patients of the health benefits of even a modest weight reduction. Furthermore, all NAFLD patients, whether obese or of normal weight, should be informed that a healthy diet has benefits beyond weight reduction. They should be advised to reduce saturated/trans fat and increase polyunsaturated fat, with special emphasize on omega-3 fatty acids. They should reduce added sugar to its minimum, try to avoid soft drinks containing sugar, including fruit juices that contain a lot of fructose, and increase their fiber intake. For the heavy meat eaters, especially those of red and processed meats, less meat and increased fish intake should be recommended. Minimizing fast food intake will also help maintain a healthy diet. PA should be integrated into behavioral therapy in NAFLD, as even small gains in PA and fitness may have significant health benefits. Potentially therapeutic dietary supplements are vitamin E and vitamin D, but both warrant further research. Unbalanced nutrition is not only strongly associated with NAFLD, but is also a risk factor that a large portion of the population is exposed to. Therefore, it is important to identify dietary patterns that will serve as modifiable risk factors for the prevention of NAFLD and its complications.


The impact of documentation on communication during patient-physiotherapist interactions: A qualitative observational study.
Documentation is an essential component of physiotherapy practice for clinical, legal, and ethical reasons. Research in other healthcare contexts suggests that documentation impacts upon communication in patient-practitioner interactions. Thus, the objective of this qualitative study was to examine how physiotherapists and their patients communicate during episodes of documentation. The research was informed by ethnomethodology and ethnography. In total, 113 patient-physiotherapist interactions were observed in Switzerland and Australia with video-recordings, audio-recordings, and field notes collected as data. Episodes of documentation within these interactions were transcribed, and both verbal and non-verbal communication were analyzed inductively. Analysis identified that communication during documentation was characterized by: pauses in conversation, pre-established order of questioning, minimal eye contact, use of direct communication, and an emphasis on objectivity. The use of documentation was observed to alter the wording of questioning as well as the sequence and flow of conversation between patient and physiotherapist. In addition, the observed communicative features seemed to restrict patient participation, and may hinder the achievement of a patient-centered approach. Recognizing the importance of documentation, we address the challenges that our research highlighted by proposing strategies to assist educators and clinicians to optimize communication with patients when incorporating documentation into practice.


Fiber orientation-dependent white matter contrast in gradient echo MRI.
Recent studies have shown that there is a direct link between the orientation of the nerve fibers in white matter (WM) and the contrast observed in magnitude and phase images acquired using gradient echo MRI. Understanding the origin of this link is of great interest because it could offer access to a new diagnostic tool for investigating tissue microstructure. Since it has been suggested that myelin is the dominant source of this contrast, creating an accurate model for characterizing the effect of the myelin sheath on the evolution of the NMR signal is an essential step toward fully understanding WM contrast. In this study, we show by comparison of the results of simulations and experiments carried out on human subjects at 7T, that the magnitude and phase of signals acquired from WM in vivo can be accurately characterized by (i) modeling the myelin sheath as a hollow cylinder composed of material having an anisotropic magnetic susceptibility that is described by a tensor with a radially oriented principal axis, and (ii) adopting a two-pool model in which the water in the sheath has a reduced T(2) relaxation time and spin density relative to its surroundings, and also undergoes exchange. The accuracy and intrinsic simplicity of the hollow cylinder model provides a versatile framework for future exploitation of the effect of WM microstructure on gradient echo contrast in clinical MRI.


The voltage dependence of depolarization-induced calcium release in isolated skeletal muscle triads.
We demonstrate for the first time in this study that triadic vesicles derived from skeletal muscle display a voltage dependence of depolarization-induced calcium release similar to that found in intact muscle. We confirm previous studies by Dunn (1989) which demonstrated that changes in extravesicular potassium induced membrane potential changes in isolated transverse tubules with the voltage sensitive dye DiSC(3)-5. Depolarization-induced calcium release was studied in isolated triadic vesicles through similar changes in extravesicular [K] while clamping extravesicular Ca++ to submicromolar concentrations. The amplitude of fast phase of calcium release, identified as depolarization-induced calcium release, varied with the percentage of transverse tubules in the preparation (determined through 3H-PN200-110 specific activity) and different levels of depolarization. Threshold activation of calcium release was obtained with a 40.5 mV potential change; maximal calcium release was obtained with a 75 to 81 mV potential change. Boltzmann fits to the normalized depolarization induced calcium release plotted against the membrane potential change yielded a voltage dependence (k = 4.5 mV per e-fold change) very similar to that found in intact muscle (k = 3-4 mV per e-fold change; Baylor, Chandler & Marshall 1978, 1983; Miledi et al., 1981). Substitution of methanesulfonate for propionate as the impermeant ion or addition of valinomycin in the depolarizing solutions had little effect on the voltage dependence of calcium release.


Prairie dog alarm calls encode labels about predator colors.
Some animals have the cognitive capacity to differentiate between different species of predators and generate different alarm calls in response. However, the presence of any addition information that might be encoded into alarm calls has been largely unexplored. In the present study, three similar-sized human females walked through a Gunnison's prairie dog (Cynomys gunnisoni) colony wearing each of three different-colored shirts: blue, green, and yellow. We recorded the alarm calls and used discriminant function analysis to assess whether the calls for the different-colored shirts were significantly different. The results showed that the alarm calls for the blue and the yellow shirts were significantly different, but the green shirt calls were not significantly different from the calls for the yellow shirt. The colors that were detected, with corresponding encoding into alarm calls, reflect the visual perceptual abilities of the prairie dogs. This study suggests that prairie dogs are able to incorporate labels about the individual characteristics of predators into their alarm calls, and that the complexity of information contained in animal alarm calls may be greater than has been previously believed.


Selenium segregation in femtosecond-laser hyperdoped silicon revealed by electron tomography.
Doping of silicon with chalcogens (S, Se, Te) by femtosecond laser irradiation to concentrations well above the solubility limit leads to near-unity optical absorptance in the visible and infrared (IR) range and is a promising route toward silicon-based IR optoelectronics. However, open questions remain about the nature of the IR absorptance and in particular about the impact of the dopant distribution and possible role of dopant diffusion. Here we use electron tomography using a high-angle annular dark-field (HAADF) detector in a scanning transmission electron microscope (STEM) to extract information about the three-dimensional distribution of selenium dopants in silicon and correlate these findings with the optical properties of selenium-doped silicon. We quantify the tomography results to extract information about the size distribution and density of selenium precipitates. Our results show correlation between nanoscale distribution of dopants and the observed sub-band gap optical absorptance and demonstrate the feasibility of HAADF-STEM tomography for the investigation of dopant distribution in highly-doped semiconductors.


Inhibitors of telomerase and poly(ADP-ribose) polymerases synergize to limit the lifespan of pancreatic cancer cells.
Imetelstat (GRN163L) is a potent and selective inhibitor of telomerase. We have previously reported that GRN163L could shorten telomeres and limit the lifespan of CD18/HPAF and CAPAN1 pancreatic cancer cells. Here, we examined the effects of GRN163L on two other pancreatic cancer cell lines: AsPC1 and L3.6pl. In both lines, chronic exposure to GRN163L led to an initial shortening of telomeres followed by a stabilization of extremely short telomeres. In AsPC1 cells, telomere attrition eventually led to the induction of crisis and the loss of the treated population. In L3.6pl cells, crisis was transient and followed by the emergence of GRN163L-resistant cells, which could grow at increasing concentrations of GRN163L. The Shelterin complex is a telomere-associated complex that limits the access of telomerase to telomeres. The telomerase inhibitory function of this complex can be enhanced by drugs that block the poly(ADP-ribosyl)ation of its TRF1 and/or TRF2 subunits. Combined treatment of the GRN163L-resistant L3.6pl cells with GRN163L and 3-aminobenzamide (3AB), a general inhibitor of poly(ADP-ribose) polymerases, led to additional telomere shortening and limited the lifespan of the resistant cells. Results from this work suggest that inhibitors of telomerase and poly(ADP-ribose) polymerases can cooperate to limit the lifespan of pancreatic cancer cells.


Understanding the conformational impact of chemical modifications on monoclonal antibodies with diverse sequence variation using hydrogen/deuterium exchange mass spectrometry and structural modeling.
Chemical modifications can potentially induce conformational changes near the modification site and thereby impact the safety and efficacy of protein therapeutics. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) has emerged as a powerful analytical technique with high spatial resolution and sensitivity in detecting such local conformational changes. In this study, we utilized HDX-MS combined with structural modeling to examine the conformational impact on monoclonal antibodies (mAbs) caused by common chemical modifications including methionine (Met) oxidation, aspartic acid (Asp) isomerization, and asparagine (Asn) deamidation. Four mAbs with diverse sequences and glycosylation states were selected. The data suggested that the impact of Met oxidation was highly dependent on its location and glycosylation state. For mAbs with normal glycosylation in the Fc region, oxidation of the two conserved Met252 and Met428 (Kabat numbering) disrupted the interface interactions between the CH2 and CH3 domains, thus leading to a significant decrease in CH2 domain thermal stability as well as a slight increase in aggregation propensity. In contrast, Met oxidation in the variable region and CH3 domain had no detectable impact on mAb conformation. For aglycosylated mAb, Met oxidation could cause a more global conformational change to the whole CH2 domain, coincident with the larger decrease in thermal stability and significant increase in aggregation rate. Unlike Met oxidation, Asn deamidation and Asp isomerization mostly had very limited effects on mAb conformation, with the exception of succiminide intermediate formation which induced a measurable local conformational change to be more solvent protected. Structural modeling suggested that the succinimide intermediate was stabilized by adjacent aromatic amino acids through ring-ring stacking interactions.


Sulphur limitation provokes physiological and leaf proteome changes in oilseed rape that lead to perturbation of sulphur, carbon and oxidative metabolisms.
The decline in industrial emissions of sulphur (S) has led to a sulphate depletion in soil resulting in an alteration of crop performance. In oilseed rape, an S deficiency dramatically reduced the seed yield and/or quality. Paradoxically, little is known about the impact of sulphate limitation on oilseed rape leaf metabolism, despite it being a key determinant of growth. In order to identify the metabolic processes involved in the oilseed rape response to S restriction, an analysis of the young leaf proteome combined with a physiological study was carried out at the vegetative stage. S limitation does not significantly reduce the total shoot biomass but inhibits growth and photosynthesis of young leaves. This photosynthesis decline is not due to a decrease in chlorophyll content, which remains similar to Control. The increase in anthocyanins and H(2)O(2) content in young leaves of S-limited plants suggests that S restriction leads to an oxidative stress. Proteomic analysis at 35 d of S limitation also revealed the induction of 12-oxophitodienoate reductase and ACC synthase, respectively involved in jasmonate and ethylene biosynthesis, two phytohormones that could be implicated in oxidative stress. Proteins involved in photosynthesis and carbon metabolism were also modulated by S restriction. In particular, the decrease in plastocyanin and ferredoxin-NADP reductase suggests that H(2)O(2) accumulation is associated with perturbation of the photosynthetic electron transport chain. The accumulation of chloroplastic Cu-Zn SOD reinforces the idea that an oxidative stress probably occurs in the chloroplast. Proteomic results suggest that the maintenance of chlorophyll in S-limited conditions is related to an accumulation of Water Soluble Chlorophyll binding Proteins, involved in the protection of chlorophyll against ROS. The accumulation of the catalytic α-subunit of chloroplastic ATP synthase suggests that energy production is maintained. S limitation leads to photosynthesis and carbon metabolism disturbances that could be responsible for the oxidative stress observed in the young leaves of oilseed rape. Despite this, induction of proteins involved in oxidative stress resistance and energy production shows that the leaf capacity to capture and use photosynthetic active radiations for ATP production remains efficient for as long as possible.


Correction of Lower Eyelid Retraction Using Acellular Human Dermis During Orbital Decompression.
To investigate the surgical outcome of grafting acellular human dermis compared with concurrent lower eyelid retractor recession during swinging eyelid orbital decompression for correction of lower eyelid retraction in patients with Graves' orbitopathy. Institutional Review Board-approved retrospective nonrandomized study of 54 Graves' orbitopathy patients (95 eyes) from 2008 to 2012. Patients who received decompression were divided into 3 groups: Group 1 with conjunctival lengthening using 0.3-mm thickness AlloDerm (36 eyes), Group 2 with inferior retractor recession (33 eyes), and Group 3 with decompression only (26 eyes). Outcome measures included lower eyelid height, inferior sclera show, cosmetic appearance, and complications. Baseline clinical characteristics and the degree of improvement of exophthalmos were not different between groups. Preoperative MRD2 was higher in group 1 (8.0 mm) than in groups 2 (6.9 mm, p < 0.001) and 3 (6.6 mm, p < 0.001). Mean improvement of MRD2 as well as postoperative improvement of inferior scleral show at postoperative 4 to 6 months were significantly higher in group 1 (2.7 mm) than in groups 2 (1.8 mm, p < 0.001) and 3 (1.2 mm, p < 0.001). Simultaneous correction of lower eyelid retraction using thin AlloDerm during swinging eyelid orbital decompression maximizes improvement of lower eyelid retraction compared with concurrent inferior retractor recession.


Living on the edge with too many mouths to feed: why dopamine neurons die.
Although genes, protein aggregates, environmental toxins, and other factors associated with Parkinson's disease (PD) are widely distributed in the nervous system and affect many classes of neurons, a consistent feature of PD is the exceptional and selective vulnerability of dopamine (DA) neurons of the SNc. What is it about these neurons, among all other neurons in the brain, that makes them so susceptible in PD? We hypothesize that a major contributory factor is the unique cellular architecture of SNc DA neuron axons. Their large, complex axonal arbour puts them under such a tight energy budget that it makes them particularly susceptible to factors that contribute to cell death, including unique molecular characteristics associated with SNc DA neurons and nonspecific, nervous-system-wide factors.


RAPA-ConvNets: Modified Convolutional Networks for Accelerated Training on Architectures With Analog Arrays.
Analog arrays are a promising emerging hardware technology with the potential to drastically speed up deep learning. Their main advantage is that they employ analog circuitry to compute matrix-vector products in constant time, irrespective of the size of the matrix. However, ConvNets map very unfavorably onto analog arrays when done in a straight-forward manner, because kernel matrices are typically small and the constant time operation needs to be sequentially iterated a large number of times. Here, we propose to parallelize the training by replicating the kernel matrix of a convolution layer on distinct analog arrays, and randomly divide parts of the compute among them. With this modification, analog arrays execute ConvNets with a large acceleration factor that is proportional to the number of kernel matrices used per layer (here tested 16-1024). Despite having more free parameters, we show analytically and in numerical experiments that this new convolution architecture is self-regularizing and implicitly learns similar filters across arrays. We also report superior performance on a number of datasets and increased robustness to adversarial attacks. Our investigation suggests to revise the notion that emerging hardware architectures that feature analog arrays for fast matrix-vector multiplication are not suitable for ConvNets.


IL-1 β and IL-6 upregulation in children with H1N1 influenza virus infection.
The role of cytokines in relation to clinical manifestations, disease severity, and outcome of children with H1N1 virus infection remains thus far unclear. The aim of this study was to evaluate interleukin IL-1 β and IL-6 plasma expressions and their association with clinical findings, disease severity, and outcome of children with H1N1 infection. We prospectively evaluated 15 children with H1N1 virus infection and 15 controls with lower respiratory tract infections (LRTI). Interleukin plasma levels were measured using immunoenzymatic assays. Significantly higher levels of IL-1 β and IL-6 were detected in all patients with H1N1 virus infection compared to controls. It is noteworthy to mention that in H1N1 patients with more severe clinical manifestations of disease IL-1 β and IL-6 expressions were significantly upregulated compared to H1N1 patients with mild clinical manifestations. In particular, IL-6 was significantly correlated with specific clinical findings, such as severity of respiratory compromise and fever. No correlation was found between interleukin expression and final outcome. In conclusion, H1N1 virus infection induces an early and significant upregulation of both interleukins IL1 β and IL-6 plasma expressions. The upregulation of these cytokines is likely to play a proinflammatory role in H1N1 virus infection and may contribute to airway inflammation and bronchial hyperreactivity in these patients.


Norepinephrine release in the immature ovary is regulated by autoreceptors and neuropeptide-Y.
Experiments were undertaken to study the role that neuropeptide-Y (NPY) and adrenergic autoreceptors may play in the regulation of norepinephrine (NE) release from the rat ovary. Ovaries from 28- to 32-day-old rats were preincubated with [3H]NE, and the release of the recently taken up catecholamine in response to electric field stimulation was assessed. The release was strictly dependent on the presence of extracellular calcium and decreased when the frequency of stimulation was increased. This drop in [3H]NE release was significantly reversed by exposure of the ovaries during stimulation to yohimbine, a selective alpha 2-adrenoreceptor blocker. The existence of prejunctional alpha 2-adrenergic autoreceptors in ovarian nerves was further suggested by the ability of exogenous NE to mimic the inhibitory effect of high frequency stimulation. NPY inhibited by 40% the release of [3H]NE induced by electrical stimulation. The specificity of this effect and its prejunctional nature were demonstrated by the finding that avian pancreatic polypeptide, a structural homolog of NPY that is not recognized by prejunctional NPY receptors of the Y2 subtype, failed to alter the induced release of [3H]NE. Neither NPY, avian pancreatic polypeptide, nor peptide-YY, another member of the pancreatic polypeptide fold family, altered progesterone or estradiol secretion from whole ovaries or granulosa cells in culture, suggesting that NPY (and its structural homologs) does not directly affect ovarian steroidogenesis. The results suggest that 1) the release of NE from ovarian sympathetic nerves is subjected to a dual modulatory influence provided by NE itself and NPY; and 2) this regulatory effect is exerted via specific prejunctional receptors. Such NE/NPY actions are likely to regulate the availability of NE to its postsynaptic receptors during ovarian development and adult function. The fact that NPY is mostly released during high frequency stimulation raises the possibility of NPY involvement in ovarian dysfunctions associated with situations of enhanced sympathetic discharge, such as strenuous exercise and psychogenic amenorrhea.


Spatial and quantitative datasets of the pancreatic β-cell mass distribution in lean and obese mice.
A detailed understanding of pancreatic β-cell mass distribution is a key element to fully appreciate the pathophysiology of models of diabetes and metabolic stress. Commonly, such assessments have been performed by stereological approaches that rely on the extrapolation of two-dimensional data and provide very limited topological information. We present ex vivo optical tomographic data sets of the full β-cell mass distribution in cohorts of obese ob/ob mice and their lean controls, together with information about individual islet β-cell volumes, their three-dimensional coordinates and shape throughout the volume of the pancreas between 4 and 52 weeks of age. These data sets offer the currently most comprehensive public record of the β-cell mass distribution in the mouse. As such, they may serve as a quantitative and topological reference for the planning of a variety of in vivo or ex vivo experiments including computational modelling and statistical analyses. By shedding light on intra- and inter-lobular variations in β-cell mass distribution, they further provide a powerful tool for the planning of stereological sampling assessments.


European position paper on rhinosinusitis and nasal polyps 2007.
Rhinosinusitis is a significant and increasing health problem which results in a large financial burden on society. This evidence based position paper describes what is known about rhinosinusitis and nasal polyps, offers evidence based recommendations on diagnosis and treatment, and considers how we can make progress with research in this area. Rhinitis and sinusitis usually coexist and are concurrent in most individuals; thus, the correct terminology is now rhinosinusitis. Rhinosinusitis (including nasal polyps) is defined as inflammation of the nose and the paranasal sinuses characterised by two or more symptoms, one of which should be either nasal blockage/obstruction/congestion or nasal discharge (anterior/posterior nasal drip), +/- facial pain/pressure, +/- reduction or loss of smell; and either endoscopic signs of polyps and/or mucopurulent discharge primarily from middle meatus and/or; oedema/mucosal obstruction primarily in middle meatus, and/or CT changes showing mucosal changes within the ostiomeatal complex and/or sinuses. The paper gives different definitions for epidemiology, first line and second line treatment and for research. Furthermore the paper describes the anatomy and (patho)physiology, epidemiology and predisposing factors, inflammatory mechanisms, evidence based diagnosis, medical and surgical treatment in acute and chronic rhinosinusitis and nasal polyposis in adults and children. Evidence based schemes for diagnosis and treatment are given for the first and second line clinicians. Moreover attention is given to complications and socio-economic cost of chronic rhinosinusitis and nasal polyps. Last but not least the relation to the lower airways is discussed.


Control by stimulus features during fading.
Sixteen children were given four successive circle-size discrimination problems with luminance as the fading stimulus. Children who were first presented with a difficult size discrimination failed to acquire this discrimination. Those who first received an easy discrimination learned the difficult discrimination. At the end of each 10-trial block, two probe stimuli were presented to monitor any shift in control from luminance to size. One probe was the same size as the positive stimulus but of different luminance; the other was the same luminance but of different size. If, in the course of fading, size and luminance both controlled responding, fading was successful. If luminance alone controlled responding until the end of fading, the size discrimination was not established. Dual control, and thus successful fading, resulted when the target stimuli were very discriminable, or when the target stimuli were subtly different provided that previous fading series had first established less subtle discriminations.


Enhanced de novo lipogenesis in the leptin-unresponsive pancreatic islets of prediabetic Zucker diabetic fatty rats: role in the pathogenesis of lipotoxic diabetes.
Overaccumulation of fat in pancreatic islets of obese ZDF fa/fa rats is believed to cause beta-cell failure and diabetes. Previously, we demonstrated that ZDF islets have an increased capacity to esterify fatty acids imported via the circulation. Here we examine the capacity of ZDF islets to synthesize fatty acids de novo. Compared with age-matched wild-type (+/+) control islets, acetyl CoA carboxylase (ACC) mRNA was fivefold and sixfold higher and fatty acid synthetase (FAS) was fourfold and sevenfold higher in prediabetic and diabetic ZDF islets, respectively. Incorporation of label from [14C]glucose into lipids was 84% higher in ZDF islets and was not suppressed normally by fatty acids. Chronic hyperleptinemia, induced by adenoviral transfer of leptin cDNA, reduced ACC and FAS mRNA in +/+ islets by 93 and 80%, respectively, but did not decrease the high ACC and FAS expression in islets of fa/fa rats. Recombinant leptin cultured with islets isolated from +/+ rats lowered ACC and FAS expression by 66 and 47%, respectively, but had no effect in fa/fa islets. We conclude that de novo lipogenesis in islets is controlled by leptin and remains low in leptin-responsive islets. It is increased in leptin-insensitive fa/fa islets, contributing to the fat overload that leads to beta-cell dysfunction and diabetes.


Fiber Bragg grating-based thermometer for drill bit temperature monitoring.
The temperature measurement of a drill bit during an implantology drilling process is proposed by using a fiber Bragg grating fitted inside the drill bit. Due to the rotational nature of the drilling process, a free-space fiber-optic rotary joint is used for interrogating the fiber Bragg grating. Due to mechanical clearances and interferometric noise induced at this rotary joint, signal integrity is strongly deteriorated and is not workable without adequate measures. These measures involve a proper fiber lensing and a signal processing in order to remove the interferometric noise. Finally, a heating measurement on an implantology drill bit is performed and discussed for drilling several holes on a pork jaw sample.


Radiotherapy of chondrosarcoma of bone.
A retrospective analysis of 31 cases of chondrosarcoma of bone treated by radiotherapy at the Princess Margaret Hospital between 1958 and 1976 is presented. In comparison with other large series, our group of patients were found to have been unfavourably selected with respect to the known prognostic factors: histology (39% mesenchymal and dedifferentiated), site (75% arising in the trunk and head and neck), adequacy of operative treatment (none having had a complete surgical excision), and presenting symptoms (two-thirds presenting with pain). Twelve patients with primary (good, moderate, and unknown differentiation) chondrosarcoma were radically irradiated; 6 of these 12 have been alive and well without tumor for periods ranging from three and half to 16 years and 3 of these are alive and well for 15 years or more following radiotherapy. The other 6 patients responded or disease stabilized following radiotherapy for periods ranging from 16 months to eight years. One poorly differentiated tumor was radically irradiated and did not respond. Eleven patients were irradiated palliatively (5 dedifferentiated and mesenchymal, 4 secondary, and 2 primary chondrosarcomas), generally with low doses of irradiation, and only 4 responded transiently for periods ranging from three to 12 months. Seven patients with mesenchymal and dedifferentiated tumors were radically irradiated. Four responded or disease stabilized, and 1 of these patients was alive and well at 3 years; 3 did not respond. Six died with distant metastasis. It is concluded that chondrosarcoma of bone is a radioresponsive tumor and the place of radiotherapy in the treatment of this disease and the reason for its being labelled a radioresistant tumor are discussed. The problems of assessing response of chondrosarcoma to therapy are also discussed. It is suggested that chemotherapy may have a role in the management of mesenchymal and dedifferentiated chondrosarcoma.


In vitro evaluation of intraluminal factors that may alter intestinal permeability in ponies with carbohydrate-induced laminitis.
To study the in vitro effects of cecal contents incubated with corn starch on colonic permeability in horses. 4 healthy adult ponies. Mucosal specimens were obtained from the right ventral colon and mounted in Ussing chambers. Changes in short circuit current, conductance, and large-molecule permeability in response to addition of cecal contents and cecal contents incubated with corn starch were evaluated for 120 minutes. Incubation of cecal contents with corn starch for 8 hours resulted in a decrease in cecal content pH and an increase in lactic acid concentration. These changes were similar to those reported in vivo for ponies given corn starch. Exposure of colonic mucosa to cecal contents incubated with corn starch resulted in an increase in tissue conductance and permeability of technetium Tc 99m pentetate, compared with mucosa exposed to cecal contents alone. In vitro exposure of colonic mucosa to cecal contents incubated with starch resulted in increased paracellular permeability. Fermentation of excessive amounts of carbohydrate in the intestinal lumen of horses may directly induce increased intestinal permeability associated with carbohydrate-induced laminitis.


Sparse spectrum model for a turbulent phase.
Monte Carlo (MC) simulation of phase front perturbations by atmospheric turbulence finds numerous applications for design and modeling of the adaptive optics systems, laser beam propagation simulations, and evaluating the performance of the various optical systems operating in the open air environment. Accurate generation of two-dimensional random fields of turbulent phase is complicated by the enormous diversity of scales that can reach five orders of magnitude in each coordinate. In addition there is a need for generation of the long "ribbons" of turbulent phase that are used to represent the time evolution of the wave front. This makes it unfeasible to use the standard discrete Fourier transform-based technique as a basis for the MC simulation algorithm. We propose a new model for turbulent phase: the sparse spectrum (SS) random field. The principal assumption of the SS model is that each realization of the random field has a discrete random spectral support. Statistics of the random amplitudes and wave vectors of the SS model are arranged to provide the required spectral and correlation properties of the random field. The SS-based MC model offers substantial reduction of computer costs for simulation of the wide-band random fields and processes, and is capable of generating long aperiodic phase "ribbons." We report the results of model trials that determine the number of sparse components, and the range of wavenumbers that is necessary to accurately reproduce the random field with a power-law spectrum.


Environmental effects on pregnancy rate in beef cattle.
Ten years of calving records were examined from Bos taurus crossbred cows (mean of 182 cows/yr) to quantify the effects of environmental conditions during the breeding season on pregnancy rate. Estimated breeding dates were determined by subtracting 283 d from the calving date. Relationships were determined between the proportion of cows bred during the periods from the beginning of the breeding season until d 21, 42, and 60 of the breeding season and the corresponding environmental variables. Weather data were compiled from a weather station located approximately 20 km from the research site. Average daily temperature and relative humidity were used to calculate daily temperature-humidity index (THI). Daily averages for each environmental variable were averaged for each period. Minimum temperature (MNTP) and THI for the first 21 and 42 d of the breeding season were negatively associated (P < 0.001) with pregnancy rate. For the 0-to 21-d, 0- to 42-d, and 0- to 60-d breeding periods, respective r2 for average temperatures were 0.32, 0.37, and 0.11, whereas r2 for MNTP were 0.45, 0.40, and 0.10 and r2 for THI were 0.38, 0.41, and 0.11, respectively, for the same breeding periods. The negative associations of temperature and THI with pregnancy rate are most pronounced during the first 21 d of the breeding season, with a -3.79 and -2.06% change in pregnancy rate for each unit of change in MNTP and THI, respectively. A combination of environmental variables increased the R2 to 0.67. In this analysis, windspeed was found to be positively associated with pregnancy rate in all equations and increased the R2 in all breeding periods. Optimum MNTP for the 0- to 21-d, 0- to 42-d, and 0- to 60-d breeding periods was 12.6, 13.5, and 14.9 degrees C, respectively. For the 0- to 60-d breeding period, optimum THI was 68.0, whereas the THI threshold, the calculated level at which cattle will adapt, was found to be 72.9. Reductions in pregnancy rate are likely when the average MNTP and THI equal or exceed 16.7 degrees C and 72.9, respectively, and for Bos taurus beef cows that are pasture bred during a 60-d spring-summer period.


Responses of the metabolism of the larvae of Pocillopora damicornis to ocean acidification and warming.
Ocean acidification and warming are expected to threaten the persistence of tropical coral reef ecosystems. As coral reefs face multiple stressors, the distribution and abundance of corals will depend on the successful dispersal and settlement of coral larvae under changing environmental conditions. To explore this scenario, we used metabolic rate, at holobiont and molecular levels, as an index for assessing the physiological plasticity of Pocillopora damicornis larvae from this site to conditions of ocean acidity and warming. Larvae were incubated for 6 hours in seawater containing combinations of CO2 concentration (450 and 950 µatm) and temperature (28 and 30°C). Rates of larval oxygen consumption were higher at elevated temperatures. In contrast, high CO2 levels elicited depressed metabolic rates, especially for larvae released later in the spawning period. Rates of citrate synthase, a rate-limiting enzyme in aerobic metabolism, suggested a biochemical limit for increasing oxidative capacity in coral larvae in a warming, acidifying ocean. Biological responses were also compared between larvae released from adult colonies on the same day (cohorts). The metabolic physiology of Pocillopora damicornis larvae varied significantly by day of release. Additionally, we used environmental data collected on a reef in Moorea, French Polynesia to provide information about what adult corals and larvae may currently experience in the field. An autonomous pH sensor provided a continuous time series of pH on the natal fringing reef. In February/March, 2011, pH values averaged 8.075 ± 0.023. Our results suggest that without adaptation or acclimatization, only a portion of naïve Pocillopora damicornis larvae may have suitable metabolic phenotypes for maintaining function and fitness in an end-of-the century ocean.


Gut microbes contribute to variation in solid organ transplant outcomes in mice.
Solid organ transplant recipients show heterogeneity in the occurrence and timing of acute rejection episodes. Understanding the factors responsible for such variability in patient outcomes may lead to improved diagnostic and therapeutic approaches. Rejection kinetics of transplanted organs mainly depends on the extent of genetic disparities between donor and recipient, but a role for environmental factors is emerging. We have recently shown that major alterations of the microbiota following broad-spectrum antibiotics, or use of germ-free animals, promoted longer skin graft survival in mice. Here, we tested whether spontaneous differences in microbial colonization between genetically similar individuals can contribute to variability in graft rejection kinetics. We compared rejection kinetics of minor mismatched skin grafts in C57BL/6 mice from Jackson Laboratory (Jax) and Taconic Farms (Tac), genetically similar animals colonized by different commensal microbes. Female Tac mice rejected skin grafts from vendor-matched males more quickly than Jax mice. We observed prolonged graft survival in Tac mice when they were exposed to Jax mice microbiome through co-housing or fecal microbiota transplantation (FMT) by gastric gavage. In contrast, exposure to Tac mice did not change graft rejection kinetics in Jax mice, suggesting a dominant suppressive effect of Jax microbiota. High-throughput sequencing of 16S rRNA gene amplicons from Jax and Tac mice fecal samples confirmed a convergence of microbiota composition after cohousing or fecal transfer. Our analysis of amplicon data associated members of a single bacterial genus, Alistipes, with prolonged graft survival. Consistent with this finding, members of the genus Alistipes were absent in a separate Tac cohort, in which fecal transfer from Jax mice failed to prolong graft survival. These results demonstrate that differences in resident microbiome in healthy individuals may translate into distinct kinetics of graft rejection, and contribute to interpersonal variability in graft outcomes. The association between Alistipes and prolonged skin graft survival in mice suggests that members of this genus might affect host physiology, including at sites distal to the gastrointestinal tract. Overall, these findings allude to a potential therapeutic role for specific gut microbes to promote graft survival through the administration of probiotics, or FMT.


Uncertainty Propagation in Nerve Impulses Through the Action Potential Mechanism.
We investigate the propagation of probabilistic uncertainty through the action potential mechanism in nerve cells. Using the Hodgkin-Huxley (H-H) model and Stochastic Collocation on Sparse Grids, we obtain an accurate probabilistic interpretation of the deterministic dynamics of the transmembrane potential and gating variables. Using Sobol indices, out of the 11 uncertain parameters in the H-H model, we unravel two main uncertainty sources, which account for more than 90 % of the fluctuations in neuronal responses, and have a direct biophysical interpretation. We discuss how this interesting feature of the H-H model allows one to reduce greatly the probabilistic degrees of freedom in uncertainty quantification analyses, saving CPU time in numerical simulations and opening possibilities for probabilistic generalisation of other deterministic models of great importance in physiology and mathematical neuroscience.


Occupational biomechanical risk factors for radial nerve entrapment in a 13-year prospective study among male construction workers.
The aim was to assess the association between occupational biomechanical exposure and the occurrence of radial nerve entrapment (RNE) in construction workers over a 13-year follow-up period. A cohort of 229 707 male construction workers who participated in a national occupational health surveillance programme (1971-1993) was examined prospectively (2001-2013) for RNE. Height, weight, age, smoking status and job title (construction trade) were obtained on health examination. RNE case status was defined by surgical release of RNE, with data from the Swedish national registry for out-patient surgery records. A job exposure matrix was developed, and biomechanical exposure estimates were assigned according to job title. Highly correlated exposures were summed into biomechanical exposure scores. Negative binomial models were used to estimate the relative risks (RR) (incidence rate ratios) of RNE surgical release for the biomechanical factors and exposure sum scores. Predicted incidence was assessed for each exposure score modelled as a continuous variable to assess exposure-response relationships. The total incidence rate of surgically treated RNE over the 13-year observation period was 3.53 cases per 100 000 person-years. There were 92 cases with occupational information. Increased risk for RNE was seen in workers with elevated hand-grip forces (RR=1.79, 95% CI 0.97 to 3.28) and exposure to hand-arm vibration (RR=1.47, 95% CI 1.08 to 2.00). Occupational exposure to forceful handgrip work and vibration increased the risk for surgical treatment of RNE.


Mutually exclusive expression of virulence genes by malaria parasites is regulated independently of antigen production.
The primary virulence determinant of Plasmodium falciparum malaria parasite-infected cells is a family of heterogeneous surface receptors collectively referred to as PfEMP1. These proteins are encoded by a large, polymorphic gene family called var. The family contains approximately 60 individual genes, which are subject to strict, mutually exclusive expression, with the single expressed var gene determining the antigenic, cytoadherent, and virulence phenotype of the infected cell. The mutually exclusive expression pattern of var genes is imperative for the parasite's ability to evade the host's immune response and is similar to the process of "allelic exclusion" described for mammalian Ig and odorant receptor genes. In mammalian systems, mutually exclusive expression is ensured by negative feedback inhibition mediated by production of a functional protein. To investigate how expression of the var gene family is regulated, we have created transgenic lines of parasites in which expression of individual var loci can be manipulated. Here we show that no such negative feedback system exists in P. falciparum and that this process is dependent solely on the transcriptional regulatory elements immediately adjacent to each gene. Transgenic parasites that are selected to express a var gene in which the PfEMP1 coding region has been replaced by a drug-selectable marker silence all other var genes in the genome, thus effectively knocking out all PfEMP1 expression and indicating that the modified gene is still recognized as a member of the var gene family. Mutually exclusive expression in P. falciparum is therefore regulated exclusively at the level of transcription, and a functional PfEMP1 protein is not necessary for viability or for proper gene regulation in cultured parasites.


An indigenous religious ritual selects for resistance to a toxicant in a livebearing fish.
Human-induced environmental change can affect the evolutionary trajectory of populations. In Mexico, indigenous Zoque people annually introduce barbasco, a fish toxicant, into the Cueva del Azufre to harvest fish during a religious ceremony. Here, we investigated tolerance to barbasco in fish from sites exposed and unexposed to the ritual. We found that barbasco tolerance increases with body size and differs between the sexes. Furthermore, fish from sites exposed to the ceremony had a significantly higher tolerance. Consequently, the annual ceremony may not only affect population structure and gene flow among habitat types, but the increased tolerance in exposed fish may indicate adaptation to human cultural practices in a natural population on a very small spatial scale.


The Presence of Clitoromegaly in the Nonclassical Form of 21-Hydroxylase Deficiency Could Be Partially Modulated by the CAG Polymorphic Tract of the Androgen Receptor Gene.
In the nonclassical form (NC), good correlation has been observed between genotypes and 17OH-progesterone (17-OHP) levels. However, this correlation was not identified with regard to the severity of hyperandrogenic manifestations, which could depend on interindividual variability in peripheral androgen sensitivity. Androgen action is modulated by the polymorphic CAG tract (nCAG) of the androgen receptor (AR) gene and by polymorphisms in 5α-reductase type 2 (SRD5A2) enzyme, both of which are involved in the severity of hyperandrogenic disorders. To analyze whether nCAG-AR and SRD5A2 polymorphisms influence the severity of the nonclassical phenotype. NC patients (n = 114) diagnosed by stimulated-17OHP ≥10 ng/mL were divided into groups according to the beginning of hyperandrogenic manifestations (pediatric and adolescent/adult) and CYP21A2 genotypes (C/C: homozygosis for mild mutations; A/C: compound heterozygosis for severe/mild mutations). CYP21A2 mutations were screened by allelic-specific PCR, MLPA and/or sequencing. HpaII-digested and HpaII-undigested DNA samples underwent GeneScan analysis to study nCAG, and the SRD5A2 polymorphisms were screened by RLFP. Mean nCAG did not differ among pediatric, adolescent/adult and asymptomatic subjects. In the C/C genotype, we observed a significantly lower frequency of longer CAG alleles in pediatric patients than in adolescent/adults (p = 0.01). In patients carrying the A/C genotype, the frequencies of shorter and longer CAG alleles did not differ between pediatric patients and adolescent/adults (p>0.05). Patients with clitoromegaly had significantly lower weighted CAG biallelic mean than those without it: 19.1±2.7 and 21.6±2.5, respectively (p = 0.007), independent of the CYP21A2 genotype's severity. The SRD5A2 polymorphisms were not associated with the variability of hyperandrogenic NC phenotypes. In this series, we observed a modulatory effect of the CAG-AR tract on clinical manifestations of the NC form. Although the NC form is a monogenic disorder, our preliminary data suggested that the interindividual variability of the hyperandrogenic phenotype could arise from polygenic interactions.


'Attending to the wound and the person' - patients' experiences and expectations of a newly established traumatic brain injury clinic.
This qualitative study aimed to gain a better understanding of how medical and social services in the UK currently support patients with Traumatic Brain Injury (TBI) in the community. Furthermore, we explored patients' wishes and expectations of a newly established TBI clinic. We conducted semi-structured interviews with 10 patients with mild-to-severe TBI. The interview schedule was designed to cover contacts with health services, information provided, post-discharge support, current social circumstances, expectations from the newly established brain injury service and participants' desires for any new service. Transcripts were analysed using a thematic analysis. Participants highlighted the importance of the human component of their care and of fostering trusting relationships. This validates patients' experience and helps them to regain confidence. Follow-up and education are important for patients and relatives through all stages of care, regardless of the severity of the injury. Patients strive for meaningful lives and need to be supported to engage in activities. They need hands-on support, particularly with the UK's bureaucratic welfare system. There is much room for improvement in the TBI community care in the UK. Our findings support the development of a holistic service that can address the multifactorial problems which the patients with TBI and their families face.


Circulating inflamma-miRs in aging and age-related diseases.
Evidence on circulating microRNAs (miRNAs) is indisputably opening a new era in systemic and tissue-specific biomarker research, highlighting new inter-cellular and inter-organ communication mechanisms. Circulating miRNAs might be active messengers eliciting a systemic response as well as non-specific "by-products" of cell activity and even of cell death; in either case they have the potential to be clinically relevant biomarkers for a number of physiopathological processes, including inflammatory responses and inflammation-related conditions. A large amount of evidence indicates that miRNAs can exert two opposite roles, activating as well as inhibiting inflammatory pathways. The inhibitory action probably relates to the need for activating anti-inflammatory mechanisms to counter potent proinflammatory signals, like the nuclear factor kappaB (NF-κB) pathway, to prevent cell and tissue destruction. MiRNA-based anti-inflammatory mechanisms may acquire a crucial role during aging, where a chronic, low-level proinflammatory status is likely sustained by the cell senescence secretome and by progressive activation of immune cells over time. This process entails age-related changes, especially in extremely old age, in those circulating miRNAs that are capable of modulating the inflammatory status (inflamma-miRs). Interestingly, a number of such circulating miRNAs seem to be promising biomarkers for the major age-related diseases that share a common chronic, low-level proinflammatory status, such as cardiovascular disease (CVD), type 2 diabetes mellitus (T2DM), Alzheimer Disease (AD), rheumatoid arthritis (RA), and cancers.


Introducing leprosy post-exposure prophylaxis into the health systems of India, Nepal and Indonesia: a case study.
Leprosy has a wide range of clinical and socio-economic consequences. India, Indonesia and Nepal contribute significantly to the global leprosy burden. After integration, the health systems are pivotal in leprosy service delivery. The Leprosy Post Exposure Prophylaxis (LPEP) program is ongoing to investigate the feasibility of providing single dose rifampicin (SDR) as post-exposure prophylaxis (PEP) to the contacts of leprosy cases in various health systems. We aim to compare national leprosy control programs, and adapted LPEP strategies in India, Nepal and Indonesia. The purpose is to establish a baseline of the health system's situation and document the subsequent adjustment of LPEP, which will provide the context for interpreting the LPEP results in future. The study followed the multiple-case study design with single units of analysis. The data collection methods were direct observation, in-depth interviews and desk review. The study was divided into two phases, i.e. review of national leprosy programs and description of the LPEP program. The comparative analysis was performed using the WHO health system frameworks (2007). In all countries leprosy services including contact tracing is integrated into the health systems. The LPEP program is fully integrated into the established national leprosy programs, with SDR and increased documentation, which need major additions to standard procedures. PEP administration was widely perceived as well manageable, but the additional LPEP data collection was reported to increase workload in the first year. The findings of our study led to the recommendation that field-based leprosy research programs should keep health systems in focus. The national leprosy programs are diverse in terms of organizational hierarchy, human resource quantity and capacity. We conclude that PEP can be integrated into different health systems without major structural and personal changes, but provisions are necessary for the additional monitoring requirements.


Effect of early versus delayed interferon beta-1b treatment on disability after a first clinical event suggestive of multiple sclerosis: a 3-year follow-up analysis of the BENEFIT study.
Several controlled studies provide evidence that treatment with interferon beta in patients with a first event suggestive of multiple sclerosis (MS) delays conversion to clinically definite MS (CDMS). Our aim was to determine whether early initiation of treatment with interferon beta prevents development of confirmed disability in MS. In the initial placebo-controlled phase of the double-blinded BENEFIT study, patients with a first event suggestive of MS and a minimum of two clinically silent lesions in MRI were randomised to receive either interferon beta-1b 250 microg (n=292) or placebo (n=176) subcutaneously every other day for 2 years, or until diagnosis of CDMS. Patients were then eligible to enter the follow-up phase with open-label interferon beta-1b. In the current prospectively planned analysis 3 years after randomisation, the effects of early interferon beta-1b treatment were compared with those of delayed treatment initiated after diagnosis of CDMS or after 2 years on the study. The primary outcomes of this ITT analysis were time to diagnosis of CDMS, time to confirmed expanded disability status scale (EDSS) progression, and score on a patient-reported functional assessment scale (FAMS-TOI). This trial is registered with ClinicalTrials.gov, number NCT00185211. Of the 468 patients originally randomised, 418 (89%) entered the follow-up phase; 392 (84%) completed 3 years' post-randomisation follow-up. After 3 years, 99 (37%) patients in the early group developed CDMS compared with 85 (51%) patients in the delayed treatment group. Early treatment reduced the risk of CDMS by 41% (hazard ratio 0.59, 95% CI 0.44-0.80; p=0.0011; absolute risk reduction 14%) compared with delayed treatment. Over 3 years, 42 (16%) patients in the early group and 40 (24%) in the delayed group had confirmed EDSS progression; early treatment reduced the risk for progression of disability by 40% compared with delayed treatment (0.60, 0.39-0.92; p=0.022; absolute risk reduction 8%). The FAMS-TOI score was high and stable in both groups over the 3-year period (p=0.31). Our data suggest that early initiation of treatment with interferon beta-1b prevents the development of confirmed disability, supporting its use after the first manifestation of relapsing-remitting MS.


The preference for dual-gold(i) catalysis in the hydro(alkoxylation vs. phenoxylation) of alkynes.
Dinuclear gold complexes and their use in catalysis have received significant recent attention, but there are few critical comparisons of mono- versus dual gold-catalysed pathways. Herein we study the hydroalkoxylation and hydrophenoxylation of alkynes using density functional theory calculations, and compare two possible mechanisms that have been proposed previously on the basis of theoretical and experimental studies, which unravel different preferences because of both the nature of the alkyne and alcohol and the non-innocent role of the counter-anion of a dual gold based catalyst. Entropy is found to have a significant effect, rendering the nucleophilic attack of the monoaurated intermediate [Au(L)(η2-alkyne)]+ difficult both kinetically and thermodynamically; this mechanism cannot easily form only the trans-alkene product that is observed experimentally. Instead, a reaction via a dual gold catalysed mechanism presents much lower barriers. In addition, for the sake of direct comparison with recent results by Belanzoni and Zuccaccia, oversimplification of the N-heterocyclic carbene (NHC) ligand in the calculations might decrease the enthalpy barrier and lead to results that are not directly applicable to experiments. Moreover, the alkylic or arylic nature of the alkyne and/or alcohol is also tested.


Continuing education in neurometabolic disorders--serine deficiency disorders.
Serine deficiency disorders comprise a new group of inborn errors of serine metabolism. Patients affected with these disorders present with major neurological symptoms including congenital microcephaly, seizures, psychomotor retardation or polyneuropathy. The diagnosis of serine deficiency is based on the detection of low concentrations of the amino acids serine and glycine in fasted plasma and cerebrospinal fluid (CSF). Amino acid analysis of cerebrospinal fluid is preferable over plasma analysis, because the deficiencies are more pronounced in CSF. Because of the interference of amino acids absorbed from the diet, diagnostic procedures have to be performed in the fasted state. Although the disorders are probably rare and not many cases have been reported, recognition of serine deficiency is important, given the fact that the disorders are potentially treatable. The clinical symptoms respond well to amino acid replacement therapy. So far, three serine deficiency disorders have been reported; 3-phosphoglycerate dehydrogenase deficiency, 3-phosphoserine phosphatase deficiency and a still unexplained serine deficiency disorder. In this paper, we will discuss the various serine deficiency disorders, their biochemical abnormalities and the results of amino acid replacement therapy.


Rapid diagnosis of Japanese encephalitis by using an immunoglobulin M dot enzyme immunoassay.
Japanese encephalitis (JE) occurs in rural settings in southern and eastern Asia, where diagnostic facilities are limited. For the diagnosis of JE virus (JEV) infection, we developed a nitrocellulose membrane-based immunoglobulin M (IgM) capture dot enzyme immunoassay (MAC DOT) that is rapid, simple to use, requires no specialized equipment, and can distinguish JEV from dengue infection. In a prospective field study in southern Vietnam, 155 cerebrospinal fluid (CSF) and 341 serum samples were collected from 111 children and 83 adults with suspected encephalitis. The JEV MAC DOT, performed on site, was scored visually from negative to strongly positive by two observers, and the results were compared subsequently with those of the standard IgM capture enzyme-linked immunosorbent assay. For the 179 patients with adequate specimens, the MAC DOT correctly identified 59 of 60 JEV-positive patients and 118 of 119 JEV-negative patients (sensitivity [95% confidence intervals], 98.3% [92.1 to 99.91%]; specificity, 99.2% [95.9 to 100.0%]; positive predictive value, 0.98; negative predictive value, 0.99). The MAC DOT also correctly identified three patients with dengue encephalopathy. Admission specimens were positive for 73% of JE patients. Interobserver agreement for MAC DOT diagnosis was excellent (kappa = 0.94). The JEV MAC DOT is a simple and reliable rapid diagnostic test for JE in rural hospitals.


Production of thrombopoietin (TPO) by rat hepatocytes and hepatoma cell lines.
Recently, we purified rat thrombopoietin (TPO) from plasma of irradiated rats (XRP) by measuring its activity that stimulated the production of megakaryocytes from megakaryocyte progenitor cells (CFU-MK) in vitro. We then cloned the cDNAs for rat and human TPO. In this study, we found the production of TPO by hepatocytes isolated with the collagenase perfusion method from both normal and thrombocytopenic rats, by a two-step fractionation of hepatocyte culture medium (CM). Subsequently, CM of rat hepatoma cell lines was screened for the presence of TPO; three cell lines, H4-II-E, McA-RH8994, and HTC, were found to produce TPO. According to the purification procedure for TPO from XRP, TPO was partially purified from 2 L CM of each of three cell lines with a six-step procedure. In the final reverse-phase column, TPO from each cell line was eluted with the same retention time as that from XRP, and the TPO fraction exhibited megakaryocyte colony-stimulating activity (Meg-CSA). TPO-active fraction eluted from the final reverse-phase column was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), extracted from the gel, and assayed. TPO activity from each cell line was found in the respective molecular weight region, indicating the heterogeneity of the TPO molecule. Using reverse transcriptase-polymerase chain reaction (RT-PCR), we detected the expression of TPO mRNA in hepatocytes, three hepatoma cell lines, normal rat liver, and X-irradiated rat liver. Northern blot analysis showed that TPO mRNA was expressed mainly in liver among the various organs tested. These data demonstrate that TPO is produced by rat hepatocytes and hepatoma cell lines and suggest that liver may be the primary organ that produces TPO.


Quantitative real-time PCR detection of insulin signalling-related genes in pancreatic islets isolated from healthy cats.
The cat has recently been proposed as a valuable model for type 2 diabetes mellitus (T2DM), because feline diabetes shares several similarities with the disease in humans. Impaired beta-cell function, decreased beta-cell mass, insulin resistance that is often related to obesity, and pancreatic amyloid deposition, are among these common features. In this study, and to further develop the cat as a model of T2DM, feline pancreatic islets were isolated and real-time PCR quantification of mRNA transcripts of genes central to beta-cell function and survival established. In particular, mRNA quantification systems were determined for insulin, the insulin enhancer pancreatic duodenal homeobox-1 (PDX-1), the insulin suppressor CCAAT/enhancer binding protein-beta (C/EBPbeta), glucose transporter isoform 2 (GLUT2), Fas receptor, the caspase-8 inhibitor FLIP (FLICE [caspase-8]-inhibitory protein) and two chemokines, interleukin (IL)-8 and monocyte chemoattractant protein-1 (MCP-1). Pancreatic islets were isolated by collagenase digestion from healthy cat donors. Partial feline mRNA sequences were determined for PDX-1, C/EBPbeta, GLUT2 and FLIP using primers identified from conserved regions of human, dog and rat mRNA. These novel and the previously available sequences (insulin, Fas receptor, IL-8 and MCP-1) were used to design feline-specific primers suitable for real-time PCR in isolated pancreatic islets. The adopted protocol of collagenase digestion yielded pancreatic islets that were frequently surrounded by acinar cells. Quantification of mRNA transcripts was simple and reproducible in healthy cats. Characterisation of genes related to insulin signalling in cats will prove useful to better understand the pathogenesis of feline diabetes and possibly of human T2DM.


Measures of lower body function and risk of mortality over 7 years of follow-up.
The study examined whether a test of walking speed provides similar predictive information on mortality risk as does a summary measure of lower body function. Data were from the Hispanic Established Population for the Epidemiologic Study of the Elderly database and included Mexican Americans aged 65 years or more (1993-2000). Primary measures included a short physical performance battery, a test of walking speed, and mortality. The average age of the sample was 72.0 years, and 58.3 percent were women. The observed hazard ratio of mortality risk was similar for the full short physical performance battery and walking speed alone, in both unadjusted and adjusted baseline models. A time-dependent walking speed measure showed a more than twofold increased risk of mortality for individuals categorized with slower walking speed. The results also showed a linear association between continuous walking speed and mortality with and without adjustment for baseline covariates. This study provides evidence that walking speed alone can provide similar information on mortality risk as does a more comprehensive summary measure of physical performance. Because walking speed is a quick and easy-to-administer test, findings have implications for clinical use, especially among underserved minority groups where cultural and language barriers may exist.


An unusual case of dyspnoea in an elderly man.
Defective closure of the posterolateral pleuroperitoneal canal during embryogenesis gives rise to a congenital hernia (usually left sided) which was originally described by Bochdalek in 1948. It manifests primarily in children with respiratory symptoms and pulmonary hypoplasia. It is exceptionally rare for this defect to present in adulthood, with just over 50 symptomatic cases being described in the literature. Most adolescent and adult cases are diagnosed incidentally during imaging for upper gastrointestinal symptoms. It is unusual for adults to present with respiratory symptoms. We describe the case of a 71-year-old man who presented with features of left ventricular failure due to an exceptionally large, right sided Bochdalek hernia. This is the oldest clinical presentation of a right sided Bochdalek hernia, which uniquely included trans-diaphragmatic herniation of the pancreas.


Use of deferred consent for severely ill children in a multi-centre phase III trial.
Voluntary participation of a subject in research respects a subject's rights, strengthens its ethical conduct, and is formalized by the informed consent process. Clinical trials of life-saving interventions for medical emergencies often necessitate enrollment of patients where prior written individual informed consent is impossible. Although there are regulations and guidelines on protecting subjects in emergency research, these have been criticised for being limited and unnecessarily restrictive. Across Europe and the United States stringent regulations have resulted in a substantial decline of clinical trials involving emergency interventions. We are conducting a trial of fluid resuscitation in children with hypovolaemic shock in six hospitals across three malaria-endemic African countries. The design is pragmatic as children are enrolled on clinical criteria alone and is being conducted in hospitals with facilities typical of many district hospitals across Africa. The trial aims to inform strategy for managing children with febrile illness and features of shock. In order to develop appropriate consent processes for the trial, we conducted a narrative review of current international recommendations for emergency consent. Practical or specific guidance was generally sparse or confusing with few examples in the literature to direct our informed consent process. For a sub-group of children who were critically sick or where parents themselves were otherwise too distressed to consider prior written consent, we opted for a modified form of deferred consent. This included verbal assent from guardians at the point of enrollment, with full written consent obtained after stabilising the child. For children who died prior to full written consent, ethical permission was received to waiver full consent. In light of the controversy around guidance and regulations in this area we report how and why we have used a modified system of deferred consent in an emergency intervention trial in children. Although approved by all relevant ethics committees and operational in 3 countries in Africa, formal research is now necessary to explore the perceptions and experiences of parents, health workers, researchers and ethics committees of the modified method of deferred consent.


Dual lentivirus infection potentiates neuroinflammation and neurodegeneration: viral copassage enhances neurovirulence.
Infection by multiple lentiviral strains is recognized as a major driving force in the human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) epidemic, but the neuropathogenic consequences of multivirus infections remain uncertain. Herein, we investigated the neurovirulence and underlying mechanisms of dual lentivirus infections with distinct viral strains. Experimental feline immunodeficiency virus (FIV) infections were performed using cultured cells and an in vivo model of AIDS neuropathogenesis. Dual infections were comprised of two FIV strains (FIV-Ch and FIV-PPR) as copassaged or superinfected viruses, with subsequent outcome analyses of host immune responses, viral load, neuropathological features, and neurobehavioral performance. Dual infections of feline macrophages resulted in greater IL-1beta (interleukin-1beta), TNF-alpha (tumor necrosis factor alpha), and IDO (indoleamine 2,3-dioxygenase) expression and associated neurotoxic properties. FIV coinfection and sequential superinfection in vivo also induced greater IL-1beta, TNF-alpha, and IDO expression in the basal ganglia (BG) and cortex (CTX), compared to the monovirus- and mock-infected groups, although viral loads were similar in single virus- and dual virus-infected animals. Immunoblot analyses disclosed lower synaptophysin immunoreactivity in the CTX resulting from FIV super- and coinfections. Cholinergic and GABAergic neuronal injury was evident in the CTX of animals with dual FIV infections. With increased glial activation and neuronal loss in dual FIV-infected brains, immunohistochemical analysis also revealed elevated detection of cleaved caspase-3 in dysmorphic neurons, which was associated with worsened neurobehavioral abnormalities among animals infected with the copassaged viruses. Dual lentivirus infections caused an escalation in neuroinflammation and ensuing neurodegeneration, underscoring the contribution of infection by multiple viruses to neuropathogenesis.


General Prediction of Peptide-MHC Binding Modes Using Incremental Docking: A Proof of Concept.
The class I major histocompatibility complex (MHC) is capable of binding peptides derived from intracellular proteins and displaying them at the cell surface. The recognition of these peptide-MHC (pMHC) complexes by T-cells is the cornerstone of cellular immunity, enabling the elimination of infected or tumoral cells. T-cell-based immunotherapies against cancer, which leverage this mechanism, can greatly benefit from structural analyses of pMHC complexes. Several attempts have been made to use molecular docking for such analyses, but pMHC structure remains too challenging for even state-of-the-art docking tools. To overcome these limitations, we describe the use of an incremental meta-docking approach for structural prediction of pMHC complexes. Previous methods applied in this context used specific constraints to reduce the complexity of this prediction problem, at the expense of generality. Our strategy makes no assumption and can potentially be used to predict binding modes for any pMHC complex. Our method has been tested in a re-docking experiment, reproducing the binding modes of 25 pMHC complexes whose crystal structures are available. This study is a proof of concept that incremental docking strategies can lead to general geometry prediction of pMHC complexes, with potential applications for immunotherapy against cancer or infectious diseases.


[Merkel cell carcinoma during treatment with TNF-alpha inhibitors: coincidence or warning?].
Increasing numbers of patients are being treated with TNF-alpha inhibitors. Two patients in our outpatient clinic developed Merkel cell carcinoma during treatment with TNF alpha inhibitors. Since this is a very rare malignancy, this is a remarkable observation. A 70-year-old male with rheumatoid arthritis had been treated with etanercept for two years when he discovered a nodule on his elbow that started growing rapidly. It was diagnosed as Merkel cell carcinoma. Despite treatment, the patient died 2 years later. Merkel cell carcinoma is an aggressive malignancy with a clinically benign aspect. Patients treated with TNF-alpha inhibitors possibly have an increased risk of developing this malignancy.


Stress vulnerability in adolescents with chronic fatigue syndrome: experimental study investigating heart rate variability and skin conductance responses.
Stress vulnerability has been implicated in adolescent chronic fatigue syndrome (CFS), but has rarely been investigated directly. This study compared psychological and physiological responses to a laboratory social performance task in adolescents with CFS with chronic illness (asthma) and healthy control groups. Adolescents with CFS (n = 60), adolescents with asthma (n = 31) and healthy adolescents (n = 78) completed questionnaires before and after a social performance task. Skin conductance responses (SCR; mean SCR and Max-Min) and heart rate variability (low frequency/high frequency; LF/HF and root mean square difference of successive RR intervals; RMSSD) was measured before, during and after the task. Baseline heart rate variability (HRV) (RMSSD) was significantly lower in the CFS and Asthma groups than the HC. During the speech, the CFS and Asthma groups had higher HRV (LF/HF) than the HC, adjusting for baseline LF/HF. Although the asthma group showed a subsequent reduction in HRV during recovery, the CFS group did not. Similarly, during recovery after the task, the CFS group showed a continued increase in skin conductance (Min-Max), unlike the Asthma and HC groups. Compared to control groups, adolescents with CFS expected to find the task more difficult, were more anxious beforehand and afterwards, rated it as more difficult, evaluated their performance more negatively and had lower observer ratings of performance. Parents of adolescents with CFS expected that their child would perform less well in the task than parents of control participants. Adolescents with CFS showed autonomic nervous system responses that are consistent with chronic stress vulnerability, difficulty coping with acute stress and slower recovery after acute stress. Self-report measures also indicated greater trait, pre- and posttask anxiety in the CFS group.


Absolute and relative densities of fast-food versus other restaurants in relation to weight status: Does restaurant mix matter?
Given the continuing epidemic of obesity, policymakers are increasingly looking for levers within the local retail food environment as a means of promoting healthy weights. To examine the independent and joint associations of absolute and relative densities of restaurants near home with weight status in a large, urban, population-based sample of adults. We studied 10,199 adults living in one of four cities in southern Ontario, Canada, who participated in the Canadian Community Health Survey (cycles 2005, 2007/08, 2009/10). Multivariate models assessed the association of weight status (obesity and body mass index) with absolute densities (numbers) of fast-food, full-service and other restaurants, and the relative density (proportion) of fast-food restaurants (FFR) relative to all restaurants within ~10-minute walk of residential areas. Higher numbers of restaurants of any type were inversely related to excess weight, even in models adjusting for a range of individual covariates and area deprivation. However, these associations were no longer significant after accounting for higher walkability of areas with high volumes of restaurants. In contrast, there was a direct relationship between the proportion of FFR relative to all restaurants and excess weight, particularly in areas with high volumes of FFR (e.g., odds ratio for obesity=2.55 in areas with 5+ FFR, 95% confidence interval: 1.55-4.17, across the interquartile range). Policies aiming to promote healthy weights that target the volume of certain retail food outlets in residential settings may be more effective if they also consider the relative share of outlets serving more and less healthful foods.


Bacteriophage tailspike protein based assay to monitor phase variable glucosylations in Salmonella O-antigens.
Non-typhoid Salmonella Typhimurium (S. Typhimurium) accounts for a high number of registered salmonellosis cases, and O-serotyping is one important tool for monitoring epidemiology and spread of the disease. Moreover, variations in glucosylated O-antigens are related to immunogenicity and spread in the host. However, classical autoagglutination tests combined with the analysis of specific genetic markers cannot always reliably register phase variable glucose modifications expressed on Salmonella O-antigens and additional tools to monitor O-antigen glucosylation phenotypes of S. Typhimurium would be desirable. We developed a test for the phase variable O-antigen glucosylation state of S. Typhimurium using the tailspike proteins (TSP) of Salmonella phages 9NA and P22. We used this ELISA like tailspike adsorption (ELITA) assay to analyze a library of 44 Salmonella strains. ELITA was successful in discriminating strains that carried glucose 1-6 linked to the galactose of O-polysaccharide backbone (serotype O1) from non-glucosylated strains. This was shown by O-antigen compositional analyses of the respective strains with mass spectrometry and capillary electrophoresis. The ELITA test worked rapidly in a microtiter plate format and was highly O-antigen specific. Moreover, TSP as probes could also detect glucosylated strains in flow cytometry and distinguish multiphasic cultures differing in their glucosylation state. Tailspike proteins contain large binding sites with precisely defined specificities and are therefore promising tools to be included in serotyping procedures as rapid serotyping agents in addition to antibodies. In this study, 9NA and P22TSP as probes could specifically distinguish glucosylation phenotypes of Salmonella on microtiter plate assays and in flow cytometry. This opens the possibility for flow sorting of cell populations for subsequent genetic analyses or for monitoring phase variations during large scale O-antigen preparations necessary for vaccine production.


Dental burs--what bur for which application? A survey of dental schools.
The purpose of this study was to survey North American dental schools regarding recommendations for rotary instrumentation for fixed prosthodontic and operative procedures at the predoctoral and postgraduate level. A 25-question survey was sent to 64 North American dental schools, of which 15 questions specifically related to rotary instrument recommendations. One questionnaire was addressed to the individual at each school having administrative responsibility for teaching tooth preparation techniques. A high response rate was assured by follow-up telephone calls and faxes. The distributions of bur recommendations for both predoctoral and advanced prosthodontic programs were analyzed by chi-squared tests at an a priori alpha = 0.05. Completed surveys were received from 58 of 64 dental schools, a response rate > 90%. Medium grit burs predominate in predoctoral education for gross tooth reduction for fixed prosthodontics, whereas coarse grit burs predominate at the graduate level (p < .05). The use of the diamond bur alone predominates for axial wall refinement, whereas the use of carbide burs or carbide burs in combination with diamond burs prevails for marginal refinement (p < .05). In predoctoral operative dentistry, recommendations for cavity outline form were similar at all dental schools (p > .05) and were principally tungsten carbide (WC) burs. Carbide burs are the instrument of choice for internal walls, but the WC bur alone or in combination with diamond burs is preferred for refining composite margins (p < .05). There is a broad consensus within North American dental schools on rotary instrumentation used by dental students. There is a greater use of coarser grit burs for gross tooth reduction in fixed prosthodontics at the postgraduate than predoctoral level.


A new method for investigating the relationship between diet and mortality: hazard analysis using dietary isotopes.
Background: The population of Roman Britain are renowned for having elevated nitrogen (δ15) stable isotope values, which have been interpreted as evidence for the increased consumption of marine products. However, such results are now understood to also reflect episodes of stress and disease, suggesting that new interpretations are warranted.Aim: To test a novel approach which combines hazard mortality analysis and stable isotope data to determine whether there is a relationship between age-at-death, elevated δ15N values and mortality risk.Subjects and methods: This study used published osteological and dietary stable isotope data for nitrogen (δ15N) and carbon (δ13C) of 659 1st-5th century AD individuals aged >12 years old excavated from Roman cemeteries in Britain. The relationship between diet and mortality risk was assessed using the Gompertz hazard model, and differences in median reported isotope values between the sexes was determined using a Mann Whitney test.Results: It was discovered that higher δ15N levels are associated with elevated risks of mortality, whereas the opposite pattern was observed for δ13C, and males had higher median δ13C and δ15N values.Conclusion: This study successfully demonstrated that stable isotope data can be integrated into hazard models, allowing one to connect diet and mortality in past populations. It supports the findings of other isotope studies which have established that individuals with childhood stress/trauma will have different isotope patterns.


Atypical teratoid/rhabdoid tumor with 26-year overall survival: case report.
Atypical teratoid/rhabdoid tumor (AT/RT) is an aggressive embryonic brain tumor predominantly seen in young children. The authors report an unusual case of a fourth ventricle AT/RT in an infant who survived for 26 years. The tumor was resected when the patient was 6 months of age, and radiation therapy (40-Gy total dose) was performed thereafter. The patient was free from the disease for 26 years until a recurrent tumor was found in the spinal cord. The spinal cord neoplasm was a "collision tumor" with 2 components: benign schwannoma and recurrent AT/RT. The patient died of dissemination of the recurrent tumor 5 months after it was excised. This is the longest survival of a patient with AT/RT ever reported and indicates that long-term survival, more than 20 years, can be achieved in infantile-onset AT/RT. Despite intensive treatment, the prognosis for AT/RT is very poor, especially in children younger than 3 years of age. The benefits of upfront radiation therapy for AT/RT should be carefully assessed with respect to its inevitable toxicity in very young children. However, early upfront radiation therapy may be of therapeutic interest to prevent aggressive progression of the disease.


Cue generation: How learners flexibly support future retrieval.
The successful use of memory requires us to be sensitive to the cues that will be present during retrieval. In many situations, we have some control over the external cues that we will encounter. For instance, learners create shopping lists at home to help remember what items to later buy at the grocery store, and they generate computer file names to help remember the contents of those files. Generating cues in the service of later cognitive goals is a complex task that lies at the intersection of metacognition, communication, and memory. In this series of experiments, we investigated how and how well learners generate external mnemonic cues. Across 5 experiments, learners generated a cue for each target word in a to-be-remembered list and received these cues during a later cued recall test. Learners flexibly generated cues in response to different instructional demands and study list compositions. When generating mnemonic cues, as compared to descriptions of target items, learners produced cues that were more distinct than mere descriptions and consequently elicited greater cued recall performance than those descriptions. When learners were aware of competing targets in the study list, they generated mnemonic cues with smaller cue-to-target associative strength but that were even more distinct. These adaptations led to fewer confusions among competing targets and enhanced cued recall performance. These results provide another example of the metacognitively sophisticated tactics that learners use to effectively support future retrieval.