A common structural basis for pH- and calmodulin-mediated regulation in plant glutamate decarboxylase.
Glutamate decarboxylase (Gad) catalyzes glutamate to gamma-aminobutyrate conversion. Plant Gad is a approximately 340 kDa hexamer, involved in development and stress response, and regulated by pH and binding of Ca(2+)/calmodulin (CaM) to the C-terminal domain. We determined the crystal structure of Arabidopsis thaliana Gad1 in its CaM-free state, obtained a low-resolution structure of the calmodulin-activated Gad complex by small-angle X-ray scattering and identified the crucial residues, in the C-terminal domain, for regulation by pH and CaM binding. CaM activates Gad1 in a unique way by relieving two C-terminal autoinhibition domains of adjacent active sites, forming a 393 kDa Gad1-CaM complex with an unusual 1:3 stoichiometry. The complex is loosely packed: thanks to the flexible linkers connecting the enzyme core with the six C-terminal regulatory domains, the CaM molecules retain considerable positional and orientational freedom with respect to Gad1. The complex thus represents a prototype for a novel CaM-target interaction mode. Thanks to its two levels of regulation, both targeting the C-terminal domain, Gad can respond flexibly to different kinds of cellular stress occurring at different pH values.


Historic cohort: outcome of chronic exertional compartment syndrome-suspected patients.
Exercise-related leg pain (ERLP) may be caused by chronic exertional compartment syndrome (CECS), occurring mainly in athletes and military recruits. In military populations, the effectiveness of surgical treatment in CECS is debated. The purpose of this study is to assess the outcome of surgical treatment for CECS in Alrijne Hospital (the Netherlands), a civilian hospital with supraregional referral function. A historic cohort study was performed on patients with ERLP who were suspected for CECS and were referred for intracompartmental pressure measurement (ICPM) from 2013 to 2017 (n=160). Patient demographics, ICPM and survey response were analysed. The mean delay before visitation was 29.0±30.3 months. When comparing surgical-treated patients with CECS with conservative-treated patients with ERLP, surgical-treated patients were more satisfied, reported better recovery towards former level of performance (2.8±2.0 vs 3.9±1.7 and 2.5±1.6 vs 3.2±1.4 on a 7-point Likert scale, respectively) and better subjective injury status (79.3±22.6 vs 63.5±27.4 using the Single Assessment Numeric Evaluation score). Treatment satisfaction was 75.0% in surgical-treated CECS versus 51.4% in conservative-treated ERLP. Civilian patients report improved functional outcomes after fasciotomy for CECS. Future research should focus on non-invasive diagnostic options and methods to determine which treatment is the most appropriate for each individual patient.


Transgenic tobacco plants expressing the potato virus X open reading frame 3 gene develop specific resistance and necrotic ring symptoms after infection with the homologous virus.
Tobacco plants were transformed with the open reading frame 3 gene from Potato virus X (PVX) coding for the p12 protein. Although the transgenic plants exhibited a normal morphological aspect, microscopic examination revealed extensive alterations in leaf tissue structure. After being challenged with PVX, the transgenic plants showed resistance to PVX infection and formation of specific leaf symptoms consisting of concentric rings encircled by necrotic borders. These novel symptoms were accompanied by biochemical changes normally associated with the hypersensitive response (HR) and were absent in noninfected transgenic plants or in PVX-infected nontransgenic plants. No equivalent virus resistance was observed after inoculation with Tobacco mosaic virus or Potato virus Y, suggesting the presence of a specific resistance mechanism. Despite development of HR-like symptoms, systemic acquired resistance was not induced in PVX-infected p12 transgenic plants. No evidence of an RNA-mediated resistance mechanism was found.


Tissue bridges predict recovery after traumatic and ischemic thoracic spinal cord injury.
To investigate the spatiotemporal evolution and predictive properties of intramedullary damage and midsagittal tissue bridges at the epicenter of a thoracic spinal cord injury (SCI) using MRI. We retrospectively assessed midsagittal T2-weighted scans from 25 patients with thoracic SCI (14 traumatic, 11 ischemic) at 1 month post-SCI. In 12 patients with SCI, linear mixed-effects models on serial MRI explored temporal trajectories of quantifiable lesion markers (area, length, and width) and tissue bridges. Using partial correlation analysis, we assessed associations between structural lesion characteristics at 1 month post-SCI and recovery at 1 year postinjury, adjusting for baseline clinical status, age, and sex. Lesion area decreased by 5.68 mm2 (p = 0.005), lesion length by 2.14 mm (p = 0.004), and lesion width by 0.13 mm (p = 0.004) per month. Width of tissue bridges increased by 0.06 mm (p = 0.019) per month, being similar in traumatic and ischemic SCI (p = 0.576). Smaller lesion area, length, width, and wider tissue bridges at 1 month post-SCI predicted better recovery at 1-year follow-up. Over time, the immediate area of cord damage shrunk while the cystic cavity became demarcated. Adjacent to the cyst, midsagittal tissue bridges became visible. The width of tissue bridges at 1 month post-SCI predicted recovery at 1 year follow-up. Measures of lesion area and tissue bridges early after traumatic and ischemic thoracic SCI therefore allow characterizing the evolution of focal cord damage and are predictive of recovery in thoracic SCI. Thus, lesion extent and tissue bridges hold potential to improve diagnosis and patient stratification in interventional trials.


Multimodal Imaging Evidence for a Frontoparietal Modulation of Visual Cortex during the Selective Processing of Conditioned Threat.
Emotionally salient cues are detected more readily, remembered better, and evoke greater visual cortical responses compared with neutral stimuli. The current study used concurrent EEG-fMRI recordings to identify large-scale network interactions involved in the amplification of visual cortical activity when viewing aversively conditioned cues. To generate a continuous neural signal from pericalcarine visual cortex, we presented rhythmic (10/sec) phase-reversing gratings, the orientation of which predicted the presence (CS+) or absence (CS-) of a cutaneous electric shock (i.e., the unconditioned stimulus). The resulting single trial steady-state visual evoked potential (ssVEP) amplitude was regressed against the whole-brain BOLD signal, resulting in a measure of ssVEP-BOLD coupling. Across all trial types, ssVEP-BOLD coupling was observed in both primary and extended visual cortical regions, the rolandic operculum, as well as the thalamus and bilateral hippocampus. For CS+ relative to CS- trials during the conditioning phase, BOLD-alone analyses showed CS+ enhancement at the occipital pole, superior temporal sulci, and the anterior insula bilaterally, whereas ssVEP-BOLD coupling was greater in the pericalcarine cortex, inferior parietal cortex, and middle frontal gyrus. Dynamic causal modeling analyses supported connectivity models in which heightened activity in pericalcarine cortex for threat (CS+) arises from cortico-cortical top-down modulation, specifically from the middle frontal gyrus. No evidence was observed for selective pericalcarine modulation by deep cortical structures such as the amygdala or anterior insula, suggesting that the heightened engagement of pericalcarine cortex for threat stimuli is mediated by cortical structures that constitute key nodes of canonical attention networks.


Prescribers' views and experiences of assessing the appropriateness of prescribed medications in a specialist addiction service.
Background Mental and physical health problems are common in people with substance misuse problems and medications are often required in their management. Given the extent of prescribing for service users who attend specialist addiction services, it is important to consider how prescribers in this setting assess the appropriateness of service users' prescribed medications. Objective To explore prescribers' views and experiences of assessing the appropriateness of medications prescribed for service users coming in for treatment as well as the differences between prescribers. Setting A specialist addiction service in the North of England. Method A phenomenological approach was adopted. Semi-structured interviews were conducted with four nurse prescribers and eight doctors. Data were analysed using thematic framework analysis. Main outcome measure Prescribers' views and experiences of assessing the appropriateness of prescribed medications. Results Assessment of the appropriateness of prescribed medications involved reviewing medications, assessing risk, history-taking, involvement of service users, and comparing guideline adherence and 'successful' prescribing. Doctors and nurse prescribers assessed the appropriateness of medications they considered to be within their competency. Doctors provided support to nurse prescribers and general practitioners (GPs) when dealing with issues around prescribing. Conclusion Assessment of the appropriateness of prescribed medications is complex. The recent reduction in medical expertise in specialist addiction services may negatively impact on the clinical management of service users. It appears that there is a need for further training of nurse prescribers and GPs so they can provide optimal care to service users.


Probing the interplay between geometric and electronic structure in a two-dimensional K-TCNQ charge transfer network.
Scanning tunnelling microscopy (STM), low energy electron diffraction (LEED), ultraviolet and soft X-ray photoelectron spectroscopy (UPS and SXPS) have been used to characterise the formation of a coadsorption phase of TCNQ and K on Ag(111), while the normal incident X-ray standing waves (NIXSW) technique has been used to obtain quantitative structural information. STM and LEED show that an ordered incommensurate phase is formed in which the K atoms are surrounded by four TCNQ molecules in a 'windmill' motif, characteristic of other metal/TCNQ phases, in which the nominal TCNQ : K stoichiometry is 1 : 1. UPS and SXPS data indicate the TCNQ is in a negatively-charged state. NIXSW results show that the carbon core of the TCNQ is essentially planar at a height above the Ag(111) surface closely similar to that found without coadsorbed K. In the presence of TCNQ the height of the K ions above the surface is significantly larger than on clean Ag(111), and the ions occupy sites above 'holes' in the TCNQ network. NIXSW data also show that the N atoms in the molecules must occupy sites with at least two different heights above the surface, which can be reconciled by a tilt or twist of the TCNQ molecules, broadly similar to the geometry that occurs in bulk TCNQ/K crystals.


Early interaction between vision and touch during binocular rivalry.
Multisensory integration is known to occur at high neural levels, but there is also growing evidence that cross-modal signals can be integrated at the first stages of sensory processing. We investigated whether touch specifically affected vision during binocular rivalry, a particular type of visual bistability that engages neural competition in early visual cortices. We found that tactile signals interact with visual signals outside of awareness, when the visual stimulus congruent with the tactile one is perceptually suppressed during binocular rivalry and when the interaction is strictly tuned for matched visuo-tactile spatial frequencies. We also found that voluntary action does not play a leading role in mediating the effect, since the interaction was observed also when tactile stimulation was passively delivered to the finger. However, simultaneous presentation of visual and tactile stimuli is necessary to elicit the interaction, and an asynchronous priming touch stimulus is not affecting the onset of rivalry. These results point to a very early cross-modal interaction site, probably V1. By showing that spatial proximity between visual and tactile stimuli is a necessary condition for the interaction, we also suggest that the two sensory spatial maps are aligned according to retinotopic coordinates, corroborating the hypothesis of a very early interaction between visual and tactile signals during binocular rivalry.


Mobile computation: spatiotemporal integration of the properties of objects in motion.
We demonstrate that, as an object moves, color and motion signals from successive, widely spaced locations are integrated, but letter and digit shapes are not. The features that integrate as an object moves match those that integrate when the eyes move but the object is stationary (spatiotopic integration). We suggest that this integration is mediated by large receptive fields gated by attention and that it occurs for surface features (motion and color) that can be summed without precise alignment but not shape features (letters or digits) that require such alignment. Rapidly alternating pairs of colors and motions were presented at several locations around a circle centered at fixation. The same two stimuli alternated at each location with the phase of the alternation reversing from one location to the next. When observers attended to only one location, the stimuli alternated in both retinal coordinates and in the attended stream: feature identification was poor. When the observer's attention shifted around the circle in synchrony with the alternation, the stimuli still alternated at each location in retinal coordinates, but now attention always selected the same color and motion, with the stimulus appearing as a single unchanging object stepping across the locations. The maximum presentation rate at which the color and motion could be reported was twice that for stationary attention, suggesting (as control experiments confirmed) object-based integration of these features. In contrast, the identification of a letter or digit alternating with a mask showed no advantage for moving attention despite the fact that moving attention accessed (within the limits of precision for attentional selection) only the target and never the mask. The masking apparently leaves partial information that cannot be integrated across locations, and we speculate that for spatially defined patterns like letters, integration across large shifts in location may be limited by problems in aligning successive samples. Our results also suggest that as attention moves, the selection of any given location (dwell time) can be as short as 50 ms, far shorter than the typical dwell time for stationary attention. Moving attention can therefore sample a brief instant of a rapidly changing stream if it passes quickly through, giving access to events that are otherwise not seen.


Sarcoma metastases to the skin: a clinicopathologic study of 65 patients.
Sarcoma metastases to the skin are relatively rare, because most involve the lung, liver, or deep soft tissues. The authors of this report examined the distribution and clinical significance of cutaneous and superficial subcutaneous sarcoma metastases. Sixty-five patients with histologically confirmed dermal and superficial subcutaneous sarcoma metastases were identified in pathology files from more than 25,000 patients with sarcoma who were evaluated at The University of Texas M. D. Anderson Cancer Center from 1989 to 2009. Pathology slides and clinical and radiological information were evaluated. Cutaneous metastases were documented histologically in <0.25% of patients. The mean patient age was 49 years (range, 16-79 years), and there was an equivalent ratio of men to women. The most common source of metastasis was leiomyosarcoma (28 of 65 patients; 43%). The most common region of first skin metastasis was head and neck (33 patients; 51%), and the scalp predominated (25 patients; 38%). The mean time from primary tumor diagnosis to skin metastasis was 48 months (range, 0-166 months). Fifty-three patients (81%) had multiple metastases (skin and other). Among the patients who had complete clinical information available, 31 patients (62%) had other metastases diagnosed before skin involvement, 17 patients (34%) had skin metastases diagnosed first, and 2 patients (4%) had simultaneous presentation. The following clinical outcomes were documented: Twenty-nine patients (45%) died of disease, 24 patients (37%) remained alive with disease, and 12 patients were lost to follow-up. The mean time to death was 80 months (range, 9-224 months) after primary diagnosis, 45 months (range, 5-94 months) after the first metastasis to any site, and 27 months (range, 5-65 months) after the first skin metastasis. Sarcoma metastases to the skin are rare. In this large study, leiomyosarcoma was the most common source, and the scalp was the most frequent site. The majority of patient with skin metastases harbored metastases elsewhere. However, skin was the initial site of metastasis in approximately 1 in 3 patients. Thus, clinical correlation is needed before establishing a diagnosis of primary cutaneous sarcoma, particularly leiomyosarcoma of scalp. Finally, the current results indicated that skin metastasis usually is a late event in sarcoma clinical progression and heralds a poor prognosis.


Untapped potential of health impact assessment.
The World Health Organization has promoted health impact assessment (HIA) for over 20 years. At the 2012 United Nations Conference on Sustainable Development (Rio+20), HIA was discussed as a critical method for linking health to "green economy" and "institutional framework" strategies for sustainable development. In countries having a high human development index (HDI), HIA has been added to the overall assessment suite that typically includes potential environmental and social impacts, but it is rarely required as part of the environmental and social impact assessment for large development projects. When they are performed, project-driven HIAs are governed by a combination of project proponent and multilateral lender performance standards rather than host country requirements. Not surprisingly, in low-HDI countries HIA is missing from the programme and policy arena in the absence of an external project driver. Major drivers of global change (e.g. population growth and urbanization, growing pressure on natural resources and climate change) inordinately affect low- and medium-HDI countries; however, in such countries HIA is conspicuously absent. If the cloak of HIA invisibility is to be removed, it must be shown that HIA is useful and beneficial and, hence, an essential component of the 21st century's sustainable development agenda. We analyse where and how HIA can become fully integrated into the impact assessment suite and argue that the impact of HIA must not remain obscure.


An interdisciplinary approach to personalized medicine: case studies from a cardiogenetics clinic.
In the genomic age, the challenges presented by various inherited conditions present a compelling argument for an interdisciplinary model of care. Cardiac arrhythmias with a genetic basis, such as long QT syndrome, require clinicians with expertise in many specialties to address the complex genetic, psychological, ethical and medical issues involved in treatment. The Montefiore-Einstein Center for CardioGenetics has been established to provide personalized, interdisciplinary care for families with a history of sudden cardiac death or an acute cardiac event. Four vignettes of patient care are presented to illustrate the unique capacity of an interdisciplinary model to address genetic, psychological, ethical and medical issues. Because interdisciplinary clinics facilitate collaboration among multiple specialties, they allow for individualized, comprehensive care to be delivered to families who experience complex inherited medical conditions. As the genetic basis of many complex conditions is discovered, the advantages of an interdisciplinary approach for delivering personalized medicine will become more evident.


Central nervous system infections in the tropics: diagnosis, treatment and prevention.
The present review is a brief discussion of some of the important new developments that have been reported relating to bacterial and viral infections of the central nervous system. For many of these conditions, certain issues remain unresolved, including what is the best diagnostic approach, what is the optimum treatment and how best to prevent such conditions. The development of improved surveillance, more effective vaccines and a greater understanding of the pathophysiology of these diseases all offer great potential benefits. The hope is that these advances, when they come, will be affordable to those who need them most.


An unexpected role of neuroligin-2 in regulating KCC2 and GABA functional switch.
GABAA receptors are ligand-gated Cl- channels, and the intracellular Cl- concentration governs whether GABA function is excitatory or inhibitory. During early brain development, GABA undergoes functional switch from excitation to inhibition: GABA depolarizes immature neurons but hyperpolarizes mature neurons due to a developmental decrease of intracellular Cl- concentration. This GABA functional switch is mainly mediated by the up-regulation of KCC2, a potassium-chloride cotransporter that pumps Cl- outside neurons. However, the upstream factor that regulates KCC2 expression is unclear. We report here that KCC2 is unexpectedly regulated by neuroligin-2 (NL2), a cell adhesion molecule specifically localized at GABAergic synapses. The expression of NL2 precedes that of KCC2 in early postnatal development. Upon knockdown of NL2, the expression level of KCC2 is significantly decreased, and GABA functional switch is significantly delayed during early development. Overexpression of shRNA-proof NL2 rescues both KCC2 reduction and delayed GABA functional switch induced by NL2 shRNAs. Moreover, NL2 appears to be required to maintain GABA inhibitory function even in mature neurons, because knockdown NL2 reverses GABA action to excitatory. Gramicidin-perforated patch clamp recordings confirm that NL2 directly regulates the GABA equilibrium potential. We further demonstrate that knockdown of NL2 decreases dendritic spines through down-regulating KCC2. Our data suggest that in addition to its conventional role as a cell adhesion molecule to regulate GABAergic synaptogenesis, NL2 also regulates KCC2 to modulate GABA functional switch and even glutamatergic synapses. Therefore, NL2 may serve as a master regulator in balancing excitation and inhibition in the brain.


Color Restoration of RGBN Multispectral Filter Array Sensor Images Based on Spectral Decomposition.
A multispectral filter array (MSFA) image sensor with red, green, blue and near-infrared (NIR) filters is useful for various imaging applications with the advantages that it obtains color information and NIR information simultaneously. Because the MSFA image sensor needs to acquire invisible band information, it is necessary to remove the IR cut-offfilter (IRCF). However, without the IRCF, the color of the image is desaturated by the interference of the additional NIR component of each RGB color channel. To overcome color degradation, a signal processing approach is required to restore natural color by removing the unwanted NIR contribution to the RGB color channels while the additional NIR information remains in the N channel. Thus, in this paper, we propose a color restoration method for an imaging system based on the MSFA image sensor with RGBN filters. To remove the unnecessary NIR component in each RGB color channel, spectral estimation and spectral decomposition are performed based on the spectral characteristics of the MSFA sensor. The proposed color restoration method estimates the spectral intensity in NIR band and recovers hue and color saturation by decomposing the visible band component and the NIR band component in each RGB color channel. The experimental results show that the proposed method effectively restores natural color and minimizes angular errors.


Use of ultrasound guidance improves central venous catheter insertion success rates among junior residents.
The purpose of this study was to determine whether junior residents had higher rates of first cannulation and overall success at central venous catheter insertions with the use of ultrasound (US) guidance compared to the landmark technique. We conducted a secondary analysis of data from a prospective randomized controlled study of junior residents from January 2007 through September 2008, which assessed the impact of simulation training on central venous catheter insertion success rates. Blinded independent raters observed in-hospital central venous catheter insertions using a procedural checklist. Success at first cannulation and successful insertion were the primary outcomes. Secondary outcomes included rates of technical errors and mechanical complications. Independent raters observed 480 central venous catheter insertions by 115 residents. Successful first cannulation occurred in 27% of landmark compared to 49% of dynamic US-guided (P < .01), and 50% of static US-guided (P = .01) cannulations. Insertion success occurred for 55% of landmark compared to 80% of dynamic US-guided (P < .01) and 80% of static US-guided (P < .01) cannulations. Dynamic US guidance was associated with increased odds of first cannulation success compared to the landmark technique (odds ratio [OR], 2.24; 95% confidence interval [CI], 1.37-3.67) and successful insertion (OR, 3.80; 95% CI, 2.34-6.19). Static US guidance was associated with increased odds of first cannulation success compared to the landmark technique (OR, 2.59; 95% CI, 1.25-5.39) and successful insertion (OR, 3.48; 95% CI, 1.54-7.87). The results were independent of central venous catheter insertion training, patient comorbidities, and resident specialties. There was no difference related to mechanical complications between the procedures. Dynamic and static US guidance during central venous catheter insertion was associated with improved in-hospital first cannulation rates and overall success rates of insertions by junior residents.


Content Validation of a Practice-Based Work Capacity Assessment Instrument Using ICF Core Sets.
A shift from providing long-term disability benefits to promoting work reintegration of people with remaining work capacity in many countries requires new instruments for work capacity assessments. Recently, a practice-based instrument addressing biopsychosocial aspects of functioning, the Social Medical Work Capacity instrument (SMWC), was developed. Our aim was to examine the content validity of the SMWC using ICF core sets. First, we conducted a systematic search to identify relevant ICF core sets for the working age population. Second the content of these core sets were mapped to assess the relevance and comprehensiveness of the SMWC. Next, we compared the content of the SMWC with the ICF-core sets. Two work-related core sets and 31 disease-specific core sets were identified. The SMWC and the two work-related core sets overlap on 47 categories. Compared to the work-related core sets, the Body Functions and Activities and Participation are well represented in the new instrument, while the component Environmental factors is under-represented. Compared to the disease-specific core sets, items related to the social and domestic environmental factors are under-represented, while the SMWC included work-related factors complementary to the ICF. The SMWC content seems relevant, but could be more comprehensive for the purpose of individual work capacity assessments. To improve assessing relevant biopsychosocial aspects, it is recommended to extend the instrument by adding personal and environmental (work- and social-related) factors as well as a more tailored use of the SMWC for assessing work capacity of persons with specific diseases or underlying illness.


Morphometric analysis of Leydig cells in the normal rat testis.
Leydig cells are thought to be the source of most, if not all, the testosterone produced by the testis. The goal of this study was to obtain quantitative information about rat Leydig cells and their organelles that might be correlated with pertinent physiological and biochemical data available either now or in the future. Morphometric analysis of Leydig cells in mature normal rats was carried out on tissue fixed by perfusion with buffered glutaraldehyde, and embedded in glycol methacrylate for light microscopy and in Epon for electron microscopy. In a whole testis, 82.4% of the volume was occupied by seminiferous tubules, 15.7% by the interstitial tissue, and 1.9% by the capsule. Leydig cells constituted 2.7% of testicular volume. Each cubic centimeter (contained approximatelyy 1 g) of rat testis contained about 22 million Leydig cells. An average Leydig cell had a volume of 1,210 micron3 and its plasma membrane had a surface area of 1,520 micron2. The smooth endoplasmic reticulum (SER), the most prominent organelle in Leydig cells and a major site of steroidogenic enzymes, had a surface area of approximately 10,500 micron2/cell, which is 6.9 times that of the plasma membrane and is 60% of the total membrane area of the cell. The total surface area of Leydig SER per cubic centimeter of testis tissue is approximately 2,300 cm2 or 0.23 m2. There were 3.0 mg of Leydig mitochondria in 1 g of testis tissue. The average Leydig cell contained approximately 622 mitochondria, measuring on the average 0.35 micron in diameter and 2.40 micron in length. The mitochondrial inner membrane (including cristae), another important site of steroidogenic enzymes, had a surface area of 2,920 micron2/cell, which is 1.9 times that of the plasma membrane. There were 644 cm2 of inner mitochondrial membrane/cm3 of testis tissue. These morphometric results can be correlated with published data on the rate of testosterone secretion to show that an average Leydig cell secretes approximately 0.44 pg of testosterone/d or 10,600 molecules of testosterone/s. The rate of testosterone production by each square centimeter of SER is 4.2 ng/d or 101 million molecules/s: the corresponding rate for each square centimeter of mitochondrial inner membrane is 15 ng testosterone/d or 362 million molecules/s.


Observation of the Unbiased Conformers of Putative DNA-Scaffold Ribosugars.
The constitution, configuration, and flexibility of the core sugars of DNA molecules alter their function in diverse roles. Conformational itineraries of the ribofuranosides (fs) have long been known to finely determine rates of processing, yet we also know that, strikingly, semifunctional DNAs containing pyranosides (ps) or other configurations can be created, suggesting sufficient but incompletely understood plasticity. The multiple conformers involved in such processes are necessarily influenced by context and environment: solvent, hosts, ligands. Notably, however, to date the unbiased, "naked" conformers have not been experimentally determined. Here, the inherent conformational biases of DNA scaffold deoxyribosides in unsolvated and solvated forms have now been defined using gas-phase microwave and solution-phase NMR spectroscopies coupled with computational analyses and exploitation of critical differences between natural-abundance isotopologues. Serial determination of precise, individual spectra for conformers of these 25 isotopologues in alpha (α-d) and beta (β-d); pyrano (p) and furano (f) methyl 2-deoxy-d-ribosides gave not only unprecedented atomic-level resolution structures of associated conformers but also their quantitative populations. Together these experiments revealed that typical 2E and 3E conformations of the sugar found in complex DNA structures are not inherently populated. Moreover, while both OH-5' and OH-3' are constrained by intramolecular hydrogen bonding in the unnatural αf scaffold, OH-3' is "born free" in the "naked" lowest lying energy conformer of natural βf. Consequently, upon solvation, unnatural αf is strikingly less perturbable (retaining 2T1 conformation in vacuo and water) than natural βf. Unnatural αp and βp ribosides also display low conformational perturbability. These first experimental data on inherent, unbiased conformers therefore suggest that it is the background of conformational flexibility of βf that may have led to its emergence out of multiple possibilities as the sugar scaffold for "life's code" and suggest a mechanism by which the resulting freedom of OH-3' (and hence accessibility as a nucleophile) in βf may drive preferential processing and complex structure formation, such as replicative propagation of DNA from 5'-to-3'.


Rates of hippocampal atrophy correlate with change in clinical status in aging and AD.
The cognitive continuum in the elderly population can be conceptually divided into those who are functioning normally (control subjects), those with a mild cognitive impairment (MCI), and those with probable AD. To test the hypothesis that the annualized rates of hippocampal atrophy differ as a function of both baseline and change in clinical group membership (control, MCI, or AD). The authors identified 129 subjects from the Mayo Clinic AD Research Center/AD Patient Registry who met established criteria for normal control subjects, MCI, or probable AD, both at entry and at the time of a subsequent clinical follow-up evaluation 3 +/- 1 years later. Each subject underwent an MRI examination of the head at the time of the initial assessment and at follow-up clinical assessment; the annualized percentage change in hippocampal volume was computed. Subjects who were classified as controls or patients with MCI at baseline could either remain cognitively stable or could decline to a lower functioning group over the period of observation. The annualized rates of hippocampal volume loss for each of the three initial clinical groups decreased progressively in the following order: AD > MC > control. Within the control and MCI groups, those who declined had a significantly greater rate of volume loss than those who remained clinically stable. The mean annualized rates of hippocampal atrophy by follow-up clinical group were: control-stable 1.73%, control-decliner 2.81%, MCI-stable 2.55%, MCI-decliner 3.69%, AD 3. 5%. Rates of hippocampal atrophy match both baseline cognitive status and the change in cognitive status over time in elderly persons who lie along the cognitive continuum from normal to MCI to AD.


The effects of toxoplasma infection on rodent behavior are dependent on dose of the stimulus.
Parasite Toxoplasma gondii blocks the innate aversion of rats for cat urine, putatively increasing the likelihood of a cat predating a rat. This is thought to reflect an adaptive behavioral manipulation, because toxoplasma can reproduce only in cat intestines. While it will be adaptive for the parasite to cause an absolute behavioral change, fitness costs associated with the manipulation itself suggest that the change is optimized and not maximized. We investigate these conflicting suggestions in the present report. Furthermore, exposure to cat odor causes long-lasting acquisition of learnt fear in the rodents. If toxoplasma manipulates emotional valence of cat odor rather than just sensory response, infection should affect learning driven by the aversive properties of the odor. As a second aim of the present study, we investigate this assertion. We demonstrate that behavioral changes in rodents induced by toxoplasma infection do not represent absolute all-or-none effects. Rather, these effects follow a non-monotonous function dependent on strength of stimulus, roughly resembling an inverted-U curve. Furthermore, infection affects conditioning to cat odor in a manner dependent upon strength of unconditioned stimulus employed. Non-monotonous relationship between behavioral manipulation and strength of cat odor agrees with the suggestion that a dynamic balance exists between benefit obtained and costs incurred by the parasite during the manipulation. This report also demonstrates that toxoplasma affects emotional valence of the cat odor as indicated by altered learned fear induced by cat odor.


[Diagnostic approach to infectious endocarditis].
Major advances in imaging and microbiological procedures have changed the diagnostic approach of infective endocarditis. New imaging tools have improved the search of local complications, systemic embolism and diagnosis of infection of cardiac devices. Moreover, microbiological diagnosis has been transformed by molecular techniques as long as these procedures have highlighted microorganisms thus far neglected by classical techniques. Despite these advances, endocarditis remained a clinical diagnosis which still depends on a high index of clinical suspicion.


Microelectromechanical-systems-driven two-layer rotary-blade-based adjustable iris diaphragm.
We report a miniature adjustable multi-blade iris diaphragm driven by microelectromechanical-systems-based electrostatic rotary microactuators. A proof-of-concept device is designed and fabricated using a silicon-on-insulator micromachining process. The diameter of the developed iris diaphragm aperture is experimentally demonstrated to be able to adjust from 0.45 to 1.56 mm. When used in a miniature camera lens system, the current prototype device is capable of providing an adjustment range of more than three full f/stops.


Viral determinants of age-dependent virulence of Sindbis virus for mice.
Many alphaviruses cause more severe disease in young animals than in older animals. The age-dependent resistance to severe disease is determined primarily by maturation of the host, but strains of virus can be selected that overcome the increased resistance of mature animals. Sindbis virus (SV) strain AR339 causes fatal encephalitis in newborn mice and nonfatal encephalitis in weanling mice, whereas NSV, a neuroadapted strain of SV, causes fatal encephalitis in weanling as well as newborn mice. We have previously shown that the E2 glycoprotein of NSV contained His-55, whereas AR339 E2 had Gln-55 (S. Lustig, A. C. Jackson, C. S. Hahn, D. E. Griffin, E. G. Strauss, and J. H. Strauss, J. Virol. 62:2329-2336, 1988) and that SV with E2 containing Gly-172 was more virulent for newborn mice than SV with E2 containing Arg-172 (P. C. Tucker and D. E. Griffin, J. Virol. 65:1551-1557, 1991). Here we tested the virulence for both newborn and older mice of SV containing a number of different amino acids at E2 position 55 (His, Gln, Lys, Arg, Glu, Gly) in combination with both Gly-172 and Arg-172. All the viruses were virulent for newborn mice, but the residues at both 55 and 172 influenced the virulence of the virus, and there were differences in virulence observed among the various viruses. However, only viruses with His-55 were fully virulent for 14-day-old mice, and this virulence was independent of the residue at position 172. Virus with Lys-55 was virulent for 7-day-old mice, although slightly attenuated relative to His-55. Viruses with His-55 grew more rapidly and to higher titer in the brains of 7- and 14-day-old mice, in N18 neuroblastoma cells, and in BHK cells. Our data suggest that His-55 is important for neurovirulence in older mice and acts by increasing the efficiency of virus replication.


Safety and effectiveness of insulin analogues in Moroccan patients with type 2 diabetes: a sub-analysis of the A₁chieve study.
To determine the safety and effectiveness of insulin analogues in the Moroccan cohort of the prospective, multinational, non-interventional, 24-week A₁chieve study. Moroccan patients with type 2 diabetes (T2D) starting biphasic insulin aspart 30, insulin detemir, and insulin aspart alone or in combination were included. The primary outcome was the evaluation of serious adverse drug reactions including major hypoglycaemic events. Secondary outcomes were changes in hypoglycaemic events, glycaemic parameters (HbA1c, fasting plasma glucose [FPG], postprandial plasma glucose [PPPG]), systolic blood pressure (SBP), body weight and lipid profile. Quality of life (QoL) was evaluated using the EQ-5D questionnaire. In this analysis, 1641 patients (923 insulin-naive, 718 insulin-experienced) having a mean age 57.1 years, mean BMI 26.8 kg/m(2) and mean diabetes duration 10.3 years, were included. Baseline HbA1c in the entire cohort was poor (9.7%, 83 mmol/mol). Insulin analogues statistically significantly improved glucose control (HbA1c, FPG and PPPG, p < 0.001) at Week 24. The rate of hypoglycaemia decreased from 9.31 to 4.71 events/patient-year (change in proportion of patients affected, p = 0.0002). A statistically significant improvement in lipid parameters (except HDL cholesterol) was observed while body weight changed minimally. Additionally, QoL was positively impacted (mean change in visual analogue scores from EQ-5D was 15.8 points, p < 0.001). Insulin analogue therapy resulted in improved glycaemic control and a significant overall decrease in hypoglycaemia in Moroccan T2D patients.


Effect of whey protein isolate and β-cyclodextrin wall systems on stability of microencapsulated vanillin by spray-freeze drying method.
Vanillin flavour is highly volatile in nature and due to that application in food incorporation is limited; hence microencapsulation of vanillin is an ideal technique to increase its stability and functionality. In this study, vanillin was microencapsulated for the first time by non-thermal spray-freeze-drying (SFD) technique and its stability was compared with other conventional techniques such as spray drying (SD) and freeze-drying (FD). Different wall materials like β-cyclodextrin (β-cyd), whey protein isolate (WPI) and combinations of these wall materials (β-cyd + WPI) were used to encapsulate vanillin. SFD microencapsulated vanillin with WPI showed spherical shape with numerous fine pores on the surface, which in turn exhibited good rehydration ability. On the other hand, SD powder depicted spherical shape without pores and FD encapsulated powder yielded larger particle sizes with flaky structure. FTIR analysis confirmed that there was no interaction between vanillin and wall materials. Moreover, spray-freeze-dried vanillin + WPI sample exhibited better thermal stability than spray dried and freeze-dried microencapsulated samples.


Functional magnetic resonance imaging of auditory cortical fields in awake marmosets.
The primate auditory cortex is organized into a network of anatomically and functionally distinct processing fields. Because of its tonotopic properties, the auditory core has been the main target of neurophysiological studies ranging from sensory encoding to perceptual decision-making. By comparison, the auditory belt has been less extensively studied, in part due to the fact that neurons in the belt areas prefer more complex stimuli and integrate over a wider frequency range than neurons in the core, which prefer pure tones of a single frequency. Complementary approaches, such as functional magnetic resonance imaging (fMRI), allow the anatomical identification of both the auditory core and belt and facilitate their functional characterization by rapidly testing a range of stimuli across multiple brain areas simultaneously that can be used to guide subsequent neural recordings. Bridging these technologies in primates will serve to further expand our understanding of primate audition. Here, we developed a novel preparation to test whether different areas of the auditory cortex could be identified using fMRI in common marmosets (Callithrix jacchus), a powerful model of the primate auditory system. We used two types of stimulation, band pass noise and pure tones, to parse apart the auditory core from surrounding secondary belt fields. In contrast to most auditory fMRI experiments in primates, we employed a continuous sampling paradigm to rapidly collect data with little deleterious effects. Here we found robust bilateral auditory cortex activation in two marmosets and unilateral activation in a third utilizing this preparation. Furthermore, we confirmed results previously reported in electrophysiology experiments, such as the tonotopic organization of the auditory core and regions activating preferentially to complex over simple stimuli. Overall, these data establish a key preparation for future research to investigate various functional properties of marmoset auditory cortex.


The integration of familiarity and recollection information in short-term recognition: modeling speed-accuracy trade-off functions.
Dual process models postulate familiarity and recollection as the basis of the recognition process. We investigated the time-course of integration of the two information sources to one recognition judgment in a working memory task. We tested 24 subjects with a response signal variant of the modified Sternberg recognition task (Oberauer, 2001) to isolate the time course of three different probe types indicating different combinations of familiarity and source information. We compared two mathematical models implementing different ways of integrating familiarity and recollection. Within each model, we tested three assumptions about the nature of the familiarity signal, with familiarity having (a) only positive values, indicating similarity of the probe with the memory list, (b) only negative values, indicating novelty, or (c) both positive and negative values. Both models provided good fits to the data. A model combining the outputs of both processes additively (Integration Model) gave an overall better fit to the data than a model based on a continuous familiarity signal and a probabilistic all-or-none recollection process (Dominance Model).


Angiography of the retina and the choroid with phase-resolved OCT using interval-optimized backstitched B-scans.
In conventional phase-resolved OCT blood flow is detected from phase changes between successive A-scans. Especially in high-speed OCT systems this results in a short evaluation time interval. This method is therefore often unable to visualize complete vascular networks since low flow velocities cause insufficient phase changes. This problem was solved by comparing B-scans instead of successive A-scans to enlarge the time interval. In this paper a detailed phase-noise analysis of our OCT system is presented in order to calculate the optimal time intervals for visualization of the vasculature of the human retina and choroid. High-resolution images of the vasculature of a healthy volunteer taken with various time intervals are presented to confirm this analysis. The imaging was performed with a backstitched B-scan in which pairs of small repeated B-scans are stitched together to independently control the time interval and the imaged lateral field size. A time interval of ≥ 2.5 ms was found effective to image the retinal vasculature down to the capillary level. The higher flow velocities of the choroid allowed a time interval of 0.64 ms to reveal its dense vasculature. Finally we analyzed depth-resolved histograms of volumetric phase-difference data to assess changes in amount of blood flow with depth. This analysis indicated different flow regimes in the retina and the choroid.


The Impact of Polymer Grafting from a Graphene Oxide Surface on Its Compatibility with a PDMS Matrix and the Light-Induced Actuation of the Composites.
Poly(dimethyl siloxane) (PDMS)-based materials with improved photoactuation properties were prepared by the incorporation of polymer-grafted graphene oxide particles. The modification of the graphene oxide (GO) surface was achieved via a surface initiated atom transfer radical polymerization (SI ATRP) of methyl methacrylate and butyl methacrylate. The modification was confirmed by thermogravimetric analysis, infrared spectroscopy and electron microscopy. The GO surface reduction during the SI ATRP was investigated using Raman spectroscopy and conductivity measurements. Contact angle measurements, dielectric spectroscopy and dynamic mechanical analyses were used to investigate the compatibility of the GO filler with the PDMS matrix and the influence of the GO surface modification on its physical properties and the interactions with the matrix. Finally, the thermal conductivity and photoactuation properties of the PDMS matrix and composites were compared. The incorporation of GO with grafted polymer chains, especially poly(n-butyl methacrylate), into the PDMS matrix improved the compatibility of the GO filler with the matrix, increased the energy dissipation due to the improved flexibility of the PDMS chains, enhanced the damping behavior and increased the thermal conductivity. All the changes in the properties positively affected the photoactuation behavior of the PDMS composites containing polymer-grafted GO.


Wingful, an extracellular feedback inhibitor of Wingless.
Secreted peptide signals control many fundamental processes during animal development. Proper responses to these signals require cognate inducible feedback antagonists. Here we report the identification of a novel Drosophila Wingless (Wg) target gene, wingful (wf), and show that it encodes a potent extracellular feedback inhibitor of Wg. In contrast to the cytoplasmic protein Naked cuticle (Nkd), the only known Wg feedback antagonist, Wf functions during larval stages, when Nkd function is dispensable. We propose that Wf may provide feedback control for the long-range morphogen activities of Wg.


Validation of Models Used to Inform Colorectal Cancer Screening Guidelines: Accuracy and Implications.
Microsimulation models synthesize evidence about disease processes and interventions, providing a method for predicting long-term benefits and harms of prevention, screening, and treatment strategies. Because models often require assumptions about unobservable processes, assessing a model's predictive accuracy is important. We validated 3 colorectal cancer (CRC) microsimulation models against outcomes from the United Kingdom Flexible Sigmoidoscopy Screening (UKFSS) Trial, a randomized controlled trial that examined the effectiveness of one-time flexible sigmoidoscopy screening to reduce CRC mortality. The models incorporate different assumptions about the time from adenoma initiation to development of preclinical and symptomatic CRC. Analyses compare model predictions to study estimates across a range of outcomes to provide insight into the accuracy of model assumptions. All 3 models accurately predicted the relative reduction in CRC mortality 10 years after screening (predicted hazard ratios, with 95% percentile intervals: 0.56 [0.44, 0.71], 0.63 [0.51, 0.75], 0.68 [0.53, 0.83]; estimated with 95% confidence interval: 0.56 [0.45, 0.69]). Two models with longer average preclinical duration accurately predicted the relative reduction in 10-year CRC incidence. Two models with longer mean sojourn time accurately predicted the number of screen-detected cancers. All 3 models predicted too many proximal adenomas among patients referred to colonoscopy. Model accuracy can only be established through external validation. Analyses such as these are therefore essential for any decision model. Results supported the assumptions that the average time from adenoma initiation to development of preclinical cancer is long (up to 25 years), and mean sojourn time is close to 4 years, suggesting the window for early detection and intervention by screening is relatively long. Variation in dwell time remains uncertain and could have important clinical and policy implications.


Assessment of the sustainability of technology by means of a thermodynamically based life cycle analysis.
Life cycle analysis is one of the tools in the assessment of the sustainability of technological options. It takes into account all effects on the ecosystem and the population which may endanger the possibilities of current and future generations. However, the main bottleneck in current LCA methodologies is the balancing of different effects, being all quantified on different scales. In this work, a methodology is proposed, which allows one to quantify different effects of the production, consumption and disposal of goods, and services on a single scale. The basis of the methodology is the second law of thermodynamics. All production, consumption and disposal processes affecting the ecosystem and the population, are quantified in terms of loss of exergy. The exergy content of a material is the maximum amount of energy which can be transformed into work at given environmental conditions. Next to the elaboration of the methodology, the new approach is illustrated by examples of the production of synthetic organic polymers, inorganic building insulation materials and different waste gas treatment options.


Organophosphorus insecticides in honey, pollen and bees (Apis mellifera L.) and their potential hazard to bee colonies in Egypt.
There is no clear single factor to date that explains colony loss in bees, but one factor proposed is the wide-spread application of agrochemicals. Concentrations of 14 organophosphorous insecticides (OPs) in honey bees (Apis mellifera) and hive matrices (honey and pollen) were measured to assess their hazard to honey bees. Samples were collected during spring and summer of 2013, from 5 provinces in the middle delta of Egypt. LC/MS-MS was used to identify and quantify individual OPs by use of a modified Quick Easy Cheap Effective Rugged Safe (QuEChERS) method. Pesticides were detected more frequently in samples collected during summer. Pollen contained the greatest concentrations of OPs. Profenofos, chlorpyrifos, malation and diazinon were the most frequently detected OPs. In contrast, ethoprop, phorate, coumaphos and chlorpyrifos-oxon were not detected. A toxic units approach, with lethality as the endpoint was used in an additive model to assess the cumulative potential for adverse effects posed by OPs. Hazard quotients (HQs) in honey and pollen ranged from 0.01-0.05 during spring and from 0.02-0.08 during summer, respectively. HQs based on lethality due to direct exposure of adult worker bees to OPs during spring and summer ranged from 0.04 to 0.1 for best and worst case respectively. It is concluded that direct exposure and/or dietary exposure to OPs in honey and pollen pose little threat due to lethality of bees in Egypt.


Characterization of grating couplers for affinity-based pesticide sensing.
The reflection grating coupler for direct affinity sensing is characterized in detail. The performance of this device and its potential in affinity sensing application are investigated with two affinity-based systems: A self-assembling protein-multilayer system based on avidin-biotin interaction was used to compare the response of the device with theoretical expectations. The analytical performance was characterized by a pesticide immunoassay carried out in an indirect test format with a covalently immobilized triazine derivative. Experimentally determined parameters were in good agreement with model calculations. During the binding of 12 protein monolayers at the surface, the change in effective refractive index Dn(eff) detected for a single layer decreased from approximately 8 x 10(-4) to less than 4 x10(-5) by more than 95%, indicating a filling of the evanescent field. By comparison with bulk refractive-index measurements, a refractive index n(D) approximately 1.38 of the protein multilayer was estimated. Fitting of the model gave a refractive index n(D) = 1.377 of the protein multilayer and an average thickness of 11 nm for a single protein layer. An average noise of Dn(eff) = 8.5 x 10(-7) was detected, corresponding to approximately 1% of the maximum response for a protein monolayer. At a triazine derivative attached to the surface through dextran-based surface chemistry, a maximum antibody loading that corresponds to an Dn(eff) of 1.5 x 10(-3) was observed. In an indirect immunoassay of the herbicide simazine, a detection limit of 0.25 mug/1 of simazine was reached with polyclonal Fab fragments in a concentration of 1 mug/ml.


Development and psychometric analysis of the PROMIS pain behavior item bank.
The measurement of pain behavior is a key component of the assessment of persons with chronic pain; however, few self-reported pain behavior instruments have been developed. We developed a pain behavior item bank as part of the Patient-Reported Outcome Measurement Information System (PROMIS). For the Wave I testing, because of the large number of PROMIS items, a complex sampling approach was used where participants were randomly assigned to either respond to two full-item banks or to multiple 7-item blocks of items. A web-based survey was designed and completed by 15,528 members of the general population and 967 individuals with different types of chronic pain. Item response theory (IRT) analysis models were used to evaluate item characteristics and to scale both items and individuals on the pain behavior domain. The pain behavior item bank demonstrated good fit to a unidimensional model (Comparative Fit Index = 0.94). Several iterations of IRT analyses resulted in a final 39-item pain behavior bank, and different IRT models were fit to the total sample and to those participants who experienced some pain. The results indicated that these items demonstrated good coverage of the pain behavior construct. Pain behavior scores were strongly related to pain intensity and moderately related to self-reported general health status. Mean pain behavior scores varied significantly by groups based on pain severity and general health status. The PROMIS pain behavior item bank can be used to develop static short-form and dynamic measures of pain behavior for clinical studies.


Brain Transcriptional Profiles of Male Alternative Reproductive Tactics and Females in Bluegill Sunfish.
Bluegill sunfish (Lepomis macrochirus) are one of the classic systems for studying male alternative reproductive tactics (ARTs) in teleost fishes. In this species, there are two distinct life histories: parental and cuckolder, encompassing three reproductive tactics, parental, satellite, and sneaker. The parental life history is fixed, whereas individuals who enter the cuckolder life history transition from sneaker to satellite tactic as they grow. For this study, we used RNAseq to characterize the brain transcriptome of the three male tactics and females during spawning to identify gene ontology (GO) categories and potential candidate genes associated with each tactic. We found that sneaker males had higher levels of gene expression differentiation compared to the other two male tactics. Sneaker males also had higher expression in ionotropic glutamate receptor genes, specifically AMPA receptors, compared to other males, which may be important for increased spatial working memory while attempting to cuckold parental males at their nests. Larger differences in gene expression also occurred among male tactics than between males and females. We found significant expression differences in several candidate genes that were previously identified in other species with ARTs and suggest a previously undescribed role for cAMP-responsive element modulator (crem) in influencing parental male behaviors during spawning.


Nerve reinnervation and itch behavior in a rat burn wound model.
In this study, we investigated whether postburn itch in rats, after a full thickness burn, is correlated to the nervous reinnervation of the burn wound area. For this purpose, we determined scratching duration (expressed as second/hour) at 24 hours, 2, 4, 8, and 12 weeks postburn and combined this with immunohistochemistry for protein gene product 9.5 (PGP9.5) to identify all nerve fibers, calcitonin gene related peptide (CGRP) to identify peptidergic fibers, tyrosine hydroxylase (TH) for sympathetic fibers, and growth-associated protein 43 (GAP-43) for regrowing fibers. We found a modest, but highly significant, increase in scratching duration of all burn wound rats from 3 to 12 weeks postburn (maximally 63 ± 9.5 second/hour compared to sham 3.1 ± 1.4 second/hour at 9 weeks). At 24 hours postburn, all nerve fibers had disappeared from the burn area. Around 4 weeks postburn PGP 9.5- and CGRP-immunoreactive nerve fibers returned to control levels. TH- and GAP-43-IR nerve fibers, which we found to be almost completely colocalized, did not regrow. No correlation was found between scratching duration and nervous reinnervation of the skin. The present results suggest that in rat, like in human, burn wound healing will induce increased scratching, which is not correlated to the appearance of nervous reinnervation.


The Effect of Language on Economic Behavior: Evidence from Savings Rates, Health Behaviors, and Retirement Assets.
Languages differ widely in the ways they encode time. I test the hypothesis that the languages that grammatically associate the future and the present, foster future-oriented behavior. This prediction arises naturally when well-documented effects of language structure are merged with models of intertemporal choice. Empirically, I find that speakers of such languages: save more, retire with more wealth, smoke less, practice safer sex, and are less obese. This holds both across countries and within countries when comparing demographically similar native households. The evidence does not support the most obvious forms of common causation. I discuss implications for theories of intertemporal choice.


Methods for Estimating Avoidable Costs of Excessive Alcohol Consumption.
Background: Alcohol is a risk factor with serious consequences for society and individuals. This study aims to present methods and approaches that might be used to estimate the costs related to excessive alcohol consumption. It emphasizes the need for general methods and approaches that are easily applicable, because the level of digitalization and data availability vary across regions. The lack of data makes many methods inapplicable and useless. The ease of applicability will help to make cost-of-illness studies and their results comparable globally. Methods: This study is based on data from the Czech Republic in 2017. Drinking alcohol results in costs of healthcare, social care, law enforcement, and administrative costs of public authorities. To quantify the cost of drinking in the Czech Republic, the top-down approach, bottom-up approach, human capital approach and attributable fractions were used. Results: In 2017, the cost related to alcohol was estimated at 0.66% of the national GDP. Lost productivity represented 54.45% of total cost related to alcohol. All cost related to alcohol is considered to be avoidable. Conclusions: The methods and approaches applied to estimate the cost of disease or any other health issue should be generalized regarding the availability of data and specifics of provided services to people who are addicted or have any kind of disability.


The Nitrogen Balancing Act: Tracking the Environmental Performance of Food Production.
Farmers, food supply-chain entities, and policymakers need a simple but robust indicator to demonstrate progress toward reducing nitrogen pollution associated with food production. We show that nitrogen balance-the difference between nitrogen inputs and nitrogen outputs in an agricultural production system-is a robust measure of nitrogen losses that is simple to calculate, easily understood, and based on readily available farm data. Nitrogen balance provides farmers with a means of demonstrating to an increasingly concerned public that they are succeeding in reducing nitrogen losses while also improving the overall sustainability of their farming operation. Likewise, supply-chain companies and policymakers can use nitrogen balance to track progress toward sustainability goals. We describe the value of nitrogen balance in translating environmental targets into actionable goals for farmers and illustrate the potential roles of science, policy, and agricultural support networks in helping farmers achieve them.


Postoperative nausea and vomiting in pediatric anesthesia.
Postoperative nausea and vomiting (PONV) has a high incidence in children and requires prophylactic and therapeutic strategies. PONV can be reduced by the avoidance of nitrous oxide, volatile anesthetics, and the reduction of postoperative opioids. The use of dexamethasone, 5-HT3 antagonists, or droperidol alone is potent, but combinations are even more effective to reduce PONV. Droperidol has a Food and Drug Administration warning. Hence, dexamethasone and 5-HT3 antagonists should be preferred as prophylactic drugs. It is further reasonable to adapt PONV prophylaxis to different risk levels. Prolonged surgery time, inpatients, types of surgery (e.g. strabismus and ear-nose-throat surgery), and patients with PONV in history should be treated as high risk, whereas short procedures and outpatients are to be treated as low risk. Concluding from the existing guidelines and data on the handling of PONV in children at least 3 years, the following recommendations are given: outpatients undergoing small procedures should receive a single prophylaxis, outpatients at high risk a double prophylaxis, inpatients with surgery time of more than 30 min and use of postoperative opioids should get double prophylaxis, and inpatients receiving a high-risk surgical procedure or with other risk factors a triple prophylaxis (two drugs and total intravenous anesthesia). Dimenhydrinate can be used as a second choice, whereas droperidol and metoclopramide can only be recommended as rescue therapy.


A chemical screen for medulloblastoma identifies quercetin as a putative radiosensitizer.
Treatment of medulloblastoma in children fails in approximately 30% of patients, and is often accompanied by severe late sequelae. Therefore, more effective drugs are needed that spare normal tissue and diminish long-term side effects. Since radiotherapy plays a pivotal role in the treatment of medulloblastoma, we set out to identify novel drugs that could potentiate the effect of ionizing radiation.Thereto, a small molecule library, consisting of 960 chemical compounds, was screened for its ability to sensitize towards irradiation. This small molecule screen identified the flavonoid quercetin as a novel radiosensitizer for the medulloblastoma cell lines DAOY, D283-med, and, to a lesser extent, D458-med at low micromolar concentrations and irradiation doses used in fractionated radiation schemes. Quercetin did not affect the proliferation of neural precursor cells or normal human fibroblasts. Importantly, in vivo experiments confirmed the radiosensitizing properties of quercetin. Administration of this flavonoid at the time of irradiation significantly prolonged survival in orthotopically xenografted mice. Together, these findings indicate that quercetin is a potent radiosensitizer for medulloblastoma cells that may be a promising lead for the treatment of medulloblastoma in patients.


Photon diffusion in a homogeneous medium bounded externally or internally by an infinitely long circular cylindrical applicator. V. Steady-state fluorescence.
As Part V in our series, this paper examines steady-state fluorescence photon diffusion in a homogenous medium that contains a homogenous distribution of fluorophores, and is enclosed by a "concave" circular cylindrical applicator or is enclosing a "convex" circular cylindrical applicator, both geometries being infinite in the longitudinal dimension. The aim is to predict by analytics and examine with the finite-element method the changing characteristics of the fluorescence-wavelength photon-fluence rate and the ratio (sometimes called the Born ratio) of it versus the excitation-wavelength photon-fluence rate, with respect to the source-detector distance. The analysis is performed for a source and a detector located on the medium-applicator interface and aligned either azimuthally or longitudinally in both concave and convex geometries. When compared to its steady-state counterparts on a semi-infinite medium-applicator interface with the same line-of-sight source-detector distance, the fluorescence-wavelength photon-fluence rate reduces faster along the longitudinal direction and slower along the azimuthal direction in the concave geometry, and conversely in the convex geometry. However, the Born ratio increases slower in both azimuthal and longitudinal directions in the concave geometry and faster in both directions in the convex geometry, respectively, when compared to that in the semi-infinite geometry.


Effects of a low-salt diet on idiopathic hypercalciuria in calcium-oxalate stone formers: a 3-mo randomized controlled trial.
A direct relation exists between sodium and calcium excretion, but randomized studies evaluating the sustained effect of a low-salt diet on idiopathic hypercalciuria, one of the main risk factors for calcium-oxalate stone formation, are still lacking. Our goal was to evaluate the effect of a low-salt diet on urinary calcium excretion in patients affected by idiopathic calcium nephrolithiasis. Patients affected by idiopathic calcium stone disease and hypercalciuria (>300 mg Ca/d in men and >250 mg Ca/d in women) were randomly assigned to receive either water therapy alone (control diet) or water therapy and a low-salt diet (low-sodium diet) for 3 mo. Twenty-four-hour urine samples were obtained twice from all patients: one sample at baseline on a free diet and one sample after 3 mo of treatment. A total of 210 patients were randomly assigned to receive a control diet (n = 102) or a low-sodium diet (n = 108); 13 patients (2 on the control diet, 11 on the low-sodium diet) withdrew from the trial. At the follow-up visit, patients on the low-sodium diet had lower urinary sodium (mean +/- SD: 68 +/- 43 mmol/d at 3 mo compared with 228 +/- 57 mmol/d at baseline; P < 0.001). Concomitant with this change, they showed lower urinary calcium (271 +/- 86 mg/d at 3 mo compared with 361 +/- 129 mg/d on the control diet, P < 0.001) and lower oxalate excretion (28 +/- 8 mg/d at 3 mo compared with 32 +/- 10 mg/d on the control diet, P = 0.001). Urinary calcium was within the normal range in 61.9% of the patients on the low-salt diet and in 34.0% of those on the control diet (difference: +27.9%; 95% CI: +14.4%, +41.3%; P < 0.001). A low-salt diet can reduce calcium excretion in hypercalciuric stone formers. This trial was registered at clinicaltrials.gov as NCT01005082.


[Evaluation of multiplex PCR assay using dual priming oligonucleotide system for detection mutation in the Duchenne muscular dystrophy gene].
Exon deletions of Duchenne muscular dystrophy (DMD) gene account for most of the alterations found in DMD and Becker muscular dystrophy (BMD). This study was to evaluate the usefulness of dual priming oligonucleotide multiplex PCR (DPO PCR) in detection of exon deletions of DMD gene. Thirty-seven DMD or BMD patients who had known exon deletions detected by conventional multiplex PCR (conventional PCR) and nine control subjects were enrolled in this study. When a discrepancy was shown between the results of conventional PCR and DPO PCR, the multiplex ligation-dependent probe amplification (MLPA) technique was performed as a confirmation test. The same deletions previously identified by conventional PCR in 32 out of 37 subjects were also detected by DPO PCR. For the five subjects (13.5%) showing discrepant results between the conventional PCR and DPO PCR, MLPA was performed and its results were found to correlate better with those of DPO PCR. The discrepancies were due to false positive or false negative results of the conventional PCR. DPO PCR shows a high agreement of results with the conventional PCR and is considered an adequate method to be used as a primary genetic test for the diagnosis of DMD. Because of an improved accuracy, especially for determining the boundaries of DMD gene deletions, DPO PCR can be very useful as a supplement to the conventional PCR.


Circulating opioid peptides during water immersion in normal man.
1. This study was designed to evaluate variations in plasma beta-endorphin, methionine-enkephalin, adrenocorticotropic hormone and serum prolactin in healthy volunteers during head-out water immersion. 2. Water immersion induced an increase in methionine-enkephalin plasma levels, which was associated with a significant fall in mean arterial pressure and heart rate. 3. Conversely, a suppression of plasma beta-endorphin, adrenocorticotropic hormone and serum prolactin was detected during water immersion. 4. We suggest that a dopaminergic inhibitory control mechanism may be involved in regulating circulating levels of beta-endorphin, adrenocorticotropic hormone and prolactin in normal subjects undergoing extracellular fluid volume expansion produced by water immersion.


National Machine Guarding Program: Part 2. Safety management in small metal fabrication enterprises.
Small manufacturing businesses often lack important safety programs. Many reasons have been set forth on why this has remained a persistent problem. The National Machine Guarding Program (NMGP) was a nationwide intervention conducted in partnership with two workers' compensation insurers. Insurance safety consultants collected baseline data in 221 business using a 33-question safety management audit. Audits were completed during an interview with the business owner or manager. Most measures of safety management improved with an increasing number of employees. This trend was particularly strong for lockout/tagout. However, size was only significant for businesses without a safety committee. Establishments with a safety committee scored higher (55% vs. 36%) on the safety management audit compared with those lacking a committee (P < 0.0001). Critical safety management programs were frequently absent. A safety committee appears to be a more important factor than business size in accounting for differences in outcome measures.


Outpatient antibiotic use in France between 2000 and 2010: after the nationwide campaign, it is time to focus on the elderly.
Antibiotic overconsumption is the main force driving the emergence of multidrug-resistant bacterial strains. To promote better antibiotic use in France, a nationwide campaign has been run every year from October to March since 2002. In 2007, it was shown that winter outpatient antibiotic consumption had decreased by 26.5% compared to the 2000-2002 baseline period. Here, we quantified outpatient antibiotic use between 2000 and 2010 as a follow-up analysis of the nationwide campaign. Reimbursed outpatient antibiotic prescriptions were extracted from computerized French National Health Insurance databases. Entire series and age group and antibiotic class analyses were computed. Time series analyses used autoregressive moving-average models with exogenous variables and intervention functions. Two periods were considered: October to March "campaign" periods and April to September "warm" periods. Compared to the precampaign (2000-2002) baseline period, the numbers of weekly antibiotic prescriptions per 1,000 inhabitants during campaign periods decreased until winter 2006 to 2007 (-30% [95% confidence interval {CI}, -36.3 to -23.8%]; P < 0.001) and then stabilized except for individuals >60 years of age, for whom prescriptions reached the precampaign level. During the warm periods from April to September, no significant differences were estimated compared to the baseline level for the entire series, but seniors had an increasing trend that became significant as of 2005, reaching +21% (95% CI, +12.9 to +29.6%) in 2009 (P < 0.0001). These results highlight the need for a better understanding of antibiotic use by the elderly, requiring research with targeted and tailored public health actions for this population.


Postprandial effects of almond consumption on human osteoclast precursors--an ex vivo study.
Consumption of almonds has been associated with increased bone mineral density, but the direct effects of almonds on bone cells are not known. We determined whether serum obtained following the consumption of a meal containing 60 g of almonds affects human osteoclast formation, function, and gene expression in vitro. Human osteoclast precursors were cultured in medium containing 10% serum obtained from 14 healthy subjects at baseline and 4 hours following the consumption of 3 test meals containing almonds, potatoes, and rice and balanced for macronutrient composition. Osteoclast formation was determined by the number of tartrate-resistant acid phosphatase (TRAP)(+) multinucleated cells, and osteoclast function was assessed by measuring TRAP activity in the culture medium and calcium released from OsteoAssay (Lonza Walkersville, Walkersville, MD, USA) plates. The expression of cathepsin K, receptor activator of nuclear factor kB, and matrix metalloproteinase-9 genes was measured by real-time reverse transcriptase-polymerase chain reaction. Compared with serum obtained at baseline, serum obtained 4 hours following the consumption of the almond meal reduced osteoclast formation by approximately 20%, TRAP activity by approximately 15%, calcium release by approximately 65%, and the expression of cathepsin K, receptor activator of nuclear factor kB, and matrix metalloproteinase-9 by 13% to 23%. No effects were observed with serum obtained from the other test meals. Serum obtained 4 hours following the consumption of an almond meal inhibits osteoclast formation, function, and gene expression in cultured human osteoclast precursors, and provides evidence for a positive effect of almonds on bone health.


Energy levels at systole vs. diastole in normal hamster hearts vs. myopathic hamster hearts.
The following studies were carried out to examine energy metabolites and cardiac performance of the failing heart (hereditary cardiomyopathy) of the Syrian hamster (strain UM-X7.1) perfused either by normal or stress conditions, and to determine whether cyclical changes in energy-related metabolites occurred in the glucose-perfused hearts of both normal and heart failure animals. Hamster hearts from 250-day-old animals with moderate heart failure were removed and perfused either as nonworking hearts (Langendorff method, an afterload pressure of 90 mm Hg and 2.5 mM calcium in the perfusate) or as working hearts with stress conditions [an afterload of 110 mm Hg, high calcium concentrations in the perfusate (3.5 mM), and 10(-8) M isoproterenol]. Mechanical parameters (developed pressure and max dP/dt) and measurements of oxygen consumption indicated that both contractility and oxygen consumption had fallen 50% in myopathic hearts, compared with those of normal hamsters perfused with either of the two conditions. By means of a specially designed stimulator-triggered freeze clamp, hearts were terminated at systole and diastole, and tissue content of ATP, ADP, AMP, adenosine, phosphocreatine, creatine, pyruvate, lactate, and inorganic phosphate were analyzed. A 50% reduction in cardiac performance of the cardiomyopathic hamster hearts was associated with a corresponding reduction in systolic ATP, adenosine, and phosphocreatine values, while inorganic phosphate and lactate increased. With glucose as the sole substrate, the high energy phosphates, ATP and phosphocreatine, reached maximum values during diastole and minimum values during systole.(ABSTRACT TRUNCATED AT 250 WORDS)


Swedish Society of Rheumatology 2018 guidelines for investigation, treatment, and follow-up of giant cell arteritis.
Objective: To develop evidence-based guidelines for the management of giant cell arteritis (GCA) as a complement to guidelines in other areas of rheumatology, issued by the Swedish Society of Rheumatology. Methods: A working group selected key areas for recommendations, reviewed the available evidence, and wrote draft guidelines. These were discussed and revised according to standard procedures within the Swedish Society of Rheumatology, including a one-day meeting open to all members. For key recommendations, the quality of evidence was assessed according to GRADE. The final guidelines were approved by the Society board in March 2018. Results: The guidelines include recommendations on diagnostic procedures, pharmacological treatment, follow-up, and adjuvant treatment. Ultrasonography is complementary to temporal artery biopsy (TAB) in the diagnostic work-up. Other imaging techniques (magnetic resonance imaging and positron emission tomography/computed tomography) are important in evaluating large-vessel involvement. Glucocorticoids (oral, or intravenous in cases with ischaemic complications) remain the first line treatment for GCA. Addition of tocilizumab is recommended for patients with relapsing disease who meet five criteria, representing active disease that has been objectively verified by TAB or imaging. Tocilizumab may also be considered in patients with newly diagnosed GCA who are at major risk of severe glucocorticoid side effects. Based on current evidence, tocilizumab treatment for > 1 year cannot be recommended. Conclusion: These guidelines are based on current evidence and consensus within Swedish rheumatology. Following major developments in diagnostics and treatment of GCA, such guidelines are important for clinical practice, and should be updated on a regular basis.


Complex relationships between substrate sequence and sensitivity to alterations in γ-secretase processivity induced by γ-secretase modulators.
γ-Secretase catalyzes the final cleavage of the amyloid precursor protein (APP), resulting in the production of amyloid-β (Aβ) peptides with different carboxyl termini. Presenilin (PSEN) and amyloid precursor protein (APP) mutations linked to early onset familial Alzheimer's disease modify the profile of Aβ isoforms generated, by altering both the initial γ-secretase cleavage site and subsequent processivity in a manner that leads to increased levels of the more amyloidogenic Aβ42 and in some circumstances Aβ43. Compounds termed γ-secretase modulators (GSMs) and inverse GSMs (iGSMs) can decrease and increase levels of Aβ42, respectively. As GSMs lower the level of production of pathogenic forms of long Aβ isoforms, they are of great interest as potential Alzheimer's disease therapeutics. The factors that regulate GSM modulation are not fully understood; however, there is a growing body of evidence that supports the hypothesis that GSM activity is influenced by the amino acid sequence of the γ-secretase substrate. We have evaluated whether mutations near the luminal border of the transmembrane domain (TMD) of APP alter the ability of both acidic, nonsteroidal anti-inflammatory drug-derived carboxylate and nonacidic, phenylimidazole-derived classes of GSMs and iGSMs to modulate γ-secretase cleavage. Our data show that point mutations can dramatically reduce the sensitivity to modulation of cleavage by GSMs but have weaker effects on iGSM activity. These studies support the concept that the effect of GSMs may be substrate selective; for APP, it is dependent on the amino acid sequence of the substrate near the junction of the extracellular domain and luminal segment of the TMD.


Selection of peripheral and intestinal T lymphocytes lacking CD3 zeta.
The CD3 zeta chain of the TCR plays a pivotal role in the activation of T cell responses toward foreign antigen and in the selection of the T cell repertoire. T lymphocytes from mice deficient in CD3 zeta (CD3 zeta/eta-/- mice) express very few cell surface TCR-CD3 complexes, and these animals have poorly developed thymuses which lack single-positive CD8 and CD4 thymocytes. Nevertheless, a substantial number of single-positive CD4+ and CD8+ T lymphocytes are found in peripheral lymphoid organs of CD3 zeta/eta-/- animals. Using double-mutant mice, generated by breeding CD3 zeta/eta-/- mice with others deficient in the expression of either class I or class II MHC molecules, we demonstrate here that positive selection of peripheral CD4+ and CD8+ T lymphocytes can occur in the absence of CD3 zeta/eta molecules. Analysis of the intestinal intra-epithelial lymphocytes from CD3 zeta/eta-/- mice revealed a novel T cell population expressing high levels of an alternative TCR alpha beta, due to the replacement of CD3 zeta by Fc epsilon RI gamma. Developmentally, these cells also depend on class I MHC expression. In contrast, TCR gamma delta/Fc epsilon RI gamma+ T cells develop independently of MHC class I or class II molecules. These experiments demonstrate that the unique subset of intestinal TCR alpha beta/Fc epsilon RI gamma+ lymphocytes is developmentally dependent on MHC expression. The restricted expression of TCR alpha beta/Fc epsilon RI gamma+ cells in the intestinal mucosa (rather than the thymus or lymph nodes) supports the hypothesis that selection of these T cells occurs extrathymically.


Challenges in international medicine: ethical dilemmas, unanticipated consequences, and accepting limitations.
While personal and organizational challenges occur in every area of health care, practitioners of international medicine face unique problems and dilemmas that are rarely discussed in training programs. Health professions schools, residency and fellowship programs, nongovernmental organizations (NGOs), and government programs have a responsibility to make those new to international medicine aware of the special circumstances that they may face and to provide methods for understanding and dealing with these circumstances. Standard "domestic" approaches to such challenges may not work in international medicine, even though these challenges may appear to be similar to those faced in other clinical settings. How should organizations ensure that well-meaning health intervention efforts do not cause adverse unintended sequelae? How should an individual balance respect for cultural uniqueness and local mores that may profoundly differ from his or her own beliefs, with the need to remain a moral agent true to one's self? When is acceptance the appropriate response to situations in which limitations of resources seem to preclude any good solution? Using a case-based approach, the authors discuss issues related to the four major international medicine domains: clinical practice (postdisaster response, resource limitations, standards of care), medical systems and systems development (prehospital care, wartime casualties, sustainable change, cultural awareness), teaching (instruction and local resources, professional preparation), and research (questionable funded studies, clinical trials, observational studies). It is hoped that this overview may help prepare those involved with international medicine for the challenges and dilemmas they may face and help frame their responses to these situations.


Habitual physical exercise has beneficial effects on telomere length in postmenopausal women.
It has been reported that women benefit from the maintenance of telomere length by estrogen. Exercise may favorably influence telomere length, although results are inconsistent regarding the duration and type of exercise and the cell type used to measure telomere length. The purpose of this study was to investigate the relationship between habitual physical exercise and telomere length in peripheral blood mononuclear cells (PBMCs) in postmenopausal women. Postmenopausal women were chosen as study participants because they are typically estrogen deficient. This experimental-control, cross-sectional study included 44 healthy, nondiabetic, nonsmoking, postmenopausal women. Habitual exercisers and sedentary participants were matched for age and body mass index. Body weight, height, blood pressure, and waist and hip circumference were measured. Mitochondrial DNA copy number and telomere length in PBMCs were determined, and biochemical tests were performed. Habitual physical exercise was defined as combined aerobic and resistance exercise performed for at least 60 minutes per session more than three times a week for more than 12 months. The mean age of all participants was 58.11 ± 6.84 years, and participants in the habitual exercise group had been exercising more than three times per week for an average of 19.23 ± 5.15 months. Serum triglyceride levels (P = 0.01), fasting insulin concentrations (P < 0.01), and homeostasis model assessment of insulin resistance (P < 0.01) were significantly lower and high-density lipoprotein cholesterol levels (P < 0.01), circulating adiponectin (P < 0.01), mitochondrial DNA copy number (P < 0.01), and telomere length (P < 0.01) were significantly higher in the habitual exercise group than in the sedentary group. In a stepwise multiple regression analysis, habitual exercise (β = 0.522, P < 0.01) and adiponectin levels (β = 0.139, P = 0.03) were the independent factors associated with the telomere length of PBMCs in postmenopausal women. Habitual physical exercise is associated with greater telomere length in postmenopausal women. This finding suggests that habitual physical exercise in postmenopausal women may reduce telomere attrition.


Growth factors mobilize multiple pools of KCa channels in developing parasympathetic neurons: role of ADP-ribosylation factors and related proteins.
In developing ciliary ganglion (CG) neurons, movement of functional large-conductance (BK type) Ca(2+)-activated K+ (K(Ca)) channels to the cell surface is stimulated by the endogenous growth factors TGF(beta)1 and beta-neuregulin-1 (NRG1). Here we show that a brief NRG1 treatment (0.5-1.5 h) mobilizes K(Ca) channels in a post-Golgi compartment, but longer treatments (>3.5 h) mobilize K(Ca) channels located in the endoplasmic reticulum or Golgi apparatus. Specifically, the effects of 3.5 h NRG1 treatment were completely blocked by treatments that disrupt Golgi apparatus function. These include inhibition of microtubules, or inhibition of the ADP-ribosylation factor-1 (ARF1) system by brefeldin A, by over-expression of dominant-negative ARF1, or over-expression of an ARF1 GTPase-activating protein that blocks ARF1 cycling between GTP- and GDP-bound states. These treatments had no effect on stimulation of K(Ca) evoked by 1.5 h treatment with NRG1, indicating that short-term responses to NRG1 do not require an intact Golgi apparatus. By contrast, both the acute and sustained effects of NRG1 were inhibited by treatments that block trafficking processes that occur close to the plasma membrane. Thus mobilization of K(Ca) was blocked by treatments than inhibit ADP-ribosylation factor-6 (ARF6) signaling, including overexpression of dominant-negative ARF6, dominant-negative ARNO, or dominant-negative phospholipase D1. TGF(beta)1, the effects of which on K(Ca) are much slower in onset, is unable to selectively mobilize channels in the post-Golgi pool, and its effects on K(Ca) are completely blocked by inhibition of microtubules, Golgi function and also by plasma membrane ARF6 and phospholipase D1 signaling.


Behavioral and physiologic indicators of pain in nonverbal patients with a traumatic brain injury: an integrative review.
The use of behavioral and physiologic indicators is recommended for pain assessment in nonverbal patients. Traumatic brain injuries (TBI) can lead to neurologic changes and affect the way patients respond to pain. As such, commonly used indicators of pain may not apply to TBI patients. This study aimed to review the literature about behavioral/physiologic indicators of pain in nonverbal TBI patients. An integrative review method was used. Medline (from 1948 to June 2011), Cinahl, and Cochrane databases were searched using any combination of the terms brain injury, behavioral indicators, behavioral scale, physiologic indicators, pain, pain assessment, and pain measurement. All articles reporting expert opinion or original data about the validity of behavioral and/or physiologic indicators of pain in TBI patients were considered. For each article included, the quality of findings/clinical recommendations was graded independently by two raters using SORT taxonomy. Eight papers were reviewed. Overall, TBI patients seemed to present a wider range of behavioral reactions to pain than other adult populations. In addition to the commonly observed grimace, agitation, and increased muscle tension, 14%-72% of TBI patients showed raising eyebrows, opening eyes, weeping eyes, and absence of muscle tension when exposed to pain. Those atypical reactions appeared to be present only in the acute phase of TBIs recovery. Similarly to other populations, vital signs were identified as potential indicators of pain in TBI patients. Further research studying TBI patients and considering changes in level of consciousness, location/severity of brain injury, and administration of analgesic/sedative is needed. Until then, nurses should follow the current clinical recommendations.


Failure of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate to enhance sister chromatid exchange, mitotic segregation, or expression of mutations in Chinese hamster cells.
The potent tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) was tested for its ability (a) to induce sister chromatid exchange, (b) to increase the rate of transition at the adenine phosphoribosyltransferase (apt) locus from the presumptive heterozygous state ((+/- to the homozygous state (-/ - or -), and (c) to enhance the frequency of mutations expressed after ultraviolet radiation mutagenesis. We have found no significant effect of TPA in any of these experiments. Sister chromatid exchange frequencies in both V79 and Chinese hamster ovary cells remained unchanged by TPA treatment under various conditions, a result inconsistent with the hypothesis that an important effect of TPA might be to increase the rate of chromosomal mitotic recombination (and hence segregation of recessive mutations) in a manner akin to increased chromatid recombination. We have also been unable to obtain evidence for mitotic recombination affecting the aprt locus in Chinese hamster ovary cells for which the rate of change to a high level of resistance to azaadenine was measured. The rate of 8.6 X 10(-7) mutation (and/or segregations) per cell generation assessed by fluctuation analysis was not increased by the continuous presence of TPA, 4 microgram/ml, in the medium. In the third set of experiments, mutant frequencies in Chinese hamster ovary cells after ultraviolet mutagenesis were measured for the markers ouabain resistance, thioguanine resistance, and azaadenine resistance, under conditions with and without pretreatment with TPA before mutant selection. No convincing enhancement in mutation expression was observed. In summary, these results argue that promotion by TPA does not proceed by a mechanism involving genetic recombination or the altered expression of newly mutated alleles.


GPI-anchored proteins are confined in subdiffraction clusters at the apical surface of polarized epithelial cells.
Spatio-temporal compartmentalization of membrane proteins is critical for the regulation of diverse vital functions in eukaryotic cells. It was previously shown that, at the apical surface of polarized MDCK cells, glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) are organized in small cholesterol-independent clusters of single GPI-AP species (homoclusters), which are required for the formation of larger cholesterol-dependent clusters formed by multiple GPI-AP species (heteroclusters). This clustered organization is crucial for the biological activities of GPI-APs; hence, understanding the spatio-temporal properties of their membrane organization is of fundamental importance. Here, by using direct stochastic optical reconstruction microscopy coupled to pair correlation analysis (pc-STORM), we were able to visualize and measure the size of these clusters. Specifically, we show that they are non-randomly distributed and have an average size of 67 nm. We also demonstrated that polarized MDCK and non-polarized CHO cells have similar cluster distribution and size, but different sensitivity to cholesterol depletion. Finally, we derived a model that allowed a quantitative characterization of the cluster organization of GPI-APs at the apical surface of polarized MDCK cells for the first time. Experimental FRET (fluorescence resonance energy transfer)/FLIM (fluorescence-lifetime imaging microscopy) data were correlated to the theoretical predictions of the model.


NMDA receptor antibody encephalitis.
N-methyl-D-aspartate receptor (NMDAR) antibody encephalitis is a recently described immunotherapy-responsive panencephalitis with characteristic features that include a psychiatric onset and a later movement disorder. This entity was first described as a paraneoplastic phenomenon in young women with ovarian teratomata. However, more recently it has become clear that the majority of patients, particularly children, do not harbor a tumor and that males can also be affected. With the development of the NMDAR antibody assay, now available worldwide, a few patients with classical limbic encephalitis and early psychosis and epilepsy have also been found to harbor these antibodies. Early diagnosis followed by immunotherapies and tumor removal, when relevant, expedite recovery from the condition. Antibody levels correlate with the clinical severity of the disease in individual patients, and the antibodies have been shown to substantially reduce NMDA receptors on hippocampal neurons both in vitro and in vivo, supporting the likely direct pathogenicity of the NMDAR antibodies.