Relationships Between Sensorimotor Inhibition and Mobility in Older Adults With and Without Parkinson's Disease.
Reduced cortical sensorimotor inhibition is associated with mobility and cognitive impairments in people with Parkinson's disease (PD) and older adults (OAs). However, there is a lack of clarity regarding the relationships among sensorimotor, cognitive, and mobility impairments. The purpose of this study was to determine how cortical sensorimotor inhibition relates to impairments in mobility and cognition in people with PD and OAs. Cortical sensorimotor inhibition was characterized with short-latency afferent inhibition (SAI) in 81 people with PD and 69 OAs. Six inertial sensors recorded single- and dual-task gait and postural sway characteristics during a 2-minute walk and a 1-minute quiet stance. Cognition was assessed across the memory, visuospatial, executive function, attention, and language domains. SAI was significantly impaired in the PD compared to the OA group. The PD group preformed significantly worse across all gait and postural sway tasks. In PD, SAI significantly correlated with single-task foot strike angle and stride length variability, sway area, and jerkiness of sway in the coronal and sagittal planes. In OAs, SAI significantly related to single-task gait speed and stride length, dual-task stride length, and immediate recall (memory domain). No relationship among mobility, cognition, and SAI was observed. Impaired SAI related to slower gait in OA and to increased gait variability and postural sway in people with PD, all of which have been shown to be related to increased fall risk.


Anoxic photocatalytic treatment of synthetic mining wastewater using TiO2 and scavengers for complexed cyanide recovery.
The wastewater from gold exploitation is well known for the toxic nature of recalcitrant cyanide metallic complexes. In this work the selectivity in the photocatalytic degradation of gold mining wastewater (Fe(CN)63-) using suspended TiO2 with alcoholic and organic acid scavengers in a mini-CPC photoreactor with a 30 W UV-A LED as an artificial source of light was evaluated. The study was done in four stages: 1. load catalyst determination, 2. combination of scavengers in a typical photocatalytic degradation, 3. evaluation of scavenger concentration and 4. kinetic study. The decomposition into CN- and Fe removal of the cyanocomplex were tracked. It was observed that formic acid (FA) and t-butanol (t-ButOH) were the best scavengers for the photocatalytic degradation under anoxic conditions. The best concentrations of acceptors used in the study were 10 mM FA and 10 mM t-ButOH at 20 W m-2 of UV-A power, reaching 80% degradation of Fe(CN)63-, 40% Fe removal and 18 ppm of free cyanide CN release to the liquid phase. The electrical efficiency of oxidation per order (EEo) was increased by about 50% with the addition of scavengers instead of traditional anoxic photocatalytic treatment. It was proved that the photocatalytic decomposition of the Fe cyanocomplex was done through the photoreduction path of the metal complex.


Crystallization of molecular systems from solution: phase diagrams, supersaturation and other basic concepts.
The aim of the tutorial review is to show that any crystallization from solution is guided by stable or metastable equilibria and thus can be rationalized by using phase diagrams. Crystallization conducted by cooling, by evaporation and by anti-solvent addition is mainly considered. The driving force of crystallization is quantified and the occurrence of transient metastable states is logically explained by looking at the pathways of crystallization and the progressive segregation which might occur in a heterogeneous system.


Comparative effectiveness and safety of homologous two-dose ChAdOx1 versus heterologous vaccination with ChAdOx1 and BNT162b2.
Small trials have suggested that heterologous vaccination with first-dose ChAdOx1 and second-dose BNT162b2 may generate a better immune response than homologous vaccination with two doses of ChAdOx1. In this cohort analysis, we use linked data from Catalonia (Spain), where those aged <60 who received a first dose of ChAdOx1 could choose between ChAdOx1 and BNT162b2 for their second dose. Comparable cohorts were obtained after exact-matching 14,325/17,849 (80.3%) people receiving heterologous vaccination to 14,325/149,386 (9.6%) receiving homologous vaccination by age, sex, region, and date of second dose. Of these, 464 (3.2%) in the heterologous and 694 (4.8%) in the homologous groups developed COVID-19 between 1st June 2021 and 5th December 2021. The resulting hazard ratio (95% confidence interval) is 0.66 [0.59-0.74], favouring heterologous vaccination. The two groups had similar testing rates and safety outcomes. Sensitivity and negative control outcome analyses confirm these findings. In conclusion, we demonstrate that a heterologous vaccination schedule with ChAdOx1 followed by BNT162b2 was more efficacious than and similarly safe to homologous vaccination with two doses of ChAdOx1. Most of the infections in our study occurred when Delta was the predominant SARS-CoV-2 variant in Spain. These data agree with previous phase 2 randomised trials.


Potentials of gum from Detarium microcarpum (DM) and Mucuna flagellipes (MF) seeds as raw beef burger stabilizers.
Raw beef burgers containing graded levels (0.25, 0.5, 0.75. and 1.0%) of polysaccharide gums extracted from Detarium microcarpum (DM) and Mucuna flagellipes (MF) were produced. Unstabilized beef burgers and beef burgers containing gum tragacanth (TR) were also produced simultaneously to serve as control. The raw beef burgers were evaluated for selected physicochemical and sensory properties so as to assess the stabilization potentials of DM and MF polysaccharide gums relative to the conventional TR. Beef burgers containing the polysaccharide gums had significantly (P < 0.05) lower shrinkage, higher water holding capacity (WHC), and better stability under ambient conditions (27 +/- 1 degrees C and RH 90.6%) then the unstabilized burgers. Among the stabilized burgers, DM gum burgers had lower (P < or = 0.05) shrinkage and higher WHC than MF and TR gums burgers. All the gum-stabilized beef burgers required 9-26% higher compression force indicating a less friable product than the unstabilized burgers. At 0.25 and 0.5% levels of gum incorporation DM gum burgers had comparable compression force with TR gum burgers and this was lower (P < or = 0.05) than the compression force of MF gum burgers. Sensory panel results indicated a lower score for finger feel in gum-stablized burgers than the unstabilized burgers. DM and TR gum burgers had comparable scores with TR gum burgers in all the sensory attributes tested. Overall acceptability score showed that all the gum-stabilized beef burgers were acceptable. It was evident from the study that polysaccharide gums from DM and MF seeds can serve as effective stabilizers in beef burgers without adverse effect on the quality of the product.


Prediction of twin-arginine signal peptides.
Proteins carrying twin-arginine (Tat) signal peptides are exported into the periplasmic compartment or extracellular environment independently of the classical Sec-dependent translocation pathway. To complement other methods for classical signal peptide prediction we here present a publicly available method, TatP, for prediction of bacterial Tat signal peptides. We have retrieved sequence data for Tat substrates in order to train a computational method for discrimination of Sec and Tat signal peptides. The TatP method is able to positively classify 91% of 35 known Tat signal peptides and 84% of the annotated cleavage sites of these Tat signal peptides were correctly predicted. This method generates far less false positive predictions on various datasets than using simple pattern matching. Moreover, on the same datasets TatP generates less false positive predictions than a complementary rule based prediction method. The method developed here is able to discriminate Tat signal peptides from cytoplasmic proteins carrying a similar motif, as well as from Sec signal peptides, with high accuracy. The method allows filtering of input sequences based on Perl syntax regular expressions, whereas hydrophobicity discrimination of Tat- and Sec-signal peptides is carried out by an artificial neural network. A potential cleavage site of the predicted Tat signal peptide is also reported. The TatP prediction server is available as a public web server at http://www.cbs.dtu.dk/services/TatP/.


The effect of progressive hearing loss on the morphology of endbulbs of Held and bushy cells.
Studies of congenital and early-onset deafness have demonstrated that an absence of peripheral sound-evoked activity in the auditory nerve causes pathological changes in central auditory structures. The aim of this study was to establish whether progressive acquired hearing loss could lead to similar brain changes that would degrade the precision of signal transmission. We used complementary physiologic hearing tests and microscopic techniques to study the combined effect of both magnitude and duration of hearing loss on one of the first auditory synapses in the brain, the endbulb of Held (EB), along with its bushy cell (BC) target in the anteroventral cochlear nucleus. We compared two hearing mouse strains (CBA/Ca and heterozygous shaker-2+/-) against a model of early-onset progressive hearing loss (DBA/2) and a model of congenital deafness (homozygous shaker-2-/-), examining each strain at 1, 3, and 6 months of age. Furthermore, we employed a frequency model of the mouse cochlear nucleus to constrain our analyses to regions most likely to exhibit graded changes in hearing function with time. No significant differences in the gross morphology of EB or BC structure were observed in 1-month-old animals, indicating uninterrupted development. However, in animals with hearing loss, both EBs and BCs exhibited a graded reduction in size that paralleled the hearing loss, with the most severe pathology seen in deaf 6-month-old shaker-2-/- mice. Ultrastructural pathologies associated with hearing loss were less dramatic: minor changes were observed in terminal size but mitochondrial fraction and postsynaptic densities remained relatively stable. These results indicate that acquired progressive hearing loss can have consequences on auditory brain structure, with prolonged loss leading to greater pathologies. Our findings suggest a role for early intervention with assistive devices in order to mitigate long-term pathology and loss of function.


Divergent roles of nitric oxide and rho kinase in vasomotor regulation of human retinal arterioles.
Although the arteriolar segment contributes to flow regulation, there is sparse information at the single microvessel level on how vasomotor function is regulated in the human retina. The authors have previously reported vasoreactivity and its underlying mechanisms in isolated porcine retinal arterioles. Herein, they studied human retinal arterioles for comparison. Retinal tissues were obtained from seven patients undergoing enucleation. Human and porcine retinal arterioles were isolated and pressurized to 55 cm H(2)O luminal pressure for vasoreactivity study using videomicroscopic techniques. Isolated human and porcine retinal arterioles developed myogenic tone and dilated dose dependently to bradykinin, adenosine, and sodium nitroprusside. Stepwise increases in luminal flow produced graded dilation with approximately 60% dilation at the highest flow tested. Nitric oxide (NO) synthase inhibitor L-NAME nearly abolished dilations to bradykinin and flow and attenuated the adenosine-induced dilation without altering the response to nitroprusside. Endothelin-1 caused dose-dependent constriction. Rho kinase (ROCK) inhibitor H-1152 blocked both myogenic tone and endothelin-1-induced constriction. Responses of retinal arterioles to all agonists and increased flow were similar between pigs and humans. Isolated human retinal arterioles dilate to bradykinin and increased flow in an NO-dependent manner. NO contributes, in part, to adenosine-induced vasodilation. Conversely, ROCK activation mediates myogenic tone and endothelin-1-induced vasoconstriction. Similarities in these vasoactive responses and the underlying mechanisms between human and porcine retinal arterioles support the latter as a viable experimental model of the human retinal microcirculation.


Gene-wise association of variants in four lysosomal storage disorder genes in neuropathologically confirmed Lewy body disease.
Variants in GBA are associated with Lewy Body (LB) pathology. We investigated whether variants in other lysosomal storage disorder (LSD) genes also contribute to disease pathogenesis. We performed a genetic analysis of four LSD genes including GBA, HEXA, SMPD1, and MCOLN1 in 231 brain autopsies. Brain autopsies included neuropathologically defined LBD without Alzheimer Disease (AD) changes (n = 59), AD without significant LB pathology (n = 71), Alzheimer disease and lewy body variant (ADLBV) (n = 68), and control brains without LB or AD neuropathology (n = 33). Sequencing of HEXA, SMPD1, MCOLN1 and GBA followed by 'gene wise' genetic association analysis was performed. To determine the functional effect, a biochemical analysis of GBA in a subset of brains was also performed. GCase activity was measured in a subset of brain samples (n = 64) that included LBD brains, with or without GBA mutations, and control brains. A lipidomic analysis was also performed in brain autopsies (n = 67) which included LBD (n = 34), ADLBV (n = 3), AD (n = 4), PD (n = 9) and control brains (n = 17), comparing GBA mutation carriers to non-carriers. In a 'gene-wise' analysis, variants in GBA, SMPD1 and MCOLN1 were significantly associated with LB pathology (p range: 0.03-4.14 x10(-5)). Overall, the mean levels of GCase activity were significantly lower in GBA mutation carriers compared to non-carriers (p<0.001). A significant increase and accumulation of several species for the lipid classes, ceramides and sphingolipids, was observed in LBD brains carrying GBA mutations compared to controls (p range: p<0.05-p<0.01). Our study indicates that variants in GBA, SMPD1 and MCOLN1 are associated with LB pathology. Biochemical data comparing GBA mutation carrier to non-carriers support these findings, which have important implications for biomarker development and therapeutic strategies.


Treatment of hospital wastewater effluent by nanofiltration and reverse osmosis.
Considerable concern exists regarding the appearance and effects of trace and ultra trace pollutants in the aquatic environment. In this context, it is necessary to identify relevant hot spot wastewater - such as hospital wastewater - and to implement specific wastewater treatment solutions. Membrane bioreactor (MBR) technology seems to be a suitable pre-treatment approach for the subsequent advanced treatment by high pressure membrane systems such as nanofiltration (NF) and reverse osmosis (RO). This paper is based upon investigations on the first full scale MBR for separate treatment of hospital wastewater in Germany. In this study an NF as well as an RO module for further treatment of the MBR filtrate were tested. The removal efficiencies were assessed using the following target compounds: bezafibrate, bisoprolol, carbamazepine, clarithromycin, ciprofloxacin, diclofenac, ibuprofen, metronidazole, moxifloxacin, telmisartan and tramadol. In summary, the results of this study confirmed that MBR technology followed by an advanced treatment for trace pollutant removal is an adequate approach for specific treatment of hot spot wastewater such as hospital wastewater. In particular, it was shown that - comparing the tested NF and RO - only (a two stage) RO is appropriate to remove pharmaceutical residues from hospital wastewater entirely. The recommended yield of the 2-stage RO is 70% which results in a retentate sidestream of 9%. Our investigations proved that RO is a very efficient treatment approach for elimination of trace pollutants.


Monotherapy may be suboptimal for severe bacteremic pneumococcal pneumonia.
Although monotherapy for pneumococcal pneumonia is standard, a survival benefit of combination beta-lactam and macrolide therapy has been suggested. Initial empirical therapy with a combination of effective antibiotic agents would have a better outcome than a single effective antibiotic agent in patients with bacteremic pneumococcal pneumonia. A review of adult bacteremic pneumococcal pneumonia within the Methodist Healthcare System, Memphis, Tenn, between January 1, 1996, and July 31, 2000. Empirical therapy was defined as all antibiotic agents received in the first 24 hours after presentation. On the basis of culture results, empirical therapy was classified as single effective therapy (SET), dual effective therapy (DET), or more than DET (MET). Acute Physiology and Chronic Health Evaluation II (APACHE II)-based predicted mortality, and Pneumonia Severity Index scores were calculated. Of the 225 patients identified, 99 were classified as receiving SET, 102 as receiving DET, and 24 as receiving MET. Compared with the other groups, patients who received MET had statistically significantly more severe pneumonia as measured by the Pneumonia Severity Index score (P =.04) and predicted mortality (P =.03). Mortality within the SET group was significantly higher than within the DET group (P =.02, odds ratio, 3.0 [95% confidence intervals, 1.2-7.6]), even when the DET and MET groups (P =.04) were combined. In a logistic regression model including antibiotic therapy and clinical risk factors for mortality, SET remained an independent predictor of mortality with a predicted mortality-adjusted odds ratio for death of 6.4 (95% confidence intervals, 1.9-21.7). All deaths occurred in patients with a Pneumonia Severity Index score higher than 90, and the predicted mortality-adjusted odds ratio for death with SET in this subgroup was 5.5 (95% confidence intervals, 1.7-17.5). We found that SET is associated with a significantly greater risk of death than DET. Therefore, monotherapy may be suboptimal for patients with severe bacteremic pneumococcal pneumonia who have Pneumonia Severity Index scores higher than 90.


Neohemigaster Malloch, 1939 and Pterogenia Bigot, 1859 (Diptera: Platystomatidae) from eastern Eurasia, with the description of four new species.
Four new species are described: Neohemigaster angustifrons sp.n. from Vietnam, Neohenigaster antropovi sp.n. from the Russian Far East, Neohemigaster tetralineata sp.n. from Taiwan and Pterogenia tenebrica sp.n. from Taiwan. In addition to Neohemigaster ussurica (Korneyev, 2001), six species formerly assigned to Pterogenia are transferred to Neoheinigaster: Neohemigaster eurysterna (Hendel, 1914) comb. nov., Neoheinigaster flavopicta (Hennig, 1940) comb. nov., Neohemigaster minuspicta (Hennig, 1940) comb. nov., Neohemigaster inonticola (Frey, 1964) comb. nov., Neohenigaster ornata (Hennig, 1940) comb. nov., and Neoheinigaster rectivena (Enderlein, 1924) comb. nov. A preliminary list of diagnostic characters separating the genera Pterogenia and Neohenigaster is compiled. A key to species of Pterogenia and Neohentigaster, known from Far East Russia, Japan, China, Taiwan, Vietnam and Burma, is provided.


Focus on TRP channels in cystic fibrosis.
The Transient Receptor Potential (TRP) protein superfamily is a group of cation channels expressed in various cell types and involved in respiratory diseases such as cystic fibrosis (CF), the genetic disease caused by CF Transmembrane conductance Regulator (CFTR) mutations. In human airway epithelial cells, there is growing evidence for a functional link between CFTR and TRP channels. TRP channels contribute to transmitting extracellular signals into the cells and, in an indirect manner, to CFTR activity via a Ca2+ rise signaling. Indeed, mutated CFTR-epithelial cells are characterized by an increased Ca2+ influx and, on the opposite, by a decreased of magnesium influx, both being mediated by TRP channels. This increasing cellular Ca2+ triggers the activation of calcium-activated chloride channels (CaCC) or CFTR itself, via adenylyl cyclase, PKA and tyrosine kinases activation, but also leads to an exaltation of the inflammatory response. Another shortcoming in mutated CFTR-epithelial cells is a [Mg2+]i decrease, associated with impaired TRPM7 functioning. This deregulation has to be taken into consideration in CF physiopathology, as Mg2+ is required for ATP hydrolysis and CFTR activity. The modulation of druggable TRP channels could supplement CF therapy either an anti-inflammatory drug or for CFTR potentiation, according to the balance between exacerbation and respite phases. The present paper focus on TRPA1, TRPC6, TRPM7, TRPV2, TRPV4, TRPV6 and ORAI 1, the proteins identified, for now, as dysfunctional channels, in CF cells.


True logarithmic amplification of frequency clock in SS-OCT for calibration.
With swept source optical coherence tomography (SS-OCT), imprecise signal calibration prevents optimal imaging of biological tissues such as coronary artery. This work demonstrates an approach using a true logarithmic amplifier to precondition the clock signal, with the effort to minimize the noises and phase errors for optimal calibration. This method was validated and tested with a high-speed SS-OCT. The experimental results manifest its superior ability on optimization of the calibration and improvement of the imaging performance. Particularly, this hardware-based approach is suitable for real-time calibration in a high-speed system where computation time is constrained.


Dynamic formation of a hot field reversed configuration with improved confinement by supersonic merging of two colliding high-β compact toroids.
A hot stable field-reversed configuration (FRC) has been produced in the C-2 experiment by colliding and merging two high-β plasmoids preformed by the dynamic version of field-reversed θ-pinch technology. The merging process exhibits the highest poloidal flux amplification obtained in a magnetic confinement system (over tenfold increase). Most of the kinetic energy is converted into thermal energy with total temperature (T{i}+T{e}) exceeding 0.5 keV. The final FRC state exhibits a record FRC lifetime with flux confinement approaching classical values. These findings should have significant implications for fusion research and the physics of magnetic reconnection.


Disclosing Gender-Based Violence During Health Care Visits: A Patient-Centered Approach.
The purpose of this study was to better understand victims' perspectives regarding decisions to disclose gender-based violence, namely, intimate partner violence (IPV) and human trafficking, to health care providers and what outcomes matter to them when discussing these issues with their provider. Twenty-five participants from racially/ethnically diverse backgrounds were recruited from a family justice center located in the southeastern United States. Two fifths had experienced human trafficking, and the remaining had experienced IPV. Upon obtaining informed consent, semistructured, in-depth interviews were conducted. Interviews were audio recorded and transcribed verbatim. Qualitative content analysis was used to examine interview data. Five primary themes emerged. Three themes focused on factors that may facilitate or impede disclosure: patient-provider connectedness, children, and social support. The fourth theme was related to ambiguity in the role of the health care system in addressing gender-based violence. The final theme focused on outcomes participants hope to achieve when discussing their experiences with health care providers. Similar themes emerged from both IPV and human trafficking victims; however, victims of human trafficking were more fearful of judgment and had a stronger desire to keep experiences private. Cultural factors also played an important role in decisions around disclosure and may interact with the general disparities racial/ethnic minority groups face within the health care system. Recognizing factors that influence patient engagement with the health care system as it relates to gender-based violence is critical. The health care system can respond to gender-based violence and its associated comorbidities in numerous ways and interventions must be driven by the patient's goals and desired outcomes of disclosure. These interventions may be better served by taking patient-centered factors into account and viewing the effectiveness of intervention programs through a behavioral, patient-centered lens.


International Consensus on Standardization of Data Collection for Complications Associated With Esophagectomy: Esophagectomy Complications Consensus Group (ECCG).
Perioperative complications influence long- and short-term outcomes after esophagectomy. The absence of a standardized system for defining and recording complications and quality measures after esophageal resection has meant that there is wide variation in evaluating their impact on these outcomes. The Esophageal Complications Consensus Group comprised 21 high-volume esophageal surgeons from 14 countries, supported by all the major thoracic and upper gastrointestinal professional societies. Delphi surveys and group meetings were used to achieve a consensus on standardized methods for defining complications and quality measures that could be collected in institutional databases and national audits. A standardized list of complications was created to provide a template for recording individual complications associated with esophagectomy. Where possible, these were linked to preexisting international definitions. A Delphi survey facilitated production of specific definitions for anastomotic leak, conduit necrosis, chyle leak, and recurrent nerve palsy. An additional Delphi survey documented consensus regarding critical quality parameters recommended for routine inclusion in databases. These quality parameters were documentation on mortality, comorbidities, completeness of data collection, blood transfusion, grading of complication severity, changes in level of care, discharge location, and readmission rates. The proposed system for defining and recording perioperative complications associated with esophagectomy provides an infrastructure to standardize international data collection and facilitate future comparative studies and quality improvement projects.


Exercise responses in patients treated for pulmonary tuberculosis by thoracoplasty.
Twenty eight subjects (mean age 64 years) who had been treated for tuberculosis by thoracoplasty in the past performed an increasing work rate exercise test, from which maximum oxygen consumption (VO2max), ventilation and heart rate were measured. VO2max was significantly lower than predicted, being 0.75 l/min in 17 subjects, 1.0 l/min in 10, and 1.5 l/min in one. Only one subject achieved a heart rate of 85% of the predicted maximum. The ratio of heart rate to oxygen consumption (HR/VO2) and heart rate at standard interpolated submaximal levels of oxygen uptake at 0.75 l/min (heart rate 0.75) and 1.0 l/min (heart rate 1.0) were normal. VO2max correlated with ventilation at maximal exercise (VE max) (r = 0.87) and FEV1 (r = 0.47). It did not correlate with resting arterial oxygen or carbon dioxide tensions, FEV1, maximum inspiratory pressure, angle of scoliosis, or number of ribs resected. The relation between ventilation and oxygen consumption (VE/VO2) and VE at the submaximal levels of oxygen consumption of 0.75 l/min (VE 0.75) and 1.0 l/min (VE 1.0) were normal. In 10 subjects a plateau of breathing frequency (fmax) was reached, after which the increase in ventilation was achieved by a further increase in tidal volume (VT). These subjects showed significantly lower values for the forced expiratory ratio, VO2max, and VEmax than those with a normal relation between tidal volume and breathing frequency. VEmax was correlated with FEV1 (r = 0.61), FVC (r = 0.46), maximum VT (r = 0.55), change in VT (r = 0.52), fmax (r = 0.56), and change in breathing frequency (r = 0.72). These results indicate that exercise in patients treated for tuberculosis by thoracoplasty is limited by ventilatory capacity and that this is due to a reduction in both dynamic lung volumes and respiratory frequency.


Dissociating task difficulty from incongruence in face-voice emotion integration.
In the everyday environment, affective information is conveyed by both the face and the voice. Studies have demonstrated that a concurrently presented voice can alter the way that an emotional face expression is perceived, and vice versa, leading to emotional conflict if the information in the two modalities is mismatched. Additionally, evidence suggests that incongruence of emotional valence activates cerebral networks involved in conflict monitoring and resolution. However, it is currently unclear whether this is due to task difficulty-that incongruent stimuli are harder to categorize-or simply to the detection of mismatching information in the two modalities. The aim of the present fMRI study was to examine the neurophysiological correlates of processing incongruent emotional information, independent of task difficulty. Subjects were scanned while judging the emotion of face-voice affective stimuli. Both the face and voice were parametrically morphed between anger and happiness and then paired in all audiovisual combinations, resulting in stimuli each defined by two separate values: the degree of incongruence between the face and voice, and the degree of clarity of the combined face-voice information. Due to the specific morphing procedure utilized, we hypothesized that the clarity value, rather than incongruence value, would better reflect task difficulty. Behavioral data revealed that participants integrated face and voice affective information, and that the clarity, as opposed to incongruence value correlated with categorization difficulty. Cerebrally, incongruence was more associated with activity in the superior temporal region, which emerged after task difficulty had been accounted for. Overall, our results suggest that activation in the superior temporal region in response to incongruent information cannot be explained simply by task difficulty, and may rather be due to detection of mismatching information between the two modalities.


Understanding Stem Cell Immunogenicity in Therapeutic Applications.
Stem cells and their differentiated progeny offer great hope for treating disease by providing an unlimited source of cells for repairing or replacing damaged tissue. Initial studies suggested that, unlike 'normal' transplants, specific characteristics of stem cells enabled them to avoid immune attack. However, recent findings have revealed that the immunogenicity of stem cells may have been underestimated. Here, we review the current understanding of the mechanisms of immune recognition associated with stem cell immunogenicity, and discuss the relevance of reprogramming and differentiation strategies used to generate cells or tissue from stem cells for implantation in eliciting an immune response. We examine the effectiveness of current strategies for minimising immune attack in light of our experience in the transplantation field and, in this context, outline important challenges moving forward.


Short-term effects on mobility, activities of daily living and health-related quality of life of a Conductive Education programme for adults with multiple sclerosis, Parkinson's disease and stroke.
To evaluate the impact of 10 sessions of Conductive Education on mobility, functional independence and health-related quality of life in adults with multiple sclerosis, Parkinson's disease and stroke. Observational, pretest-posttest design. The National Institute of Conductive Education, Birmingham, UK. One hundred and twenty-nine self-referred community-living individuals with moderate disability. Following an individual consultation, participants attended 10 diagnostic-specific group sessions of Conductive Education, scheduled daily or less commonly weekly. All participants were assessed using the Barthel Index and the Nottingham Extended Activities of Daily living Index. Stroke, multiple sclerosis and Parkinson's disease patients also completed the Short Form 36 (SF-36) Health Survey, the Multiple Sclerosis Quality of Life 54 questionnaire (MSQoL-54) and the Parkinson's Disease Questionnaire (PDQ-39) respectively. One hundred and five individuals completed the programme as well as all pre and post-assessments. Of these, 34 had suffered a stroke, 55 had Parkinson's disease and 16 had multiple sclerosis. Stroke patients demonstrated statistically significant improvements in the Nottingham Extended Activities of Daily living Index (mean change 3.4, P<0.001) and the SF-36 mental health subsection (mean change 7.8, P<0.001). Non-significant trends towards improved physical and mental function were demonstrated by other outcome measures across all three diagnoses. The results emphasize potential benefits of Conductive Education for individuals with stroke. The results will inform further randomized comparisons of the effects of Conductive Education in neurologically disabled people.


Tetratricopeptide repeat protein-associated proteins contribute to the virulence of Porphyromonas gingivalis.
Porphyromonas gingivalis is one of the most etiologically important microorganisms in periodontal disease. We found in a previous study that PG1385 (TprA) protein, a tetratricopeptide repeat (TPR) protein, was upregulated in P. gingivalis wild-type cells placed in a mouse subcutaneous chamber and that a tprA mutant was clearly less virulent in the mouse subcutaneous abscess model (M. Yoshimura et al., Oral Microbiol. Immunol. 23:413-418, 2008). In the present study, we investigated the gene expression profile of tprA mutant cells placed in a mouse subcutaneous chamber and found that 9 genes, including PG2102 (tapA), PG2101 (tapB), and PG2100 (tapC) genes, were downregulated in the tprA mutant compared with those in the wild type. Expression of a cluster of tapA, tapB, and tapC genes of the mutant was also downregulated in an in vitro culture with enriched brain heart infusion medium. The TprA protein has three TPR motifs known as a protein-protein interaction module. Yeast two-hybrid system analysis and in vitro protein binding assays with immunoprecipitation and surface plasmon resonance detection revealed that the TprA protein could bind to TapA and TapB proteins. TprA and TapB proteins were located in the periplasmic space, whereas TapA, which appeared to be one of the C-terminal domain family proteins, was located at the outer membrane. We constructed tapA, tapB, and tapC single mutants and a tapA-tapB-tapC deletion mutant. In the mouse subcutaneous infection experiment, all of the mutants were less virulent than the wild type. These results suggest that TprA, TapA, TapB, and TapC are cooperatively involved in P. gingivalis virulence.


Implementation and validation of a Raman lidar measurement of middle and upper tropospheric water vapor.
Implementation of a Raman lidar measurement of middle and upper tropospheric water vapor is described for a system that uses a 532-nm exciting wavelength, fiber-optic signal transfer, and Q-branch selection. Particular attention is given to the minimization of systematic biases introduced by fluorescent reemission of energy associated with elastic backscatter returns. We compare lidar profiles with collocated radiosonde measurements by using the Vaisala H-Humicap capacitive captor. The variations in the water-vapor concentrations on vertical scales of the order of 1 km in the upper troposphere observed by the two instruments present significant differences. Independent characterization of random and systematic lidar measurement errors and radiosonde sensor response characteristics lead to the conclusion that these differences are due to radiosonde sensor response. These intercomparisons indicate that the lidar measurement can provide important information on water-vapor distributions in the radiatively important 8-11-km region.


PSD-95 and PKC converge in regulating NMDA receptor trafficking and gating.
Neuronal NMDA receptors (NMDARs) colocalize with postsynaptic density protein-95 (PSD-95), a putative NMDAR anchoring protein and core component of the PSD, at excitatory synapses. PKC activation and PSD-95 expression each enhance NMDAR channel opening rate and number of functional channels at the cell surface. Here we show in Xenopus oocytes that PSD-95 and PKC potentiate NMDA gating and trafficking in a nonadditive manner. PSD-95 and PKC each enhance NMDA channel activity, with no change in single-channel conductance, reversal potential or mean open time. PSD-95 and PKC each potentiate NMDA channel opening rate (k(beta)) and number of functional channels at the cell surface (N), as indicated by more rapid current decay and enhanced charge transfer in the presence of the open channel blocker MK-801. PSD-95 and PKC each increase NMDAR surface expression, as indicated by immunofluorescence. PKC potentiates NMDA channel function and NMDAR surface expression to the same final absolute values in the absence or presence of PSD-95. Thus, PSD-95 partially occludes PKC potentiation. We further show that Ser-1462, a putative phosphorylation target within the PDZ-binding motif of the NR2A subunit, is required for PSD-95-induced potentiation and partial occlusion of PKC potentiation. Coimmunoprecipitation experiments with cortical neurons in culture indicate that PKC activation promotes assembly of NR2 with NR1, and that the newly assembled NMDARs are not associated with PSD-95. These findings predict that synaptic scaffolding proteins and protein kinases convergently modulate NMDAR gating and trafficking at synaptic sites.


Determination of ibogaine and noribogaine in biological fluids and hair by LC-MS/MS after Tabernanthe iboga abuse Iboga alkaloids distribution in a drowning death case.
Tabernanthe iboga belongs to the Apocynaceae family. In this study, we report the case of a 37-year-old black male working as a security agent in Paris and found dead naked on the beach in Gabon after consumption of iboga. Autopsy revealed a drowning fatality and a myocardial abnormality (myocardial bridging). Samples of blood, urine, bile, gastric content, liver, lungs, vitreous, spleen and hair were taken. Biological fluids were liquid-liquid extracted with saturated NH4Cl pH 9.5 and methylene chloride/isopropanol (95/5, v/v) in presence of clonazepam-d(4), used as internal standard. After decontamination with dichloromethane, hair was cut into small pieces then sonicated for 2h in saturated NH4Cl pH 9.5 before extraction by methylene chloride/isopropanol (95/5, v/v). After evaporation the residues were reconstituted in methanol/ACN/formate buffer pH 3, from which 10 microL were injected into an ODB Uptisphere C(18) column (150 mm x 2.1mm, 5 microm) and eluted with a gradient of acetonitrile and formate buffer delivered at a flow rate of 200 microL/min. A Quantum Ultra triple-quadrupole mass spectrometer was used for analyses. Ionization was achieved using electrospray in the positive ionization mode (ESI). For each compound, detection was related to three daughter ions (ibogaine: m/z 311.4-->122.1, 174.1 and 188.1; noribogaine: m/z 297.4-->122.1, 159.1 and 160.1; clonazepam-d(4): m/z 319.9-->218.1, 245.1 and 274.1). Ibogaine and noribogaine were detected in all autopsy samples. Hair segmentation was not possible as hair was very short and frizzy. Concentrations of 1.2 and 2.5 ng/mg, respectively were detected. Neither other licit or illicit drugs nor alcohol were found. The presence of ibogaine and noribogaine in all autopsy samples was consistent with the recent absorption of Tabernanthe iboga, which was assumed to be responsible of the drowning fatality. The history of exposure, regarding hair analysis, is discussed. LC-MS/MS appears to be the best method for analyzing complex and poorly volatile alkaloids in autopsy samples and particularly in hair, due to the presence of a nitrogen ring and the relatively low concentrations to be measured.


Early infant diagnosis of HIV in three regions in Tanzania; successes and challenges.
By the end of 2009 an estimated 2.5 million children worldwide were living with HIV-1, mostly as a consequence of vertical transmission, and more than 90% of these children live in sub-Saharan Africa. In 2008 the World Health Organization (WHO), recommended early initiation of Highly Active Antiretroviral Therapy (HAART) to all HIV infected infants diagnosed within the first year of life, and since 2010, within the first two years of life, irrespective of CD4 count or WHO clinical stage. The study aims were to describe implementation of EID programs in three Tanzanian regions with differences in HIV prevalences and logistical set-up with regard to HIV DNA testing. Data were obtained by review of the prevention from mother to child transmission of HIV (PMTCT) registers from 2009-2011 at the Reproductive and Child Health Clinics (RCH) and from the databases from the Care and Treatment Clinics (CTC) in all the three regions; Kilimanjaro, Mbeya and Tanga. Statistical tests used were Poisson regression model and rank sum test. During the period of 2009 - 2011 a total of 4,860 exposed infants were registered from the reviewed sites, of whom 4,292 (88.3%) were screened for HIV infection. Overall proportion of tested infants in the three regions increased from 77.2% in 2009 to 97.8% in 2011. A total of 452 (10.5%) were found to be HIV infected (judged by the result of the first test). The prevalence of HIV infection among infants was higher in Mbeya when compared to Kilimanjaro region RR = 1.872 (95%CI = 1.408 - 2.543) p < 0.001. However sample turnaround time was significantly shorter in both Mbeya (2.7 weeks) and Tanga (5.0 weeks) as compared to Kilimanjaro (7.0 weeks), p=<0.001. A substantial of loss to follow-up (LTFU) was evident at all stages of EID services in the period of 2009 to 2011. Among the infants who were receiving treatment, 61% were found to be LFTU during the review period. The study showed an increase in testing of HIV exposed infants within the three years, there is large variations of HIV prevalence among the regions. Challenges like; sample turnaround time and LTFU must be overcome before this can translate into the intended goal of early initiation of lifelong lifesaving antiretroviral therapy for the infants.


Tyrosinase as a dual reporter gene for both photoacoustic and magnetic resonance imaging.
Reporter genes are useful scientific tools for analyzing promoter activity, transfection efficiency, and cell migration. The current study has validated the use of tyrosinase (involved in melanin production) as a dual reporter gene for magnetic resonance and photoacoustic imaging. MCF-7 cells expressing tyrosinase appear brown due to melanin. Magnetic resonance imaging of tyrosinase-expressing MCF-7 cells in 300 μL plastic tubes displayed a 34 to 40% reduction in T1 compared to normal MCF-7 cells when cells were incubated with 250 μM ferric citrate. Photoacoustic imaging of tyrosinase-expressing MCF-7 cells in 700 μm plastic tubes displayed a 20 to 57-fold increase in photoacoustic signal compared to normal MCF-7 cells. The photoacoustic signal from tyrosinase-expressing MCF-7 cells was significantly greater than blood at 650 nm, suggesting that tyrosinase-expressing cells can be differentiated from the vasculature with in vivo photoacoustic imaging. The imaging results suggest that tyrosinase is a useful reporter gene for both magnetic resonance and photoacoustic imaging.


Endothelial progenitor cells: therapeutic perspective for ischemic stroke.
Endothelial progenitor cells (EPCs), which can be cultured in vitro from mononuclear cells in peripheral blood or bone marrow, express both hematopoietic stem cell and endothelial cell markers on their surface. They are believed to participate in endothelial repair and postnatal angiogenesis due to their abilities of differentiating into endothelial cells and secreting protective cytokines and growth factors. Mounting evidence suggests that circulating EPCs are reduced and dysfunctional in various diseases including hypertension, diabetes, coronary heart disease, and ischemic stroke. Therefore, EPCs have been documented to be a potential biomarker for vascular diseases and a hopeful candidate for regenerative medicine. Ischemic stroke, as the major cause of disability and death, still has limited therapeutics based on the approaches of vascular recanalization or neuronal protection. Emerging evidence indicates that transplantation of EPCs is beneficial for the recovery of ischemic cerebral injury. EPC-based therapy could open a new avenue for ischemic cerebrovascular disease. Currently, clinical trials for evaluating EPC transfusion in treating ischemic stroke are underway. In this review, we summarize the general conceptions and the characteristics of EPCs, and highlight the recent research developments on EPCs. More importantly, the rationale, perspectives, and strategies for using them to treat ischemic stroke will be discussed.


Effects of similar intakes of marine n-3 fatty acids from enriched food products and fish oil on cardiovascular risk markers in healthy human subjects.
There is convincing evidence that consumption of fish and fish oil rich in long-chain (LC) n-3 PUFA (n-3 LCPUFA), EPA (20 : 5n-3) and DHA (22 : 6n-3) reduce the risk of CHD. The aim of the present study was to investigate whether n-3 LCPUFA-enriched food products provide similar beneficial effects as fish oil with regard to incorporation into plasma lipids and effects on cardiovascular risk markers. A parallel 7-week intervention trial was performed where 159 healthy men and women were randomised to consume either 34 g fish pâté (n 44), 500 ml fruit juice (n 38) or three capsules of concentrated fish oil (n 40), all contributing to a daily intake of approximately 1 g EPA and DHA. A fourth group did not receive any supplementation or food product and served as controls (n 37). Plasma fatty acid composition, serum lipids, and markers of inflammation and oxidative stress were measured. Compared with the control group, plasma n-3 LCPUFA and EPA:arachidonic acid ratio increased equally in all intervention groups. However, no significant changes in blood lipids and markers of inflammation and oxidative stress were observed. In conclusion, enriched fish pâté and fruit juice represent suitable delivery systems for n-3 LCPUFA. However, although the dose given is known to reduce the risk of CVD, no significant changes were observed on cardiovascular risk markers in this healthy population.


What's distressing about having type 1 diabetes? A qualitative study of young adults' perspectives.
Diabetes distress is a general term that refers to the emotional burdens, anxieties, frustrations, stressors and worries that stem from managing a severe, complex condition like Type 1 diabetes. To date there has been limited research on diabetes-related distress in younger people with Type 1 diabetes. This qualitative study aimed to identify causes of diabetes distress in a sample of young adults with Type 1 diabetes. Semi-structured interviews with 35 individuals with Type 1 diabetes (23-30 years of age). This study found diabetes related-distress to be common in a sample of young adults with Type 1 diabetes in the second phase of young adulthood (23-30 years of age). Diabetes distress was triggered by multiple factors, the most common of which were: self-consciousness/stigma, day-to-day diabetes management difficulties, having to fight the healthcare system, concerns about the future and apprehension about pregnancy. A number of factors appeared to moderate distress in this group, including having opportunities to talk to healthcare professionals, attending diabetes education programmes and joining peer support groups. Young adults felt that having opportunities to talk to healthcare professionals about diabetes distress should be a component of standard diabetes care. Some aspects of living with diabetes frequently distress young adults with Type 1 diabetes who are in their twenties. Clinicians should facilitate young adults' attendance at diabetes education programmes, provide them with opportunities to talk about their diabetes-related frustrations and difficulties and, where possible, assist in the development of peer-support networks for young adults with diabetes.


Risk factors for incident age-related macular degeneration: pooled findings from 3 continents.
To examine risk factors for incident age-related macular degeneration (AMD) after combining data from 3 population-based cohort studies. Population-based cohort study. A population of 9523 adults (age range, 43-95 years at baseline) living in Australia, The Netherlands, and the United States who participated in a baseline examination and a follow-up examination on average 5 or 6 years later. Similar procedures were used at all study sites. Examinations included a standardized questionnaire, pupillary dilation, and stereoscopic color fundus photography. Fundus photographs were graded for lesions associated with AMD using the Wisconsin and International Age-Related Maculopathy Grading Systems. Senior investigators from each site adjudicated all photos graded as late AMD. Incidence of late AMD. Among studies, distributions for most risk factors differed, and overall incidence rates were similar. In the Beaver Dam Eye Study, total serum cholesterol was inversely associated with incident neovascular AMD. In the Blue Mountains Eye Study, current smoking (defined as smoking at the time of the baseline examination) was associated with an increased risk of incident geographic atrophy and late AMD; increased total serum cholesterol, having diabetes, and older age at menopause were positively associated with incident geographic atrophy; and an increase in high-density lipoprotein serum cholesterol was inversely related to incident geographic atrophy. In the Rotterdam Study, current smoking was associated with an increased risk of incident geographic atrophy, neovascular AMD, and late AMD; past smoking was associated with an increased risk of incident neovascular AMD and late AMD; and an increased number of years between menarche and menopause was directly related to incident geographic atrophy. After pooling data, the only statistically significant relationships found were between smoking and total serum cholesterol and incident AMD. Current smoking was associated with an increased incidence of geographic atrophy and late AMD (odds ratios [ORs] relative to nonsmokers: 2.83 and 2.35, respectively; ORs relative to past smokers: 2.80 and 1.82, respectively), and total serum cholesterol was associated directly with incident geographic atrophy (OR: 1.08 per 10 mg/dl) and inversely with incident neovascular AMD (OR: 0.94 per 10 mg/dl). Pooled data support a growing body of evidence indicating that smoking is related to an increased risk of incident AMD. Current smokers were at higher risk of incident AMD than both past smokers and those who never smoked. The relationships found in this study between total serum cholesterol and incident geographic atrophy and neovascular AMD are not readily explained.


Truncation of exon 1 from the c-myc gene results in prolonged c-myc mRNa stability.
The human c-myc gene consists of three exons transcribed from two distinct promoters and the function of the first, noncoding exon is unknown. In COLO 320 cells, there co-exist normal and truncated (i.e., lacking exon 1) c-myc genes, both of which are transcribed. Studies on the turnover of c-myc mRNA show that the normal mRNA has an in vivo half-life of approximately 30 min which is approximately similar to the turnover time of the mRNA in lymphoblastoid cells. However, the truncated mRNA was found to be substantially more stable. This observation was also made with a Burkitt's lymphoma cell line which has a translocated, truncated c-myc gene. Therefore truncation of the c-myc gene can cause the mRNA to be more stable than the full size product suggesting that this can be a crucial factor in the activation of the c-myc oncogene, by exon 1 loss, in chromosomal translocation. The results also suggest a role for exon 1 in the c-myc mRNA degradative mechanism.


Automated annotation of chemical names in the literature with tunable accuracy.
A significant portion of the biomedical and chemical literature refers to small molecules. The accurate identification and annotation of compound name that are relevant to the topic of the given literature can establish links between scientific publications and various chemical and life science databases. Manual annotation is the preferred method for these works because well-trained indexers can understand the paper topics as well as recognize key terms. However, considering the hundreds of thousands of new papers published annually, an automatic annotation system with high precision and relevance can be a useful complement to manual annotation. An automated chemical name annotation system, MeSH Automated Annotations (MAA), was developed to annotate small molecule names in scientific abstracts with tunable accuracy. This system aims to reproduce the MeSH term annotations on biomedical and chemical literature that would be created by indexers. When comparing automated free text matching to those indexed manually of 26 thousand MEDLINE abstracts, more than 40% of the annotations were false-positive (FP) cases. To reduce the FP rate, MAA incorporated several filters to remove "incorrect" annotations caused by nonspecific, partial, and low relevance chemical names. In part, relevance was measured by the position of the chemical name in the text. Tunable accuracy was obtained by adding or restricting the sections of the text scanned for chemical names. The best precision obtained was 96% with a 28% recall rate. The best performance of MAA, as measured with the F statistic was 66%, which favorably compares to other chemical name annotation systems. Accurate chemical name annotation can help researchers not only identify important chemical names in abstracts, but also match unindexed and unstructured abstracts to chemical records. The current work is tested against MEDLINE, but the algorithm is not specific to this corpus and it is possible that the algorithm can be applied to papers from chemical physics, material, polymer and environmental science, as well as patents, biological assay descriptions and other textual data.


Mental health in HIV-positive pregnant women: results from Angola.
Our objective was to assess the mental health status of pregnant women who are HIV-positive, compared with other groups of pregnant women. We evaluated pregnant HIV-positive women attending the Medecins Sans Frontiers (MSF) HIV clinic in Malanje, Angola (N = 23). The control group consisted of pregnant women coming for antenatal clinic consultations who were not known to be HIV-positive (N=134). To assess mental health, the 12-item General Health Questionnaire (GHQ-12) was used. A score of three or greater was considered to indicate significant emotional distress. We also examined determinants of emotional distress in logistic multivariate regression models. We found that the mean score on the GHQ-12 for the HIV-positive group was more than twice the mean score of the controls, indicating poorer mental health in the HIV-positive group. Two-thirds of HIV-positive women had significant emotional distress, more than twice that in the control group. As well as HIV status, marital status was a strong independent predictor of mental health status, with married women experiencing less emotional distress. Thus, in our sample, pregnant women who were HIV-positive had a much poorer mental health status than the controls. Strategies to improve the mental health of HIV-positive mothers must be implemented and evaluated; efforts to decrease the levels of stigma and discrimination in this population are of key importance.


Functional consequence of fibulin-4 missense mutations associated with vascular and skeletal abnormalities and cutis laxa.
Fibulin-4 is a 60kDa calcium binding glycoprotein that has an important role in development and integrity of extracellular matrices. It interacts with elastin, fibrillin-1 and collagen IV as well as with lysyl oxidases and is involved in elastogenesis and cross-link formation. To date, several mutations in the fibulin-4 gene (FBLN4/EFEMP2) are known in patients whose major symptoms are vascular deformities, aneurysm, cutis laxa, joint laxity, or arachnodactyly. The pathogenetic mechanisms how these mutations translate into the clinical phenotype are, however, poorly understood. In order to elucidate these mechanisms, we expressed fibulin-4 mutants recombinantly in HEK293 cells, purified the proteins in native forms and analyzed alterations in protein synthesis, secretion, matrix assembly, and interaction with other proteins in relation to wild type fibulin-4. Our studies show that different mutations affect these properties in multiple ways, resulting in fibulin-4 deficiency and/or impaired ability to form elastic fibers. The substitutions E126K and C267Y impaired secretion of the protein, but not mRNA synthesis. Furthermore, the E126K mutant showed less resistance to proteases, reduced binding to collagen IV and fibrillin-1, as well as to LTBP1s and LTBP4s. The A397T mutation introduced an extra O-glycosylation site and deleted binding to LTBP1s. We show that fibulin-4 binds stronger than fibulin-3 and -5 to LTBP1s, 3, and 4s, and to the lysyl oxidases LOX and LOXL1; the binding of fibulin-4 to the LOX propeptide was strongly reduced by the mutation E57K. These findings show that different mutations in the fibulin-4 gene result in different molecular defects affecting secretion rates, protein stability, LOX-induced cross-linking, or binding to other ECM components and molecules of the TGF-β pathway, and thus illustrate the complex role of fibulin-4 in connective tissue assembly.


Prostaglandin E1 influences pulsatile preservation characteristics and early graft function in expanded criteria donor kidneys.
Unlike simple cold storage, machine preservation allows dynamic assessment and manipulation of the donor organ prior to transplantation. We prospectively compared the effects of five pharmacological agents added to the perfusate during machine preservation of expanded criteria donor (ECD) kidneys in order to (1) describe their influence on perfusion parameters and (2) determine their influence on early graft outcome. Two hundred seventy-five consecutive ECD kidneys were preserved in our laboratory between 1/1/94 and 12/31/97 by either machine perfusion (MP) or cold storage (CS). ECD kidneys were defined as those requiring pretransplant biopsy. ECD kidneys were divided by method of preservation and MP kidneys were randomized to receive prostaglandin E1 (MP+PGE1), trifluoperazine (TFP), verapamil (VER), papaverine (PAP), mannitol (MAN), or no intervention during the period of machine perfusion. CS kidneys were randomized to receive PGE1 (CS+PGE1), TFP, VER, PAP, or no intervention. All MP kidneys were preserved by continuous hypothermic pulsatile perfusion using Belzer II solution and perfusion parameters were measured every hour during pulsatile perfusion. All CS kidneys were stored in 1.0 L of University of Wisconsin (UW) solution. The addition of PGE1 to machine perfusate increased renal flow and decreased renal resistance. Moreover, the MP+PGE1-treated group was associated with improved early graft function compared to all other groups. The addition of VER, TFP, PAP, or MAN influenced neither the perfusion characteristics nor the incidence of early graft function in MP kidneys. Similarly, the addition of VER, TFP, or PAP did not influence early graft function in the CS kidneys. The CS+PGE1 group exhibited a significantly lower incidence of early graft function than did the MP+PGE1 group. PGE1 treatment during machine preservation improves hydrostatic perfusion parameters and reduces the incidence of delayed graft function in ECD kidneys. Moreover, the addition of PGE1, TFP, VER, or PAP to UW does not influence early graft function in the CS kidney.


Non-homologous end-joining partners in a helical dance: structural studies of XLF-XRCC4 interactions.
XRCC4 (X-ray cross-complementation group 4) and XLF (XRCC4-like factor) are two essential interacting proteins in the human NHEJ (non-homologous end-joining) pathway that repairs DNA DSBs (double-strand breaks). The individual crystal structures show that the dimeric proteins are homologues with protomers containing head domains and helical coiled-coil tails related by approximate two-fold symmetry. Biochemical, mutagenesis, biophysical and structural studies have identified the regions of interaction between the two proteins and suggested models for the XLF-XRCC4 complex. An 8.5 Å (1 Å = 0.1 nm) resolution crystal structure of XLF-XRCC4 solved by molecular replacement, together with gel filtration and nano-ESI (nano-electrospray ionization)-MS results, demonstrates that XLF and XRCC4 dimers interact through their head domains and form an alternating left-handed helical structure with polypeptide coiled coils and pseudo-dyads of individual XLF and XRCC4 dimers at right angles to the helical axis.


Alcohol use and church attendance among seventh through twelfth grade students, Dominican Republic, 2011.
Excessive alcohol consumption increases the years of life lost to premature death and disability worldwide. Religion is a mitigating factor in alcohol consumption. A survey in the Dominican Republic showed increasing church attendance by middle and high school students (N = 3,478) was associated with a delay in age at first alcoholic drink, fewer students who had consumed alcohol in the past month (current drinkers), lower alcohol consumption levels, fewer episodes of inebriation, and less heavy episodic alcohol consumption (all P < 0.0001). The results suggested that it may be useful to conceive of church-attending youth as a subset of the adolescent social network when planning primary alcohol prevention programs for young people.


Prostaglandin E2 inhibits production of Th1 lymphokines but not of Th2 lymphokines.
PGE2 is known to inhibit IL-2 and IFN-gamma production from Th cells and is widely viewed as a general immunosuppressant. However, PGE2 was found not to inhibit IL-4 production from Th2 clones, and IL-5 production from these clones was slightly enhanced. The same results were obtained with short term T cell lines, which indicates that the lack of inhibition of IL-4 and IL-5 production by PGE2 is a general phenomenon. PGE2 functions by increasing cAMP levels through activation of adenylate cyclase. Despite its failure to inhibit lymphokine release, PGE2 was capable of increasing cAMP levels in Th2 cells, and forskolin, a direct activator of adenylate cyclase, also did not inhibit IL-4 or IL-5 production. These data indicate that the failure of PGE2 to inhibit IL-4 and IL-5 production was not due to an inability of PGE2 to induce an increase in intracellular cAMP, and suggested instead that the expression of IL-4 and IL-5 in Th2 cells is insensitive to elevated cAMP levels. When Th0 clones were examined, PGE2 was again found to differentially affect IL-2 and IL-4 production in three of five clones tested. In two additional Th0 clones, both IL-2 and IL-4 production were inhibited. These data suggest that lymphokine production may be regulated on two different levels. First, Th1- and Th2-associated lymphokines may be differentially sensitive to intracellular signals such as cAMP. Second, T cell subsets may exist, including subsets of Th0 cells, with different signaling pathways. In addition, our data suggest that PGE2 may play an important role in regulating the development of a response dominated by Th1- or Th2-associated lymphokines.


Noninvasive diagnosis of fetal aneuploidy by shotgun sequencing DNA from maternal blood.
We directly sequenced cell-free DNA with high-throughput shotgun sequencing technology from plasma of pregnant women, obtaining, on average, 5 million sequence tags per patient sample. This enabled us to measure the over- and underrepresentation of chromosomes from an aneuploid fetus. The sequencing approach is polymorphism-independent and therefore universally applicable for the noninvasive detection of fetal aneuploidy. Using this method, we successfully identified all nine cases of trisomy 21 (Down syndrome), two cases of trisomy 18 (Edward syndrome), and one case of trisomy 13 (Patau syndrome) in a cohort of 18 normal and aneuploid pregnancies; trisomy was detected at gestational ages as early as the 14th week. Direct sequencing also allowed us to study the characteristics of cell-free plasma DNA, and we found evidence that this DNA is enriched for sequences from nucleosomes.


Decreased plasma tryptophan concentration in major depression: relationship to melancholia and weight loss.
Plasma total tryptophan (TRP) concentration was significantly lower in 31 patients with major depression compared to a healthy control group. The ratio of plasma TRP concentration to that of other branch chain amino acids (the TRP:BCAA ratio) was also decreased. Further analysis revealed that the decrease in plasma TRP and TRP:BCAA ratio was most apparent in patients with major depression and melancholia. Overall, women but not men had significantly decreased plasma tryptophan concentrations, perhaps because of a contributory effect of weight loss; this latter effect, however, could not be distinguished clearly from a diagnosis of melancholia. Our data suggest that in some depressed patients, reductions in plasma tryptophan availability may contribute to abnormalities in brain 5-hydroxytryptamine function.


Animal models to investigate fungal biofilm formation.
Microbial biofilms play an essential role in several infectious diseases and are defined as extensive communities of sessile organisms irreversibly associated with a surface, encased within a polysaccharide-rich extracellular matrix (ECM), and exhibiting enhanced resistance to antimicrobial drugs. Forming a biofilm provides the microbes protection from environmental stresses due to contaminants, nutritional depletion, or imbalances, but is dangerous to human health due to their inherent robustness and elevated resistance.The use of indwelling medical devices (e.g., central venous catheters, CVCs) in current therapeutic practice is associated with 80-90 % of hospital-acquired bloodstream and deep tissue infections. Most cases of catheter-related bloodstream infections (CRBSIs) involve colonization of microorganisms on catheter surfaces where they form a biofilm. Additionally, Fusarium solani and F. oxysporum were the causative organisms of the 2005/2006 outbreak of contact lens-associated fungal keratitis in the United States, Europe, the UK, and Singapore, and these infections involved formation of biofilms on contact lens. Fungal biofilm formation is studied using a number of techniques, involving the use of a wide variety of substrates and growth conditions. In vitro techniques involving the use of confocal scanning laser/scanning electron microscopy, metabolic activity assay, dry weight measurements, and antifungal susceptibility assays are increasingly used by investigators to quantify and evaluate biofilm morphology. However, there are not many in vivo models used to validate biofilm-associated infections. In this protocol, we describe a clinically relevant rabbit model of C. albicans biofilm-associated catheter infection to evaluate the morphology, topography, and architecture of fungal biofilms. We also describe a murine model of contact lens-associated Fusarium keratitis.Evaluation of the formation of fungal biofilms on catheters in vivo, their analysis using scanning electron microscopy (SEM) and quantitative catheter culture (QCC), and treatment of biofilms using antimicrobial lock therapy can be completed in ~20-25 days using the described methods. The rabbit model has utility in evaluating the efficacy of lock solutions. In addition, the murine model of contact lens-associated Fusarium keratitis enables characterizing/comparing the formation of Fusarium biofilms on contact lenses in vitro and determining their role in vivo.


Interactions in the native state of monellin, which play a protective role against aggregation.
A series of recent studies have provided initial evidence about the role of specific intra-molecular interactions in maintaining proteins in their soluble state and in protecting them from aggregation. Here we show that the amino acid sequence of the protein monellin contains two aggregation-prone regions that are prevented from initiating aggregation by multiple non-covalent interactions that favor their burial within the folded state of the protein. By investigating the behavior of single-chain monellin and a series of five of its mutational variants using a variety of biochemical, biophysical and computational techniques, we found that weakening of the non-covalent interaction that stabilizes the native state of the protein leads to an enhanced aggregation propensity. The lag time for fibrillation was found to correlate with the apparent midpoint of thermal denaturation for the series of mutational variants, thus showing that a reduced thermal stability is associated with an increased aggregation tendency. We rationalize these findings by showing that the increase in the aggregation propensity upon mutation can be predicted in a quantitative manner through the increase in the exposure to solvent of the amyloidogenic regions of the sequence caused by the destabilization of the native state. Our findings, which are further discussed in terms of the structure of monellin and the perturbation by the amino acid substitutions of the contact surface between the two subdomains that compose the folded state of monellin, provide a detailed description of the specific intra-molecular interactions that prevent aggregation by stabilizing the native state of a protein.


Normal viability and altered pharmacokinetics in mice lacking mdr1-type (drug-transporting) P-glycoproteins.
The mdr1-type P-glycoproteins (P-gps) confer multidrug resistance to cancer cells by active extrusion of a wide range of drugs from the cell. To study their physiological roles, we have generated mice genetically deficient in the mdr1b gene [mdr1b (-/-) mice] and in both the mdr1a and mdr1b genes [mdr1a/1b (-/-) mice]. In spite of the host of functions speculatively attributed to the mdrl-type P-gps, we found no physiological abnormalities in either strain. Viability, fertility, and a range of histological, hematological, serum-chemical, and immunological parameters were not abnormal in mdr1a/1b (-/-) mice. The high level of mdrlb P-gp normally present in the pregnant uterus did not protect fetuses from a drug (digoxin) in the bloodstream of the mother, although the protein did reduce drug accumulation in the adrenal gland and ovaries. Pharmacologically, mdr1a/1b (-/-) mice behaved similarly to the previously analyzed mdr1a (-/-) mice, displaying, for instance, increased brain penetration and reduced elimination of digoxin. However, both mdr1a and mdr1b P-gps contributed to the extrusion of rhodamine from hematopoietic progenitor cells, suggesting a potential role for the endogenous mdr1-type P-gps in protection of bone marrow against cytotoxic anticancer drugs. This, and the normal viability of mdr1a/1b (-/-) mice, has implications for the use of P-gp-blocking agents in cancer and other chemotherapy. mdr1a/1b (-/-) mice should provide a useful model system to further test the pharmacological roles of the drug-transporting P-gps and to analyze the specificity and effectivity of P-gp-blocking drugs.


Systems biology of pro-angiogenic therapies targeting the VEGF system.
Vascular endothelial growth factor (VEGF) is a family of cytokines for which the dysregulation of expression is involved in many diseases; for some, excess VEGF causes pathological hypervascularization, while for others VEGF-induced vascular remodeling may alleviate ischemia and/or hypoxia. Anti-angiogenic therapies attacking the VEGF pathway have begun to live up to their promise for treatment of certain cancers and of age-related macular degeneration. However, the corollary is not yet true: in coronary artery disease and peripheral artery disease, clinical trials of pro-angiogenic VEGF delivery have not, so far, proven successful. The VEGF and VEGF-receptor system is complex, with at least five ligand genes, some encoding multiple protein isoforms and five receptor genes. A systems biology approach for designing pro-angiogenic therapies, using a combination of quantitative experimental approaches and detailed computational models, is essential to deal with this complexity and to understand the effects of drugs targeting the system. This approach allows us to learn from unsuccessful clinical trials and to design and test novel single therapeutics or combinations of therapeutics. Among the parameters that can be varied in order to determine optimal strategy are dosage, timing of multiple doses, route of administration, and the molecular target.


Classifying perinatal mortality using verbal autopsy: is there a role for nonphysicians?
Because of a physician shortage in many low-income countries, the use of nonphysicians to classify perinatal mortality (stillbirth and early neonatal death) using verbal autopsy could be useful. To determine the extent to which underlying perinatal causes of deaths assigned by nonphysicians in Guatemala, Pakistan, Zambia, and the Democratic Republic of the Congo using a verbal autopsy method are concordant with underlying perinatal cause of death assigned by physician panels. Using a train-the-trainer model, 13 physicians and 40 nonphysicians were trained to determine cause of death using a standardized verbal autopsy training program. Subsequently, panels of two physicians and individual nonphysicians from this trained cohort independently reviewed verbal autopsy data from a sample of 118 early neonatal deaths and 134 stillbirths. With the cause of death assigned by the physician panel as the reference standard, sensitivity, specificity, positive and negative predictive values, and cause-specific mortality fractions were calculated to assess nonphysicians' coding responses. Robustness criteria to assess how well nonphysicians performed were used. Causes of early neonatal death and stillbirth assigned by nonphysicians were concordant with physician-assigned causes 47% and 57% of the time, respectively. Tetanus filled robustness criteria for early neonatal death, and cord prolapse filled robustness criteria for stillbirth. There are significant differences in underlying cause of death as determined by physicians and nonphysicians even when they receive similar training in cause of death determination. Currently, it does not appear that nonphysicians can be used reliably to assign underlying cause of perinatal death using verbal autopsy.


Decisive influence of substitution positions in molecular self-assembly.
Molecular self-assembly provides a versatile tool for creating functional molecular structures at surfaces. A rational design of molecular structure formation requires not only an in-depth understanding of the subtle balance between intermolecular and molecule-surface interactions, but might also involve considering chemical changes of the molecules, such as deprotonation. Here, we present a systematic investigation of a comparatively simple class of molecules, namely dihydroxybenzoic acid, which, nevertheless, enables creating a rich variety of structures when deposited onto calcite (10.4) held at room temperature. Based on non-contact atomic force microscopy measurements in ultra-high vacuum, our study demonstrates the decisive impact of the positions of the hydroxyl groups on the structure formation. Six isomers of dihydroxybenzoic acid exist which form six different molecular structures on the calcite surface. Surprisingly, only two isomers arrange into stable, ordered structures at sub-monolayer coverage: 2,5-dihydroxybenzoic acid forms a commensurate (1 × 5) structure, composed of deprotonated molecules. A double-row structure consisting of protonated molecules is observed for 3,5-dihydroxybenzoic acid. The positions of the functional groups steer the molecular self-assembly of dihydroxybenzoic acids in three distinct ways, namely by (a) affecting the deprotonation tendency of the acid group, (b) influencing the intermolecular interaction as already indicated by greatly different bulk structures and (c) altering the molecule-substrate matching. Our results, thus, shed light on the impact of rather small changes in the molecular structure on the structural variety in molecular self-assembly on surfaces.


Evolution of US maize (Zea mays L.) root architectural and anatomical phenes over the past 100 years corresponds to increased tolerance of nitrogen stress.
Increasing the nitrogen use efficiency of maize is an important goal for food security and agricultural sustainability. In the past 100 years, maize breeding has focused on yield and above-ground phenes. Over this period, maize cultivation has changed from low fertilizer inputs and low population densities to intensive fertilization and dense populations. The authors hypothesized that through indirect selection the maize root system has evolved phenotypes suited to more intense competition for nitrogen. Sixteen maize varieties representing commercially successful lines over the past century were planted at two nitrogen levels and three planting densities. Root systems of the most recent material were 7 º more shallow, had one less nodal root per whorl, had double the distance from nodal root emergence to lateral branching, and had 14% more metaxylem vessels, but total mextaxylem vessel area remained unchanged because individual metaxylem vessels had 12% less area. Plasticity was also observed in cortical phenes such as aerenchyma, which increased at greater population densities. Simulation modelling with SimRoot demonstrated that even these relatively small changes in root architecture and anatomy could increase maize shoot growth by 16% in a high density and high nitrogen environment. The authors concluded that evolution of maize root phenotypes over the past century is consistent with increasing nitrogen use efficiency. Introgression of more contrasting root phene states into the germplasm of elite maize and determination of the functional utility of these phene states in multiple agronomic conditions could contribute to future yield gains.


Radiology Information Systems: DREAM. patient management.
Until recently Radiology Information Systems (RISs) were considered a set of separate, often heterogeneous applications at the functional, technological and architectural level. In last years, the need for an integrated methodological approach and an engineering vision of the system design has become the most important aspect in the implementation of RISs. Based on this approach, the fundamental organizational, operational and clinical objectives of an advanced RIS together with the major critical factors of success, are presented. Such objectives and requirements have constituted the starting basis for the implementation of the DREAM (Distributed radiological environment advanced and multimedia) system, the result of the collaboration of the "Università Cattolica del S. Cuore, Policlinico A. Gemelli" of Rome and "GESI-Gestione Sistemi per l'Informatica". An overview of the main functional characteristics, the most qualifying aspects of the system and the methodological approach followed in its implementation, are reported.


Influence of daylight and noise current on cloud and aerosol observations by spaceborne elastic scattering lidar.
The influence of daylight and noise current on cloud and aerosol observations by realistic spaceborne lidar was examined by computer simulations. The reflected solar radiations, which contaminate the daytime return signals of lidar operations, were strictly and explicitly estimated by accurate radiative transfer calculations. It was found that the model multilayer cirrus clouds and the boundary layer aerosols could be observed during the daytime and the nighttime with only a few laser shots. However, high background noise and noise current make it difficult to observe volcanic aerosols in middle and upper atmospheric layers. Optimal combinations of the laser power and receiver field of view are proposed to compensate for the negative influence that is due to these noises. For the computer simulations, we used a realistic set of lidar parameters similar to the Experimental Lidar in-Space Equipment of the National Space Development Agency of Japan.


Optic vesicle-like structures derived from human pluripotent stem cells facilitate a customized approach to retinal disease treatment.
Differentiation methods for human induced pluripotent stem cells (hiPSCs) typically yield progeny from multiple tissue lineages, limiting their use for drug testing and autologous cell transplantation. In particular, early retina and forebrain derivatives often intermingle in pluripotent stem cell cultures, owing to their shared ancestry and tightly coupled development. Here, we demonstrate that three-dimensional populations of retinal progenitor cells (RPCs) can be isolated from early forebrain populations in both human embryonic stem cell and hiPSC cultures, providing a valuable tool for developmental, functional, and translational studies. Using our established protocol, we identified a transient population of optic vesicle (OV)-like structures that arose during a time period appropriate for normal human retinogenesis. These structures were independently cultured and analyzed to confirm their multipotent RPC status and capacity to produce physiologically responsive retinal cell types, including photoreceptors and retinal pigment epithelium (RPE). We then applied this method to hiPSCs derived from a patient with gyrate atrophy, a retinal degenerative disease affecting the RPE. RPE generated from these hiPSCs exhibited a disease-specific functional defect that could be corrected either by pharmacological means or following targeted gene repair. The production of OV-like populations from human pluripotent stem cells should facilitate the study of human retinal development and disease and advance the use of hiPSCs in personalized medicine.


Individual variations in platelet reactivity towards ADP, epinephrine, collagen and nitric oxide, and the association to arterial function in young, healthy adults.
Platelet aggregation and secretion can be induced by a large number of endogenous activators, such as collagen, adenosine diphosphate (ADP) and epinephrine. Conversely, the blood vessel endothelium constitutively release platelet inhibitors including nitric oxide (NO) and prostacyclin. NO and prostacyclin are also well-known vasodilators and contribute to alterations in local blood flow and systemic blood pressure. In this study we investigated individual variations in platelet reactivity and arterial functions including blood pressure and flow-mediated vasodilation (FMD) in 43 young, healthy individuals participating in the Lifestyle, Biomarkers and Atherosclerosis (LBA) study. Platelet aggregation and dense granule secretion were measured simultaneously by light transmission and luminescence. FMD was measured with ultrasound. The platelet function assay showed inter-individual differences in platelet reactivity. Specifically, a sub-group of individuals had platelets with an increased response to low concentrations of ADP and epinephrine, but not collagen. When the NO-donor S-nitroso-N-acetyl-DL-penicillamine (SNAP) was combined with high doses of these platelet activators, the results indicated for sub-groups of NO-sensitive and NO-insensitive platelets. The individuals with NO-sensitive platelets in response to SNAP in combination with collagen had a higher capacity of FMD of the arteria brachialis. Platelet reactivity towards ADP, epinephrine and NO differs between young, healthy individuals. Some individuals have a more effective response towards NO, both in the aspect of platelet inhibition ex vivo, as well as vasodilation in vivo.


Oncogenic mutations in GNAQ occur early in uveal melanoma.
Early/initiating oncogenic mutations have been identified for many cancers, but such mutations remain unidentified in uveal melanoma (UM). An extensive search for such mutations was undertaken, focusing on the RAF/MEK/ERK pathway, which is often the target of initiating mutations in other types of cancer. DNA samples from primary UMs were analyzed for mutations in 24 potential oncogenes that affect the RAF/MEK/ERK pathway. For GNAQ, a stimulatory alpha(q) G-protein subunit which was recently found to be mutated in UMs, resequencing was expanded to include 67 primary UMs and 22 peripheral blood samples. GNAQ status was analyzed for association with clinical, pathologic, chromosomal, immunohistochemical, and transcriptional features. Activating mutations at codon 209 were identified in GNAQ in 33 (49%) of 67 primary UMs, including 2 (22%) of 9 iris melanomas and 31 (54%) of 58 posterior UMs. No mutations were found in the other 23 potential oncogenes. GNAQ mutations were not found in normal blood DNA samples. Consistent with GNAQ mutation being an early or initiating event, this mutation was not associated with any clinical, pathologic, or molecular features associated with late tumor progression. GNAQ mutations occur in about half of UMs, representing the most common known oncogenic mutation in this cancer. The presence of this mutation in tumors at all stages of malignant progression suggests that it is an early event in UM. Mutations in this G-protein-coupled receptor provide new insights into UM pathogenesis and could lead to new therapeutic possibilities.


e-Prescribing, efficiency, quality: lessons from the computerization of UK family practice.
Nearly all general practice physicians (GPs) in the United Kingdom (UK) have electronic health record (EHR) systems in their practices compared with perhaps 15% of primary care physicians in the United States (U.S.). Based on interviews of 13 general GPs and review of current literature, the authors argue that the historical experience of widespread electronic health record uptake in the UK provides insight into features that might motivate broad adoption in the United States. These features include electronic prescribing, improved quality and consistency of care, practice efficiencies that have both timesaving and revenue generating effects, and potential shielding from malpractice claims.


Squeaking with a sliding joint: mechanics and motor control of sound production in palinurid lobsters.
The origin of arthropod sound-producing morphology typically involves modification of two translating body surfaces, such as the legs and thorax. In an unusual structural rearrangement, I show that one lineage of palinurid lobsters lost an antennal joint articulation, which transformed this joint from moving with one degree of freedom into a sliding joint with multiple degrees of freedom. With this sliding joint, 'stick-and-slip' sounds are produced by rubbing the base of each antenna against the antennular plate. To understand the musculo-skeletal changes that occurred during the origin and evolutionary variation of this sound-producing mechanism, I examined joint morphology and antennal muscle anatomy across sound-producing and non-sound-producing palinurids. Plectrum movement and antennal muscle activity were measured in a sound-producing species, Panulirus argus. The promotor muscle pulls the plectrum over the file during sound-producing and non-sound-producing movements; a higher intensity of muscle activity is associated with sound production. The promotor muscle is larger and attaches more medially in sound-producing palinurids than in non-sound producers. In Panulirus argus, each shingle on the file has an additional ridge; in Palinurus elephas, the shingle surfaces are smooth. These differences in shingle surface features suggest variation in the stick-and-slip properties of the system. Translational motion permitted by the sliding joint is necessary for sound production; hence, the construction of a sliding joint is a key modification in the origin of this sound-producing mechanism.


Lack of evidence of sexual transmission of hepatitis C among monogamous couples: results of a 10-year prospective follow-up study.
The risk of sexual transmission of hepatitis C virus (HCV) infection was evaluated among 895 monogamous heterosexual partners of HCV chronically infected individuals in a long-term prospective study, which provided a follow-up period of 8,060 person-years. Seven hundred and seventy-six (86.7%) spouses were followed for 10 yr, corresponding to 7,760 person-years of observation. One hundred and nineteen (13.3%) spouses (69 whose infected partners cleared the virus following treatment and 50 who ended their relationship or were lost at follow-up) contributed an additional 300 person-years. All couples denied practicing anal intercourse or sex during menstruation, as well as condom use. The average weekly rate of sexual intercourse was 1.8. Three HCV infections were observed during follow-up corresponding to an incidence rate of 0.37 per 1,000 person-years. However, the infecting HCV genotype in one spouse (2a) was different from that of the partner (1b), clearly excluding sexual transmission. The remaining two couples had concordant genotypes, but sequence analysis of the NS5b region of the HCV genome, coupled with phylogenetic analysis showed that the corresponding partners carried different viral isolates, again excluding the possibility of intraspousal transmission of HCV. Our data indicate that the risk of sexual transmission of HCV within heterosexual monogamous couples is extremely low or even null. No general recommendations for condom use seem required for individuals in monogamous partnerships with HCV-infected partners.


Admission handoff communications: clinician's shared understanding of patient severity of illness and problems.
Communication errors are a leading cause of medical mistakes. Handoff communications during the admission of a patient are a critical point of communication during which patient care is transferred from one clinician to another. The transmission of the patient's current severity of illness and active problems is integral to this communication. Our objective was to determine if this information is conveyed by the current handoff process between resident physicians. We recorded admission handoff communications between residents and then asked each resident to independently rate the patient's severity of illness and to list the patient's problems. The scores and lists were compared for agreement. Attending physicians listened to the recordings and also assessed severity of illness and patient's problem lists. Three quarters of the handoffs had agreement about the severity of the patient's illness. However, there was low agreement about the most severe problem and the total problem lists between residents involved in the handoff communication. Attending physicians were able to identify more patient problems. We conclude that information needed to assess the patient's severity of illness and problems may have been present in the handoff communications but may not have been fully received and integrated by the residents. In addition, attending physicians may have an additional capacity to "infer"" information, perhaps because of prior clinical experience or expertise. This study implies that residents may need more formal education, training, and evaluation of their handoffs to improve patient safety.


Simultaneous kidney-pancreas transplantation without antilymphocyte induction.
The introduction of potent new immunosuppressive agents may allow simultaneous kidney-pancreas transplantation to be performed without antilymphocyte induction. We analyzed 30 simultaneous kidney-pancreas transplantations receiving tacrolimus, mycophenolate mofetil, and steroids without without antilymphocyte induction. Eighteen patients underwent pancreas transplantation with portal-enteric (P-E) drainage and the remaining 12 had systemic bladder (S-B) drainage. Target 12 hr trough tacrolimus levels for the first 3 months after simultaneous kidney-pancreas transplantation were 15-20 ng/ml. The oral mycophenolate mofetil dose was 2-3 g/day begun immediately posttransplant in two to four divided doses. Steroids were tapered according to protocol. All patients experienced immediate function of both kidney and pancreas grafts. One-year actuarial patient, kidney, and pancreas graft survival rates are 93, 93, and 90%, respectively. Nine patients (30%) had a total of 13 rejection episodes (12 biopsy proven) including 4 within 2 weeks, 6 between 2 weeks and 3 months, and 3 beyond 3 months after simultaneous kidney-pancreas transplantation. Three rejection episodes were treated with steroids alone and 10 were treated with antilymphocyte therapy (5 OKT3 and 5 ATGAM). A total of seven patients (23%) received antilymphocyte therapy. Three patients (10%) had more than one rejection episode. Two pancreas grafts (7%) and one kidney graft (3%) were lost from rejection. Four patients (13%) developed cytomegalovirus infection, but none had tissue-invasive cytomegalovirus. At present, 22 surviving patients (81%) remain on triple immunosuppression with tacrolimus, mycophenolate mofetil, and prednisone with excellent dual graft function. Tacrolimus, mycophenolate mofetil, and prednisone immunosuppression without without antilymphocyte induction is safe and effective after simultaneous kidney-pancreas transplantation.


The impact of splicing on protein domain architecture.
Many proteins are composed of protein domains, functional units of common descent. Multidomain forms are common in all eukaryotes making up more than half of the proteome and the evolution of novel domain architecture has been accelerated in metazoans. It is also becoming increasingly clear that alternative splicing is prevalent among vertebrates. Given that protein domains are defined as structurally, functionally and evolutionarily distinct units, one may speculate that some alternative splicing events may lead to clean excisions of protein domains, thus generating a number of different domain architectures from one gene template. However, recent findings indicate that smaller alternative splicing events, in particular in disordered regions, might be more prominent than domain architectural changes. The problem of identifying protein isoforms is, however, still not resolved. Clearly, many splice forms identified through detection of mRNA sequences appear to produce 'nonfunctional' proteins, such as proteins with missing internal secondary structure elements. Here, we review the state of the art methods for identification of functional isoforms and present a summary of what is known, thus far, about alternative splicing with regard to protein domain architectures.


Itraconazole, a new triazole that is orally active in aspergillosis.
Itraconazole is a new orally active triazole derivative with broad-spectrum antifungal activity. This drug is effective in experimental aspergillosis and possesses in vitro activity against various species and strains of Aspergillus. Morphological destruction of inoculated hyphae and complete inhibition of hyphal outgrowth in culture is obtained from 0.07 micrograms ml-1 (10(-7)M) onward. These properties make itraconazole a likely candidate for clinical evaluation in disseminated aspergillosis.


Equilibrium free energies from nonequilibrium measurements using maximum-likelihood methods.
We present a maximum likelihood argument for the Bennett acceptance ratio method, and derive a simple formula for the variance of free energy estimates generated using this method. This derivation of the acceptance ratio method, using a form of logistic regression, a common statistical technique, allows us to shed additional light on the underlying physical and statistical properties of the method. For example, we demonstrate that the acceptance ratio method yields the lowest variance for any estimator of the free energy which is unbiased in the limit of large numbers of measurements.