Redox activity and chemical interactions of metal oxide nano- and micro-particles with dithiothreitol (DTT).
The wide application and production of nanotechnology have increased the interest in studying the toxicity of nano- and micro-sized particles escaping into air from various aspects of the production process. Metal oxides (MOs) are one particular class of particles that exist abundantly in ambient PM. Studies show an emphasis on biological mechanisms by which inhalation exposure to MOs leads to disease. However, different biological assays provide different redox activity rankings making it difficult to assess the contributions of various MOs to measures of aggregate toxicity in multi-pollutant systems such as ambient PM. Therefore, research to evaluate the chemical interaction between these particles and molecules that are relevant to cellular redox activity can help in establishing indicators of reactivity. In particular, this study assesses the redox activity of six MOs mainly emitted from anthropogenic industrial activities using the dithiothreitol (DTT) assay. DTT is commonly used in acellular assays due to its analogous structure to cellular glutathione. The structural and chemical behaviors between active MOs and DTT were elucidated using FTIR, NMR, and BET methods. The results indicate that the health risk (redox activity) associated with MOs is mainly a function of their surface reactivity demonstrated by the ability of the oxidized (S-H) bond in DTT to form a stable bond with the MO surface.


Young Athletes With Quadriceps Femoris Strength Asymmetry at Return to Sport After Anterior Cruciate Ligament Reconstruction Demonstrate Asymmetric Single-Leg Drop-Landing Mechanics.
Young athletes who have had anterior cruciate ligament (ACL) reconstruction demonstrate suboptimal rates of return to sport, high rates of second ACL injuries, and persistent movement asymmetries. Therefore, the influence of musculoskeletal impairments on movement mechanics in this population needs to be further evaluated. The primary hypothesis was that among young athletes who have had ACL reconstruction, those with greater quadriceps strength asymmetry would demonstrate altered single-leg drop-landing mechanics at return to sport compared with individuals with more symmetric quadriceps strength and also compared with healthy controls (ie, those with no ACL reconstruction). A second hypothesis was that quadriceps strength symmetry would predict single-leg drop-landing symmetry in individuals who have undergone ACL reconstruction. Controlled laboratory study. The study entailed a total of 103 participants (age, 17.4 years) at the time of return to sport after ACL reconstruction and 47 control participants (age, 17.0 years). The quadriceps index (QI) was calculated for isometric quadriceps strength, which was then used to divide the ACL reconstruction participants into high-quadriceps (QI ≥90%; n = 52) and low-quadriceps (QI <85%; n = 41) subgroups. Biomechanical data were collected by use of 3-dimensional motion analysis during a single-leg drop-landing task. The LSI was calculated for kinematic and kinetic sagittal-plane variables of interest during landing. Group differences were compared by use of 1-way analysis of variance and linear regression analyses (α < .05). Both the low- and high-quadriceps groups demonstrated greater limb asymmetry during landing compared with the control group in knee flexion excursion (mean LSI ± SD: low quadriceps, 85.8% ± 15.5% [P < .001]; high quadriceps, 94.2% ± 15.6% [P = .019]; control, 102.7% ± 14.1%), peak trunk flexion angle (low quadriceps, 129.2% ± 36.6% [P < .001]; high quadriceps, 110.5% ± 22.6% [P = .03]; control, 95.5% ± 26.2%), and peak knee extension moment (low quadriceps, 79.5% ± 25.2% [P < .001]; high quadriceps, 89.9% ± 19.8% [P = .005]; control, 102.2% ± 10.9%). Compared with the high-quadriceps group, the low-quadriceps group also demonstrated greater asymmetry during landing in knee flexion excursion (P = .026), peak trunk flexion angle (P = .006), and peak knee extension moment (P = .034). In the ACL reconstruction group, quadriceps strength symmetry predicted symmetry in knee flexion excursion, peak trunk flexion, and peak knee extension moment (all P < .001) and predicted symmetry in peak trunk flexion angle (P < .001) after controlling for graft type, knee-related pain, function with activities of daily living, and sport function. At the time of return to sport, athletes who had undergone ACL reconstruction, including those in both the high- and low-quadriceps groups, demonstrated asymmetry during a single-leg drop-landing task compared with controls. Compensations included increased trunk flexion, decreased knee flexion excursion, and decreased knee extension moments on the involved limb. In addition, individuals in the low-quadriceps group demonstrated greater movement asymmetry compared with individuals in the high-quadriceps group. Restoration of symmetric quadriceps strength after ACL reconstruction is associated with more symmetric mechanics during a single-leg drop-landing movement. However, this appears to be multifactorial, as the high-quadriceps group also demonstrated landing asymmetries. Restoration of symmetric quadriceps strength may improve postoperative athletic participation; however, future study is warranted.


Yeast deubiquitinase Ubp3 interacts with the 26 S proteasome to facilitate Rad4 degradation.
Deubiquitinating enzymes (DUBs) function in a variety of cellular processes by removing ubiquitin moieties from substrates, but their role in DNA repair has not been elucidated. Yeast Rad4-Rad23 heterodimer is responsible for recognizing DNA damage in nucleotide excision repair (NER). Rad4 binds to UV damage directly while Rad23 stabilizes Rad4 from proteasomal degradation. Here, we show that disruption of yeast deubiquitinase UBP3 leads to enhanced UV resistance, increased repair of UV damage and Rad4 levels in rad23Δ cells, and elevated Rad4 stability. A catalytically inactive Ubp3 (Ubp3-C469A), however, is unable to affect NER or Rad4. Consistent with its role in down-regulating Rad4, Ubp3 physically interacts with Rad4 and the proteasome, both in vivo and in vitro, suggesting that Ubp3 associates with the proteasome to facilitate Rad4 degradation and thus suppresses NER.


Process-based quality management for clinical implementation of adaptive radiotherapy.
Intensity-modulated adaptive radiotherapy (ART) has been the focus of considerable research and developmental work due to its potential therapeutic benefits. However, in light of its unique quality assurance (QA) challenges, no one has described a robust framework for its clinical implementation. In fact, recent position papers by ASTRO and AAPM have firmly endorsed pretreatment patient-specific IMRT QA, which limits the feasibility of online ART. The authors aim to address these obstacles by applying failure mode and effects analysis (FMEA) to identify high-priority errors and appropriate risk-mitigation strategies for clinical implementation of intensity-modulated ART. An experienced team of two clinical medical physicists, one clinical engineer, and one radiation oncologist was assembled to perform a standard FMEA for intensity-modulated ART. A set of 216 potential radiotherapy failures composed by the forthcoming AAPM task group 100 (TG-100) was used as the basis. Of the 216 failures, 127 were identified as most relevant to an ART scheme. Using the associated TG-100 FMEA values as a baseline, the team considered how the likeliness of occurrence (O), outcome severity (S), and likeliness of failure being undetected (D) would change for ART. New risk priority numbers (RPN) were calculated. Failures characterized by RPN ≥ 200 were identified as potentially critical. FMEA revealed that ART RPN increased for 38% (n = 48/127) of potential failures, with 75% (n = 36/48) attributed to failures in the segmentation and treatment planning processes. Forty-three of 127 failures were identified as potentially critical. Risk-mitigation strategies include implementing a suite of quality control and decision support software, specialty QA software/hardware tools, and an increase in specially trained personnel. Results of the FMEA-based risk assessment demonstrate that intensity-modulated ART introduces different (but not necessarily more) risks than standard IMRT and may be safely implemented with the proper mitigations.


Reality monitoring and visual hallucinations in Parkinson's disease.
Between 8 and 40% of Parkinson disease (PD) patients will have visual hallucinations (VHs) during the course of their illness. Although cognitive impairment has been identified as a risk factor for hallucinations, more specific neuropsychological deficits underlying such phenomena have not been established. Research in psychopathology has converged to suggest that hallucinations are associated with confusion between internal representations of events and real events (i.e. impaired-source monitoring). We evaluated three groups: 17 Parkinson's patients with visual hallucinations, 20 Parkinson's patients without hallucinations and 20 age-matched controls, using tests of visual imagery, visual perception and memory, including tests of source monitoring and recollective experience. The study revealed that Parkinson's patients with hallucinations appear to have intact visual imagery processes and spatial perception. However, there were impairments in object perception and recognition memory, and poor recollection of the encoding episode in comparison to both non-hallucinating Parkinson's patients and healthy controls. Errors were especially likely to occur when encoding and retrieval cues were in different modalities. The findings raise the possibility that visual hallucinations in Parkinson's patients could stem from a combination of faulty perceptual processing of environmental stimuli, and less detailed recollection of experience combined with intact image generation.


HERFD XAS/ATR-FTIR batch reactor cell.
An autoclave reactor was modified to perform simultaneously high energy resolution fluorescence detected X-ray absorption spectroscopy (HERFD XAS) and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy measurements without altering the reactor design. This operando cell allows one to follow changes in the electronic and geometric structure of the catalyst with HERFD XAS and relate them to the simultaneously detected activity and reaction species with ATR-FTIR formed during the reaction in the liquid phase. The capability of the cell is demonstrated by two studies. The first shows the reduction of Au/CeO(2) precursors in different solvents. The second shows that mainly Au(0) is present in the hydrogenation of nitrobenzene over Au/CeO(2).


Implications of excess weight on mental wellbeing.
To assess differences in perceived health-related quality of life among individuals who are normal weight, overweight and obese in the general population, with particular emphasis on mental health. A cross-sectional study was conducted among the general adult population in Queensland, Australia. Participants (n = 1212) were selected randomly for computer-assisted telephone interview in July 2007. The sample ranged between 18 and 93 years, with a mean age of 51.10 years (s.d. = 15.92). Demographic and physical and mental health (SF-12) data were collected. Self-reported height and weight were used to classify participants into three groups based on their body mass index: normal weight; overweight; obese. The associations between body mass index categories and SF-12 scores were investigated. In this population sample, excess weight was associated with poorer physical health. In addition, significant associations were observed between excess weight and poor mental health for particular age groups. Obesity had a significant association with poor mental wellbeing for individuals who are aged 45 to 54 years. No sex differences were observed. The results provide additional evidence of the relationship between excess weight and mental wellbeing and highlight the need for health professionals to be cognisant of the potential for individuals who are obese to have a higher risk of experiencing mental health problems.


Concomitant administration of hapten and IL-2-toxin (DAB486-IL-2) results in specific deletion of antigen-activated T cell clones.
During the proliferative burst after Ag recognition, T cells express cell-surface, high-affinity IL-2R. The importance of IL-2R+ T cells in supporting/mediating tissue injury has been documented by the ability of mAb anti-IL2R therapies to prevent allograft rejection and autoimmunity. The delayed-type hypersensitivity (DTH) response, an experimental model of T-dependent immunity, offers the possibility of studying responses mounted against defined Ag. We previously reported that the chimeric IL-2 toxin (DAB486-IL-2) prevents DTH responses and selectively eliminates activated IL-2R bearing CD4 and CD8 T cells from lymph nodes draining the site of inflammation. We have examined the duration of immunosuppression and relative specificity of action of DAB486-IL-2 and anti-CD3 mAb for Ag-activated clones in a murine model of DTH using two different noncross-reacting haptens. Treatment with DAB486-IL-2 generates a state of selective unresponsiveness to subsequent challenge with the hapten introduced during the therapeutic period. Immediately after cessation of DAB486-IL-2 therapy, immunization with an unrelated hapten induces a normal vigorous immune response. By comparison, anti-CD3 mAb treatment causes a broad immunosuppression because unrelated haptens introduced after anti-CD3 therapy do not evoke a vigorous immune response. After cessation of DAB486-IL-2 toxin treatment response to the hapten is eventually restored probably by cells trafficking from the thymus, because thymectomized hosts remain unresponsive to the hapten. Taken together these data reinforce the role of the IL-2R as an important target for immunosuppression in T cell-mediated immune reactions. DAB-486-IL-2 treatment confers highly selective immunosuppression.


Spectral Doppler sonography: a noninvasive method for predicting dyspermia.
Sonography is a noninvasive, office-based diagnostic tool often used for evaluation of subfertile men. Previous studies have suggested that a resistive index (RI) greater than 0.6 is associated with impaired spermatogenesis. We sought to validate this threshold in a urologic patient population presenting for infertility evaluation. We retrospectively reviewed 99 consecutive patients seen for nonobstructive male infertility at our institution. Patient demographics, semen analysis parameters, hormone profiles, lipid profiles, and penile and scrotal sonographic measurements were recorded. The RI was calculated from measurements of the peak systolic velocity and end-diastolic velocity. Ninety-one patients fit the inclusion criteria and were subsequently divided into 2 groups based on RI: group 1 with RI values of 0.6 or less (n = 49) and group 2 with RI values greater than 0.6 (n = 42). Variables that were significantly different between the groups included age, total sperm count, percent motile sperm, total motile sperm, follicle-stimulating hormone, high-density lipoprotein, and testis volume. On the other hand, body mass index, forward progression, World Health Organization score, total testosterone, free testosterone, estradiol, total cholesterol, low-density lipoprotein, and triglycerides were not significantly different between the groups. A receiver operating characteristic curve revealed an area under the curve of 0.64 (confidence interval, 0.52-0.75; P = .025). At the threshold of greater than 0.6, the RI had specificity of 63.27% and a 1.56 likelihood ratio to predict total motile sperm less than 20 × 10(6) at spermatogenesis. An intratesticular RI greater than 0.6 is associated with impaired spermatogenesis. This finding supports the use of testicular spectral Doppler sonography as a noninvasive tool for evaluation of testicular function.


Metabolic changes associated with second-generation antipsychotic use in Alzheimer's disease patients: the CATIE-AD study.
The second-generation antipsychotics are associated with metabolic abnormalities in patients with schizophrenia. Elderly patients with Alzheimer's disease are frequently treated with these antipsychotics, but limited data are available on their metabolic effects. The authors assessed 186 male and 235 female Alzheimer's disease outpatients from the Clinical Antipsychotic Trials of Intervention Effectiveness-Alzheimer's Disease (CATIE-AD) for changes in weight, waist circumference, blood pressure, fasting glucose, and lipids in relation to duration of second-generation antipsychotic use (i.e., olanzapine, quetiapine, and risperidone) throughout the 36-week trial, using logistic regression and mixed-effects models. Women showed significant weight gain (0.14 lb/week of use) while change was nonsignificant in men. Clinically significant weight gain (i.e., > or = 7% of body weight) was seen among patients with antipsychotic use < or = 12 weeks (odds ratio [OR]=1.56, 95% CI=0.53 to 4.58), between 12 and 24 weeks (OR=2.89, 95% CI=0.97 to 8.64), and > 24 weeks (OR=3.38, 95% CI=1.24 to 9.23) relative to patients who did not use antipsychotics during the trial. Olanzapine and quetiapine treatments were significantly associated with weight gain (0.12 and 0.14 lb/week, respectively). In addition, olanzapine was significantly associated with decreases in HDL cholesterol (-0.19 mg/dl/week) and increased girth (0.07 inches/week) relative to the placebo group. No treatment effects were noted for changes in blood pressure, glucose, and triglycerides. Second-generation antipsychotic use was associated with weight gain in women, with olanzapine and quetiapine in particular, and with unfavorable change in HDL cholesterol and girth with olanzapine. The potential consequences of these effects suggest that patients with Alzheimer's disease treated with second-generation antipsychotics should be monitored closely.


Exploring brain function from anatomical connectivity.
The intrinsic relationship between the architecture of the brain and the range of sensory and behavioral phenomena it produces is a relevant question in neuroscience. Here, we review recent knowledge gained on the architecture of the anatomical connectivity by means of complex network analysis. It has been found that cortico-cortical networks display a few prominent characteristics: (i) modular organization, (ii) abundant alternative processing paths, and (iii) the presence of highly connected hubs. Additionally, we present a novel classification of cortical areas of the cat according to the role they play in multisensory connectivity. All these properties represent an ideal anatomical substrate supporting rich dynamical behaviors, facilitating the capacity of the brain to process sensory information of different modalities segregated and to integrate them toward a comprehensive perception of the real world. The results here exposed are mainly based on anatomical data of cats' brain, but further observations suggest that, from worms to humans, the nervous system of all animals might share these fundamental principles of organization.


Left atrial appendage occlusion by invagination and double suture technique.
Left atrial appendage (LAA) plays a crucial role as a source of atrial thrombus in patients with atrial fibrillation (AF). Thus, the need to close LAA becomes evident in patients with AF who undergo concomitant mitral valve surgery. Unfortunately, it has been reported a high rate of unsuccessful LAA occlusion, regardless of the technique employed.We propose a safe and simple method for LAA occlusion consisting in invagination of the appendage into the left atrium, followed by two sutures (purse string suture around the base of the LAA and a reinforce running suture).


Randomized double-blind trial of beta-carotene and vitamin C in women with minor cervical abnormalities.
A double-blind, placebo-controlled, randomized, factorial study using a daily oral administration of 30 mg beta-carotene and/or 500 mg vitamin C was conducted in 141 women with colposcopically and histologically confirmed minor squamous atypia or cervical intra-epithelial neoplasia (CIN) I. Over approximately 2 years of follow-up, 43 lesions regressed to normal and 13 progressed to CIN II. The regression rate was slightly higher, but not significantly so, in those randomized to beta-carotene compared to no beta-carotene (hazard ratio = 1.58, 95% CI: 0.86-2.93, P = 0.14) and slightly lower, but not statistically significant, for those randomized to vitamin C compared to no vitamin C (hazard ratio = 0.65, 95% CI: 0.35-1.21, P = 0.17). In a model with no interaction, the progression rate was slightly higher in those randomized to beta-carotene (hazard ratio = 1.75, 95% CI: 0.57-5.36, P = 0.32) and also in those randomized to vitamin C (hazard ratio = 2.40, 95% CI: 0.74-7.80, P = 0.13). Neither of these were statistically significant. However, there was some evidence of an interaction effect of the two compounds on the progression rate (P = 0.052), with seven of the progressed lesions occurring in those randomized to both vitamins compared to a total of six in the three other groups. The currently available evidence from this and other trials suggests that high doses of these compounds are unlikely to increase the regression or decrease the progression of minor atypia and CIN I.


Managing acute stress response to major trauma.
In this article, the authors review the current empiric literature on early interventions. Findings on the effects, course, help-seeking, and recovery from disasters are first reviewed, with recommendations given that are pertinent to intervention following mass casualties. In reviewing the most commonly used interventions, it is clear that evidence from well-controlled studies showing that early intervention can help prevent longer-term problems is limited. The authors discuss the approaches that have received the most attention or empiric support as early interventions following trauma, which include psychologic debriefing, cognitive-behavioral interventions, eye movement desensitization and processing (EMDR) and other neoteric approaches, and psychopharmacology. At this time, the most promising results for prevention of psychopathology have been achieved with brief four- or five-session cognitive-behavioral therapy. In contrast, randomized clinical trials on psychologic debriefing currently suggest that this approach is either ineffective at preventing psychopathology, or contributive to post-traumatic stress disorder symptoms. Research support is currently lacking for EMDR and pharmacotherapy as early interventions. A major challenge to the field is to integrate the practical experience and knowledge of professional responders with well-controlled, timely intervention research, and to effectively disseminate these findings to practitioners in the field.


A Pilot Study on Automatic Three-Dimensional Quantification of Barrett's Esophagus for Risk Stratification and Therapy Monitoring.
Barrett's epithelium measurement using widely accepted Prague C&M classification is highly operator dependent. We propose a novel methodology for measuring this risk score automatically. The method also enables quantification of the area of Barrett's epithelium (BEA) and islands, which was not possible before. Furthermore, it allows 3-dimensional (3D) reconstruction of the esophageal surface, enabling interactive 3D visualization. We aimed to assess the accuracy of the proposed artificial intelligence system on both phantom and endoscopic patient data. Using advanced deep learning, a depth estimator network is used to predict endoscope camera distance from the gastric folds. By segmenting BEA and gastroesophageal junction and projecting them to the estimated mm distances, we measure C&M scores including the BEA. The derived endoscopy artificial intelligence system was tested on a purpose-built 3D printed esophagus phantom with varying BEAs and on 194 high-definition videos from 131 patients with C&M values scored by expert endoscopists. Endoscopic phantom video data demonstrated a 97.2% accuracy with a marginal ± 0.9 mm average deviation for C&M and island measurements, while for BEA we achieved 98.4% accuracy with only ±0.4 cm2 average deviation compared with ground-truth. On patient data, the C&M measurements provided by our system concurred with expert scores with marginal overall relative error (mean difference) of 8% (3.6 mm) and 7% (2.8 mm) for C and M scores, respectively. The proposed methodology automatically extracts Prague C&M scores with high accuracy. Quantification and 3D reconstruction of the entire Barrett's area provides new opportunities for risk stratification and assessment of therapy response.


Propolis and amnion reepithelialise second-degree burns in rats.
Burns are serious consequences of trauma in terms of both imminent mortality and prolonged periods of morbidity. They are often accompanied by unsatisfactory cosmetic as well as functional and psychological outcomes. These complications emphasise the need for stronger efforts in achieving greater diversity and effectiveness in the treatment of skin burns. This study aimed to verify the effectiveness of gross and microscopic epidermal and dermal responses in the process of regenerative repair or healing of burns in rats that were treated either daily with 5% propolis ointment or by autologous amnion graft. Second-degree burns were inflicted in the neck region of female rats by contact with a hot metal (at 130 °C) for 5 s. Propolis treatment accelerated the process of tissue repair and led to decreased local inflammation, which indicates that treatment with propolis was successful in the initial period (7 days) and stimulated the production of collagen fibre (assessed by morphometry) in all the periods evaluated (14 and 21 days). Amnion treatment inhibited local inflammation (assessed macroscopically), stimulated local epithelial regeneration (assessed microscopically) and stimulated the production of collagen fibre (assessed by morphometry) in the days following burn. These treatments offer new therapeutic strategies for treating severe skin burns; these strategies may allow the minimisation of scar formation, a more rapid return of function and, ultimately, a better quality of life for burn patients.


Signal Increase on Unenhanced T1-Weighted Images in the Rat Brain After Repeated, Extended Doses of Gadolinium-Based Contrast Agents: Comparison of Linear and Macrocyclic Agents.
In this prospective preclinical study, we evaluated T1-weighted signal intensity in the deep cerebellar nuclei (CN) and globus pallidus (GP) up to 24 days after repeated administration of linear and macrocyclic gadolinium-based contrast agents (GBCAs) using homologous imaging and evaluation methods as in the recently published retrospective clinical studies. In a second part of the study, cerebrospinal fluid (CSF) spaces were evaluated for contrast enhancement by fluid-attenuated magnetic resonance imaging (MRI). Sixty adult male Wistar-Han rats were randomly divided into a control and 5 GBCA groups (n = 10 per group). The administered GBCAs were gadodiamide, gadopentetate dimeglumine, and gadobenate dimeglumine (linear GBCAs) as well as gadobutrol and gadoterate meglumine (macrocyclic GBCAs) and saline (control). Over a period of 2 weeks, the animals received 10 intravenous injections at a dose of 2.5 mmol Gd/kg body weight, each on 5 consecutive days per week. Before GBCA administration, as well as 3 and 24 days after the last injection, a whole-brain MRI was performed using a standard T1-weighted 3-dimensional turbo spin echo sequence on a clinical 1.5 T scanner. The ratios of signal intensities in deep CN to pons (CN/Po) and GP to thalamus (GP/Th) were determined. For the evaluation of the CSF spaces, 18 additional rats were randomly divided into 6 groups (n = 3 per group) that received the same GBCAs as in the first part of the study. After MR cisternography for anatomical reference, a fluid-attenuated inversion recovery sequence was performed before and 1 minute after intravenous injection of a dose of 1 mmol Gd/kg body weight GBCA or saline. A significantly increased signal intensity ratio of CN/Po was observed 3 and 24 days after the last injection of gadodiamide and gadobenate dimeglumine. No significant changes were observed between the 2 time points. Gadopentetate dimeglumine injection led to a moderately elevated but statistically not significant CN/Po signal intensity ratio. No increased CN/Po signal intensity ratios were determined in the MRI scans of rats that received macrocyclic GBCAs gadobutrol and gadoterate meglumine or saline. The ratio of signal intensity in GP/Th was not elevated in any group injected with GBCAs or saline. Enhanced signal intensities of CSF spaces were observed in the postcontrast fluid-attenuated inversion recovery images of all animals receiving GBCAs but not for saline. In this animal study in rats, increased signal intensity in the CN was found up to 24 days after multiple, extended doses of linear GBCAs. However, in contrast to clinical reports, the signal enhancement in the GP was not reproduced, demonstrating the limitations of this animal experiment. The elevated signal intensities remained persistent over the entire observation period. In contrast, no changes of signal intensities in either the CN or the GP were observed for macrocyclic GBCAs. However, all GBCAs investigated were able to pass the blood-CSF barrier in rats to a certain, not yet quantified extent.


A Multicenter International Randomized Controlled Manikin Study on Different Protocols of Cardiopulmonary Resuscitation for Laypeople: The MANI-CPR Trial.
Compression-only cardiopulmonary resuscitation (CPR) is a suggested technique for laypeople facing out-of-hospital cardiac arrest (OHCA). However, it is difficult performing high-quality CPR until emergency medical services arrival with this technique. We aimed to verify whether incorporating intentional interruptions of different frequency and duration increases laypeople's CPR quality during an 8-minute scenario compared with compression-only CPR. We performed a multicenter randomized manikin study selecting participants from 2154 consecutive laypeople who followed a basic life support/automatic external defibrillation course. People who achieved high-quality CPR in 1-minute test on a computerized manikin were asked to participate. Five hundred seventy-six were enrolled, and 59 were later excluded for technical reasons or incorrect test recording. Participants were randomized in an 8-minute OHCA scenario using 3 CPR protocols (30 compressions and 2-second pause, 30c2s; 50 compressions and 5-second pause, 50c5s; 100 compressions and 10-second pause, 100c10s) or compression-only technique. The main outcome was the percentage of chest compressions with adequate depth. Five hundred seventeen participants were evaluated. There was a statistically significant difference regarding the percentage of compressions with correct depth among the groups (30c2s, 96%; 50c5s, 96%; 100c10s, 92%; compression only, 79%; P = 0.006). Post hoc comparison showed a significant difference for 30c2s (P = 0.023) and for 50c5s (P = 0.003) versus compression only. Regarding secondary outcome, there were a higher chest compression fraction in the compression-only group and a higher rate of pauses longer than 10 seconds in the 100c10s. In a simulated OHCA, 30c2s and 50c5s protocols were characterized by a higher rate of chest compressions with correct depth than compression only. This could have practical consequences in laypeople CPR training and recommendations.Clinical Trial Registration: NCT02632500.


B7H3 regulates differentiation and serves as a potential biomarker and theranostic target for human glioblastoma.
B7H3 (CD276), a co-stimulator molecule of the cell surface B7 protein superfamily, is expressed on glioblastomas (GBM). Recently, B7H3 functions beyond immune costimulation have been demonstrated. However, the mechanisms underlying B7H3 function are diverse and not well understood. GBM tumors contain undifferentiated self-renewing cells, which confound therapeutic attempts. We investigated the role of B7H3 in the regulation of GBM cell differentiation and the regulatory pathways involved. Analysis of public databases (TCGA, Rembrandt, and GEO NCBI) and RNA sequencing were performed to explore the role of B7H3 in GBM. Knockdown and overexpression of B7H3, were used to verify the downstream pathway in vitro. Further studies in vivo were performed to support the new finding. Bioinformatics analysis identified a correlation between the expression of B7H3, the expression of glioma self-renewing cell (GSC)-related genes, and MYC expression. These observations were verified by RNA-sequencing analyses in primary GBM cell lines. In vitro knockdown of B7H3-induced differentiation, associated with downregulation of SMAD6 (a TGF-β pathway inhibitor) and enhancement of SMAD1 phosphorylation-induced SMAD4 expression. Importantly, activation of the TGF-β pathway resulted in downregulation of MYC expression. In vivo assays conducted in a human GBM cell line xenograft mouse model demonstrated that B7H3 knockdown decreased MYC expression and inhibited tumor growth. B7H3 knockdown could regulate GBM differentiation by modulating MYC expression. So, B7H3 could serve as a potential theranostic target for the treatment of patients with GBM.


Exome sequencing reveals mutated SLC19A3 in patients with an early-infantile, lethal encephalopathy.
To accomplish a diagnosis in patients with a rare unclassified disorder is difficult. In this study, we used magnetic resonance imaging pattern recognition analysis to identify patients with the same novel heritable disorder. Whole-exome sequencing was performed to discover the mutated gene. We identified seven patients sharing a previously undescribed magnetic resonance imaging pattern, characterized by initial swelling with T2 hyperintensity of the basal nuclei, thalami, cerebral white matter and cortex, pons and midbrain, followed by rarefaction or cystic degeneration of the white matter and, eventually, by progressive cerebral, cerebellar and brainstem atrophy. All patients developed a severe encephalopathy with rapid deterioration of neurological functions a few weeks after birth, followed by respiratory failure and death. Lactate was elevated in body fluids and on magnetic resonance spectroscopy in most patients. Whole-exome sequencing in a single patient revealed two predicted pathogenic, heterozygous missense mutations in the SLC19A3 gene, encoding the second thiamine transporter. Additional predicted pathogenic mutations and deletions were detected by Sanger sequencing in all six other patients. Pathology of brain tissue of two patients demonstrated severe cerebral atrophy and microscopic brain lesions similar to Leigh's syndrome. Although the localization of SLC19A3 expression in brain was similar in the two investigated patients compared to age-matched control subjects, the intensity of the immunoreactivity was increased. Previously published patients with SLC19A3 mutations have a milder clinical phenotype, no laboratory evidence of mitochondrial dysfunction and more limited lesions on magnetic resonance imaging. In some, cerebral atrophy has been reported. The identification of this new, severe, lethal phenotype characterized by subtotal brain degeneration broadens the phenotypic spectrum of SLC19A3 mutations. Recognition of the associated magnetic resonance imaging pattern allows a fast diagnosis in affected infants.


Probable causes of increasing brucellosis in free-ranging elk of the Greater Yellowstone Ecosystem.
While many wildlife species are threatened, some populations have recovered from previous overexploitation, and data linking these population increases with disease dynamics are limited. We present data suggesting that free-ranging elk (Cervus elaphus) are a maintenance host for Brucella abortus in new areas of the Greater Yellowstone Ecosystem (GYE). Brucellosis seroprevalence in free-ranging elk increased from 0-7% in 1991-1992 to 8-20% in 2006-2007 in four of six herd units around the GYE. These levels of brucellosis are comparable to some herd units where elk are artificially aggregated on supplemental feeding grounds. There are several possible mechanisms for this increase that we evaluated using statistical and population modeling approaches. Simulations of an age-structured population model suggest that the observed levels of seroprevalence are unlikely to be sustained by dispersal from supplemental feeding areas with relatively high seroprevalence or an older age structure. Increases in brucellosis seroprevalence and the total elk population size in areas with feeding grounds have not been statistically detectable. Meanwhile, the rate of seroprevalence increase outside the feeding grounds was related to the population size and density of each herd unit. Therefore, the data suggest that enhanced elk-to-elk transmission in free-ranging populations may be occurring due to larger winter elk aggregations. Elk populations inside and outside of the GYE that traditionally did not maintain brucellosis may now be at risk due to recent population increases. In particular, some neighboring populations of Montana elk were 5-9 times larger in 2007 than in the 1970s, with some aggregations comparable to the Wyoming feeding-ground populations. Addressing the unintended consequences of these increasing populations is complicated by limited hunter access to private lands, which places many ungulate populations out of administrative control. Agency-landowner hunting access partnerships and the protection of large predators are two management strategies that may be used to target high ungulate densities in private refuges and reduce the current and future burden of disease.


Lectin-resistant CHO cells: selection of seven new mutants resistant to ricin.
In attempts to isolate new CHO glycosylation mutants, selection protocols using plant lectins that bind galactose residues of cell surface carbohydrates were applied to mutagenized CHO populations. The lectins were used alone or in combination to obtain seven ricin-resistant phenotypes. Each mutant had distinctive properties compared with previously described ricin-resistant CHO cells. One of the new phenotypes was dominant in somatic cell hybrids, and the others were recessive. Complementation analyses between related lectin-resistant (LecR) phenotypes indicated that each new isolate represented a novel genotype. Five of the mutants had properties typical of new CHO glycosylation mutants. The remaining two mutants were not readily categorized. Although they did not appear to be ricin-internalization or protein-synthesis mutants, they also did not display the marked alterations in sensitivity to several lectins of different sugar specificity expected for glycosylation mutants. The seven new LecR mutants described in these studies brings the total number of different LecR CHO mutants isolated by this and other laboratories to about 40. Criteria for identifying new LecR mutations in CHO cells are discussed.


Batrachochytrium dendrobatidis in the live frog trade of Telmatobius (Anura: Ceratophryidae) in the tropical Andes.
Species of frogs in the genus Telmatobius are traded and sold for human consumption in the Andes and in coastal cities of Peru and Bolivia. These frogs are harvested from wild populations. We report high prevalence of infection by the pathogenic fungus Batrachochytrium dendrobatidis in live frogs purchased at the main market in Cusco, Peru, from January 2008 to January 2010. We suggest that the transport of native anurans through the live frog trade could facilitate the spread of this fungus among Andean frogs. The tropical Andes are the most important biodiversity hotspot for amphibians. Because many neotropical taxa are known to be susceptible to chytridiomycosis, the presence of a large reservoir of infection in the frog trade poses a significant threat to amphibian conservation.


eCAT: Online electronic lab notebook for scientific research.
eCAT is an electronic lab notebook (ELN) developed by Axiope Limited. It is the first online ELN, the first ELN to be developed in close collaboration with lab scientists, and the first ELN to be targeted at researchers in non-commercial institutions. eCAT was developed in response to feedback from users of a predecessor product. By late 2006 the basic concept had been clarified: a highly scalable web-based collaboration tool that possessed the basic capabilities of commercial ELNs, i.e. a permissions system, controlled sharing, an audit trail, electronic signature and search, and a front end that looked like the electronic counterpart to a paper notebook. During the development of the beta version feedback was incorporated from many groups including the FDA's Center for Biologics Evaluation & Research, Uppsala University, Children's Hospital Boston, Alex Swarbrick's lab at the Garvan Institute in Sydney and Martin Spitaler at Imperial College. More than 100 individuals and groups worldwide then participated in the beta testing between September 2008 and June 2009. The generally positive response is reflected in the following quote about how one lab is making use of eCAT: "Everyone uses it as an electronic notebook, so they can compile the diverse collections of data that we generate as biologists, such as images and spreadsheets. We use to it to take minutes of meetings. We also use it to manage our common stocks of antibodies, plasmids and so on. Finally, perhaps the most important feature for us is the ability to link records, reagents and experiments." By developing eCAT in close collaboration with lab scientists, Axiope has come up with a practical and easy-to-use product that meets the need of scientists to manage, store and share data online. eCAT is already being perceived as a product that labs can continue to use as their data management and sharing grows in scale and complexity.


Amino acid residues involved in determining the processivity of the 3'-5' exonuclease activity in a family B DNA polymerase from the thermoacidophilic archaeon Sulfolobus solfataricus.
Herein, we report on the mutational analysis of a 70-amino acid segment (region 1, residues 438-508) of family B DNA polymerase from the thermoacidophilic archaeon Sulfolobus solfataricus (Sso DNA pol). Region 1, which lies between the Exo III sequence and the similarity motif D- -SLYP, connects the exonuclease and polymerase domains of Sso DNA pol. Two C-terminally deleted forms of the enzyme, proteins N438 (residues 1-438) and N508 (residues 1-508), were overproduced in the recombinant form and biochemically characterized. They contain the three evolutionarily conserved Exo motifs, but differ in the extent of the C-terminal deletion, since only N508 includes region 1. Both have been found to retain a Mn2+-dependent 3'-5' exonuclease activity, whose thermal stability appears to be increased in comparison to that of the full-sized enzyme. Assays for processive 3'-5' exonuclease activity, carried out with the heparin trap method on a 24-base oligonucleotide, have revealed that protein N508, as well as the full-length Sso DNA pol, retains a level of processivity of the degradative function substantially higher than that for protein N438. In addition, six site-specific mutations have been introduced at the highly conserved Y-GG/A motif, which has been found within Sso DNA pol region 1. All mutant proteins (Lys491Ile, Tyr495Ser, Lys496Ile, Gly497Ala, and Ala498Val) display increased processivity of their 3'-5' exonuclease activity, with the exception of protein Tyr495Phe. By a steady-state kinetic analysis of the exonucleolytic reaction on a 24-base oligonucleotide, the above site-specific mutations have been found to affect Km values consistently with the observed differences in the processivity values, whereas the effect on the kcat values seems to be less important. The results from this mutational analysis indicate that region 1 is involved in determining the processivity of the proofreading function, directly interacting with the nucleic acid substrate.


Sleep Pattern of Adolescents in a School in Delhi, India: Impact on their Mood and Academic Performance.
To examine the sleep pattern and observe differences in sleep routines, phase preferences, mood, attendance, and academic performance among different adolescent age students. Secondly, to observe the age at which sleep phase transition and changes in sleep requirement become evident. A cross-sectional study was conducted among 501 students (aged 11-15 y) of a school in Delhi, India. Students were evaluated for their sleep patterns, sleep duration, habits of napping, quality of sleep, sleepiness, depression, phase preferences by self-reported school sleep habits survey questionnaire along with school performance and attendance. Significant differences were found in sleep pattern of students aged 11-12 y and 13-15 y. Bedtime shifted to a later time with increasing age but early morning schools kept the wake time same, leading to a decline in total sleep duration of older adolescents. Older adolescents had higher depression but poor attendance and academic performance. Prevalence of sleep deprivation increased with age, from 83.7% to 87.1% in 11-12 y to 90.5% to 92.5% in 13-15 y. The study clearly identifies 12-13 y as age of transition of sleep pattern among adolescents. Though significant differences were found in the academic performance, mood and attendance among preteens and teens but no direct association was seen between academic performances and sleep pattern. A complex multifactorial association between sleep patterns, attendance, mood and academic performance which may change over days, months, or years should be explored further in a longitudinal follow up study.


A novel recombinase polymerase amplification (RPA) assay for the rapid isothermal detection of Neospora caninum in aborted bovine fetuses.
The development of a method to rapidly diagnose Neospora caninum infection is highly desirable. Recombinase polymerase amplification (RPA), combined with lateral flow (LF) strips, is a novel approach to rapidly amplify and visualize DNA. We have developed a prototype LF-RPA assay, using primers and a probe that targeted a specific sequence in the N. caninum NC-5 gene. The N. caninum-specific LF-RPA assay was first tested on purified DNA from oocysts and amplified N. caninum DNA to detectable levels in 10 min, at a constant temperature and without the need for an expensive thermocycler. The designed RPA primers and probe displayed 100% specificity for detecting N. caninum without any cross-reaction with DNA from nine related protozoan spp. (eg Toxoplasma gondii, Sarcocystis gigantean, Sarcocystis zuoi, Hammondia hammondi, Hammondia heydorni, Eimeria cylindrica, Plasmodium falciparum, Theileria annulata and Babesia bigemina). Although, LF-RPA assay detected amounts as low as 50 fg of N. caninum DNA, it was nearly 5-fold less sensitive than previously published qPCR and nested PCR assays. We tested the diagnostic performance of the LF-RPA assay for the detection of N. caninum DNA in aborted bovine fetal tissue samples, and compared the results with those obtained from nested PCR. Out of the 75 samples examined, 18 (24%) and 17 (22.6%) tested positive using LF-RPA and nested PCR, respectively. Our results indicate that LF-RPA is a suitable assay for the rapid and reliable detection of N. caninum.


EphA4 defines a class of excitatory locomotor-related interneurons.
Relatively little is known about the interneurons that constitute the mammalian locomotor central pattern generator and how they interact to produce behavior. A potential avenue of research is to identify genetic markers specific to interneuron populations that will assist further exploration of the role of these cells in the network. One such marker is the EphA4 axon guidance receptor. EphA4-null mice display an abnormal rabbit-like hopping gait that is thought to be the result of synchronization of the normally alternating, bilateral locomotor network via aberrant crossed connections. In this study, we have performed whole-cell patch clamp on EphA4-positive interneurons in the flexor region (L2) of the locomotor network. We provide evidence that although EphA4 positive interneurons are not entirely a homogeneous population, most of them fire in a rhythmic manner. Moreover, a subset of these interneurons provide direct excitation to ipsilateral motor neurons as determined by spike-triggered averaging of the local ventral root DC trace. Our findings substantiate the role of EphA4-positive interneurons as significant components of the ipsilateral locomotor network and describe a group of putative excitatory central pattern generator neurons.


Antimicrobial activity of linear lipopeptides derived from BP100 towards plant pathogens.
A collection of 36 lipopeptides were designed from the cecropin A-melittin hybrid peptide BP100 (H-Lys-Lys-Leu-Phe-Lys-Lys-Ile-Leu-Lys-Tyr-Leu-NH2) previously described with activity against phytopathogenic bacteria. These lipopeptides were synthesized on solid-phase and screened for their antimicrobial activity, toxicity and proteolytic stability. They incorporated a butanoyl, a hexanoyl or a lauroyl group at the N-terminus or at the side chain of a lysine residue placed at each position of the sequence. Their antimicrobial activity and hemolysis depended on the fatty acid length and its position. In particular, lipopeptides containing a butanoyl or a hexanoyl chain exhibited the best biological activity profile. In addition, we observed that the incorporation of the acyl group did not induce the overexpression of defense-related genes in tomato. Best lipopeptides were BP370, BP378, BP381, BP387 and BP389, which were highly active against all the pathogens tested (minimum inhibitory concentration of 0.8 to 12.5 μM), low hemolytic, low phytotoxic and significantly stable to protease degradation. This family of lipopeptides might be promising functional peptides useful for plant protection.


Comparison of Customized Abutments Made from Titanium and a Machinable Precious Alloy.
To investigate the clinical usefulness, a customized abutment produced with the Pd-Ag-In alloy was compared with a customized abutment produced with the conventional titanium alloy for discoloration and mechanical accuracy. Discoloration and resistance to corrosion of the Pd-Ag-In alloy were evaluated using chemical solutions. Marginal adaptation of internal-type implants and abutments was compared using 10 titanium abutments and 10 Pd-Ag-In abutments using a surface measuring system. A detorque test was performed on 12 implant-abutment complexes of each control and experimental group to investigate screw joint stability. Cyclic loading simulating a human's mastication movement for 1 year was applied after 30 Ncm initial tightening, and the removal torque was measured using a digital torque gauge. The noninferiority test was conducted to compare the Pd-Ag-In alloy with a titanium abutment with a 10% margin. The Pd-Ag-In alloy had a warm yellow color and displayed stable resistance to discoloration and corrosion, resulting in an advantageous esthetic property. When compared to the titanium alloy, it did not show noninferiority with respect to the gap between the implant and the abutment; the gap was approximately 13.3 μm on average, which was not significantly different from those observed in previous studies. After long-term use, it displayed statistically significant noninferiority in the removal torque value compared to the titanium group. The Pd-Ag-In alloy-based customized abutment had good mechanical properties of the implant-abutment complex as well as a superior esthetic property, and can provide favorable outcomes in anterior implant restoration.


CT-Colonography vs. Colonoscopy for Detection of High-Risk Sessile Serrated Polyps.
Sessile serrated polyps (SSPs) are suggested to be the precursors of 15-30% of all colorectal cancers (CRCs). Therefore, CRC screening modalities should also be designed to detect high-risk SSPs. We compared computed tomography colonography (CTC) with colonoscopy-based screening for the detection of high-risk SSPs in average-risk individuals. Data from a randomized controlled trial that compared CTC with colonoscopy for population screening were used for the analysis. Individuals diagnosed at CTC with a lesion ≥10 mm in size were referred for colonoscopy. Individuals with only 6-9 mm lesions were offered surveillance CTC. This surveillance CTC was followed by a colonoscopy when a lesion ≥6 mm was detected. Yield of both was accumulated to mimic current American College of Radiology CTC referral strategy (referral of individuals with any lesion ≥6 mm). Per participant detection of ≥1 high-risk (dysplastic and/or ≥10 mm) SSP was compared with colonoscopy using multiple logistic regression analysis. In total, 8,844 individuals were invited to participate (in 2:1 allocation), of which 1,276 colonoscopy and 982 CTC invitees participated in the study. In the colonoscopy arm, 4.3% of individuals were diagnosed with ≥1 high-risk SSP, compared with 0.8% in the CTC arm (odds ratio (OR) 5.5; 95% confidence interval (CI) 2.6-11.6; P<0.001). In total, 3.1% of individuals in the colonoscopy arm were diagnosed with high-risk SSPs as most advanced lesion, compared with 0.4% in the CTC arm (OR 7.7; 95% CI 2.7-21.6; P<0.001). The current CTC strategy showed a marked lower detection for especially flat high-risk SSPs (17 vs. 0), high-risk SSP located in the proximal colon (32 vs. 1), and SSPs with dysplasia (30 vs. 1). In a randomized controlled setting, the detection rate of high-risk SSPs was significantly higher with colonoscopy than CTC. These results might have implications for CTC as a CRC modality for opportunistic screening in average-risk adults.


Distribution of DNA polymerase alpha during nuclear division cycles in Drosophila melanogaster embryo.
An immunocytochemical method using a specific monoclonal antibody was employed to detect DNA polymerase alpha in Drosophila melanogaster embryos during the first 13 nuclear division cycles after fertilization. The anti-DNA polymerase alpha antibody stained the ooplasm of the unfertilized egg, indicating that DNA polymerase alpha is maternally stored. Strong nuclear staining with the antibody over the weaker staining of the cytoplasm was observed at interphase throughout the 13 nuclear division cycles. The staining of the cytoplasmic regions surrounding the nucleus was much stronger than the other region of the syncytial cytoplasm until cycle 10. Although prophase nuclei were stained with the antibody, metaphase chromosomes were never stained throughout the 13 cycles. The chromosomal (nuclear) staining reappeared at anaphase until cycle 11 and at telophase in later cycles. The staining of the syncytial cytoplasm except for the cortical region became faint by cycle 13, suggesting the consumption of the maternal storage by this cycle. These results suggest that DNA polymerase alpha dissociates from chromosomes at the beginning of metaphase; then in later mitotic phases, it is transported from the syncytial cytoplasm into nuclei to participate in formation of the active DNA replication enzyme complex.


Normal volumetric and T1 relaxation time values at 1.5 T in segmented pediatric brain MRI using a MP2RAGE acquisition.
This study introduced a tailored MP2RAGE-based brain acquisition for a comprehensive assessment of the normal maturing brain. Seventy normal patients (35 girls and 35 boys) from 1 to 16 years of age were recruited within a prospective monocentric study conducted from a single University Hospital. Brain MRI examinations were performed at 1.5 T using a 20-channel head coil and an optimized 3D MP2RAGE sequence with a total acquisition time of 6:36 min. Automated 38 region segmentation was performed using the MorphoBox (template registration, bias field correction, brain extraction, and tissue classification) which underwent a major adaptation of three age-group T1-weighted templates. Volumetry and T1 relaxometry reference ranges were established using a logarithmic model and a modified Gompertz growth respectively. Detailed automated brain segmentation and T1 mapping were successful in all patients. Using these data, an age-dependent model of normal brain maturation with respect to changes in volume and T1 relaxometry was established. After an initial rapid increase until 24 months of life, the total intracranial volume was found to converge towards 1400 mL during adolescence. The expected volumes of white matter (WM) and cortical gray matter (GM) showed a similar trend with age. After an initial major decrease, T1 relaxation times were observed to decrease progressively in all brain structures. The T1 drop in the first year of life was more pronounced in WM (from 1000-1100 to 650-700 ms) than in GM structures. The 3D MP2RAGE sequence allowed to establish brain volume and T1 relaxation time normative ranges in pediatrics. • The 3D MP2RAGE sequence provided a reliable quantitative assessment of brain volumes and T1 relaxation times during childhood. • An age-dependent model of normal brain maturation was established. • The normative ranges enable an objective comparison to a normal cohort, which can be useful to further understand, describe, and identify neurodevelopmental disorders in children.


Modulating mitochondrial intracellular location as a redox signal.
Mitochondria have various essential functions in metabolism and in determining cell fate during apoptosis. In addition, mitochondria are also important nodes in a number of signaling pathways. For example, mitochondria can modulate signals transmitted by second messengers such as calcium. Because mitochondria are also major sources of reactive oxygen species (ROS), they can contribute to redox signaling--for example, by the production of ROS such as hydrogen peroxide that can reversibly modify cysteine residues and thus the activity of target proteins. Mitochondrial ROS production is thought to play a role in hypoxia signaling by stabilizing the oxygen-sensitive transcription factor hypoxia-inducible factor-1α. New evidence has extended the mechanism of mitochondrial redox signaling in cellular responses to hypoxia in interesting and unexpected ways. Hypoxia altered the microtubule-dependent transport of mitochondria so that the organelles accumulated in the perinuclear region, where they increased the intranuclear concentration of ROS. The increased ROS in turn enhanced the expression of hypoxia-sensitive genes such as VEGF (vascular endothelial growth factor) not by reversibly oxidizing a protein, but by oxidizing DNA sequences in the hypoxia response element of the VEGF promoter. This paper and other recent work suggest a new twist on mitochondrial signaling: that the redistribution of mitochondria within the cell can be a component of regulatory pathways.


Occult glove perforations: frequency during interventional radiologic procedures.
Because the intact surgical glove is an important barrier against exposure to blood and other body fluids, occult glove perforations occurring during procedures may have implications for infection control and protection of health care workers. Although the frequency of occult glove perforations during surgery has been studied, no such data have been reported for interventional radiology. This study was designed to determine the frequency of occult glove perforations in interventional radiology. Gloves used during interventional radiologic procedures were collected and tested for occult perforations according to previously accepted methods. The frequency of occult defects in unused gloves was also determined. A form completed for each pair of gloves indicated length of use, awareness of perforations, and information about the procedure. The anatomic distribution of perforations was also studied. Occult perforations were found in 49 (10%) of 492 used gloves. The frequency of defects in unused gloves was 1% (2/160). Gloves worn fewer than 2 hr had a perforation rate of 7% (29/406), and those worn 2 hr or longer had a rate of 23% (20/86; p less than .0001). Perforations were noticed by the wearer in only three (6%) of the 49 perforations. The perforation rate for inner gloves of a double-glove set was 3% (1/34; the corresponding outer glove also was perforated). Although the 10% rate of occult glove perforation during interventional radiologic procedures found in this series is lower than rates described with surgical procedures, it is not insignificant. The significant increase in the rate of perforation when gloves are worn more than 2 hr suggests that gloves should routinely be changed at or before 2 hr of wear, particularly in procedures involving high-risk patients.


Mammography promotion in the emergency department: a pilot study.
The objective of this pilot study is to assess the need, desire, and applicability of a mammography promotion project in the emergency department (ED). A convenience sample from the ED of a public University hospital was surveyed to determine their mammography status, interest in a program to promote mammography, and barriers to mammography. The survey included demographics information, health care access, including health insurance and primary care provider, mammography status and date of mammogram, as well as a checklist of potential barriers. Participants were also asked whether they would be interested in mammography promotion in this setting. More than 15% of the 197 women surveyed had never received a mammogram, and more than half had not received 1 in the past year. The most common barriers to mammography were competing demands and money. Three quarters of the women said they would be interested in mammography promotion while waiting for care in the ED. This study provides promise that mammography promotion activities may be appropriately placed in the ED and provides a solid platform from which researchers and nurses may launch efforts to develop preventive health interventions in innovative public health care settings.


The accuracy of dietary recall of infant feeding and food allergen data.
Research investigating the association of infant dietary factors with later health outcomes often relies on maternal recall. It is unclear what the effect of recall bias is on the accuracy of the information obtained. The present study aimed to determine the extent of recall bias on the accuracy of infant feeding and food allergen data collected 10 years later. Mothers were recruited from a prospective birth cohort from the Isle of Wight. When their child was 10 years of age (2011/2012), mothers were requested to complete a retrospective infant feeding questionnaire asking the same questions as those solicited in 2001/2002. In total, 125 mothers participated. There was substantial agreement for recollection of any breastfeeding (κ = 0.79) and the duration of breastfeeding from 10 years earlier (r = 0.84). Some 94% of mothers recalled accurately that their child had received formula milk. The exact age at which formula milk was first given was reliably answered (r = 0.63). The brand of formula milk was poorly recalled. Recall of age of introduction of solid food was not reliable (r = 0.16). The age of introduction of peanuts was the only food allergen that was recalled accurately (86%). The present study highlights the importance of maternal recall bias of infant feeding practices over 10 years. Recall related to breastfeeding and formula feeding were reliable, whereas recalls related to age of introduction of solid or allergenic foods, apart from peanut, were not. Caution should be applied when interpreting studies relying on dietary recall.


The 2-phase case-control design: an efficient way to use expert-time.
The objective of this paper is to show the benefits of using a 2-phase case-control (2PCC) design in identifying dose-response relationships between cumulative occupational exposure as assessed by experts and lung cancer incidence in an actual study. A population-based case-control study including 246 cases and 531 controls was conducted in an area with high lung cancer rates in Northeast France. Detailed occupational and personal risk factors were obtained in face-to-face interviews. Cumulative expert-based exposure scores were obtained from a subset of 215 cases and 269 controls stratified on smoking and a prior algorithmic exposure score for asbestos, crystalline silica, and polycyclic aromatic hydrocarbons (PAH) in the framework of a 2PCC design. This subset deliberately under-sampled large strata among controls but not among cases. Logistic regression models adapted to 2PCC studies were applied and corresponding computations of attributable fractions and their confidence intervals developed. Based on this 2PCC design, statistically significant dose-response relationships were obtained for asbestos, crystalline silica, PAH, and diesel motor exhaust. Simulations within this study showed that 2PCC studies were always more powerful than random samples. The 2PCC design may be the design of choice when resources allow only a limited number of subjects with a full expert-based exposure assessment.


Synergy between the Host Immune System and Bacteriophage Is Essential for Successful Phage Therapy against an Acute Respiratory Pathogen.
The rise of multi-drug-resistant (MDR) bacteria has spurred renewed interest in the use of bacteriophages in therapy. However, mechanisms contributing to phage-mediated bacterial clearance in an animal host remain unclear. We investigated the effects of host immunity on the efficacy of phage therapy for acute pneumonia caused by MDR Pseudomonas aeruginosa in a mouse model. Comparing efficacies of phage-curative and prophylactic treatments in healthy immunocompetent, MyD88-deficient, lymphocyte-deficient, and neutrophil-depleted murine hosts revealed that neutrophil-phage synergy is essential for the resolution of pneumonia. Population modeling of in vivo results further showed that neutrophils are required to control both phage-sensitive and emergent phage-resistant variants to clear infection. This "immunophage synergy" contrasts with the paradigm that phage therapy success is largely due to bacterial permissiveness to phage killing. Lastly, therapeutic phages were not cleared by pulmonary immune effector cells and were immunologically well tolerated by lung tissues.


Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5.
Large plasmids from Agrobacterium tumefaciens, Salmonella typhimurium, Escherichia coli, Pseudomonas putida, and Pseudomonas aeruginosa were routinely and consistently isolated using a procedure which does not require ultracentrifugation but includes steps designed to separate large-plasmid DNA from the bacterial folded chromosome. It also selectively removes fragments of broken chromosome. A variety of large plasmids was readily visualized with agarose gel electorphoresis, including five between 70 and 85 megadaltons (Mdal) in size, six between 90 and 143 Mdal, one that was larger than 200 Mdal, and one that was larger than 300 Mdal. This isolation procedure allowed initial estimation of the molecular sizes of the two IncP2 plasmids, pMG1 and pMG5, which were 312 and 280 Mdal, respectively. A standard curve for size determination by gel electrophoresis including plasmids between 23 and 143 Mdal in size did not extrapolate linearly for plasmids of the 300-Mdal size range. Unique response of different plasmids to the isolation procedure included sensitivity of IncP1 plasmids to high pH and the co-isolation of a 20-Mdal "cryptic" plasmid in conjunction.


Cdc25A is a novel phosphatase functioning early in the cell cycle.
The cdc25+ tyrosine phosphatase is a key mitotic inducer of the fission yeast Schizosaccharomyces pombe, controlling the timing of the initiation of mitosis. Mammals contain at least three cdc25+ homologues called cdc25A, cdc25B and cdc25C. In this study we investigate the biological function of cdc25A. Although very potent in rescuing the S.pombe cdc25 mutant, cdc25A is less structurally related to the S.pombe enzyme. Northern and Western blotting detection reveals that unlike cdc25B, cdc25C and cdc2, cdc25A is predominantly expressed in late G1. Moreover, immunodepletion of cdc25A in rat cells by microinjection of a specific antibody effectively blocks their cell cycle progression from G1 into the S phase, as determined by laser scanning single cell cytometry. These results indicate that cdc25A is not a mitotic regulator but a novel phosphatase that plays a crucial role in the start of the cell cycle. In view of its strong ability to activate cdc2 kinase and its specific expression in late G1, cdc2-related kinases functioning early in the cell cycle may be targets for this phosphatase.


Single surgeon experience with laparoscopic surgery in pediatric patients with inflammatory bowel disease.
Laparoscopic management of inflammatory bowel disease (IBD) has been well established in adults. Its efficacy and safety are less defined in pediatrics. We reviewed our single surgeon laparoscopic experience with pediatric IBD patients. All consecutive patients 17 years old or younger with IBD who underwent initial laparoscopic surgery for management of disease from May 2002 to May 2011 were identified. Patients were analyzed retrospectively with respect to age, gender, weight, American Society of Anesthesiologists classification, history of prior abdominal surgery, operative time, need for conversion to open procedure, estimated blood loss, need for blood transfusion, length of hospitalization, need for intensive care unit monitoring, and patient outcomes within 30 days. In total, 44 pediatric patients underwent 52 laparoscopic surgeries. Forty-two patient cases (80.8%) had had no prior abdominal surgery. Mean operative time was 132 minutes. Two patients (3.8%) underwent conversion from a laparoscopic to open approach. Mean estimated blood loss was 64 mL. The average hospitalization was 5.5 days. Postoperative complications occurred in 10 patients (19.2%): 2 were treated for wound infection or abscess, 7 required re-admission, and 1 required return to the operating room for exploratory laparotomy. Laparoscopic management of IBD has not routinely been implemented in the pediatric population. We demonstrate acceptable perioperative morbidity and rate of conversion to open with an overall successful laparoscopic approach in pediatric patients. This may have particular long-term benefits in a population faced with longer life expectancy. An initial laparoscopic approach is a feasible and safe option for pediatric IBD patients.


The (111) surface of NaAu2: structure, composition, and stability.
The (111) surface of single-crystal NaAu(2) is a model for catalytically active, powdered NaAu(2). We prepare and characterize this surface with a broad suite of techniques. Preparation in ultrahigh vacuum consists of the traditional approach of ion bombardment (to remove impurities) and thermal annealing (to restore surface order). Both of these steps, however, cause loss of sodium (Na), and repeated treatments eventually trigger conversion of the surface and near-surface regions to crystalline gold. The bulk has a limited ability to repopulate the surface Na. Under conditions where Na depletion is minimized, electron diffraction patterns are consistent with the bulk-terminated structure, and scanning tunneling microscopy reveals mesa-like features with lateral dimensions of a few tens of nanometers. The tops of the mesas do not possess fine structure characteristic of a periodic lattice, suggesting that the surface layer is disordered under the conditions of these experiments.


Investigation of the effects of intravenous magnesium sulphate on cardiac rhythm in acute myocardial infarction.
To examine the effect of doubling serum magnesium concentration on the incidence of arrhythmias in patients with suspected acute myocardial infarction. Randomised double blind clinical trial. Coronary care unit of a teaching hospital. Clinical data were collected on 2316 randomised patients with suspected acute myocardial infarction. Holter monitoring was performed in a subgroup of 70 patients and analysed in 48 patients in whom acute myocardial infarction was confirmed. By random allocation, patients received either an intravenous loading dose of 8 mmol magnesium sulphate over five minutes plus 65 mmol over the next 24 hours, or equal volumes of saline. (a) Clinically documented arrhythmias; (b) use of antiarrhythmic treatments, cardioversion, and insertion of a pacemaker; (c) incidence of all abnormal rhythms during Holter monitoring. In the main trial the incidence of rhythm disturbance while in the coronary care unit (expressed as the odds ratio (OR) for magnesium: placebo and its 95% confidence interval) was not significantly different between treatment groups for ventricular fibrillation (OR 0.74; 0.46 to 1.20), ventricular tachycardia (OR 0.87; 0.63 to 1.20), supraventricular tachycardia (OR 0.69; 0.38 to 1.26), atrial fibrillation (OR 0.92; 0.69 to 1.23), or heart block of any degree (OR 1.17; 0.83 to 1.65). Sinus bradycardia was significantly more common in the magnesium group (OR 1.38; 1.03 to 1.85; p = 0.02). These findings were corroborated by the use of treatments for rhythm disturbance and the data from Holter monitoring. The regimen of intravenous magnesium sulphate used here had no significant effect on arrhythmia in acute myocardial infarction. The reduction in mortality that has been shown with this form of treatment is not attributable to suppression of life threatening rhythm disturbances.


Comparison of Natural Language Processing Rules-based and Machine-learning Systems to Identify Lumbar Spine Imaging Findings Related to Low Back Pain.
To evaluate a natural language processing (NLP) system built with open-source tools for identification of lumbar spine imaging findings related to low back pain on magnetic resonance and x-ray radiology reports from four health systems. We used a limited data set (de-identified except for dates) sampled from lumbar spine imaging reports of a prospectively assembled cohort of adults. From N = 178,333 reports, we randomly selected N = 871 to form a reference-standard dataset, consisting of N = 413 x-ray reports and N = 458 MR reports. Using standardized criteria, four spine experts annotated the presence of 26 findings, where 71 reports were annotated by all four experts and 800 were each annotated by two experts. We calculated inter-rater agreement and finding prevalence from annotated data. We randomly split the annotated data into development (80%) and testing (20%) sets. We developed an NLP system from both rule-based and machine-learned models. We validated the system using accuracy metrics such as sensitivity, specificity, and area under the receiver operating characteristic curve (AUC). The multirater annotated dataset achieved inter-rater agreement of Cohen's kappa > 0.60 (substantial agreement) for 25 of 26 findings, with finding prevalence ranging from 3% to 89%. In the testing sample, rule-based and machine-learned predictions both had comparable average specificity (0.97 and 0.95, respectively). The machine-learned approach had a higher average sensitivity (0.94, compared to 0.83 for rules-based), and a higher overall AUC (0.98, compared to 0.90 for rules-based). Our NLP system performed well in identifying the 26 lumbar spine findings, as benchmarked by reference-standard annotation by medical experts. Machine-learned models provided substantial gains in model sensitivity with slight loss of specificity, and overall higher AUC.


Impact of COVID-19 national lockdown on asthma exacerbations: interrupted time-series analysis of English primary care data.
The impact of COVID-19 and ensuing national lockdown on asthma exacerbations is unclear. We conducted an interrupted time-series (lockdown on 23 March 2020 as point of interruption) analysis in asthma cohort identified using a validated algorithm from a national-level primary care database, the Optimum Patient Care Database. We derived asthma exacerbation rates for every week and compared exacerbation rates in the period: January to August 2020 with a pre-COVID-19 period and January to August 2016-2019. Exacerbations were defined as asthma-related hospital attendance/admission (including accident and emergency visit), or an acute course of oral corticosteroids with evidence of respiratory review, as recorded in primary care. We used a generalised least squares modelling approach and stratified the analyses by age, sex, English region and healthcare setting. From a database of 9 949 387 patients, there were 100 165 patients with asthma who experienced at least one exacerbation during 2016-2020. Of 278 996 exacerbation episodes, 49 938 (17.9%) required hospital visit. Comparing pre-lockdown to post-lockdown period, we observed a statistically significant reduction in the level (-0.196 episodes per person-year; p<0.001; almost 20 episodes for every 100 patients with asthma per year) of exacerbation rates across all patients. The reductions in level in stratified analyses were: 0.005-0.244 (healthcare setting, only those without hospital attendance/admission were significant), 0.210-0.277 (sex), 0.159-0.367 (age), 0.068-0.590 (region). There has been a significant reduction in attendance to primary care for asthma exacerbations during the pandemic. This reduction was observed in all age groups, both sexes and across most regions in England.


Detection of promoter activity by flow cytometric analysis of GFP reporter expression.
Low efficiency of transfection is often the limiting factor for acquiring conclusive data in reporter assays. It is especially difficult to efficiently transfect and characterize promoters in primary human cells. To overcome this problem we have developed a system in which reporter gene expression is quantified by flow cytometry. In this system, green fluorescent protein (GFP) reporter constructs are co-transfected with a reference plasmid that codes for the mouse cell surface antigen Thy-1.1 and serves to determine transfection efficiency. Comparison of mean GFP expression of the total transfected cell population with the activity of an analogous luciferase reporter showed that the sensitivity of the two reporter systems is similar. However, because GFP expression can be analyzed at the single-cell level and in the same cells the expression of the reference plasmid can be monitored by two-color fluorescence, the GFP reporter system is in fact more sensitive, particularly in cells which can only be transfected with a low efficiency.


Charge states of y ions in the collision-induced dissociation of doubly charged tryptic peptide ions.
Bonds that break in collision-induced dissociation (CID) are often weakened by a nearby proton, which can, in principle, be carried away by either of the product fragments. Since peptide backbone dissociation is commonly charge-directed, relative intensities of charge states of product y- and b-ions depend on the final location of that proton. This study examines y-ion charge distributions for dissociation of doubly charged peptide ions, using a large reference library of peptide ion fragmentation generated from ion-trap CID of peptide ions from tryptic digests. Trends in relative intensities of y(2+) and y(1+) ions are examined as a function of bond cleavage position, peptide length (n), residues on either side of the bond and effects of residues remote from the bond. It is found that y(n-2)/b(2) dissociation is the most sensitive to adjacent amino acids, that y(2+)/y(1+) steadily increase with increasing peptide length, that the N-terminal amino acid can have a major influence in all dissociations, and in some cases other residues remote from the bond cleavage exert significant effects. Good correlation is found between the values of y(2+)/y(1+) for the peptide and the proton affinities of the amino acids present at the dissociating peptide bond. A few deviations from this correlation are rationalized by specific effects of the amino acid residues. These correlations can be used to estimate trends in y(2+)/y(1+) ratios for peptide ions from amino acid proton affinities.


School-Based Preventive Dental Program in Rural Communities of the Republic of Armenia.
Objectives: This paper describes a school-based preventive dental program implemented in 14 rural schools within nine villages of Armenia. As part of the program, school-based toothbrushing stations (called Brushadromes) were installed in the participating schools. The intervention included school-based supervised toothbrushing with fluoride toothpaste and oral hygiene education. Methods: The study evaluates the prevalence and levels of dental caries among rural schoolchildren in 2013 (before the implementation of the preventive program, referred to as a pre-intervention group) and 2017 (4 years after the start of the program, referred to as an intervention group) in two randomly selected villages where the program was implemented. A repeated cross-sectional study design was used. The prevalence of caries and the number of decayed, missing, and filled teeth in permanent dentition (DMFT) and primary dentition (dmft) were recorded among 6-7 and 10-11-year-old schoolchildren in 2013 (n = 166) and 2017 (n = 148). The pre-intervention and intervention groups include different children in the same age range, from the same villages, examined at different time points. In both instances, they represented over 95% of the 6-7 and 10-11-year-old student populations of the studied villages. Pearson Chi-square, Fisher's Exact test, independent t-test, and quasi-likelihood Poisson regression were utilized for data analysis. Results: Schoolchildren involved in the intervention had significantly less decay levels compared to same-age pre-intervention groups. For 10-11-year-old schoolchildren involved in the program, the mean number of permanent teeth with caries was lower by a factor of 0.689 (lower by 31.1%), p = 0.008, 95% CI, 0.523; 0.902, compared to the 10-11-year-old pre-intervention group, after controlling for age, sex, child's socio-economic vulnerability status, the village of residence, and the number of permanent teeth with fillings. Conclusions: The study indicates a significantly lower level of caries among schoolchildren in the studied two villages where the intervention was implemented. The described intervention is particularly suitable in rural settings where water fluoridation is not available and homes have limited availability of running water.


Aneurysms with persistent filling after failed treatment with the Pipeline embolization device.
The Pipeline embolization device (PED) has become a valuable tool in the treatment of cerebral aneurysms. Although failures with PED treatment have been reported, the characteristics and course of these aneurysms remain a topic of uncertainty. Electronic medical records and imaging studies were reviewed for all patients treated with the PED between July 2010 and March 2015 to identify characteristics of patients and aneurysms with residual filling after PED treatment. Of 316 cases treated at a single institution, 281 patients had a long-term follow-up. A total of 52 (16.4%) aneurysms with residual filling were identified and constituted the study population. The mean patient age in this population was 58.8 years. The mean aneurysm size was 10.1 mm ± 7.15 mm. Twelve aneurysms were fusiform (23%). Of the aneurysms with residual filling, there were 20 carotid ophthalmic (CO) aneurysms (20% of all CO aneurysms treated), 10 other paraclinoid aneurysms (16.4% of all paraclinoid aneurysms), 7 posterior communicating artery (PCoA) aneurysms (21.9% of all PCoA aneurysms), 7 cavernous internal carotid artery (ICA) aneurysms (14.9% of all cavernous ICA aneurysms), 4 vertebrobasilar (VB) junction aneurysms (14.8% of all VB junction aneurysms), and 3 middle cerebral artery (MCA) aneurysms (25% of all MCA aneurysms). Eleven patients underwent placement of more than one PED (21.2%), with a mean number of devices of 1.28 per case. Eight of 12 aneurysms were previously treated with a stent (15.4%). Nineteen patients underwent re-treatment (36.5%); the 33 patients who did not undergo re-treatment (63.5%) were monitored by angiography or noninvasive imaging. In multivariate analysis, age older than 65 years (OR 2.65, 95% CI 1.33-5.28; p = 0.05), prior stent placement across the target aneurysm (OR 2.94, 95% CI 1.15-7.51; p = 0.02), aneurysm location in the distal anterior circulation (MCA, PCoA, and anterior choroidal artery: OR 2.72, 95% CI 1.19-6.18; p = 0.017), and longer follow-up duration (OR 1.06, 95% CI 1.03-1.09; p < 0.001) were associated with incomplete aneurysm occlusion. While the PED can allow for treatment of large, broad-necked aneurysms with high efficacy, treatment failures do occur (16.4%). Aneurysm size, shape, and previous treatment may influence treatment outcome.


Transcranial alternating current stimulation at 10 Hz modulates response bias in the Somatic Signal Detection Task.
Ongoing, pre-stimulus oscillatory activity in the 8-13 Hz alpha range has been shown to correlate with both true and false reports of peri-threshold somatosensory stimuli. However, to directly test the role of such oscillatory activity in behaviour, it is necessary to manipulate it. Transcranial alternating current stimulation (tACS) offers a method of directly manipulating oscillatory brain activity using a sinusoidal current passed to the scalp. We tested whether alpha tACS would change somatosensory sensitivity or response bias in a signal detection task in order to test whether alpha oscillations have a causal role in behaviour. Active 10 Hz tACS or sham stimulation was applied using electrodes placed bilaterally at positions CP3 and CP4 of the 10-20 electrode placement system. Participants performed the Somatic Signal Detection Task (SSDT), in which they must detect brief somatosensory targets delivered at their detection threshold. These targets are sometimes accompanied by a light flash, which could also occur alone. Active tACS did not modulate sensitivity to targets but did modulate response criterion. Specifically, we found that active stimulation generally increased touch reporting rates, but particularly increased responding on light trials. Stimulation did not interact with the presence of touch, and thus increased both hits and false alarms. TACS stimulation increased reports of touch in a manner consistent with our observational reports, changing response bias, and consistent with a role for alpha activity in somatosensory detection.


The sol-gel transition of ultra-low solid content TEMPO-cellulose nanofibril/mixed-linkage β-glucan bionanocomposite gels.
We present the preparation, morphological analysis, and rheological characterization of ultra-low solid content gels prepared by physically cross-linking TEMPO-oxidized cellulose nanofibrils (TEMPO-CNF) with the soluble plant-cell-wall polysaccharide, mixed-linkage β-glucan (MLG). Of particular note, gel formation was rapidly induced by very small amounts of MLG (e.g. 0.125% w/v) at extremely low TEMPO-CNF concentration (0.05% w/v), which independently were otherwise fluid and thus easily handled. Rheology of these bionanocomposite gel systems as a function of MLG and TEMPO-CNF concentrations revealed that the critical gel concentration of MLG and TEMPO-CNF followed a power-law relation of the concentration of the other component. Surprisingly, these systems also exhibited an additional transition to thick gels at high TEMPO-CNF and MLG concentrations that was visible only at low frequencies. Cryogenic scanning electron microscopy (cryo-SEM) imaging of admixture solutions and gels revealed increased network crowding with increasing MLG amounts. The data are consistent with the hypothesis that non-covalent cellulose-MLG interactions, analogous to those occurring within plant cell walls, drive gel formation. The ability to tune gel physical properties simply by controlling CNF (a promising forest bioproduct) and MLG (a readily available agricultural polysaccharide) fractions at very low solid and polymer content opens new possibilities for material applications in diverse industries.


A possible initial folding intermediate: the C-terminal proteolytic domain of tryptophan synthase beta chains folds in less than 4 milliseconds into a condensed state with non-native-like secondary structure.
The isolated F2-V8 peptide corresponding to the 101 C-terminal residues of Escherichia coli tryptophan synthase beta chains folds into a heat-stable, yet fluctuating, condensed state that contains a lot of secondary structure. However, this state has non-native-like secondary and supersecondary structures [Chaffotte, A., Guillou, Y., Delepierre, M., Hinz, H.-J., & Goldberg, M. E. (1991) Biochemistry 30, 8067-8074]. To characterize the rate of appearance of this state, stopped-flow studies on the far-ultraviolet circular dichroism (CD) and on the binding of 1-anilino-8-naphthalenesulfonate (ANS) have been conducted during the folding of guanidine-unfolded F2-V8. It was shown that both the CD signal at 222 nm and the ANS binding properties of folded isolated F2-V8 were regained, at 20 degrees C, within the dead time of the stopped-flow apparatus, which was 4 ms. At 12 degrees C, the binding of ANS was also completed within this dead time, but the ellipticity showed some minor later changes. After a rapid overshoot of the CD signal that occurred during the 4-ms dead time, a small readjustment of the ellipticity to the final value occurred more slowly and was completed after about 25 ms. Thus, even at 12 degrees C, the hydrophobic core and most of the secondary structure of folded F2-V8 were formed in less than 4 ms. These observations strongly suggest that the previously described condensed non-native-like state of F2-V8 results from a very rapid, nonspecific, hydrophobic collapse. It is proposed that such a state may be a general early intermediate in protein folding.


Comparison of two methods of bacterial DNA extraction from human fecal samples contaminated with Clostridium perfringens, Staphylococcus aureus, Salmonella Typhimurium, and Campylobacter jejuni.
In this study, 2 methods of DNA extraction were evaluated for use in conjunction with the screening system Rapid Foodborne Bacterial Screening 24 (RFBS24), which employs multiplex real-time SYBR Green polymerase chain reaction (SG-PCR) and can simultaneously detect 24 target genes of foodborne pathogens in fecal DNA samples. The QIAamp DNA Stool mini kit (Qkit) and Ultra Clean Fecal DNA Isolation Kit (Ukit) were used for bacterial DNA extraction from fecal samples artificially inoculated with Clostridium perfringens, Staphylococcus aureus, Salmonella Typhimurium, and Campylobacter jejuni. SG-PCR and simplex real-time quantitative PCR (S-qPCR) analyses revealed higher copy numbers (8-234 times) of DNA in samples obtained using Ukit compared with those obtained using Qkit, resulting in lower cycle threshold values for the Ukit samples of the 4 bacteria on SG-PCR analysis. Fecal DNA samples from patients infected during foodborne outbreaks of Salmonella and Campylobacter were also prepared by Qkit and Ukit methods and subjected to RFBS24 analyses. Higher numbers of RFBS24 bacterial target genes were detected in DNA samples obtained using Ukit compared with those obtained using Qkit. Thus, the higher DNA extraction efficiency of the Ukit method compared with Qkit renders the former more useful in achieving improved detection rates of these 4 bacteria in fecal samples using SG-PCR.


Cognitive-behavioral mediators of changing multiple behaviors: smoking and a sedentary lifestyle.
A significant percentage of the U.S. population has multiple poor health behaviors. Understanding the relationship among these behavioral risk factors is important for designing effective multiple risk factor interventions. While there is some evidence suggesting that participation in physical exercise may have a positive impact on smoking cessation, there is much to be learned about the relationships between cognitive-behavioral (self-efficacy, decisional-balance) and motivational mechanisms (stage of change) which have been shown to mediate changes in both exercise and smoking behavior. The sample comprised 332 smokers employed at two workplaces-a government agency and a medical center-recruited as part of a larger worksite health promotion project and who completed questionnaires on their smoking and exercise behaviors. The results revealed significant relationships between smoking variables and exercise variables. Smokers who rated as important the positive benefits of smoking also rated as important the costs associated with increased physical activity. Similarly, the negative consequences of smoking were significantly associated with the positive benefits of physical activity. Self-efficacy for one behavior was significantly associated with self-efficacy for the other. Significant differences by exercise and smoking stage of change were found on the cross-behavior sets of variables (self-efficacy, pros, cons). Smokers who were contemplating a more active lifestyle reported the negative consequences of smoking to be significantly more important to them than smokers who were not considering adoption of a more active lifestyle. Smokers who were exercising regularly reported significantly more confidence in their ability to refrain from smoking than smokers not exercising regularly. Finally, smokers preparing for quitting reported less confidence in their ability to exercise than smokers who had already taken action to change their smoking behavior. The cognitive mechanisms associated with changes in smoking behavior are related to the cognitive variables which have been shown to predict changes in exercise behavior. Significant relationships in mediating mechanisms including decisional balance and self-efficacy between smoking and exercise provide preliminary information on how change in one risk behavior may relate to change in another. These associations have implications for future intervention research and for methods research on multiple risk factors interactions.


Molecular basis of the attenuated phenotype of human APOBEC3B DNA mutator enzyme.
The human APOBEC3A and APOBEC3B genes (A3A and A3B) encode DNA mutator enzymes that deaminate cytidine and 5-methylcytidine residues in single-stranded DNA (ssDNA). They are important sources of mutations in many cancer genomes which show a preponderance of CG->TA transitions. Although both enzymes can hypermutate chromosomal DNA in an experimental setting, only A3A can induce double strand DNA breaks, even though the catalytic domains of A3B and A3A differ by only 9% at the protein level. Accordingly we sought the molecular basis underlying A3B attenuation through the generation of A3A-A3B chimeras and mutants. It transpires that the N-terminal domain facilitates A3B activity while a handful of substitutions in the catalytic C-terminal domain impacting ssDNA binding serve to attenuate A3B compared to A3A. Interestingly, functional attenuation is also observed for the rhesus monkey rhA3B enzyme compared to rhA3A indicating that this genotoxic dichotomy has been selected for and maintained for some 38 million years. Expression of all human ssDNA cytidine deaminase genes is absent in mature sperm indicating they contribute to somatic mutation and cancer but not human diversity.


Carbonodraco lundi gen et sp. nov., the oldest parareptile, from Linton, Ohio, and new insights into the early radiation of reptiles.
Redescription of the holotype specimen of Cephalerpeton ventriarmatum Moodie, 1912, from the Middle Pennsylvanian (Moscovian) Francis Creek Shale of Mazon Creek, Illinois, confirms that it is a basal eureptile with close postcranial similarities to other protorothyridids, such as Anthracodromeus and Paleothyris. The skull is long and lightly built, with large orbits and a dorsoventrally short mandible similar to most basal eureptiles. Two specimens referred previously to Cephalerpeton cf. C. ventriarmatum from the approximately coeval Linton, Ohio, locality differ significantly from the holotype in cranial and mandibular proportions and tooth morphology. This material and an additional Linton specimen compare favourably to 'short-faced' parareptiles, such as Colobomycter and Acleistorhinus, and justify recognition of an acleistorhinid parareptile in the Linton assemblage. The new binomen is thus the oldest known parareptile.


Diagnosis and treatment of psychogenic parkinsonism.
Parkinson's disease (PD) is the most common movement disorder encountered in movement disorder clinics but psychogenic parkinsonism (PP) is relatively rare. Based on a study of 32 patients, coupled with a comprehensive review of the literature, the diagnosis, clinical phenomenology, epidemiology, natural history, imaging and treatment, this article provides a critical review of PP. In addition to other PD-like symptoms, patients with PP often exhibit abrupt onset, typically in response to a stressful event, followed by a fluctuating course, early disability, bilateral shaking and slowness, non-decremental slowness when performing repetitive movements, voluntary resistance against passive movement without cogwheel rigidity, distractibility, generalised and 'give-way' weakness, stuttering speech, bizarre gait and a variety of behavioural, sexual and other motor and non-motor symptoms. Levodopa related motor complications do not occur even in the setting of chronic, high dose therapy. In some cases, differentiation between PD and PP is very difficult, and in those cases the response to placebo may be helpful. A comprehensive, multidisciplinary assessment of patients with PP, when combined with insight oriented psychotherapy, physiotherapy and treatment of comorbid depression, is often helpful, although the prognosis for this group of disorders remains relatively poor.