Application of the symplectic finite-difference time-domain method to light scattering by small particles.
A three-dimensional fourth-order finite-difference time-domain (FDTD) program with a symplectic integrator scheme has been developed to solve the problem of light scattering by small particles. The symplectic scheme is nondissipative and requires no more storage than the conventional second-order FDTD scheme. The total-field and scattered-field technique is generalized to provide the incident wave source conditions in the symplectic FDTD (SFDTD) scheme. The perfectly matched layer absorbing boundary condition is employed to truncate the computational domain. Numerical examples demonstrate that the fourth-order SFDTD scheme substantially improves the precision of the near-field calculation. The major shortcoming of the fourth-order SFDTD scheme is that it requires more computer CPU time than a conventional second-order FDTD scheme if the same grid size is used. Thus, to make the SFDTD method efficient for practical applications, one needs to parallelize the corresponding computational code.


Duplex ultrasound investigation of the veins in chronic venous disease of the lower limbs--UIP consensus document. Part II. Anatomy.
Duplex ultrasound investigation has become the reference standard in assessing the morphology and haemodynamics of the lower limb veins. The project described in this paper was an initiative of the Union Internationale de Phlébologie (UIP). The aim was to obtain a consensus of international experts on the methodology to be used for assessment of anatomy of superficial and perforating veins in the lower limb by ultrasound imaging. The authors performed a systematic review of the published literature on duplex anatomy of the superficial and perforating veins of the lower limbs; afterwards they invited a group of experts from a wide range of countries to participate in this project. Electronic submissions from the authors and the experts (text and images) were made available to all participants via the UIP website. The authors prepared a draft document for discussion at the UIP Chapter meeting held in San Diego, USA in August 2003. Following this meeting a revised manuscript was circulated to all participants and further comments were received by the authors and included in subsequent versions of the manuscript. Eventually, all participants agreed the final version of the paper. The experts have made detailed recommendations concerning the methods to be used for duplex ultrasound examination as well as the interpretation of images and measurements obtained. This document provides a detailed methodology for complete ultrasound assessment of the anatomy of the superficial and perforating veins in the lower limbs. The authors and a large group of experts have agreed a methodology for the investigation of the lower limbs venous system by duplex ultrasonography, with specific reference to the anatomy of the main superficial veins and perforators of the lower limbs in healthy and varicose subjects.


Data Resource Profile: Clinical Practice Research Datalink (CPRD).
The Clinical Practice Research Datalink (CPRD) is an ongoing primary care database of anonymised medical records from general practitioners, with coverage of over 11.3 million patients from 674 practices in the UK. With 4.4 million active (alive, currently registered) patients meeting quality criteria, approximately 6.9% of the UK population are included and patients are broadly representative of the UK general population in terms of age, sex and ethnicity. General practitioners are the gatekeepers of primary care and specialist referrals in the UK. The CPRD primary care database is therefore a rich source of health data for research, including data on demographics, symptoms, tests, diagnoses, therapies, health-related behaviours and referrals to secondary care. For over half of patients, linkage with datasets from secondary care, disease-specific cohorts and mortality records enhance the range of data available for research. The CPRD is very widely used internationally for epidemiological research and has been used to produce over 1000 research studies, published in peer-reviewed journals across a broad range of health outcomes. However, researchers must be aware of the complexity of routinely collected electronic health records, including ways to manage variable completeness, misclassification and development of disease definitions for research.


Influence of sensory integration procedures on language development.
The relationship between language development and sensory integration was explored through single case experimental studies of one female and three male aphasic children ranging in age from 4 years, 0 months to 5 years, 3 months. Other agencies had assessed all the children in the area of language development at least 6 months before the start of occupational therapy. Three of the four children had received either speech therapy, special education specific to aphasia, or both, before starting occupational therapy. Additional baseline data on language expression and comprehension, as well as on sensory integrative functioning, were gathered before beginning a year of occupational therapy that involved sensory integration procedures. Inspection of rate of language growth before and after starting occupational therapy showed a consistent increase in rate of growth in language comprehensive concomitant with occupational therapy compared to previous growth rate. The two children with depressed postrotary nystagmus demonstrated notable gains on expressive language measures.


All-Inside Anterior Cruciate Ligament Reconstruction-A Systematic Review of Techniques, Outcomes, and Complications.
The all-inside technique (AIT) for anterior cruciate ligament reconstruction (ACLR) is gaining popularity as a more anatomic, less invasive, technique with the potential for more rapid recovery. This systematic review aims to critically assess components of the technique, its safety profile, outcomes, and complications. PUBMED, EMBASE, and MEDLINE were searched for studies discussing primary ACLR using the AIT. Article screening, quality assessment, and data abstraction were completed in duplicate, and a minimal clinically important difference (MCID) was used to frame the descriptive results in a clinically significant context. A total of 13 studies satisfied the inclusion criteria. Five-hundred twenty six patients (mean age 31.9 ± 5.9 years) underwent ACLR using the AIT. The mean follow-up was 18.2 ± 7.7 months. Graft choice included autograft (73.8%) and allograft (26.2%). Drilling techniques for femoral sockets were outside-in (53.4%) and through the anteromedial portal (35.4%), whereas tibial sockets were drilled outside-in (35.0%) and through the superomedial portal (3.8%). The rehabilitation protocol had an immediate focus on obtaining full knee extension, jogging permitted 2 to 4 months postoperatively in 61.5% of studies, and return to cutting and pivoting sports 6 to 9 months postoperatively in 69.2% of studies. A MCID was reached for subjective International Knee Documentation Committee scores at 6, 12, and 24 months follow-up and Lysholm knee score at 24 months follow-up. An improvement in outcomes was most notably between 6 and 12 months postoperatively. There was a total of 31 complications (5.89%) and included graft rerupture (2.47%), loss of extension of 1° to 10° (1.14%), and cartilage or meniscus injuries on the operated knee (0.760%). Complications related to the surgical technique were not reported. The AIT for ACLR shows potential as a minimally invasive approach given the low graft failure rates and short-term improvements in knee function and stability, pain and patient important outcomes from this approach. Comparative studies with large sample sizes and a long-term follow-up are required to assess the proposed advantages of this technique. This is a Level IV study.


Glyoxal oxidase from Phanerochaete chrysosporium is a new radical-copper oxidase.
A free radical-coupled copper complex has been identified as the catalytic structure in the active site of glyoxal oxidase from Phanerochaete chrysosporium based on a combination of spectroscopic and biochemical studies. The native (inactive) enzyme is activated by oxidants leading to the elimination of the cupric EPR signal consistent with formation of an antiferromagnetically coupled radical-copper complex. Oxidation also leads to the appearance of a substoichiometric free radical EPR signal with an average g value (gav = 2.0055) characteristic of phenoxyl tau-radicals arising from a minority apoenzyme fraction. Optical absorption, CD, and spectroelectrochemical measurements on the active enzyme reveal complex spectra extending into the near IR and define the redox potential for radical formation (E 1/2 = 0.64 V versus NHE, pH 7.0). Resonance Raman spectra have identified the signature of a modified (cysteinyl-tyrosine) phenoxyl in the vibrational spectra of the active complex. This radical-copper motif has previously been found only in galactose oxidase, with which glyoxal oxidase shares many properties despite lacking obvious sequence identity, and catalyzing a distinct reaction. The enzymes thus represent members of a growing class of free radical metalloenzymes based on the radical-copper catalytic motif and appear to represent functional variants that have evolved to distinct catalytic roles.


Emotion perception and electrophysiological correlates in Huntington's disease.
This study aimed to characterise, emotion perception deficits in symptomatic Huntington's disease (HD) via the use of event-related potentials (ERPs). ERP data were recorded during a computerised facial expression task in 11 HD participants and 11 matched controls. Expression (scrambled, neutral, happy, angry, disgust) classification accuracy and intensity were assessed. Relationships between ERP indices and clinical disease characteristics were also examined. Accuracy was significantly lower for HD relative to controls, due to reduced performance for neutral, angry and disgust (but not happy) faces. Intensity ratings did not differ between groups. HD participants displayed significantly reduced visual processing amplitudes extending across pre-face (P100) and face-specific (N170) processing periods, whereas subsequent emotion processing amplitudes (N250) were similar across groups. Face-specific and emotion-specific derivations of the N170 and N250 ('neutral minus scrambled' and 'each emotion minus neutral', respectively) did not differ between groups. Our data suggest that the facial emotion recognition performance deficits in HD are primarily related to neural degeneration underlying 'generalised' visual processing, rather than face or emotional specific processing. ERPs are a useful tool to separate functionally discreet impairments in HD, and provide an important avenue for biomarker application that could more-selectively track disease progression.


Temporal dynamics of type 2 deiodinase expression after melatonin injections in Syrian hamsters.
In many species living in temperate zones, reproduction is controlled by the photoperiod. Recent findings have clarified that type 2 iodothyronine deiodinase (Dio2) plays a significant role in the photoperiodic response of gonads in the mediobasal hypothalamus, converting the prohormone T(4) into bioactive T(3). In mammals, Dio2 expression is suppressed by long-term melatonin injections, although the signal transduction pathways that link the melatonin signal to Dio2 expression are unknown. As a first step to approach the problem, we have here investigated the temporal dynamics of the melatonin effect on Dio2 expression using male Syrian hamsters. Dio2 mRNA levels were found to show diurnal rhythms under long-day conditions in an area adjacent to the tuberoinfundibular sulcus and in the ependymal cell layer lining the ventrobasal walls of the third ventricle. Daily sc melatonin injections given in the late afternoon under long-day condition suppressed the Dio2 mRNA levels already at the first day after the onset of the treatment in the ependymal cell layer lining the ventrobasal walls of the third ventricle, and 1 d later in an area adjacent to the tuberoinfundibular sulcus. These suppressive effects were sustained for at least 2 d after a single injection. Furthermore, we examined the temporal changes of the Dio2 expression after the onset of the treatment, showing that the suppression did not occur until midday of the next day. These data suggest that melatonin is involved in the signal transduction mechanisms controlling the photoperiodic response of gonads by acting on Dio2 expression rather rapidly through indirect pathways.


High Y-chromosomal Differentiation Among Ethnic Groups of Dir and Swat Districts, Pakistan.
The ethnic groups that inhabit the mountainous Dir and Swat districts of northern Pakistan are marked by high levels of cultural and phenotypic diversity. To obtain knowledge of the extent of genetic diversity in this region, we investigated Y-chromosomal diversity in five population samples representing the three main ethnic groups residing within these districts, including Gujars, Pashtuns and Kohistanis. A total of 27 Y-chromosomal short tandem repeats (Y-STRs) and 331 Y-chromosomal single nucleotide polymorphisms (Y-SNPs) were investigated. In the Y-STRs, we observed very high and significant levels of genetic differentiation in nine of the 10 pairwise between-group comparisons (RST 0.179-0.746), and the differences were mirrored in the Y-SNP haplogroup frequency distribution. No genetic differences were found between the two Pashtun subethnic groups Tarklanis and Yusafzais (RST = 0.000). Utmankhels, also considered Pashtuns culturally, were not closely related to any of the other population samples (RST 0.451-0.746). Thus, our findings provide examples of both associations and dissociations between cultural and genetic legacies. When analyzed within a larger continental-scale context, these five ethnic groups fall mostly outside the previously characterized Y-chromosomal gene pools of the Indo-Pakistani subcontinent. Male founder effects, coupled with culturally and topographically based constraints upon marriage and movement, are likely responsible for the high degree of genetic structure in this region.


Role of PH Monitoring in Laryngopharyngeal Reflux Patients with Voice Disorders.
Laryngopharyngeal reflux (LPR) is considered an important cause of voice disorder. We aimed to determine the frequency of LPR in patients with voice disorder and the association between Koufman Reflux Symptom Index (RSI), Reflux Finding Score (RFS), gastroesophageal reflux disease (GERD), and proximal acid reflux in these patients. We performed a prospective study in patients aged more than 18 years presenting at the ear, nose, and throat (ENT) clinic with a change in voice lasting more than 3 weeks. All patients underwent nasopharyngolaryngoscopy and a dual-probe esophageal pH study. LPR was diagnosed by a Koufman RSI of >13 and/or RFS of >7. GERD was diagnosed according to a DeMeester Johnson score of >14.7. Proximal acid reflux was diagnosed if acid exposure time was >0.02% in a proximal pH probe. The study included 30 patients with a voice disorder. The mean age of participants was 38.5 years and 40% of patients were female. Using either of the two criteria, LPR was present in 46.7% of patients, half of whom had GERD. Among the remaining 53.3% patients with a voice disorder and no evidence of LPR, GERD was present in 25%. There was no significant association between the presence of LPR based on RSI (P=1) and GERD or RFS and GERD (P=0.06). Proximal acid reflux was present in only 10% patients with a voice disorder, and there was no significant association of this test with RFS (P=1) or RSI (P=1). Approximately half of the patients with a voice disorder have LPR, and only a subset of these patients have evidence of GERD. Fiberoptic laryngoscopic findings (RFS) complementing RSI appears to be important in diagnosing possible reflux etiology in voice disorders and can be an indicator for instituting anti-reflux therapy. However, there is no significant association between RSI, RFS, and GERD suggesting that these tests evaluate different features of the disease. Proximal acid reflux is uncommon in patients with voice disorder based on current measurement criteria. Acid exposure time as measured in the proximal probe of a 24-hour dual pH probe may need to be re-evaluated as one of the diagnostic criteria for LPR.


Long-term outcomes of over 8,000 medial Oxford Phase 3 Unicompartmental Knees-a systematic review.
Background and purpose - There is debate as to the relative merits of unicompartmental and total knee arthroplasty (UKA, TKA). Although the designer surgeons have achieved good results with the Oxford UKA there is concern over the reproducibility of these outcomes. Therefore, we evaluated published long-term outcomes of the Oxford Phase 3 UKA. Patients and methods - We searched databases to identify studies reporting ≥10 year outcomes of the medial Oxford Phase 3 UKA. Revision, non-revision, and re-operation rates were calculated per 100 component years (% pa). Results - 15 studies with 8,658 knees were included. The annual revision rate was 0.74% pa (95% CI 0.67-0.81, n = 8,406) corresponding to a 10-year survival of 93% and 15-year survival of 89%. The non-revision re-operation rate was 0.19% pa (95% CI 0.13-0.25, n = 3,482). The re-operation rate was 0.89% pa (95% CI 0.77-1.02, n = 3,482). The most common causes of revision were lateral disease progression (1.42%), aseptic loosening (1.25%), bearing dislocation (0.58%), and pain (0.57%) (n = 8,658). Average OKS scores were 40 at 10 years (n = 3,417). The incidence of medical complications was 0.83% (n = 1,443). Interpretation - Very good outcomes were achieved by both designer and non-designer surgeons. The PROMs, medical complication rate, and non-revision re-operation rate were better than those found in meta-analyses and publications for TKA but the revision rate was higher. However, if failure is considered to be all re-operations and not just revisions, then the failure rate of UKA was less than that of TKA.


Emergency preparedness for the home healthcare nurse.
Recent events are quickly becoming the hard teacher: giving tests of preparedness first, then allowing for lessons afterward. However, each emergency teaches valuable lessons about assessment and preparation, and each lesson learned can be integrated into assessment of and preparation for the next event. This article outlines key steps that home care nurses can take to ensure patient safety during emergencies or disasters.


Working memory and new learning following pediatric traumatic brain injury.
Working memory (WM), the ability to monitor, process and maintain task relevant information on-line to respond to immediate environmental demands, is controlled by frontal systems (D'Esposito et al., 2006), which are particularly vulnerable to damage from a traumatic brain injury (TBI). This study employed the adult-based Working Memory model of Baddeley and Hitch (1974) to examine the relationship between working memory function and new verbal learning in children with TBI. A cross-sectional sample of 36 school-aged children with a moderate to severe TBI was compared to age-matched healthy Controls on a series of tasks assessing working memory subsystems: the Phonological Loop (PL) and Central Executive (CE). The TBI group performed significantly more poorly than Controls on the PL measure and the majority of CE tasks. On new learning tasks, the TBI group consistently produced fewer words than Controls across the learning and delayed recall phases. Results revealed impaired PL function related to poor encoding and acquisition on a new verbal learning task in the TBI group. CE retrieval deficits in the TBI group contributed to general memory dysfunction in acquisition, retrieval and recognition memory. These results suggest that the nature of learning and memory deficits in children with TBI is related to working memory impairment.


Multifunctional Optical Tomography System With High-Fidelity Surface Extraction Based on a Single Programmable Scanner and Unified Pinhole Modeling.
Macroscopic optical tomography is a non-invasive method that can visualize the 3D distribution of intrinsic optical properties or exogenous fluorophores, making it highly attractive for small animal imaging. However, reconstructing the images requires prior knowledge of surface information. To address this, existing systems often use additional hardware components or integrate multimodal information, which is expensive and introduces new issues such as image registration. Our goal is to develop a multifunctional optical tomography system that can extract surface information using a concise hardware design. Our proposed system uses a single programmable scanner to implement both surface extraction and optical tomography functions. A unified pinhole model is used to describe both the illumination and detection procedures for capturing 3D point cloud. Line-shaped scanning is adopted to improve both spatial resolution and speed of surface extraction. Finally, we integrate the extracted surface information into the optical tomographic reconstruction to more accurately map the fluorescence distribution. Comprehensive phantom experiments with different levels of complexity were designed to evaluate the performance of surface extraction and fluorescence tomography. We also imaged the axillary lymph nodes in living mice after injection of fluorophore, demonstrating the proposed system facilitates more reliable fluorescence tomography. We have successfully developed a versatile optical tomography system by leveraging concise hardware design and unified pinhole modeling. Phantom validation demonstrates that our system provides high-precision surface information with a maximum error of 0.1 mm, while the surface-guided FMT reconstruction is more reliable than the blind reconstruction using simplified surface geometry, elevating several quantitative metrics including RMSE, CNR, and Dice. Our work explores the feasibility of obtaining additional surface information using existing components of standalone optical tomography. This makes the optical tomographic technique more accurate and more accessible to biomedical researchers.


NOAEL-dose of a neonicotinoid pesticide, clothianidin, acutely induce anxiety-related behavior with human-audible vocalizations in male mice in a novel environment.
Neonicotinoids are novel systemic pesticides acting as agonists on the nicotinic acetylcholine receptors (nAChRs) of insects. Experimental studies have revealed that neonicotinoids pose potential risks for the nervous systems of non-target species, but the brain regions responsible for their behavioral effects remain incompletely understood. This study aimed to assess the neurobehavioral effects of clothianidin (CTD), a later neonicotinoid developed in 2001 and widely used worldwide, and to explore the target regions of neonicotinoids in the mammalian brain. A single-administration of 5 or 50mg/kg CTD to male C57BL/6N mice at or below the no-observed-adverse-effect level (NOAEL) induced an acute increase in anxiety during the elevated plus-maze test. In addition, mice in the CTD-administered group spontaneously emitted human-audible vocalizations (4-16kHz), which are behavioral signs of aversive emotions, and showed increased numbers of c-fos immunoreactive cells in the paraventricular thalamic nucleus and dentate gyrus of the hippocampus. In conclusion, mice exposed to NOAEL-dose CTD would be rendered vulnerable to a novel environment via the activation of thalamic and hippocampal regions related to stress responses. These findings should provide critical insight into the neurobehavioral effects of neonicotinoids on mammals.


The role of research in setting priorities for health care.
Research of all types plays a fundamental role in setting health care priorities, in part by informing the development of robust clinical practice guidelines. Today's investments in research may make tomorrow's choices about who gets what kinds of care easier, not more difficult. The converse also holds: today's investments in providing care, if made at the significant expense of research, will not make tomorrow's choices about care any easier. In making choices and balancing priorities, the centrality of health services research must be understood. The information necessary to make the hard choices about priorities in health care today may be generated as much, if not more, from investments in this field of inquiry as from equivalent investments in other arenas of scientific endeavour, because of the growing need for information about the effectiveness of health care and in making better use of health resources.


Total parenteral nutrition and bowel rest modify the metabolic response to endotoxin in humans.
Intestinal mucosal atrophy, as induced by total parenteral nutrition (TPN) and/or prolonged bowel rest, is hypothesized to enhance bowel endotoxin (LPS) translocation and may alter host responses to infection. To examine the effect of TPN-induced bowel atrophy on the response to LPS, 12 healthy volunteers were randomized to receive either enteral feedings (ENT, n = 6) or seven days of TPN without oral intake (TPN, n = 6). Enteral or TPN feedings were terminated 12 hours before the study period when a constant dextrose infusion (50 mg/kg/hour) was initiated and continued throughout the subsequent study period. After placement of arterial, hepatic vein, and femoral vein catheters, metabolic parameters were determined before and for six hours after an intravenous E. coli LPS challenge (20 U/kg). Subsequent peak levels of arterial glucagon (ENT, 189 +/- 39 pg/mL; TPN, 428 +/- 48; p less than 0.01), arterial epinephrine (ENT, 236 +/- 52 pg/mL; TPN, 379 +/- 49; p less than 0.05) and hepatic venous cachectin/tumor necrosis factor (cachectin/TNF) (ENT, 250 +/- 56 pg/mL; TPN, 479 +/- 136; p less than 0.05) were significantly higher in the TPN group than in the ENT group. The extremity efflux of lactate (ENT, -16 +/- 4 micrograms/min-100cc tissue; TPN, -52 +/- 13; t = 2 hours; p less than 0.05) and of amino acids (ENT, -334 +/- 77 nmol/min-100cc tissue; TPN, -884 +/- 58; t = 4 hours; p less than 0.05) were higher in the TPN subjects after the endotoxin challenge. Circulating C-reactive Protein (CRP) levels measured 24 hours postendotoxin were also significantly higher in the TPN subjects (ENT, 1.7 +/- 0.2 mg/dL; TPN, 3.2 +/- 0.3; p less than 0.01). Hence the counter-regulatory hormone and splanchnic cytokine responses to LPS were enhanced after TPN and bowel rest. This is associated with a magnified acute-phase response, peripheral amino acid mobilization, and peripheral lactate production. Thus antecedent TPN may influence the metabolic alterations seen in infection and sepsis via both an exaggerated counter-regulatory hormone response as well as an enhanced systemic and splanchnic production of cytokines.


Outcomes from the Delphi process of the Thoracic Robotic Curriculum Development Committee.
As the adoption of robotic procedures becomes more widespread, additional risk related to the learning curve can be expected. This article reports the results of a Delphi process to define procedures to optimize robotic training of thoracic surgeons and to promote safe performance of established robotic interventions as, for example, lung cancer and thymoma surgery. In June 2016, a working panel was spontaneously created by members of the European Society of Thoracic Surgeons (ESTS) and European Association for Cardio-Thoracic Surgery (EACTS) with a specialist interest in robotic thoracic surgery and/or surgical training. An e-consensus-finding exercise using the Delphi methodology was applied requiring 80% agreement to reach consensus on each question. Repeated iterations of anonymous voting continued over 3 rounds. Agreement was reached on many points: a standardized robotic training curriculum for robotic thoracic surgery should be divided into clearly defined sections as a staged learning pathway; the basic robotic curriculum should include a baseline evaluation, an e-learning module, a simulation-based training (including virtual reality simulation, Dry lab and Wet lab) and a robotic theatre (bedside) observation. Advanced robotic training should include e-learning on index procedures (right upper lobe) with video demonstration, access to video library of robotic procedures, simulation training, modular console training to index procedure, transition to full-procedure training with a proctor and final evaluation of the submitted video to certified independent examiners. Agreement was reached on a large number of questions to optimize and standardize training and education of thoracic surgeons in robotic activity. The production of the content of the learning material is ongoing.


Red Blood Cell Distribution Width Is Associated with Severity of Leukoaraiosis.
Red blood cell distribution width (RDW) is one of the routine hematologic parameters reported in the complete blood count test, which has been recognized as strong prognostic marker for various medical conditions, especially cardiovascular disease. We evaluated that RDW was also associated with the leukoaraiosis; common radiological finding of brain and that has been strongly associated with risk of stroke and dementia. In the present study, we included 1006 non-stroke individuals who underwent brain MRI and routine complete blood count test including RDW. Fazekas scale was used to measure the severity of leukoaraiosis based on fluid-attenuated inversion recovery image, and the severity was dichotomized to mild-degree (Fazekas scale: 0-1) and severe-degree leukoaraiosis (Fazekas scale: 2-3). Univariate and multivariate logistic regression models were constructed to evaluate independent risk factor for severe-degree of leukoaraiosis. Mean age of 1006 subjects was 64.34 ± 9.11 year, and mean of RDW was 12.97 ± 0.86%. The severe-degree of leukoaraiosis (Fazekas scale ≥ 2) was found in 28.83%. In the multivariate logistic regression, 4th quartile of RDW (> 13.3%) were significantly associated with the presence of severe-degree of leukoaraiosis (adjusted odds ratio, 1.87; 95% confidence interval, 1.20-2.92) compared to the 1st quartile of RDW (< 12.5%). The significance was not changed after adjustments for hemoglobin and other hematologic indices. These findings suggest that RDW is independently associated with severity of leukoaraiosis.


Changes in physical activity during the retirement transition: a series of novel n-of-1 natural experiments.
Existing evidence about the impact of retirement on physical activity (PA) has primarily focused on the average change in PA level after retirement in group-based studies. It is unclear whether findings regarding the direction of PA change after retirement from group-based studies apply to individuals. This study aimed to explore changes in PA, PA determinants and their inter-relationships during the retirement transition at the individual level. A series of n-of-1 natural experiments were conducted with seven individuals who were aged 55-76 years and approaching retirement. PA was measured by tri-axial accelerometry. Twice-daily self-report and ecological momentary assessments of evidence- and theory-based determinants of PA (e.g. sleep length/quality, happiness, tiredness, stress, time pressure, pain, intention, perceived behavioural control, priority, goal conflict and goal facilitation) were collected via a questionnaire for a period of between 3 and 7 months, which included time before and after the participant's retirement date. A personalised PA determinant was also identified by each participant and measured daily for the duration of the study. Dynamic regression models for discrete time binary data were used to analyse data for each individual participant. Two participants showed a statistically significant increase in the probability of engaging in PA bouts after retirement and two participants showed a significant time trend for a decrease and increase in PA bouts over time during the pre- to post-retirement period, respectively. There was no statistically significant change in PA after retirement for the remaining participants. Most of the daily questionnaire variables were significantly associated with PA for one or more participants but there were no consistent pattern of PA predictors across participants. For some participants, the relationship between questionnaire variables and PA changed from pre- to post-retirement. The findings from this study demonstrate the impact of retirement on individual PA trajectories. Using n-of-1 methods can provide information about unique patterns and determinants of individual behaviour over time, which has been obscured in previous research. N-of-1 methods can be used as a tool to inform personalised PA interventions for individuals within the retirement transition.


Role of the renin-angiotensin system in hepatic fibrosis and portal hypertension.
The renin-angiotensin system (RAS) is an important regulator of cirrhosis and portal hypertension. As hepatic fibrosis progresses, levels of the RAS components angiotensin (Ang) II, Ang-(1-7), angiotensin-converting enzyme (ACE), and Ang II type 1 receptor (AT1R) are increased. The primary effector Ang II regulates vasoconstriction, sodium homoeostasis, fibrosis, cell proliferation, and inflammation in various diseases, including liver cirrhosis, through the ACE/Ang II/AT1R axis in the classical RAS. The ACE2/Ang-(1-7)/Mas receptor and ACE2/Ang-(1-9)/AT2R axes make up the alternative RAS and promote vasodilation, antigrowth, proapoptotic, and anti-inflammatory effects; thus, countering the effects of the classical RAS axis to reduce hepatic fibrogenesis and portal hypertension. Patients with portal hypertension have been treated with RAS antagonists such as ACE inhibitors, Ang receptor blockers, and aldosterone antagonists, with very promising hemodynamic results. In this review, we examine the RAS, its roles in hepatic fibrosis and portal hypertension, and current therapeutic approaches based on the use of RAS antagonists in patients with portal hypertension.


SMF-1, SMF-2 and SMF-3 DMT1 orthologues regulate and are regulated differentially by manganese levels in C. elegans.
Manganese (Mn) is an essential metal that can exert toxic effects at high concentrations, eventually leading to Parkinsonism. A major transporter of Mn in mammals is the divalent-metal transporter (DMT1). We characterize here DMT1-like proteins in the nematode C. elegans, which regulate and are regulated by Mn and iron (Fe) content. We identified three new DMT1-like genes in C. elegans: smf-1, smf-2 and smf-3. All three can functionally substitute for loss of their yeast orthologues in S. cerevisiae. In the worm, deletion of smf-1 or smf-3 led to an increased Mn tolerance, while loss of smf-2 led to increased Mn sensitivity. smf mRNA levels measured by QRT-PCR were up-regulated upon low Mn and down-regulated upon high Mn exposures. Translational GFP-fusions revealed that SMF-1 and SMF-3 strongly localize to partially overlapping apical regions of the gut epithelium, suggesting a differential role for SMF-1 and SMF-3 in Mn nutritional intake. Conversely, SMF-2 was detected in the marginal pharyngeal epithelium, possibly involved in metal-sensing. Analysis of metal content upon Mn exposure in smf mutants revealed that SMF-3 is required for normal Mn uptake, while smf-1 was dispensable. Higher smf-2 mRNA levels correlated with higher Fe content, supporting a role for SMF-2 in Fe uptake. In smf-1 and smf-3 but not in smf-2 mutants, increased Mn exposure led to decreased Fe levels, suggesting that both metals compete for transport by SMF-2. Finally, SMF-3 was post-translationally and reversibly down-regulated following Mn-exposure. In sum, we unraveled a complex interplay of transcriptional and post-translational regulations of 3 DMT1-like transporters in two adjacent tissues, which regulate metal-content in C. elegans.


Caspase activation in the terminal differentiation of human epidermal keratinocytes.
The epidermis is a multilayered squamous epithelium in which dividing basal cells withdraw from the cell cycle and progressively differentiate as they are displaced toward the skin surface. Eventually, the cells lose their nucleus and other organelles to become flattened squames, which are finally shed from the surface as bags of cross-linked keratin filaments enclosed in a cornified envelope [1]. Although keratinocytes can undergo apoptosis when stimulated by a variety of agents [2], it is not known whether their normal differentiation programme uses any components of the apoptotic biochemical machinery to produce the cornified cell. Differentiating keratinocytes have been reported to share some features with apoptotic cells, such as DNA fragmentation, but these features have not been seen consistently [3]. Apoptosis involves an intracellular proteolytic cascade, mainly mediated by members of the caspase family of cysteine proteases, which cleave one another and various key intracellular target proteins to kill the cell neatly and quickly [4]. Here, we show for the first time that caspases are activated during normal human keratinocyte differentiation and that this activation is apparently required for the normal loss of the nucleus.


Fibroblast growth factor receptors 1 and 2 in keratinocytes control the epidermal barrier and cutaneous homeostasis.
Fibroblast growth factors (FGFs) are master regulators of organogenesis and tissue homeostasis. In this study, we used different combinations of FGF receptor (FGFR)-deficient mice to unravel their functions in the skin. Loss of the IIIb splice variants of FGFR1 and FGFR2 in keratinocytes caused progressive loss of skin appendages, cutaneous inflammation, keratinocyte hyperproliferation, and acanthosis. We identified loss of FGF-induced expression of tight junction components with subsequent deficits in epidermal barrier function as the mechanism underlying the progressive inflammatory skin disease. The defective barrier causes activation of keratinocytes and epidermal gammadelta T cells, which produce interleukin-1 family member 8 and S100A8/A9 proteins. These cytokines initiate an inflammatory response and induce a double paracrine loop through production of keratinocyte mitogens by dermal cells. Our results identify essential roles for FGFs in the regulation of the epidermal barrier and in the prevention of cutaneous inflammation, and highlight the importance of stromal-epithelial interactions in skin homeostasis and disease.


Predicting Synergism of Cancer Drug Combinations Using NCI-ALMANAC Data.
Drug combinations are of great interest for cancer treatment. Unfortunately, the discovery of synergistic combinations by purely experimental means is only feasible on small sets of drugs. In silico modeling methods can substantially widen this search by providing tools able to predict which of all possible combinations in a large compound library are synergistic. Here we investigate to which extent drug combination synergy can be predicted by exploiting the largest available dataset to date (NCI-ALMANAC, with over 290,000 synergy determinations). Each cell line is modeled using primarily two machine learning techniques, Random Forest (RF) and Extreme Gradient Boosting (XGBoost), on the datasets provided by NCI-ALMANAC. This large-scale predictive modeling study comprises more than 5,000 pair-wise drug combinations, 60 cell lines, 4 types of models, and 5 types of chemical features. The application of a powerful, yet uncommonly used, RF-specific technique for reliability prediction is also investigated. The evaluation of these models shows that it is possible to predict the synergy of unseen drug combinations with high accuracy (Pearson correlations between 0.43 and 0.86 depending on the considered cell line, with XGBoost providing slightly better predictions than RF). We have also found that restricting to the most reliable synergy predictions results in at least 2-fold error decrease with respect to employing the best learning algorithm without any reliability estimation. Alkylating agents, tyrosine kinase inhibitors and topoisomerase inhibitors are the drugs whose synergy with other partner drugs are better predicted by the models. Despite its leading size, NCI-ALMANAC comprises an extremely small part of all conceivable combinations. Given their accuracy and reliability estimation, the developed models should drastically reduce the number of required in vitro tests by predicting in silico which of the considered combinations are likely to be synergistic.


Role of ULK-FIP200 complex in mammalian autophagy: FIP200, a counterpart of yeast Atg17?
The yeast serine threonine kinase Atg1 appears to be a key regulator of autophagy and its kinase activity is crucial for autophagy induction. Recent reports have indicated that a mammalian Atg1 homolog, UNC-51-like kinase (ULK) 1, is required for autophagy. We found that ULK1 localizes to the autophagic isolation membrane and its kinase activity is important for autophagy induction. Furthermore, we identified a focal adhesion kinase (FAK) family interacting protein of 200 kD (FIP200) as a ULK-interacting protein. FIP200 also localizes to the isolation membrane together with ULK. Using FIP200-deficient cells, we found that FIP200 is essential for autophagosome formation and the proper function of ULK. Here, we discuss the role of the ULK-FIP200 complex in autophagy and the possibility that FIP200 functions as a mammalian counterpart of Atg17.


Biochemical and mutational analysis of a novel nicotinamidase from Oceanobacillus iheyensis HTE831.
Nicotinamidases catalyze the hydrolysis of nicotinamide to nicotinic acid and ammonia, an important reaction in the NAD(+) salvage pathway. This paper reports a new nicotinamidase from the deep-sea extremely halotolerant and alkaliphilic Oceanobacillus iheyensis HTE831 (OiNIC). The enzyme was active towards nicotinamide and several analogues, including the prodrug pyrazinamide. The enzyme was more nicotinamidase (kcat/Km = 43.5 mM(-1)s(-1)) than pyrazinamidase (kcat/Km = 3.2 mM(-1)s(-1)). Mutational analysis was carried out on seven critical amino acids, confirming for the first time the importance of Cys133 and Phe68 residues for increasing pyrazinamidase activity 2.9- and 2.5-fold, respectively. In addition, the change in the fourth residue involved in the ion metal binding (Glu65) was detrimental to pyrazinamidase activity, decreasing it 6-fold. This residue was also involved in a new distinct structural motif DAHXXXDXXHPE described in this paper for Firmicutes nicotinamidases. Phylogenetic analysis revealed that OiNIC is the first nicotinamidase described for the order Bacillales.


Describing synchronization and topological excitations in arrays of magnetic spin torque oscillators through the Kuramoto model.
The collective dynamics in populations of magnetic spin torque oscillators (STO) is an intensely studied topic in modern magnetism. Here, we show that arrays of STO coupled via dipolar fields can be modeled using a variant of the Kuramoto model, a well-known mathematical model in non-linear dynamics. By investigating the collective dynamics in arrays of STO we find that the synchronization in such systems is a finite size effect and show that the critical coupling-for a complete synchronized state-scales with the number of oscillators. Using realistic values of the dipolar coupling strength between STO we show that this imposes an upper limit for the maximum number of oscillators that can be synchronized. Further, we show that the lack of long range order is associated with the formation of topological defects in the phase field similar to the two-dimensional XY model of ferromagnetism. Our results shed new light on the synchronization of STO, where controlling the mutual synchronization of several oscillators is considered crucial for applications.


Nf1 heterozygous mice recapitulate the anthropometric and metabolic features of human neurofibromatosis type 1.
Neurofibromatosis type 1 (NF1) is a heritable cancer predisposition syndrome resulting from mutations in the NF1 tumor suppressor gene. Genotype-phenotype correlations for NF1 are rare due to the large number of NF1 mutations and role of modifier genes in manifestations of NF1; however, emerging reports suggest that persons with NF1 display a distinct anthropometric and metabolic phenotype featuring short stature, low body mass index, increased insulin sensitivity, and protection from diabetes. Nf1 heterozygous (Nf1+/-) mice accurately reflect the dominant inheritance of NF1 and are regularly employed as a model of NF1. Here, we sought to identify whether Nf1+/- mice recapitulate the anthropometric and metabolic features identified in persons with NF1. Littermate 16-20 week-old male wildtype (WT) and Nf1+/- C57B/6J mice underwent nuclear magnetic resonance (NMR), indirect calorimetry, and glucose/insulin/pyruvate tolerance testing. In some experiments, tissues were harvested for NMR and histologic characterization. Nf1+/- mice are leaner with significantly reduced visceral and subcutaneous fat mass, which corresponds with an increased density of small adipocytes and reduced leptin levels. Additionally, Nf1+/- mice are highly reliant on carbohydrates as an energy substrate and display increased glucose clearance and insulin sensitivity, but normal response to pyruvate suggesting enhanced glucose utilization and preserved gluconeogenesis. Finally, WT and Nf1+/- mice subjected to high glucose diet were protected from diet-induced obesity and hyperglycemia. Our data suggest that Nf1+/- mice closely recapitulate the anthropometric and metabolic phenotype identified in persons with NF1, which will impact the interpretation of previous and future translational studies of NF1.


Trimer effects in fragment molecular orbital-linear combination of molecular orbitals calculation of one-electron orbitals for biomolecules.
The fragment molecular orbital (FMO)-linear combination of molecular orbitals (LCMO) method incorporates as an efficient post-process calculation of one-electron orbitals of the whole system after the FMO total energy calculation. A straightforward way to increase the accuracy is inclusion of the trimer effect. Here, we derive a comprehensive formulation called the FMO3-LCMO method. To keep the computational costs of the trimer term low enough, we use a matrix-size reduction technique. We evaluated the accuracy and efficiency of the FMO3-LCMO scheme in model biological systems (alanine oligomer and chignolin). The results show that delocalized electronic orbitals with covalent and hydrogen bonds are better described at the trimer level, and the FMO3-LCMO method is applicable to quantitative evaluations of a wide range of frontier orbitals in large biosystems.


High-density functional diffuse optical tomography based on frequency-domain measurements improves image quality and spatial resolution.
Measurements of dynamic near-infrared (NIR) light attenuation across the human head together with model-based image reconstruction algorithms allow the recovery of three-dimensional spatial brain activation maps. Previous studies using high-density diffuse optical tomography (HD-DOT) systems have reported improved image quality over sparse arrays. These HD-DOT systems incorporated multidistance overlapping continuous wave measurements that only recover differential intensity attenuation. We investigate the potential improvement in reconstructed image quality due to the additional incorporation of phase shift measurements, which reflect the time-of-flight of the measured NIR light, within the tomographic reconstruction from high-density measurements. To evaluate image reconstruction with and without the additional phase information, we simulated point spread functions across a whole-scalp field of view in 24 subject-specific anatomical models using an experimentally derived noise model. The addition of phase information improves the image quality by reducing localization error by up to 59% and effective resolution by up to 21% as compared to using the intensity attenuation measurements alone. Furthermore, we demonstrate that the phase data enable images to be resolved at deeper brain regions where intensity data fail, which is further supported by utilizing experimental data from a single subject measurement during a retinotopic experiment.


The Nurse Practitioner Role: A pilot study based on the Interview Schedule from the AUSPRAC Research Toolkit.
Abstract Within any professional practice, knowledge developments to support service delivery and to understand roles inherent within that practice context are critical. The purpose of this article is to present findings from case study research that used the AUSPRAC Research Toolkit Interview Schedule and to propose an additional theme to the Interview Schedule. Case Study method was used to explore the role of a nurse practitioner within a specific context of practice in an Australian Healthcare institution. Three semi-structured interviews with a Nurse Practitioner using the AUSPRAC Research Toolkit Interview Schedule and one additional interview were employed. Data was analysed where initial free coding, then theme generation contributed to knowledge development. The AUSPRAC Research Toolkit Interview Schedule generated knowledge about the nurse practitioner role. Themes identified for interviews in the Schedule were; the organisation of care, team functioning and patient service. Analysis of data from these themes identified that information related to ongoing development of professional practice was not forthcoming from the participant. The authors recommend adding a fourth theme to the Interview Schedule to enable exploration of the professional elements of the Nurse Practitioner role.


Unifying Research on Social-Ecological Resilience and Collapse.
Ecosystems influence human societies, leading people to manage ecosystems for human benefit. Poor environmental management can lead to reduced ecological resilience and social-ecological collapse. We review research on resilience and collapse across different systems and propose a unifying social-ecological framework based on (i) a clear definition of system identity; (ii) the use of quantitative thresholds to define collapse; (iii) relating collapse processes to system structure; and (iv) explicit comparison of alternative hypotheses and models of collapse. Analysis of 17 representative cases identified 14 mechanisms, in five classes, that explain social-ecological collapse. System structure influences the kind of collapse a system may experience. Mechanistic theories of collapse that unite structure and process can make fundamental contributions to solving global environmental problems.


Chromosome-scale mega-haplotypes enable digital karyotyping of cancer aneuploidy.
Genomic instability is a frequently occurring feature of cancer that involves large-scale structural alterations. These somatic changes in chromosome structure include duplication of entire chromosome arms and aneuploidy where chromosomes are duplicated beyond normal diploid content. However, the accurate determination of aneuploidy events in cancer genomes is a challenge. Recent advances in sequencing technology allow the characterization of haplotypes that extend megabases along the human genome using high molecular weight (HMW) DNA. For this study, we employed a library preparation method in which sequence reads have barcodes linked to single HMW DNA molecules. Barcode-linked reads are used to generate extended haplotypes on the order of megabases. We developed a method that leverages haplotypes to identify chromosomal segmental alterations in cancer and uses this information to join haplotypes together, thus extending the range of phased variants. With this approach, we identified mega-haplotypes that encompass entire chromosome arms. We characterized the chromosomal arm changes and aneuploidy events in a manner that offers similar information as a traditional karyotype but with the benefit of DNA sequence resolution. We applied this approach to characterize aneuploidy and chromosomal alterations from a series of primary colorectal cancers.


Evaluation of scintigraphy to assess incorporation of intercalary cortical bone grafts in sheep.
To evaluate scintigraphy, radiography, and histomorphometric analysis for assessing incorporation of intercalary bone grafts and to compare incorporation of cortical autografts and allografts by the recipient. 12 skeletally mature sheep. A 5-cm tibial defect was filled with a cortical allograft (n = 6) or autograft (6) and stabilized with an interlocking nail. Radiography, scintigraphy, and fluorochrome bone labeling were performed every 3 months for 24 months. Radiographic evaluation included grading of the host and graft union and assessment of implants and grafts. Technetium-99m-hydroxymethylene diphosphonate radionuclide uptake was measured. Sheep were euthanatized 24 months after surgery, and bone formation was evaluated via histomorphometric analysis of fluorochrome labeling. Complete union was detected on radiographs by 21 months in all sheep but developed earlier in sheep that received an autograft versus in those that received an allograft. Radionuclide uptake peaked at 3 months and returned to presurgical values at 12 months. Histomorphometric analysis revealed fluorochrome labeling corresponding to each time point, with most bone formation at 9 through 15 months. Scintigraphy findings did not correlate well with fluorochrome labeling of newly formed bone. Although bone production around cortical bone grafts was detected by use of scintigraphy, this method did not provide accurate assessment of graft incorporation in sheep. Furthermore, bone produced by activated periosteum could not be distinguished scintgraphically from bone that replaced the graft. Intercalary autografts healed more rapidly and had greater incorporation into the host bone, compared with findings for allografts.


What can imaging tell us about cognitive impairment and dementia?
Dementia is a contemporary global health issue with far reaching consequences, not only for affected individuals and their families, but for national and global socio-economic conditions. The hallmark feature of dementia is that of irreversible cognitive decline, usually affecting memory, and impaired activities of daily living. Advances in healthcare worldwide have facilitated longer life spans, increasing the risks of developing cognitive decline and dementia in late life. Dementia remains a clinical diagnosis. The role of structural and molecular neuroimaging in patients with dementia is primarily supportive role rather than diagnostic, American and European guidelines recommending imaging to exclude treatable causes of dementia, such as tumor, hydrocephalus or intracranial haemorrhage, but also to distinguish between different dementia subtypes, the commonest of which is Alzheimer's disease. However, this depends on the availability of these imaging techniques at individual centres. Advanced magnetic resonance imaging (MRI) techniques, such as functional connectivity MRI, diffusion tensor imaging and magnetic resonance spectroscopy, and molecular imaging techniques, such as 18F fluoro-deoxy glucose positron emission tomography (PET), amyloid PET, tau PET, are currently within the realm of dementia research but are available for clinical use. Increasingly the research focus is on earlier identification of at risk preclinical individuals, for example due to family history. Intervention at the preclinical stages before irreversible brain damage occurs is currently the best hope of reducing the impact of dementia.


Microbiota and Neurodevelopmental Trajectories: Role of Maternal and Early-Life Nutrition.
Pregnancy and early life are characterized by marked changes in body microbial composition. Intriguingly, these changes take place simultaneously with neurodevelopmental plasticity, suggesting a complex dialogue between the microbes that inhabit the gastrointestinal tract and the brain. The purpose of this chapter is to describe the natural trajectory of microbiota during pregnancy and early life, as well as review the literature available on its interaction with neurodevelopment. Several lines of evidence show that the gut microbiota interacts with diet, drugs and stress both prenatally and postnatally. Clinical and preclinical studies are illuminating how these disruptions result in different developmental outcomes. Understanding the role of the microbiota in neurodevelopment may lead to novel approaches to the study of the pathophysiology and treatment of neuropsychiatric disorders.


Breast Cancer Cells in the Blood: A Pilot Study.
The goal of this pilot study was to determine in patients with operable breast cancer the incidence of breast cancer cells present in the blood, the clearance rate after surgical resection of the primary tumor, and the incidence of patients with persistent cancer cells in the blood after the primary tumor was removed. Twenty-one patients with operable breast cancer had 15 ml venous blood obtained twice prior to surgery and after surgery at 2, 4, 8, 12, 24, and 48 hours and also on days 7 and 14. Immunomagnetic selection of malignant cells was performed on each sample. Cells were then fixed on slides and immunocytochemistry performed on the collected cells. Cells that had a rosette of magnetic beads, cytoplasmic staining for keratin, and malignant morphology were counted as breast cancer cells. Eighteen of 19 of patients had cancer cells detected in at least one of the two blood samples preceding surgical removal of the primary tumor. The incidence of cancer cells in the blood of patients rapidly declined during the 48 hours postsurgery. The incidence of cancer cells in the blood remained stable in approximately 30% of patients to 14 days. The majority of breast cancer patients in this pilot study (even with small tumors and negative nodes) had detectable cancer cells in the blood prior to resection of the primary tumor. These findings justify further investigation. Successful application of this methodology may serve as a powerful indicator of which patients need systemic adjuvant therapy, the effectiveness of systemic adjuvant therapy, tumor recurrence, and early detection of breast cancer.


Nitrogen, phosphorus, and potassium uptake by wheat and their distribution in soil following successive, annual compost applications.
The overall objective of the present study was to determine the loading limits of composts that should be applied annually to irrigated wheat. We conducted a container experiment in a greenhouse during four years. It included eight treatments: sewage sludge compost (SSC) and cattle manure compost (CMC), each applied annually to a sandy soil, at rates equivalent to 3, 6, and 12 kg m(-2), and two controls, one fertilized and one unfertilized. Total dry matter (DM), grain production, and the amount of N, P, and K taken up by plants increased with increasing compost rate. Nitrogen uptake by the plants of the fertilized control was much higher than by the plants of the highest compost rate. Phosphorus and K uptake by the plants amended with the highest compost rate was much higher than by the fertilized control plants. Inorganic N quantity in the soil increased with increasing compost rate and with successive applications. The net N mineralization during the first year of wheat growth was very low, less than 3.5% of the applied organic N under all compost application rates. The contribution of the organic N mineralization increased during the second and third years. Most of the N increase in the compost treatment was found in the upper layer of 0 to 15 cm, whereas in the fertilized treatment N accumulated from the surface to the bottom of the container, 0 to 55 cm. The successive application of high rates of composts resulted in P and K accumulation in the soil profile.


Smartphone viewing distance and sleep: an experimental study utilizing motion capture technology.
There are studies reporting the negative impact of smartphone utilization on sleep. It is considered that reduction of melatonin secretion under the blue light exposure from smart-phone displays is one of the causes. The viewing distance may cause sleep disturbance, because the viewing distance determines the screen illuminance and/or asthenopia. However, to date, there has been no study closely investigating the impact of viewing distance on sleep; therefore, we sought to determine the relationship between smartphone viewing distance and subjective sleep status. Twenty-three nursing students (mean age ± standard deviation of 19.7±3.1 years) participated in the study. Subjective sleep status was assessed using the Pittsburgh Sleep Quality Index, morningness-eveningness questionnaire, and the Epworth sleepiness scale. We used the distance between the head and the hand while holding a smartphone to measure the viewing distance while using smartphones in sitting and lying positions. The distance was calculated using the three-dimensional coordinates obtained by a noncontact motion-sensing device. The viewing distance of smartphones in the sitting position ranged from 13.3 to 32.9 cm among participants. In the lying position, it ranged from 9.9 to 21.3cm. The viewing distance was longer in the sitting position than in the lying position (mean ± standard deviation: 20.3±4.7 vs 16.4±2.7, respectively, P<0.01). We found that the short viewing distance in the lying position had a positive correlation to a poorer sleep state (R2=0.27, P<0.05), lower sleep efficiency (R2=0.35, P<0.05), and longer sleep latency (R2=0.38, P<0.05). Moreover, smartphone viewing distances in lying position correlated negatively with subjective sleep status. Therefore, when recommending ideal smartphone use in lying position, one should take into account the viewing distances.


Interferon α-stimulated natural killer cells from patients with acute hepatitis C virus (HCV) infection recognize HCV-infected and uninfected hepatoma cells via DNAX accessory molecule-1.
Natural killer (NK) cells are an important component of the innate immune defense against viruses, including hepatitis C virus (HCV). The cell culture system using HCV-permissive Huh-7.5 cells make studies on interaction of NK cells and HCV-infected target cells possible. We used this system to characterize interactions of HCV-infected Huh-7.5 cells and NK cells from healthy controls and patients with acute HCV infection. IFNα- and IL-2 stimulated NK cells were cultured with HCV-infected hepatoma cells and subsequently analyzed (for degranulation and cytokine production) via multicolour flow cytometry. Luciferase assyas have been used to study inhibition of HCV replication. Further, PBMC from patients with acute hepatitis C as well as HCV-infected Huh7.5 cells have been analyzed via flow cytometry for expression of NK cell receptors and ligands, respectively. After interferon (IFN) α stimulation, NK cells from healthy controls and patients with acute hepatitis C efficiently recognized both HCV-infected and uninfected hepatoma cells. Subsequent dissection of receptor-ligand interaction revealed a dominant role for DNAM-1 and a complementary contribution of NKG2D for NK cell activation in this setting. Furthermore, IFN-α-stimulated NK cells effectively inhibited HCV replication in a DNAM-1-dependent manner. Human NK cells recognize HCV-infected hepatoma cells after IFN-α stimulation in a DNAM-1-dependent manner. Furthermore, interaction of IFN-α-stimulated NK cells with HCV-infected hepatoma cells efficiently reduced HCV replication. This study opens up future studies of NK cell interaction with HCV-infected hepatocytes to gain further insight into the pathogenesis of human HCV infection and the therapeutic effects of IFN-α.


Identification of the human cytochromes P450 responsible for the in vitro formation of the major oxidative metabolites of the antipsychotic agent olanzapine.
The formation kinetics of 2-hydroxymethyl olanzapine (2-OH olanzapine), 4'-N-oxide olanzapine (N-O olanzapine) and 4'-N-desmethyl olanzapine (NdM olanzapine) were analyzed in vitro. Biphasic kinetics were observed for formation of 2-OH and NdM olanzapine. The high-affinity enzyme responsible for 2-OH olanzapine formation by two human liver samples exhibited an intrinsic clearance (CLint) of 0.2 microliter/min/mg. NdM olanzapine formation by two human liver samples exhibited a CLint of 1.0 microliter/min/mg for the high affinity enzyme. The formation of N-O olanzapine was linear up to 300 microM olanzapine, yielding a CLint of 0.32 to 1.70 microliters/min/mg. The formation of 7-hydroxy olanzapine (7-OH olanzapine) exhibited an apparent Km of 24.2 microM. The rates of 2-OH olanzapine formation correlated with CYP2D6 levels and activity, and it was formed to the greatest extent by cDNA-expressed CYP2D6. N-O olanzapine formation correlated with human liver flavin-containing monooxygenase (FMO3) levels and activity. NdM olanzapine and 7-OH olanzapine formation correlated with CYP1A2 catalytic activities and they were formed to the greatest extent by expressed CYP1A2. These results suggest that CYP1A2 catalyzes NdM olanzapine and 7-OH olanzapine formation, CYP2D6 catalyzes 2-OH olanzapine formation and FMO3 catalyzes N-O olanzapine formation.


Thermo-optic properties of epitaxial Sr0.6Ba0.4Nb2O6 waveguides and their application as optical modulator.
A prism-coupler technique was introduced to determine the refractive indices and thermo-optic coefficients of epitaxial Sr(0.6)Ba(0.4)Nb(2)O(6) (SBN) waveguides, in a temperature range covering the ferroelectric-paraelectric phase transition. A strong enhancement in the TO coefficient is observed near T(c). This strong enhancement is related to the critical change of the polarization. The values of dn(e)/dT are significantly larger than dn(o)/dT due to the larger quadratic electro-optic coefficient in TM polarization. In TM mode, the refractive index of SBN is increased by 1.3% as the temperature is increased to 160 degrees C. Our results suggest that SBN waveguide is a potential candidate for thermo-optic modulators and switches.


Burst-firing activity of presumed 5-HT neurones of the rat dorsal raphe nucleus: electrophysiological analysis by antidromic stimulation.
We recently reported raphe neurones which frequently fired spikes in short bursts. However, the action potentials were broad and the neurones fired in a slow and regular pattern, suggesting they were an unusual type of 5-hydroxytryptamine (5-HT) neurone. In the present study, we investigated whether these putative burst-firing 5-HT neurones project to the forebrain and whether all spikes fired in bursts propagate along the axon. In anaesthetised rats, electrical stimulation of the medial forebrain bundle evoked antidromic spikes in both burst-firing neurones and in single-spiking, classical 5-HT neurones recorded in the dorsal raphe nucleus. Although the antidromic spike latency of the single-spiking and burst-firing neurones showed a clear overlap, burst-firing neurones had a significantly shorter latency than single-spiking neurones. For both burst-firing neurones and classical 5-HT neurones, antidromic spikes made collisions with spontaneously occurring spikes. Furthermore, in all burst-firing neurones tested, first, second and third order spikes in a burst could be made to collide with antidromic spike. Interestingly, in a small number of burst-firing neurones, antidromic stimulation evoked spike doublets, similar to those recorded spontaneously. From these data we conclude that burst-firing neurones in the dorsal raphe nucleus project to the forebrain, and each spike generated by the burst propagates along the axon and could thereby release transmitter (5-HT).


Genetic heterogeneity in hereditary haemorrhagic telangiectasia: possible correlation with clinical phenotype.
Hereditary haemorrhagic telangiectasia (HHT) or Osler-Weber-Rendu syndrome is an autosomal dominant vascular dysplasia characterised by recurrent haemorrhage. Our initial linkage studies found an HHT gene to be localised to 9q3 in two large kindreds. In the present study, we examine an additional five unrelated HHT families. Linkage analysis in this region resulted in a peak multipoint location score of 13.03, 10 cM proximal of D9S60. We found significant evidence for heterogeneity of HHT. Multipoint analysis supports the family specific two point studies with odds of 3,000,000:1 showing linkage and heterogeneity over linkage and homogeneity. Four of the seven families give a posterior probability of > 99% of being of the linked type, and three families appear unlinked to this region of 9q, and by multipoint analysis completely exclude the candidate region for HHT. Two new crossovers in affected persons in one of the linked families further define the proximal border of the candidate region on 9q3. A possible correlation in clinical phenotype between the 9q3 linked families and unlinked families is described. Although six of the seven families clearly meet the clinical criteria for HHT diagnosis, a significant absence of pulmonary arteriovenous malformations is seen in all three 9q3 unlinked families. Genetic heterogeneity of HHT and its potential correlation with a clinical phenotype may have a significant impact on the clinical management and treatment of HHT patients.


Why WAIT? Preliminary evaluation of the weight assistance and intervention techniques (WAIT) group.
Individuals who attempt to lose weight may struggle because they lack skills to address problematic eating behaviors. There are multiple programs that have taught patients some of these behavioral strategies; however, it is not clear which strategies patients find to be the most useful. The purpose of this study was to examine preliminary outcomes after completion of a six-week integrative group for weight management. Retrospective chart reviews were conducted of 51 patients who completed an integrative, psychological weight management group. Patients were mailed surveys 1-2 years after completion of the group assessing for current problematic eating behaviors (i.e. emotional eating and food addiction), satisfaction with treatment, and skills they continue to use. The majority of patients lost weight, were satisfied with the group, found the group to be helpful, and felt confident they could maintain behavior changes. The strategies patients most commonly continued to use post-group included mindful eating, keeping a food diary, carrying out an exercise plan, regular weigh-ins, and planning for social eating. The number of food addiction symptoms decreased from pre- to post-group. An integrative psychological weight management group may provide patients with skills and confidence to assist with managing problematic eating behaviors and weight loss.


The prevalence of silent kidney stones--an ultrasonographic screening study.
Silent and not yet discovered stones of the upper urinary tract are potentially dangerous, since in due course they may cause infection, obstruction and renal damage. The aim of this study was to determine the prevalence of such silent kidney stones in a representative Pakistani population of Karachi. We studied 201 consecutive subjects at our hospital who underwent additional kidney screening whilst undergoing abdominal ultrasound. All these subjects did not have a history or symptoms of urolithiasis. We found silent kidney stones in 3% of subjects. All stone bearers were males. Most stones were in the left kidney. Notably, multiple stones and stones of a considerable size went unnoticed. In addition to the usual figures of incidence and prevalence of stone disease drawn from patient data, there is a prevalence of 3% silent stones that may only be discovered incidentally or by screening. This is true for a "'stone country" like Pakistan. Figures for other regions have yet to be determined. Due to socioeconomic reasons, we believe that a general kidney screening for urolithiasis is, however, not indicated, at least in our country.


Up-Regulation of ninjurin expression in human hepatocellular carcinoma associated with cirrhosis and chronic viral hepatitis.
The cDNA clones, which were highly expressed in liver tissues of hepatocellular carcinoma (HCC) patients, were identified using dot hybridization and reconfirmed by Northern blot analysis. One of the clones, ninjurin (nerve injury induced protein), showed a much higher expression level in the liver tissue of HCC patients than in normal liver tissue. Interestingly, the presence of ninjurin mRNA transcripts was detected with high intensity in HCC tissues when combined with viral infection and cirrhosis, but not with a normal liver or HCC tissue unrelated with viral infection and cirrhosis. We produced a N-terminal part of recombinant ninjurin protein, as well as a monoclonal antibody specific to this polypeptide. The intensity of immunohistochemical staining of the liver tumor tissue, and regenerating tissue for the ninjurin protein, was stronger than that of normal liver tissue. These results suggest that ninjurin may play an important role in the development of HCC combined with cirrhosis and viral infection.


Metastases from renal cell carcinoma-Report of three unpredictable cases and literature review.
The capacity to metastasize after long periods of discovery and resection of the primary renal lesion or to present as metastasis with undisclosed primary lesions grant renal cell carcinomas (RCC) a formidable and unpredictable behavior. We report three unusual cases of metastasis from RCC. The first patient presented with metastatic clear cell adenocarcinoma in the right shoulder muscles, with unknown primary and revealed an undetected clear cell right RCC on radiology. The second patient presented with a hemorrhagic nasal metastasis of RCC. A history of nephrectomy for RCC, 10 years prior to the development of nasal metastasis was elicited. The third patient presented with axillary and later abdominal wall metastasis of papillary RCC. He had a history of RCC 3 years prior to the discovery of metastasis. RCC has the propensity to appear as a metastatic carcinoma with unknown primary, requiring exclusion of several diagnostic pathologic entities. The appearance of metastasis many years after disease quiescence is challenging and clinicians need to be sensitized to the need for long-term surveillance in RCC. Though the prognosis remains poor, immune checkpoint inhibitors are currently the modalities of choice in such cases.


Do the abilities of children with cerebral palsy explain their activities and participation?
The aim of this study was to use family-assessed instruments and details of children's impairments to explore factors affecting the activities and participation of children with cerebral palsy (CP). A postal survey was conducted with families of a geographically defined population of children with CP aged 6 to 12 years. Family-assessed indices of children's activities and participation were the Activities Scale for Kids (ASK) and Lifestyle Assessment Questionnaire (LAQ-CP). Families also assessed children's abilities using the Gross Motor Function and Manual Ability Classification Systems (GMFCS; MACS). Details of children's impairments were available from the 4Child epidemiological database and used with the GMFCS and MACS as explanatory variables in multiple regression analyses to identify their effect on children's activities and participation. Families of 175/314 (56%) children returned an assessment using the GMFCS and 129 (41%) children participated fully by returning all the questionnaires. Full participants (72 males, 57 females) did not differ from those who did not take part by their age, sex, CP characteristics, or associated impairments: GMFCS Level I-25, Level II-43, Level III-15, Level IV-14, Level V-23; MACS Level I-14, Level II-30, Level III-18, Level IV-13, Level V-13. Scores for the ASK and LAQ-CP Physical Independence and Mobility domains were predicted well by children's movement, manual, and intellectual disability, and also, to some extent, by the presence of seizures or speech problems. LAQ-CP domains for Economic and Clinical Burden and Social Integration were not well explained by children's abilities and impairments. Family assessment, therefore, offers a useful method for measuring children's activities and participation; however, currently available instruments do not fully represent all the domains in the International Classification of Functioning, Disability and Health. Children's abilities only partially explain their activities and participation.


Creating a three-parent child: an educational paradigm for the responsible conduct of research.
The field of assisted reproduction is renowned for its remarkable advances and constant pushing forward of research boundaries in an effort to offer innovative and effective methods for enhancing fertility. Accompanying these advances, however, are physiological, psychological, and bioethical consequences that must be considered. These concomitant advances and consequences make assisted reproduction an excellent educational paradigm for inculcating responsible conduct in both research and clinical practice. Ultimately, responsible conduct rests on the ethical researcher and clinician. Here, we present the as-yet unapproved, contentious assisted reproductive technology of mitochondrial replacement transfer (MRT) as an ideal educational platform to foster the responsible conduct of research by advancing dialogue among multi-disciplinary scholars, researchers, and students. Using a likely future case, we present the basic science, legal, and ethical considerations, and the pedagogical principles and strategies for using MRT as an effective educational paradigm. Society will benefit when the ethical issues inherent in creating children with three genetic parents as well as germline interference are discussed across multiple academic levels that include researchers, legal experts, bioethicists, and government-appointed commissions. Furthermore, undergraduate and graduate students should be included because they will likely determine the ethical fates of these biotechnologies. While emerging assisted reproduction technologies such as MRT are highly complex and will take years to be readily available for patients in need, now is the time to consider their scientific, legal, ethical, and cultural/religious implications for ensuring the responsible conduct of research.


Comparison of four antibiotics with indigenous marine Bacillus spp. in controlling pathogenic bacteria from shrimp and Artemia.
Use of antibiotics for the control of bacterial diseases in shrimp culture has caused several adverse impacts to the industry. This has resulted in the search for alternative environment friendly approaches to overcome bacterial infections. This study was conducted to investigate the use of beneficial bacteria as an alternative to antibiotics. Ten pathogenic bacterial species isolated from shrimp, Penaeus monodon, and Artemia cysts were tested for susceptibility to indigenous marine Bacillus subtilis AB65, Bacillus pumilus AB58, Bacillus licheniformis AB69 and compared with oxytetracycline, chloramphenicol, gentamicin and bacitracin, which are common antibiotics used in Asian aquaculture. The Bacillus spp. were isolated from the local marine environment for bioremediation use in shrimp hatcheries and were proven to reduce total ammonium nitrogen. The pathogenic bacterial isolates were 90% susceptible to B. subtilis AB65, 70% susceptible to B. pumilus AB58 and B. licheniformis AB69 and 100% susceptible to oxytetracycline, chloramphenicol and gentamicin but only 40% to bacitracin. Two representative isolates of the vibrio group, Vibrio alginolyticus VaM11 and Vibrio parahaemolyticus VpM1, when tested for competitive exclusion by a common broth method using the marine Bacillus spp., showed decreased viable counts from 10(8) to 10(2) cfu mL(-1). The results suggest that the action of the marine bacteria appears to be significant in protecting the host shrimp against pathogenic bacteria. In addition to the alternative use of antibiotics, the selected marine bacteria had additional bioremediation properties of reducing ammonia.


The relationship between fatigue and sleep in cancer patients: a review.
Fatigue is a major complaint among cancer patients, yet it is unknown whether cancer-related fatigue experienced during the day relates to sleep/wake cycles or to the quality and quantity of sleep obtained at night. Although it is not well defined or well understood at present, cancer-related fatigue is generally regarded as a form of tiredness that does not improve following rest or sleep. Objectively recorded sleep and biological rhythms have not been well investigated in these patients, but it appears that most cancer patients may in fact not be getting a good night's sleep. Evidence is accumulating that sleep is often disturbed in cancer patients, probably owing to a variety of causes. We posit that some degree of cancer-related fatigue experienced during the day may relate to sleep/wake cycles or to the quality and quantity of sleep obtained at night. Different components or dimensions of fatigue (physical, attentional/cognitive, emotional/affective, etc.) are probably associated in some way with disrupted sleep and desynchronized sleep/wake rhythms. These associations may change in measurable ways prior to treatment, during treatment and after treatment completion. In cancer patients, as in other medically ill patients, sleep that is inadequate or unrefreshing may be important not only to the expression of fatigue, but to the patients' quality of life and their tolerance to treatment, and may influence the development of mood disorders and clinical depression. This review summarizes the state of the literature on fatigue, sleep and circadian rhythms.


Prospective comparison of prediction rules of mortality risk for CAP in a developing country.
Aga Khan University Hospital, Karachi, Pakistan. To prospectively compare 30-day mortality with CURB65 and CRB65 scores and the three mortality risk groups (low, intermediate and high) based on these scores in hospitalised patients with community-acquired pneumonia (CAP). Longitudinal observational cohort study of adult in-patients fulfilling the definition of CAP between October 2006 and May 2007. A total of 137 patients was included. The 30-day mortality was 13.1%. Areas under the receiver operating characteristic curve for CURB65 and CRB65 scores were respectively 0.863 and 0.835. Odds of death among patients in the high mortality risk group of CURB65 score was 15.4 and those of CRB65 was 11.1 compared with the low and intermediate mortality risk groups combined as reference. CURB65 and CRB65 scores classified 46% and 24.8% patients, respectively, into the low mortality risk group. Length of hospital stay increased with the CURB65 score-based mortality risk groups but not with those based on the CRB65 score. CURB65 and CRB65 scores showed no significant difference in predicting 30-day mortality. Both scores, and the CURB65 score in particular, categorised reasonable proportions of patients into the low mortality risk group who could be discharged from the emergency room and managed as out-patients.


Ascorbic acid protects against restraint stress-induced memory deficits in Wistar rats.
Chronic stress has been shown to cause oxidative damage in the central nervous system. Although stress-induced impairments in learning and memory have been studied extensively, very few studies have investigated possible ways to prevent their ill effects. The present work was designed to study the protective effects of ascorbic acid in memory loss induced by chronic restraint stress. Adult male Wistar rats were designated into the following groups: (i) Normal control, (ii) Ascorbic acid treatment, (iii) Vehicle control, (iv) Restraint stress, (v) Restraint stress + vehicle, and (vi) Restraint stress + ascorbic acid treatment. At the end of 21 days, animals of all groups were subjected to memory tests using Morris water maze and passive avoidance apparatus. Then, the results obtained were compared between the experimental groups. Rats exposed to restraint stress alone and those pretreated with vehicle solution before restrained stress showed deficits in learning and impaired memory retention in the memory tests when compared to animals in other experimental groups. Animals pretreated with ascorbic acid before restraining showed significant improvement in memory retention in the same memory tests. Results of this study suggest the possibility of using ascorbic acid as a dietary supplement to prevent stress-induced memory impairments.


Screening for depression and anxiety disorder in children with headache.
The purpose of this study was to investigate the importance of initial screening tests for depression and anxiety disorders in children with headache. In addition, this study evaluated whether the Children's Depression Inventory (CDI) and Revised Children's Manifest Anxiety Scale (RCMAS) are suitable for screening symptoms of depression and anxiety. A retrospective chart review was conducted of 720 children aged 7-17 years who had visited a pediatric neurology clinic for headaches and were referred to a pediatric psychiatric clinic for psychiatric symptoms from January 2010 to December 2011. All patients completed the CDI and RCMAS. Among them, charts of patients with clinically significant total scores (cutoff>15) for psychiatric symptoms, as defined by the CDI and RCMAS scoring scales, were reviewed. Nineteen patients had headaches and clinically significant total scores for psychiatric symptoms. The mean age at headache diagnosis was 11.7 years, and 57% were male. Mean duration of headache was 11.5 months. Two point eight percent of the patients were diagnosed with psychiatric disorders including major depression (1.7%), generalized anxiety disorder (1.1%), and bipolar disorder (0.1%). Four patients (0.6%) were diagnosed with attention deficit/hyperactivity disorder (ADHD). Total mean CDI and RCMAS scores of patients referred to the psychiatric clinic were 18.8 and 22.2, respectively. There was no correlation between CDI or RCMAS total scores and headache frequency, duration, or severity. We recommend that all patients with headache should be screened for depression and anxiety by CDI and RCMAS scores.


D2 dopamine receptors modulate calcium channel currents and catecholamine secretion in bovine adrenal chromaffin cells.
Although dopamine is known to be present in sympathetic ganglia, its role and mode of action as a peripheral neurotransmitter are still poorly understood. Dopaminergic agonists have been shown to inhibit adrenal catecholamine release and calcium uptake. However, the specific dopamine receptor subtype mediating these effects and the receptor transduction mechanism remain unknown. We now provide evidence demonstrating 1) that slowly inactivating, voltage-gated calcium channels serve as a target site for dopaminergic modulation of chromaffin cell function and 2) that it is the D2 receptor subtype which mediates dopaminergic inhibitory effects on catecholamine secretion, 45Ca uptake and voltage-gated calcium currents. Whole cell patch clamp electrophysiological techniques were used to monitor directly voltage-gated Ca++ channels. The D2 agonist apomorphine but not the D1 agonist SKF 38393 reduced reversibly a slowly inactivating, voltage-gated calcium current in cultured chromaffin cells and this effect was blocked by the D2 receptor antagonist haloperidol. The presence of D2 but not D1 dopamine receptors on chromaffin cell membranes was demonstrated by radioligand binding methods, using the specific D1 and D2 receptor radioligands, [3H]SCH23390 and [3H]N-methylspiperone, respectively. Nicotine- and KCl (60 mM)-evoked catecholamine secretion and 45Ca uptake were inhibited by the D2 agonist, apomorphine, but not by the D1 agonist, SKF 38393. These inhibitory effects were prevented by the D2 antagonist, sulpiride, but not by the D1 antagonist, SCH 23390. D2 dopamine receptors appear to function as inhibitory modulators of adrenal catecholamine secretion with a mode of action involving inhibition of calcium channel currents.


Project Reach: A Pilot Cross-Sectional Study of the Telemedical Capabilities of Special Operations Medical Personnel.
While the US military has relied on the "Golden Hour" for casualty evacuation in Iraq and Afghanistan during most of the last two decades, Special Operations Forces (SOF) personnel have found themselves operating further outside of this established medical infrastructure. This has required prolonged casualty care beyond doctrinal timelines. Telemedicine is increasingly used by medical personnel to bridge this gap, augmenting the local staff with the expertise of a distant consultant. This pilot study was launched to establish a baseline of current SOF telemedical capabilities. This is a cross-sectional study of a 292 Department of Defense (DoD) SOF medical personnel via an online questionnaire during the period of October 2018-May 2019. Approximately 16.1% of the 292 respondents stated they received telemedicine equipment, 51.1% of respondents who received telemedicine equipment reported also receiving training on their equipment. Overall, 40.6% of respondents were "actively looking to add telemedicine to their practice," with prolonged field care as the primary intended use. Ideal characteristics of telemedicine equipment were described as a device weighing 6 lb or less, with real-time video and ultrasound transmission capabilities. The data demonstrated a gap between provider demand for telemedicine capabilities and their comfort to employ these capabilities downrange. This study suggests a need to increase the self-reported telemedicine competency of deployed local providers. Authors believe that this is best accomplished through incorporation of early integration of teleconsulting systems into mission rehearsals and unit exercises. Further study should be considered to investigate the efficacy of ongoing acquisition and training programs, along with how telemedicine is being incorporated in the unit during the predeployment training period.


Strand invasion by oligonucleotide--nuclease conjugates.
Conjugates consisting of staphylococcal nuclease crosslinked to oligonucleotides hybridize to supercoiled duplex DNA by Watson--Crick base-pairing. Here we describe this strand invasion. Affinity cleavage by these conjugates provides a probe for the local topology of the DNA duplex and is most efficient at a target DNA sequence known to form a cruciform. Additional supercoiling of the substrate DNA increases selective cleavage at other sequences. Hybridization of the conjugate to duplex DNA is temperature dependent and is stable over time. Affinity cleavage is not substantially inhibited by a 200-fold excess of the analogous unmodified oligonucleotide, demonstrating that hybridization of the unmodified oligonucleotide must be less favored and that the nuclease is involved in substrate binding. Surprisingly, affinity cleavage is also not effectively inhibited by complementary oligonucleotides unless they contain an extended 5'-sequence capable of separate interactions with the nuclease domain of the conjugate. These results suggest that the oligonucleotide-nuclease conjugate prefers to hybridize to target sequences which will allow interactions with both the oligonucleotide and the nuclease domains. Affinity cleavage by oligonucleotide-nuclease conjugates provides general insights for the design of oligonucleotides and their conjugates for strand invasion and affords a convenient competition assay for their hybridization.


Associations of Prenatal Urinary Bisphenol A Concentrations with Child Behaviors and Cognitive Abilities.
Prenatal bisphenol A (BPA) exposure has been associated with adverse neurodevelopment in epidemiological studies. However, prior studies had limited statistical power to examine sex-specific effects, and few examined child cognition. We estimated the association between prenatal BPA exposure and child neurobehavior at 3 y of age in a prospective cohort of 812 mothers and their children. We measured BPA concentration in urine samples collected at ∼12 wk gestation among women enrolled in a 10-city Canadian cohort study. At approximately 3 y of age, we assessed children's cognitive abilities with the Wechsler Primary and Preschool Scale of IntelligenceTM–III (WPPSI-III) and two scales of the Behavior Rating Inventory of Executive Function–Preschool (BRIEF-P). Parents reported children's behavior using the Behavior Assessment System for Children–2 (BASC-2) and the Social Responsiveness ScaleTM–2 (SRS-2). We estimated covariate-adjusted differences in neurobehavioral outcomes with a doubling in BPA concentration and sex-specific associations. BPA was not associated with WPPSI-III scores; child sex did not modify these associations. The association between BPA and BRIEF-P scores was modified by child sex (BPA×sex p-values≤0.03). For example, a doubling of BPA concentration was associated with 1-point (95% CI: 0.3, 1.7) poorer working memory in boys and 0.5-point (95% CI: −1.1, 0.1) better scores in girls. BPA was not associated with most BASC-2 scales; however, it was associated with more internalizing and somatizing behaviors in boys, but not in girls (BPA×sex p&-values≤0.08). A doubling of BPA concentration was associated with poorer SRS-2 scores [β=0.3 ( 95% CI: 0, 0.7)]; this association was not modified by sex. Prenatal urinary BPA concentration was associated with some aspects of child behavior in this cohort, and some associations were stronger among boys. https://doi.org/10.1289/EHP984.


Dissecting the effects of concentrated carbohydrate solutions on protein diffusion, hydration, and internal dynamics.
We present herein a thorough description of the effects of high glucose concentrations on the diffusion, hydration and internal dynamics of ubiquitin, as predicted from extensive molecular dynamics simulations on several systems described at fully atomistic level. We observe that the protein acts as a seed that speeds up the natural propensity of glucose to cluster at high concentration; the sugar molecules thus aggregate around the protein trapping it inside a dynamic cage. This process extensively dehydrates the protein surface, restricts the motions of the remaining water molecules, and drags the large-scale, collective motions of protein atoms slowing down the rate of exploration of the conformational space despite only a slight dampening of fast, local dynamics. We discuss how these effects could be relevant to the function of sugars as preservation agents in biological materials, and how crowding by small sticky molecules could modulate proteins across different reaction coordinates inside the cellular cytosol.


Systems of Power: A Spatial Envirotechnical Approach to Water Power and Industrialization in the Po Valley of Italy, ca.1880-1970.
Fossil fuels, more than water, are immediately associated with the industrial energy transition. Preindustrial energy sources, however, continued to play a role in industrial energy regimes. This was the case in the industrializing Po Valley, where water power remained crucial until the 1960s. The article analyzes the ways in which the technological and spatial features of mechanical hydropower and hydroelectricity combined with the spatial and environmental features of the Po watershed, as well as the different envirotechnical regimes associated with these configurations. Through this approach, it sheds lights on the radical change that hydropower underwent during industrialization: from discontinuous to continuous energy production, from diffusion to concentration of access to water resources, and from small- to large-scale, basin-wide system building and interdependency. Concentration, continuity, and large-scale interdependency transformed profoundly the social distribution of access to hydropower, and scaled up, without eliminating, the influence of environmental factors on water power production.


Proteomic analyses of the SMYD family interactomes identify HSP90 as a novel target for SMYD2.
The SMYD (SET and MYND domain) family of lysine methyltransferases (KMTs) plays pivotal roles in various cellular processes, including gene expression regulation and DNA damage response. Initially identified as genuine histone methyltransferases, specific members of this family have recently been shown to methylate non-histone proteins such as p53, VEGFR, and the retinoblastoma tumor suppressor (pRb). To gain further functional insights into this family of KMTs, we generated the protein interaction network for three different human SMYD proteins (SMYD2, SMYD3, and SMYD5). Characterization of each SMYD protein network revealed that they associate with both shared and unique sets of proteins. Among those, we found that HSP90 and several of its co-chaperones interact specifically with the tetratrico peptide repeat (TPR)-containing SMYD2 and SMYD3. Moreover, using proteomic and biochemical techniques, we provide evidence that SMYD2 methylates K209 and K615 on HSP90 nucleotide-binding and dimerization domains, respectively. In addition, we found that each methylation site displays unique reactivity in regard to the presence of HSP90 co-chaperones, pH, and demethylation by the lysine amine oxidase LSD1, suggesting that alternative mechanisms control HSP90 methylation by SMYD2. Altogether, this study highlights the ability of SMYD proteins to form unique protein complexes that may underlie their various biological functions and the SMYD2-mediated methylation of the key molecular chaperone HSP90.


Effects of a Small Inclination Misalignment in Talbot Interferometry by Use of Gratings with Arbitrary Line Orientation. II. Experimental Verification.
The theoretical results of the effects of a small inclination misalignment, which is formed by rotation of the beam-splitter grating around the axis on the grating plane when the axis has an arbitrary angle with respect to the line direction of the grating, between the two grating planes on the moiré fringes in the Talbot interferometry are verified by experiment. The experimental results coincide well with theoretical ones. Consequently, the effect of the small arbitrary inclination on practical measurements based on the Talbot interferometry is further explained by an example that examines the beam collimation of a lens, and the advantages and limitations of the effect are also discussed.