Estradiol treatment, physical activity, and muscle function in ovarian-senescent mice.
Estradiol (E(2)) treatment in young adult, ovariectomized mice increases physical activity and reverses deleterious effects on skeletal muscle. Here we test the hypothesis that E(2) treatment improves muscle function and physical activity in aged, ovarian-senescent mice. Plasma E(2) levels and vaginal cytology confirmed ovarian senescence in 20-month-old C57BL/6 mice. Mice were then randomly divided into activity groups, having access to a running wheel or not, and further into those receiving E(2) or placebo. Placebo-treated mice wheel ran more than E(2)-treated mice (P=0.03), with no difference between treatment groups in cage activities such as time spent being active and ambulation distance (P≥0.55). Soleus muscles from aged mice that wheel ran adapted by getting larger and stronger, irrespective of E(2) status (P≤0.02). Soleus muscle fatigue resistance was greater in mice treated with E(2) (P=0.02), but maximal isometric tetanic force was not affected (P≥0.79). Because E(2) treatment did not improve physical activity or overall muscle function in the aged, ovarian-senescent mice as predicted, a second study was initiated to examine E(2) treatment of young adult mice prematurely ovarian senescent from exposure to the chemical, 4-vinylcyclohexene diepoxide (VCD). Four-month-old C57BL/6 female mice were dosed with oil (control) or VCD. Vaginal cytology confirmed ovarian senescence in all mice treated with VCD 63 days after the onset of dosing, and then a subset of the VCD mice received E(2) (VCD+E(2)). Wheel running distance did not differ among control, VCD, and VCD+E(2) mice (P≥0.34). Soleus muscle concentric, isometric, and eccentric in vitro forces were greater in VCD+E(2) than in VCD mice (P<0.04), indicating beneficial estrogenic effects on muscle function. In general, aged and young mice with senescent ovaries were less responsive to E(2) treatment, in terms of physical activities and muscle function, than what has previously been shown for young, ovariectomized mice. These results bring forth the possibility that some component of the residual, follicle-depleted ovarian tissue influences physical activity in mice or that aging diminishes the responsiveness of skeletal muscle and related tissues to E(2) treatment.


Effectiveness of the PLAYgrounds programme on PA levels during recess in 6-year-old to 12-year-old children.
Worldwide levels of daily physical activity (PA) in children are low. This has negative health consequences. Schools have been recognised as key settings to promote PA. This study evaluates the effectiveness of the playground programme PLAYgrounds on increasing PA. PLAYgrounds was evaluated by a controlled trial, with a follow-up during one school year (10 months). Accelerometer data were collected on 1500 children in total, divided over 19 sampling moments (every 2 weeks). SOPLAY data were collected at nine sampling moments (once a month). Four intervention and four control schools were matched for playground size, number of pupils and PA levels at baseline. The intervention consisted of restructuring the playground by playground markings and by encouragement of the active use of the playground, through the provision of play equipment and educational measures such as adult encouragement and supporting physical education classes. Multilevel regression analyses were performed to analyse the effects of the intervention. PA levels in the intervention group (moderate PA) were significantly different (p<0.001) from the control group (light PA). During the intervention on an average 77.3% of the children engaged in moderate-to-vigorous physical activity in the intervention group and 38.7% in the control group. The effect of the intervention was significantly stronger for girls than for boys (p<0.001). The PLAYgrounds programme was effective in increasing PA levels in children during recess over the course of one school year. Thus, the programme could be used to provide structured PA promotion.


Electronic health records (EHRs) in clinical research and platform trials: Application of the innovative EHR-based methods developed by EU-PEARL.
Electronic Health Record (EHR) systems are digital platforms in clinical practice used to collect patients' clinical information related to their health status and represents a useful storage of real-world data. EHRs have a potential role in research studies, in particular, in platform trials. Platform trials are innovative trial designs including multiple trial arms (conducted simultaneously and/or sequentially) on different treatments under a single master protocol. However, the use of EHRs in research comes with important challenges such as incompleteness of records and the need to translate trial eligibility criteria into interoperable queries. In this paper, we aim to review and to describe our proposed innovative methods to tackle some of the most important challenges identified. This work is part of the Innovative Medicines Initiative (IMI) EU Patient-cEntric clinicAl tRial pLatforms (EU-PEARL) project's work package 3 (WP3), whose objective is to deliver tools and guidance for EHR-based protocol feasibility assessment, clinical site selection, and patient pre-screening in platform trials, investing in the building of a data-driven clinical network framework that can execute these complex innovative designs for which feasibility assessments are critically important. ISO standards and relevant references informed a readiness survey, producing 354 criteria with corresponding questions selected and harmonised through a 7-round scoring process (0-1) in stakeholder meetings, with 85% of consensus being the threshold of acceptance for a criterium/question. ATLAS cohort definition and Cohort Diagnostics were mainly used to create the trial feasibility eligibility (I/E) criteria as executable interoperable queries. The WP3/EU-PEARL group developed a readiness survey (eSurvey) for an efficient selection of clinical sites with suitable EHRs, consisting of yes-or-no questions, and a set-up of interoperable proxy queries using physicians' defined trial criteria. Both actions facilitate recruiting trial participants and alignment between study costs/timelines and data-driven recruitment potential. The eSurvey will help create an archive of clinical sites with mature EHR systems suitable to participate in clinical trials/platform trials, and the interoperable proxy queries of trial eligibility criteria will help identify the number of potential participants. Ultimately, these tools will contribute to the production of EHR-based protocol design.


Prevalence and correlates of tobacco use amongst junior collegiates in twin cities of western Nepal: a cross-sectional, questionnaire-based survey.
College students are vulnerable to tobacco addiction. Tobacco industries often target college students for marketing. Studies about prevalence of tobacco use and its correlates among college students in Nepal are lacking. A cross-sectional survey was carried out in two cities of western Nepal during January-March, 2007. A pre-tested, anonymous, self-administered questionnaire (in Nepali) adapted from Global Youth Tobacco Survey (GYTS) and a World Bank study was administered to a representative sample of 1600 students selected from 13 junior colleges by two-stage stratified random sampling. Overall prevalence of 'ever users' of tobacco products was 13.9%. Prevalence among boys and girls was 20.5% and 2.9% respectively. Prevalence of 'current users' was 10.2% (cigarette smoking: 9.4%, smokeless products: 6.5%, and both forms: 5.7%). Median age at initiation of cigarette smoking and chewable tobacco was 16 and 15 years respectively. Among the current cigarette smokers, 58.7% (88/150) were smoking at least one cigarette per day. Most (67.8%) 'Current users' purchased tobacco products by themselves from stores or got them from friends. Most of them (66.7%) smoked in tea stalls or restaurants followed by other public places (13.2%). The average daily expenditure was 20 Nepalese rupees (approximately 0.3 USD) and most (59%) students reported of having adequate money to buy tobacco products. Majority (82%) of the students were exposed to tobacco advertisements through magazines/newspapers, and advertising hoardings during a period of 30 days prior to survey. The correlates of tobacco use were: age, gender, household asset score and knowledge about health risks, family members, teachers and friends using tobacco products, and purchasing tobacco products for family members. School/college-based interventions like counseling to promote cessation among current users and tobacco education to prevent initiation are necessary. Enforcement of legislations to decrease availability, accessibility and affordability of tobacco products and policies to change social norms of tobacco use among parents and teachers are necessary to curb the tobacco use among college students.


Friction between finger flexor tendons and the pulley system in the crimp grip position.
Disruption of the finger flexor tendon pulleys are the most often occurring injury in rock climbers due to bowstringing of tendons during crimp grip position. The aim of this study was to quantify friction between the flexor tendons and pulleys and the influence of high load and speed of movement as a potential factor of pulley disruption. Friction between the flexor tendons and pulleys of eight human cadaver fingers was indirectly determined using an isokinetic movement device. During flexion and extension movement with rotational speed from 30 to 210 deg/s in the proximal interphalangeal joint and with load from 20 to 100 N to the flexor tendons the flexion force at the tip of the finger was measured. With 40 N loaded flexor tendons the force at the fingertip was 14.5 N (SD1.5) during extension and 12.6N (SD1.3) during flexion movement. Corresponding force difference of 12.9% and 3.77 N (SD0.6) force of friction can be calculated. Friction peaked at 85.8 degrees (SD2.05) of flexion of the proximal interphalangeal joint. Different speed of motion and load to the flexor tendons did not influence force difference other than linear. Considerable friction between flexor tendons and pulleys is apparent and therefore may have an influence on pulley injuries. Particularly during the crimp grip position where the proximal interphalangeal joint is flexed about 90 degrees shows the greatest amount of friction. However there was no change of friction during high speed motion and no other than linear increase during high load.


Streptococcus agalactiae-Induced Soft Tissue Infection in a Nonpregnant Adult After a Gynecological Procedure.
BACKGROUND We present a case of a 24-year-old woman with type 1- diabetes mellitus who developed necrotizing fasciitis (NF) due to Streptococcus agalactiae after a recent colposcopy. Literature review suggests this as the first case to be reported. CASE REPORT The patient initially presented to the emergency department (ED) with right lower neck pain and spasm of the right sternocleidomastoid muscle (SCM), with decreased range of motion. She was diagnosed with torticollis and was sent home on a nonsteroidal anti-inflammatory drug and spasmolytic. She returned 5 days later because of a lack of response. Magnetic resonance imaging of her neck revealed edema and inflammatory changes in the distal portion of her right SCM; an oral-systemic steroid was added to her treatment. However, she presented to the ED 3 days after her second visit with worsening symptoms. Her complaints of severe pain involving the right chest wall, development of fever, and the findings on imaging studies prompted the diagnosis of necrotizing soft-tissue infection and NF. She promptly underwent successful surgical debridement. Tissue cultures grew abundant Streptococcus agalactiae. Her antibiotics were readjusted and she was discharged to rehabilitation. Retrospective analysis of the case was notable for colposcopy with cervical biopsy and endocervical curettage for chronic cervicitis and low-grade squamous intraepithelial lesion within a week of her first ED visit. CONCLUSIONS NF caused by Streptococcus agalactiae should be suspected in patients who have had recent genitourinary/gastrointestinal procedures.


T lymphocytes control microbial composition by regulating the abundance of Vibrio in the zebrafish gut.
Dysbiosis of the intestinal microbial community is considered a risk factor for development of chronic intestinal inflammation as well as other diseases such as diabetes, obesity and even cancer. Study of the innate and adaptive immune pathways controlling bacterial colonization has however proven difficult in rodents, considering the extensive cross-talk between bacteria and innate and adaptive immunity. Here, we used the zebrafish to study innate and adaptive immune processes controlling the microbial community. Zebrafish lack a functional adaptive immune system in the first weeks of life, enabling study of the innate immune system in the absence of adaptive immunity. We show that in wild type zebrafish, the initial lack of adaptive immunity associates with overgrowth of Vibrio species (a group encompassing fish and human pathogens), which is overcome upon adaptive immune development. In Rag1-deficient zebrafish (lacking adaptive immunity) Vibrio abundance remains high, suggesting that adaptive immune processes indeed control Vibrio species. Using cell transfer experiments, we confirm that adoptive transfer of T lymphocytes, but not B lymphocytes into Rag1-deficient recipients suppresses outgrowth of Vibrio. In addition, ex vivo exposure of intestinal T lymphocytes to Rag1-deficient microbiota results in increased interferon-gamma expression by these T lymphocytes, compared to exposure to wild type microbiota. In conclusion, we show that T lymphocytes control microbial composition by effectively suppressing the outgrowth of Vibrio species in the zebrafish intestine.


Expression of enzymes involved in thyroid hormone metabolism during the early development of Xenopus tropicalis.
There are significant indications that amphibians require TH (thyroid hormones) prior to their involvement in the regulation of metamorphosis and before the development of a functional thyroid. In order to investigate the potential role for TH in pre-metamorphic Xenopus tropicalis we have cloned cDNAs for, and analysed the expression of, TPO (thyroid peroxidase), 5'DII (type II iodothyronine deiodinase) and 5DIII (type III iodothyronine deiodinase), enzymes involved in TH metabolism. Zygotic expression of TPO was detected in neurula stage embryos. Expression was observed in the notochord and later in the thyroid. The notochord was also a common site of expression for 5'DII and 5DIII. Other sites of 5'DII expression are the otic vesicles, retina, liver, blood-forming region, branchial arches and brain. 5DIII is also expressed in the brain, retina, liver, developing pro-nephros, blood-forming region and branchial arches. Embryos exposed to the TPO inhibitor methimazole showed a distinctive dose-dependent phenotype of a crimped notochord and shortened axis, together with alterations in (125)I(-) uptake. These data suggest a novel extrathyroidal role for TH during early development, and support the proposal that embryos require thyroid signalling for normal development prior to metamorphosis.


Impact of Inflammation on Myeloproliferative Neoplasm Symptom Development.
Myeloproliferative neoplasms (essential thrombocythemia, ET; polycythemia vera, PV; myelofibrosis, MF) are monoclonal malignancies associated with genomic instability, dysregulated signaling pathways, and subsequent overproduction of inflammatory markers. Acknowledged for their debilitating symptom profiles, recent investigations have aimed to determine the identity of these markers, the upstream sources stimulating their development, their prevalence within the MPN population, and the role they play in symptom development. Creation of dedicated Patient Reported Outcome (PRO) tools, in combination with expanded access to cytokine analysis technology, has resulted in a surge of investigations evaluating the potential associations between symptoms and inflammation. Emerging data demonstrates clear relationships between individual MPN symptoms (fatigue, abdominal complaints, microvascular symptoms, and constitutional symptoms) and cytokines, particularly IL-1, IL-6, IL-8, and TNF-α. Information is also compiling on the role symptoms paradoxically play in the development of cytokines, as in the case of fatigue-driven sedentary lifestyles. In this paper, we explore the symptoms inherent to the MPN disorders and the potential role inflammation plays in their development.


Impaired development of V gamma 3 dendritic epidermal T cells in p56lck protein tyrosine kinase-deficient and CD45 protein tyrosine phosphatase-deficient mice.
To determine whether p56lck protein tyrosine kinase and CD45 protein tyrosine phosphatase are involved in the signal transduction during intrathymic differentiation of gamma/delta T cells, we have examined the development of T cells expressing V gamma 3 T cell receptor (TCR) in mice deficient for either protein. The skin from both mice contained significantly reduced numbers of dendritic epidermal T cells expressing decreased levels of V gamma 3 TCR at the cell surface. Analysis of the fetal thymus from these mice suggested that maturation of V gamma 3 thymocytes was blocked at the immature stage that was characterized by the low level of V gamma 3 TCR and the high level of heat stable antigen. These results imply that both p56lck and CD45 are involved in the signal transduction during maturation of V gamma 3 T cells in the fetal thymus.


Plasmodium falciparum circumsporozoite antibody prevalence in Madagascar: a longitudinal study in three different epidemiologic areas.
A longitudinal study on the prevalence of Plasmodium falciparum circumsporozoite antibodies (Ab-CS) was carried out in 15 villages of three epidemiologic areas of Madagascar: the highlands, east coast, and the southwest region. A total of 3,967 blood samples were collected from November 1989 to April 1991 from cohorts of resident schoolchildren. The prevalence of Ab-CS in the examined population varied greatly according to the different ecoepidemiologic areas of the country. A correlation analysis was made between Ab-CS and P. falciparum parasite prevalence in the same population. High Ab-CS prevalence rates (25-75%) and levels (optical density = 0.28-0.76) were observed in the villages of the east coast (mesoendemic stable malaria). The Ab-CS prevalence rates varied from 0 to 37% in the highlands and southwest region villages (unstable malaria). The use of Ab-CS prevalence is proposed to be a useful and reliable seroepidemiologic marker of malaria endemicity in those areas of Madagascar in which malaria transmission is high and continues for more than four months a year.


Cognitive, affective, and conative theory of mind (ToM) in children with traumatic brain injury.
We studied three forms of dyadic communication involving theory of mind (ToM) in 82 children with traumatic brain injury (TBI) and 61 children with orthopedic injury (OI): Cognitive (concerned with false belief), Affective (concerned with expressing socially deceptive facial expressions), and Conative (concerned with influencing another's thoughts or feelings). We analyzed the pattern of brain lesions in the TBI group and conducted voxel-based morphometry for all participants in five large-scale functional brain networks, and related lesion and volumetric data to ToM outcomes. Children with TBI exhibited difficulty with Cognitive, Affective, and Conative ToM. The perturbation threshold for Cognitive ToM is higher than that for Affective and Conative ToM, in that Severe TBI disturbs Cognitive ToM but even Mild-Moderate TBI disrupt Affective and Conative ToM. Childhood TBI was associated with damage to all five large-scale brain networks. Lesions in the Mirror Neuron Empathy network predicted lower Conative ToM involving ironic criticism and empathic praise. Conative ToM was significantly and positively related to the package of Default Mode, Central Executive, and Mirror Neuron Empathy networks and, more specifically, to two hubs of the Default Mode Network, the posterior cingulate/retrosplenial cortex and the hippocampal formation, including entorhinal cortex and parahippocampal cortex.


The Effect of Anodal Transcranial Direct Current Stimulation Over Left and Right Temporal Cortex on the Cardiovascular Response: A Comparative Study.
Background: Stimulation of the right and left anterior insular cortex, increases and decreases the cardiovascular response respectively, thus indicating the brain's lateralization of the neural control of circulation. Previous experiments have demonstrated that transcranial direct current stimulation (tDCS) modulates the autonomic cardiovascular control when applied over the temporal cortex. Given the importance of neural control for a normal hemodynamic response, and the potential for the use of tDCS in the treatment of cardiovascular diseases, this study investigated whether tDCS was capable of modulating autonomic regulation. Methods: Cardiovascular response was monitored during a post-exercise muscle ischemia (PEMI) test, which is well-documented to increase sympathetic drive. A group of 12 healthy participants performed a PEMI test in a control (Control), sham (Sham) and two different experimental sessions where the anodal electrode was applied over the left temporal cortex and right temporal cortex with the cathodal electrode placed over the contralateral supraorbital area. Stimulation lasted 20 min at 2 mA. The hemodynamic profile was measured during a PEMI test. The cardiovascular parameters were continuously measured with a transthoracic bio-impedance device both during the PEMI test and during tDCS. Results: None of the subjects presented any side effects during or after tDCS stimulation. A consistent cardiovascular response during PEMI test was observed in all conditions. Statistical analysis did not find any significant interaction and any significant main effect of condition on cardiovascular parameters (all ps > 0.316) after tDCS. No statistical differences regarding the hemodynamic responses were found between conditions and time during tDCS stimulation (p > 0.05). Discussion: This is the first study comparing the cardiovascular response after tDCS stimulation of left and right TC both during exercise and at rest. The results of the current study suggest that anodal tDCS of the left and right TC does not affect functional cardiovascular response during exercise PEMI test and during tDCS. In light of the present and previous findings, the effect of tDCS on the cardiovascular response remains inconclusive.


To what extent does climate explain variations in reported malaria cases in early 20th century Uganda?
Malaria case statistics were analysed for the period 1926 to 1960 to identify inter-annual variations in malaria cases for the Uganda Protectorate. The analysis shows the mid-to-late 1930s to be a period of increased reported cases. After World War II, malaria cases trend down to a relative minimum in the early 1950s, before increasing rapidly after 1953 to the end of the decade. Data for the Western Province confirm these national trends, which at the time were attributed to a wide range of causes, including land development and management schemes, population mobility, interventions and misdiagnosis. Climate was occasionally proposed as a contributor to enhanced case numbers, and unusual precipitation patterns were held responsible; temperature was rarely, if ever, considered. In this study, a dynamical malaria model was driven with available precipitation and temperature data from the period for five stations located across a range of environments in Uganda. In line with the historical data, the simulations produced relatively enhanced transmission in the 1930s, although there is considerable variability between locations. In all locations, malaria transmission was low in the late 1940s and early 1950s, steeply increasing after 1954. Results indicate that past climate variability explains some of the variations in numbers of reported malaria cases. The impact of multiannual variability in temperature, while only on the order of 0.5°C, was sufficient to drive some of the trends observed in the statistics and thus the role of climate was likely underestimated in the contemporary reports. As the elimination campaigns of the 1960s followed this partly climate-driven increase in malaria, this emphasises the need to account for climate when planning and evaluating intervention strategies.


Carbonyl stress and a combination of astaxanthin/vitamin C induce biochemical changes in human neutrophils.
The purpose of the present study was to find out whether co-treatment of human neutrophils with high glucose and methylglyoxal (MGO) can alter the biochemical parameters of human neutrophils. We also examined if astaxanthin associated with vitamin C can improve those biochemical parameters. Neutrophils from healthy subjects were treated with 20mM of glucose and 30 μM MGO followed or not by the addition of the antioxidants astaxanthin (2 μM) and vitamin C (100 μM). MGO/high glucose treatment reduced the phagocytic capacity and the G6PDH, total/SOD and GR activities. Additionally, there was an increase in the activity of myeloperoxidase (MPO) with consequent increase in the hypochlorous acid production, CAT activity and in the release of IL-6 cytokine without changes in intracellular calcium mobilization. Our study also shows that the association of astaxanthin with vitamin C greatly improved neutrophil phagocytic capacity, decreasing all reactive oxygen species measured, pro-inflammatory IL-1β and TNF-α release, MPO activity and HClO production. The combination of astaxanthin with vitamin C alone has more antioxidant and anti-inflammatory effects than when they were in the presence of MGO/high glucose. Injury to the function of neutrophils due to high glucose and methylglyoxal appears not to involve oxidative stress or calcium release. The association of antioxidants astaxanthin and vitamin C promoted a significant improvement in the function of neutrophils and in the redox status.


Towards prognostic functional brain biomarkers for cervical myelopathy: A resting-state fMRI study.
Recently, there has been increasing interest in strategies to predict neurological recovery in cervical myelopathy (CM) based on clinical images of the cervical spine. In this study, we aimed to explore potential preoperative brain biomarkers that can predict postoperative neurological recovery in CM patients by using resting-state functional magnetic resonance imaging (rs-fMRI) and functional connectivity (FC) analysis. Twenty-eight patients with CM and 28 age- and sex-matched healthy controls (HCs) underwent rs-fMRI (twice for CM patients, before and six months after surgery). A seed-to-voxel analysis was performed, and the following three statistical analyses were conducted: (i) FC comparisons between preoperative CM and HC; (ii) correlation analysis between preoperative FCs and clinical scores; and (iii) postoperative FC changes in CM. Our analyses identified three FCs between the visual cortex and the right superior frontal gyrus based on the conjunction of the first two analyses [(i) and (ii)]. These FCs may act as potential biomarkers for postoperative gain in the 10-second test and might be sufficient to provide a prediction formula for potential recovery. Our findings provide preliminary evidence supporting the possibility of novel predictive measures for neurological recovery in CM using rs-fMRI.


Dopamine D2L receptors stimulate Na+/K(+)-ATPase activity in murine LTK- cells.
Ion transport can be regulated by dopamine receptors. D1-like receptors inhibit both Na+/H+ exchange (NHE) and Na+/K(+)-ATPase activity, whereas D2-like receptors stimulate NHE. However, the effect of D2-like receptors on Na+/K(+)-ATPase activity is controversial. In renal proximal tubular cells, where several D1-like and D2-like receptors are expressed, D2 agonists have been reported either to have no effect or to act in concert with D1 agonists to inhibit Na+/K(+)-ATPase activity. We therefore studied the effect of D2 receptors on Na+/K(+)-ATPase activity in LTK- cells transfected with a rat D2Long receptor cDNA (maximum receptor density = 0.91 +/- 0.26 pmol/mg protein, dissociation constant = 2.39 +/- 0.79 nM, seven experiments). The activation of D2 receptors in these transfected cells by the selective D2 agonist LY171555 led to the inhibition of forskolin-stimulated cAMP accumulation. In the D2Long-transfected, but not in nontransfected cells, LY171555 caused a concentration-dependent stimulation of Na+/K(+)-ATPase activity (EC50 = 0.55 +/- 0.2 microM, Emax = 28 +/- 6%, six experiments), which was completely blocked by the D2-selective antagonist (-)-sulpiride. The D2-stimulated Na+/K(+)-ATPase activity was not secondary to D2 receptor activation of K+ channels or NHE activity since LY171555 stimulated Na+/K(+)-ATPase activity in D2Long-transfected cells, even when K+ channels were blocked by CsCl and intracellular Na+ was clamped by monensin. The D2-stimulated Na+/K(+)-ATPase activity was blocked by pertussis toxin and mimicked by dideoxyadenosine. We conclude that agonist occupancy of D2Long dopamine receptors stimulates Na+/K(+)-ATPase activity; this effect is mediated by the inhibition of cAMP production and is independent of intracellular Na+ and K+ concentration.


Cryophotolysis of caged compounds: a technique for trapping intermediate states in protein crystals.
Caged compounds in combination with protein crystallography represent a valuable tool in studies of enzyme reaction intermediates. To date, photochemical triggering of reactions has been performed close to room temperature. Synchronous reaction initiation has only been achieved with enzymes of relatively slow turnover (<0.1 s(-1)) and caged compounds of high quantum yield. Here X-ray crystallography and microspectrophotometry were used to provide evidence that (nitrophenyl)ethyl (NPE) ester bonds can be photolyzed by UV light at cryotemperatures. NPE-caged ATP in flash-cooled crystals of Mycobacterium tuberculosis thymidylate kinase was photolyzed successfully at 100-150 K as assessed by the structural observation of ATP-dependent enzymatic conversion of TMP to TDP after temporarily warming the crystals to room temperature. A new method is proposed in which cryo-photolysis combined with temperature-controlled protein crystallography can be used to trap reaction intermediates even in some of the fastest enzymes and/or when only compounds of low quantum yield are available. Raising the temperature after cryophotolysis may allow a transition barrier to be passed and an intermediate to accumulate in the crystal. A comparable method has only been used so far with proteins displaying endogenous photosensitivity. The approach described here opens the way to studying the reaction mechanisms of a much larger number of crystalline enzymes. Furthermore, it is shown that X-ray-induced radiolysis of caged compounds occurs if high-intensity synchrotron beamlines are used. This caveat should be taken into account when deriving data-collection protocols. It could also be used potentially as a way to trigger reactions.


Inflammatory Monocytes Promote Perineural Invasion via CCL2-Mediated Recruitment and Cathepsin B Expression.
Perineural invasion (PNI) is an ominous event strongly linked to poor clinical outcome. Cells residing within peripheral nerves collaborate with cancer cells to enable PNI, but the contributing conditions within the tumor microenvironment are not well understood. Here, we show that CCR2-expressing inflammatory monocytes (IM) are preferentially recruited to sites of PNI, where they differentiate into macrophages and potentiate nerve invasion through a cathepsin B-mediated process. A series of adoptive transfer experiments with genetically engineered donors and recipients demonstrated that IM recruitment to nerves was driven by CCL2 released from Schwann cells at the site of PNI, but not CCL7, an alternate ligand for CCR2. Interruption of either CCL2-CCR2 signaling or cathepsin B function significantly impaired PNI in vivo Correlative studies in human specimens demonstrated that cathepsin B-producing macrophages were enriched in invaded nerves, which was associated with increased local tumor recurrence. These findings deepen our understanding of PNI pathogenesis and illuminate how PNI is driven in part by corruption of a nerve repair program. Further, they support the exploration of inhibiting IM recruitment and function as a targeted therapy for PNI. Cancer Res; 77(22); 6400-14. ©2017 AACR.


Does This Patient Have Acute Mountain Sickness?: The Rational Clinical Examination Systematic Review.
Acute mountain sickness (AMS) affects more than 25% of individuals ascending to 3500 m (11 500 ft) and more than 50% of those above 6000 m (19 700 ft). AMS may progress from nonspecific symptoms to life-threatening high-altitude cerebral edema in less than 1% of patients. It is not clear how to best diagnose AMS. To systematically review studies assessing the accuracy of AMS diagnostic instruments, including the visual analog scale (VAS) score, which quantifies the overall feeling of sickness at altitude (VAS[O]; various thresholds), Acute Mountain Sickness-Cerebral score (AMS-C; ≥0.7 indicates AMS), and the clinical functional score (CFS; ≥2 indicates AMS) compared with the Lake Louise Questionnaire Score (LLQS; score of ≥5). Searches of MEDLINE and EMBASE from inception to May 2017 identified 1245 publications of which 91 were suitable for prevalence analysis (66 944 participants) and 14 compared at least 2 instruments (1858 participants) using a score of 5 or greater on the LLQS as a reference standard. To determine the prevalence of AMS for establishing the pretest probability of AMS, a random-effects meta-regression was performed based on the reported prevalence of AMS as a function of altitude. AMS prevalence, likelihood ratios (LRs), sensitivity, and specificity of screening instruments. The final analysis included 91 articles (comprising 66 944 study participants). Altitude predicted AMS and accounted for 28% of heterogeneity between studies. For each 1000-m (3300-ft) increase in altitude above 2500 m (8200 ft), AMS prevalence increased 13% (95% CI, 9.5%-17%). Testing characteristics were similar for VAS(O), AMS-C, and CFS vs a score of 5 or greater on the LLQS (positive LRs: range, 3.2-8.2; P = .22 for comparisons; specificity range, 67%-92%; negative LRs: range, 0.30-0.36; P = .50 for comparisons; sensitivity range, 67%-82%). The CFS asks a single question: "overall if you had any symptoms, how did they affect your activity (ordinal scale 0-3)?" For CFS, moderate to severe reduction in daily activities had a positive LR of 3.2 (95% CI, 1.4-7.2) and specificity of 67% (95% CI, 37%-97%); no reduction to mild reduction in activities had a negative LR of 0.30 (95% CI, 0.22-0.39) and sensitivity of 82% (95% CI, 77%-87%). The prevalence of acute mountain sickness increases with higher altitudes. The visual analog scale for the overall feeling of sickness at altitude, Acute Mountain Sickness-Cerebral, and clinical functional score perform similarly to the Lake Louise Questionnaire Score using a score of 5 or greater as a reference standard. In clinical and travel settings, the clinical functional score is the simplest instrument to use. Clinicians evaluating high-altitude travelers who report moderate to severe limitations in activities of daily living (clinical functional score ≥2) should use the Lake Louise Questionnaire Score to assess the severity of acute mountain sickness.


Increased Oxidative and Nitrative Stress Accelerates Aging of the Retinal Vasculature in the Diabetic Retina.
Hyperglycemia-induced retinal oxidative and nitrative stress can accelerate vascular cell aging, which may lead to vascular dysfunction as seen in diabetes. There is no information on whether this may contribute to the progression of diabetic retinopathy (DR). In this study, we have assessed the occurrence of senescence-associated markers in retinas of streptozotocin-induced diabetic rats at 8 and 12 weeks of hyperglycemia as compared to normoglycemic aging (12 and 14 months) and adult (4.5 months) rat retinas. We have found that in the diabetic retinas there was an up-regulation of senescence-associated markers SA-β-Gal, p16INK4a and miR34a, which correlated with decreased expression of SIRT1, a target of miR34a. Expression of senescence-associated factors primarily found in retinal microvasculature of diabetic rats exceeded levels measured in adult and aging rat retinas. In aging rats, retinal expression of senescence associated-factors was mainly localized at the level of the retinal pigmented epithelium and only minimally in the retinal microvasculature. The expression of oxidative/nitrative stress markers such as 4-hydroxynonenal and nitrotyrosine was more pronounced in the retinal vasculature of diabetic rats as compared to normoglycemic aging and adult rat retinas. Treatments of STZ-rats with the anti-nitrating drug FeTPPS (10mg/Kg/day) significantly reduced the appearance of senescence markers in the retinal microvasculature. Our results demonstrate that hyperglycemia accelerates retinal microvascular cell aging whereas physiological aging affects primarily cells of the retinal pigmented epithelium. In conclusion, hyperglycemia-induced retinal vessel dysfunction and DR progression involve vascular cell senescence due to increased oxidative/nitrative stress.


Multiple discrete encephalitogenic epitopes of the autoantigen myelin basic protein include a determinant for I-E class II-restricted T cells.
Immunization with the autoantigen myelin basic protein (MBP) causes experimental allergic encephalomyelitis (EAE). Initial investigations indicated that encephalitogenic murine determinants of MBP were located only within MBP 1-37 and MBP 89-169. Encephalitogenic T cell epitopes within these fragments have been identified. Each epitope is recognized by T cells in association with separate allelic I-A molecules. A hybrid I-E-restricted T cell clone that recognizes intact mouse (self) MBP has been examined. The epitope recognized by this clone includes MBP residues 35-47. When tested in vivo, p35-47 causes EAE. T cell recognition of p35-47 occurs only in association with I-E molecules. These results provide the first clear example that antigen-specific T cells restricted by I-E class II molecules participate in murine autoimmune disease. Furthermore, it is clear that there are multiple (at least three) discrete encephalitogenic T cell epitopes of this autoantigen, each recognized in association with separate allelic class II molecules. These results may be relevant to human autoimmune diseases whose susceptibility is associated with more than one HLA-D molecule.


Complete genome sequence of the ethanol-producing Zymomonas mobilis subsp. mobilis centrotype ATCC 29191.
Zymomonas mobilis is an ethanologenic bacterium that has been studied for use in biofuel production. Of the sequenced Zymomonas strains, ATCC 29191 has been described as the phenotypic centrotype of Zymomonas mobilis subsp. mobilis, the taxon that harbors the highest ethanol-producing Z. mobilis strains. ATCC 29191 was isolated in Kinshasa, Congo, from palm wine fermentations. This strain is reported to be a robust levan producer, while in recent years it has been employed in studies addressing Z. mobilis respiration. Here we announce the finishing and annotation of the ATCC 29191 genome, which comprises one chromosome and three plasmids.


Construction and first characterization of two reciprocal hybrids between LamB from Escherichia coli K12 and Klebsiella pneumoniae.
The LamB proteins from Klebsiella pneumoniae and Escherichia coli K12 were previously shown to be highly homologous. The most conserved parts correspond to the N-proximal third and to the transmembranous portions of the molecule, while the variability occurred essentially within regions exposed to the cell surface or to the periplasm. Since the two proteins displayed identical in vitro trimer stability and in vivo pore properties, we tested whether the N-terminal parts of the two proteins could be exchanged and still allow the formation of stable and functional maltoporins. For that purpose, we expressed the LamB protein from K. pneumoniae in E. coli K12, and constructed two reciprocal hybrids between LamB from E. coli K12 and LamB from K. pneumoniae. The first hybrid (LamBE.c.-K.p.) is composed of residues 1 to 183 from LamBE.c. followed by residues 184 to 404 from LamBK.p. The second one comprises residues 1 to 183 from LamBK.p., followed by residues 184 to 421 from LamBE.c. (LamBK.p.-E.c.). Both hybrid proteins were correctly incorporated in the outer membrane of E. coli K12. Like the two parental LamB proteins, the two hybrids could be purified by affinity chromatography on a starch-sepharose column. The LamBE.c.-K.p. hybrid formed highly stable trimers, but was strongly impaired in its in vivo maltose transport function (15% of the wild-type level). The trimers formed by LamBK.p.-E.c. hybrid were less stable, but could be detected on the surface of intact cells by four anti-LamBE.c. monoclonal antibodies. This hybrid was also affected in its in vivo maltose transport function (30% of the wild-type level). As expected from the location of the residues critical for phage adsorption, both proteins had lost the phage receptor activity of the E. coli K12 LamB protein. We also examined whether LamBE.c. could form heterotrimers with LamBK.p., LamBK.p.-E.c., and LamBE.c.-K.p. In no case were heterotrimers detected, indicating that both terminal parts of the LamB protein are involved in homotrimer formation. All these data suggest that trimer formation and activity involve rare variable residues in the conserved regions and/or variable regions.


Itch: more than skin deep.
Recent research in neurophysiology of itch has indicated the existence of itch-dedicated nociceptor neurones. The perception of itch is regulated by tonically inhibitory descending neuronal pathways and nociceptor spinal neuronal circuits. There is at present no convincing evidence of an 'itch centre' in the brain. A classification of itch has been proposed, based on neurophysiological considerations, which stresses the importance of neurogenic and neuropathic itch, and assists in differential diagnosis and selection of treatment. However, more than one class of itch can occur concurrently in the same patient. The importance of cross- talk between dermal mast cells and nociceptor nerve terminals, involving cleavage of proteinase-activated receptor 2 by mast cell tryptase, is highlighted. The pruritus of cholestasis is mediated at least in part by opioid peptides synthesized by the liver, and elevated levels of these mediators are found in the plasma and skin of patients with itch due to cholestasis. The combined use of both mu-receptor antagonists and kappa-receptor agonists (anti-pruritic) is worth exploring.


A numerical study of microparticle acoustophoresis driven by acoustic radiation forces and streaming-induced drag forces.
We present a numerical study of the transient acoustophoretic motion of microparticles suspended in a liquid-filled microchannel and driven by the acoustic forces arising from an imposed standing ultrasound wave: the acoustic radiation force from the scattering of sound waves on the particles and the Stokes drag force from the induced acoustic streaming flow. These forces are calculated numerically in two steps. First, the thermoacoustic equations are solved to first order in the imposed ultrasound field taking into account the micrometer-thin but crucial thermoviscous boundary layer near the rigid walls. Second, the products of the resulting first-order fields are used as source terms in the time-averaged second-order equations, from which the net acoustic forces acting on the particles are determined. The resulting acoustophoretic particle velocities are quantified for experimentally relevant parameters using a numerical particle-tracking scheme. The model shows the transition in the acoustophoretic particle motion from being dominated by streaming-induced drag to being dominated by radiation forces as a function of particle size, channel geometry, and material properties.


Deciphering the fossil record of early bilaterian embryonic development in light of experimental taphonomy.
Experimental analyses of decay in a tunicate deuterostome and three lophotrochozoans indicate that the controls on decay and preservation of embryos, identified previously based on echinoids, are more generally applicable. Four stages of decay are identified regardless of the environment of death and decay. Embryos decay rapidly in oxic and anoxic conditions, although the gross morphology of embryos is maintained for longer under anoxic conditions. Under anoxic reducing conditions, the gross morphology of the embryos is maintained for the longest period of time, compatible with the timescale required for bacterially mediated mineralization of soft tissues. All four stages of decay were encountered under all environmental conditions, matching the spectrum of preservational qualities encountered in all fossil embryo assemblages. The preservation potential of embryos of deuterostomes and lophotrochozoans is at odds with the lack of such embryos in the fossil record. Rather, the fossil record of embryos, as sparse as it is, is dominated by forms interpreted as ecdysozoans, cnidarians, and stem-metazoans. The dearth of deuterostome and lophotrochozoan embryos may be explained by the fact that ecdysozoans, at least, tend to deposit their eggs in the sediment rather than through broadcast spawning. However, fossil embryos remain very rare and the main controlling factor on their fossilization may be the unique conspiracy of environmental conditions at a couple of sites. The preponderance of fossilized embryos of direct developers should not be used in evidence against the existence of indirect development at this time in animal evolutionary history.


Investigating a socially anxious-impulsive subgroup of adolescents: a prospective community study.
Previous research has identified a subgroup of socially anxious adults who are both anxious and impulsive. To date, however, this subgroup has not been identified in adolescence. Therefore, in this study we aimed to identify this subgroup in a sample of adolescents. In addition, we hypothesized that this subgroup would be higher on problem behaviors, and that these processes would be moderated by gender. We used longitudinal data from 714 adolescents who were in the 7th and 8th grades at Time 1. They were followed annually for three years. Cluster analyses identified an anxious-inhibited subgroup as well as an anxious-impulsive subgroup in early adolescence (Time 1). The socially anxious-impulsive adolescent boys were generally higher on both intoxication frequency and delinquency compared with all other adolescents in all clusters at each time point. Findings suggest that social anxiety subgroups may differ on problem behavior, and that early detection of an anxious-impulsive subgroup may be important to prevent maladjustment, especially for adolescent boys.


Glomerular expression of biglycan and decorin and urinary levels of decorin in primary glomerular disease.
Recent studies have suggested that small leucine-rich proteoglycans (SLRP) of the extracellular matrix play a major role in modulating the activity of growth factors and in regulating the deposition of collagens. In this study, the expression of the SLRPs biglycan and decorin in the glomeruli of patients with primary glomerular disease (minimal change disease, IgA nephropathy, and membranous nephropathy) and urine immunoreactive levels examined. Renal biopsy specimens were obtained from patients with minimal change disease, IgA nephropathy and membranous nephropathy. Immunohistochemical staining was performed on fresh-frozen samples using anti-biglycan and anti-decorin antibodies. Examination of urine proteoglycan excretion from a total of 26 patients and 8 normal volunteers was performed by indirect ELISA. In normal kidney samples, biglycan and decorin expression was found predominantly in the intrarenal arteries and tubulointerstitium, with only minimal expression in the glomeruli. Glomerular expression of these proteoglycans in glomerular disease was unchanged in all of the 4 patients examined with minimal change disease. In the case of IgA nephropathy or membranous nephropathy, some of the patients showed minimally increased immunostaining of either biglycan or decorin, but there were no signs of simultaneous upregulation of both proteoglycans. To further examine the changes in proteoglycan expression, ELISA was performed on urine samples. Urine biglycan levels were below detection levels, but high values of urine decorin immunoreactivity were found in the patients with glomerular disease. A significant negative correlation was found between urine decorin and creatinine clearance. These results suggest that distinct changes in the expression of the SLRPs biglycan and decorin may be seen in patients with primary glomerular disease. Moreover, the negative relationship between urine decorin levels and renal function supports the hypothesis that decorin may be involved in the pathophysiology of renal dysfunction in humans.


The relative timing of VMO and VL in the aetiology of anterior knee pain: a systematic review and meta-analysis.
Anterior knee pain (AKP) is a common musculoskeletal complaint. It has been suggested that one factor that may contribute to the presence of AKP is a delay in the recruitment of the vastus medialis oblique muscle (VMO) relative to the vastus lateralis muscle (VL). There is however little consensus within the literature regarding the existence or nature of any such delay in the recruitment of the VMO within the AKP population. The purpose of this systematic review and meta-analysis was to examine the relative timing of onset of the VMO and VL in those with AKP in comparison to the asymptomatic population. The bibliographic databases AMED, British Nursing Index, CINAHL, EMBASE, Ovid Medline, PEDro, Pubmed and the Cochrane Library were searched for studies comparing the timing of EMG onset of the VMO and VL in those with AKP versus the asymptomatic population. Studies fulfilling the inclusion criteria were independently assessed. Heterogeneity across the studies was measured. A meta-analysis of results was completed for those studies where adequate data was supplied. Where comparable methodologies had been used, results were pooled and analysed. Fourteen studies met the inclusion criteria; one prospective and thirteen observational case control. Eleven compared VMO and VL EMG onset times during voluntary active tasks while four investigated reflex response times. All used convenience sampling and did not state blinding of the assessor. Study methodologies/testing and assessment procedures varied and there was considerable heterogeneity within individual samples. Whilst a trend was identified towards a delay in onset of VMO relative to the VL in the AKP population during both voluntary active tasks and reflex activity, a substantial degree of heterogeneity across the pooled studies was identified (I2 = 69.9-93.4%, p < 0.01). Findings are subject to substantial and unexplained heterogeneity. A trend was demonstrated towards a delayed onset of VMO relative to VL in those with AKP in comparison to those without. However not all AKP patients demonstrate a VMO-VL dysfunction, and this is compounded by normal physiological variability in the healthy population. The clinical and therapeutic significance is therefore difficult to assess.


Virtual communication curbs creative idea generation.
COVID-19 accelerated a decade-long shift to remote work by normalizing working from home on a large scale. Indeed, 75% of US employees in a 2021 survey reported a personal preference for working remotely at least one day per week1, and studies estimate that 20% of US workdays will take place at home after the pandemic ends2. Here we examine how this shift away from in-person interaction affects innovation, which relies on collaborative idea generation as the foundation of commercial and scientific progress3. In a laboratory study and a field experiment across five countries (in Europe, the Middle East and South Asia), we show that videoconferencing inhibits the production of creative ideas. By contrast, when it comes to selecting which idea to pursue, we find no evidence that videoconferencing groups are less effective (and preliminary evidence that they may be more effective) than in-person groups. Departing from previous theories that focus on how oral and written technologies limit the synchronicity and extent of information exchanged4-6, we find that our effects are driven by differences in the physical nature of videoconferencing and in-person interactions. Specifically, using eye-gaze and recall measures, as well as latent semantic analysis, we demonstrate that videoconferencing hampers idea generation because it focuses communicators on a screen, which prompts a narrower cognitive focus. Our results suggest that virtual interaction comes with a cognitive cost for creative idea generation.


Nontargeted virus sequence discovery pipeline and virus clustering for metagenomic data.
The analysis of large microbiome data sets holds great promise for the delineation of the biological and metabolic functioning of living organisms and their role in the environment. In the midst of this genomic puzzle, viruses, especially those that infect microbial communities, represent a major reservoir of genetic diversity with great impact on biogeochemical cycles and organismal health. Overcoming the limitations associated with virus detection directly from microbiomes can provide key insights into how ecosystem dynamics are modulated. Here, we present a computational protocol for accurate detection and grouping of viral sequences from microbiome samples. Our approach relies on an expanded and curated set of viral protein families used as bait to identify viral sequences directly from metagenomic assemblies. This protocol describes how to use the viral protein families catalog (∼7 h) and recommended filters for the detection of viral contigs in metagenomic samples (∼6 h), and it describes the specific parameters for a nucleotide-sequence-identity-based method of organizing the viral sequences into quasi-species taxonomic-level groups (∼10 min).


A prospective analysis of 211 robotic-assisted surgical procedures.
The Academic Robotics Group prospectively studied 211 robotically assisted operations to assess the safety and utility of robotically assisted surgery. All operations took place at one of four member institutions between June 2000 and June 2001 using the recently FDA-approved daVinci robotic system. A variety of procedures were undertaken, including antireflux surgery (69), cholecystectomy (36), Heller myotomy (26), bowel resection (17), donor nephrectomy (15), left internal mammery artery mobilization (14), gastric bypass (seven), splenectomy (seven), adrenalectomy (six), exploratory laparoscopy (three), pyloroplasty (four), gastrojejunostomy (two), distal pancreatectomy (one), duodenal polypectomy (one), esophagectomy (one), gastric mass resection (one), and lysis of adhesions (one). Average operating room time was 188 min (range 45 to 387, SD = 83), surgical time 143 min (range 35 to 462, SD = 63), and robot time 90 min (range 12 to 235, SD = 47). Median length of stay was 1 day (range 0 to 37). There were 8 (4%) technical complications during procedures, five minor (four hook cautery dislodgement, one slipped robotic trocar) and three major (system malfunctions, two of which required conversion to standard laparoscopy). In all cases, technical problems caused only delay, without apparent altered outcome. There were medical/surgical complications in nine patients (4%). Six (3%) were considered major, including one death unrelated to the robotic procedure. The results of robotic-assisted surgery compare favorably with those of conventional laparoscopy with respect to mortality, complications, and length of stay. Robotic-assisted surgery is safe and effective and is a new reality for American surgery. The role of these devices in surgery will expand as the technology evolves.


Proteome analysis of differential protein expression in allergen-induced asthmatic mice lung after dexamethasone treatment.
Asthma has become substantially more prevalent in recent decades and is one of the foremost contributors to morbidity and mortality in industrialized countries. Corticosteroids are among the most effective medications for the treatment of asthma, but some patients do not respond well to corticosteroid treatment. In this study, we characterized the responses to an allergen and identified potential molecular targets of dexamethasone (Dex) treatment in acute asthma. Female BALB/c mice sensitized to ovalbumin (OVA) were challenged with aerosolized OVA for 1 week. During the challenge period, mice were treated daily with Dex by intraperitoneal injection. Phosphate-buffered saline treated and non-challenged mice served as control. Histological evaluation of OVA-induced mice revealed airway inflammation and goblet cell hyperplasia. In addition, interleukin 4 levels and interferon-gamma levels were increased and decreased, respectively. These changes were moderated by Dex treatment. Protein expression profiles were compared in each experimental group by two-dimensional gel electrophoresis and identified using matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry. Some proteins were increased, while others were decreased by Dex treatment. These results indicated that the regulation of protein expression might play a role in the immunological and pathological development of asthma and could be targeted for therapeutic intervention. These results may assist in the development of quantitative diagnostic markers to monitor disease progression or responses to therapy using proteomic approaches.


A kernelisation approach for multiple d-Hitting Set and its application in optimal multi-drug therapeutic combinations.
Therapies consisting of a combination of agents are an attractive proposition, especially in the context of diseases such as cancer, which can manifest with a variety of tumor types in a single case. However uncovering usable drug combinations is expensive both financially and temporally. By employing computational methods to identify candidate combinations with a greater likelihood of success we can avoid these problems, even when the amount of data is prohibitively large. Hitting Set is a combinatorial problem that has useful application across many fields, however as it is NP-complete it is traditionally considered hard to solve exactly. We introduce a more general version of the problem (α,β,d)-Hitting Set, which allows more precise control over how and what the hitting set targets. Employing the framework of Parameterized Complexity we show that despite being NP-complete, the (α,β,d)-Hitting Set problem is fixed-parameter tractable with a kernel of size O(αdk(d)) when we parameterize by the size k of the hitting set and the maximum number α of the minimum number of hits, and taking the maximum degree d of the target sets as a constant. We demonstrate the application of this problem to multiple drug selection for cancer therapy, showing the flexibility of the problem in tailoring such drug sets. The fixed-parameter tractability result indicates that for low values of the parameters the problem can be solved quickly using exact methods. We also demonstrate that the problem is indeed practical, with computation times on the order of 5 seconds, as compared to previous Hitting Set applications using the same dataset which exhibited times on the order of 1 day, even with relatively relaxed notions for what constitutes a low value for the parameters. Furthermore the existence of a kernelization for (α,β,d)-Hitting Set indicates that the problem is readily scalable to large datasets.


Proximal-distal leg development in Drosophila requires the apterous gene and the Lim1 homologue dlim1.
Proximal-distal leg development in Drosophila involves a battery of genes expressed and required in specific proximal-distal (PD) domains of the appendage. Here we report the characterisation of a new gene of this type, dlim1, a member of the Lhx family of genes whose proteins contain two Lim domains and a homeodomain. We show that the Lhx gene apterous (ap) is also required for PD leg development, and we study the functional interactions between ap, dlim1 and other PD genes during leg development. Our results show that a regulatory network formed by ap and dlim1 plus the homeobox genes aristaless and Bar specifies distal leg cell fates in Drosophila.


Neuromuscular regeneration by buccal motoneuron B15 after peripheral nerve crush in Aplysia californica.
1. We studied regeneration of neuromuscular connections by identified buccal motoneuron B15 after axotomy produced by crushing nerve 4; the intact contralateral nerve 4 served as control. Electrophysiological recordings, intracellular dye injections, and light and electron microscopy were used to characterize the nature and time course of neuromuscular reinnervation as well as the fate of the isolated distal stump of the motor axon. 2. Axonal outgrowth or sprouting in the form of numerous "regenerites" occurred from the proximal stump of the transected B15 axon, and these regenerites projected through the crush site along the length of the nerve to innervate target muscles at the periphery. 3. Reinnervation of one of the target muscles, the accessory radula closer (I5), was first detected 3 wk after nerve crush. Neuromuscular excitatory postsynaptic potentials measured in individual I5 muscle fibers were initially small and approached control amplitudes by 8 wk postlesion. Newly regenerated neuromuscular synapses displayed facilitation and depression to repeated B15 stimulation with properties similar to those of control synapses, even at early times postlesion. 4. Reinnervation of other buccal muscles by B15, such as I4, appeared slightly delayed relative to that observed for I5. No evidence of abnormal or enlarged fields of innervation were observed, and as in control preparations, regenerated neuromuscular connections were strictly limited to muscles ipsilateral to the B15 cell body. 5. Physiological evidence suggested that the distal axon stumps of B15, although isolated from their cell bodies, survive for several weeks after axotomy. In addition, several large axon profiles indicative of motor axons were seen in cross-sections of nerve 4 taken close to the muscle and distal to the crush site, indicating survival of distal axon stumps. 6. When B15 was selectively stimulated, the newly formed regenerites failed to fire the distal axon stump of B15, demonstrating that the regenerites do not reinnervate the distal stump. 7. Degeneration of axons in nerve 4 distal to the crush site was observed in cross-sections of the nerve at 8 wk postlesion; using ultrathin sections we found cellular debris in individual axon profiles as well as large acellular masses within nerve 4, the latter likely representing the concretion of many axons. Additional evidence for such degenerative changes appeared in the form of autofluorescing spherical bodies or "spheroids" both in individual axons and the nerve distal to the crush site.(ABSTRACT TRUNCATED AT 400 WORDS)


Development of a single nucleotide polymorphism barcode to genotype Plasmodium vivax infections.
Plasmodium vivax, one of the five species of Plasmodium parasites that cause human malaria, is responsible for 25-40% of malaria cases worldwide. Malaria global elimination efforts will benefit from accurate and effective genotyping tools that will provide insight into the population genetics and diversity of this parasite. The recent sequencing of P. vivax isolates from South America, Africa, and Asia presents a new opportunity by uncovering thousands of novel single nucleotide polymorphisms (SNPs). Genotyping a selection of these SNPs provides a robust, low-cost method of identifying parasite infections through their unique genetic signature or barcode. Based on our experience in generating a SNP barcode for P. falciparum using High Resolution Melting (HRM), we have developed a similar tool for P. vivax. We selected globally polymorphic SNPs from available P. vivax genome sequence data that were located in putatively selectively neutral sites (i.e., intergenic, intronic, or 4-fold degenerate coding). From these candidate SNPs we defined a barcode consisting of 42 SNPs. We analyzed the performance of the 42-SNP barcode on 87 P. vivax clinical samples from parasite populations in South America (Brazil, French Guiana), Africa (Ethiopia) and Asia (Sri Lanka). We found that the P. vivax barcode is robust, as it requires only a small quantity of DNA (limit of detection 0.3 ng/μl) to yield reproducible genotype calls, and detects polymorphic genotypes with high sensitivity. The markers are informative across all clinical samples evaluated (average minor allele frequency > 0.1). Population genetic and statistical analyses show the barcode captures high degrees of population diversity and differentiates geographically distinct populations. Our 42-SNP barcode provides a robust, informative, and standardized genetic marker set that accurately identifies a genomic signature for P. vivax infections.


Hydroxyapatite-coated total hip femoral components in patients less than fifty years old. Clinical and radiographic results after five to eight years of follow-up.
One hundred and thirty-three patients (152 hips) who were an average of thirty-nine years old (range, sixteen to forty-nine years old) received a proximally hydroxyapatite-coated femoral prosthesis as part of a total hip arthroplasty and were followed for a minimum of five years (average, 6.4 years; range, five to 8.3 years) or until revision. The average Harris hip score was 47 points (range, 22 to 77 points) preoperatively and 93 points (range, 49 to 100 points) at the time of the latest clinical evaluation. Two patients who had a well fixed femoral implant had activity-limiting pain in the thigh at the time of the most recent examination. Radiographic changes consistent with bone-remodeling (cortical hypertrophy and bone condensation) typically were seen around the mid-part of the shaft of the prosthesis. Forty-eight (32 per cent) of the 148 hips that were included in the radiographic analysis demonstrated a small amount of erosive scalloping in either zone 1 or zone 7 of Gruen et al., and intramedullary osteolysis was suspected in only one hip. All stems were radiographically osseointegrated according to a modification of the criteria described by Engh et al. Four stems were revised, but none of the revisions were performed because of mechanical failure (two stems were revised in conjunction with a revision of the cup because of pain; one, because of an infection; and one, after a traumatic femoral fracture that occurred six years postoperatively). Thus, the rates of aseptic and mechanical failure were both 0 per cent. The combined rate of failure, which included the two stems that were revised because of pain and the two stems that were associated with pain that limited activity, was 2.6 per cent (four of 152 stems). The over-all clinical results associated with hydroxyapatite-coated femoral components were excellent in this group of young patients after intermediate-term follow-up. A review of serial radiographs showed mechanically stable implants with osseous ingrowth, evidence of stress transmission at the middle part of the stem, and minimum endosteal osteolysis.


Efficacy of Low-Pressure Inflation of Oversized Drug-Coated Balloon for Coronary Artery Disease.
This study sought to assess the efficacy of oversized drug-coated balloon (DCB) inflation at low pressure for the prevention of acute dissections and late restenosis. The major limitation of DCB coronary angioplasty is the occurrence of severe dissection after inflation of DCB. Between 2014 and 2018, 273 consecutive patients were retrospectively studied. 191 lesions (154 patients) treated by oversized DCB inflation at low pressure (<4 atm, 2.4 ± 1.2 atm, DCB/artery ratio 1.14 ± 0.22; LP group) were compared with 135 lesions (119 patients) treated by the standard DCB technique (7.1 ± 2.2 atm, DCB/artery ratio 1.03 ± 0.16; SP group). Although the lesions in the LP group were more complex than those in the SP group (smaller reference diameter (2.38 mm vs. 2.57 mm, P=0.011), longer lesions (11.7 mm vs. 10.5 mm, P=0.10), and more frequent use of rotational atherectomy (45.0% vs. 28.1%, P=0.003), there was no significant difference in the NHLBI type of dissections between the two groups (11.5%, 12.0%, 5.2% vs. 12.6%, 12.6%, 2.2% in type A, B, and C, P=0.61), and no bailout stenting was required. In 125 well-matched lesion pairs after propensity score analysis, the cumulative incidence of target lesion revascularization at 3 years was 4.5% vs. 7.0%, respectively (P=0.60). Late lumen loss (-0.00 mm vs. -0.01 mm, P=0.94) and restenosis rates (7.4% vs. 7.1%, P=1.0) were similar in both of the groups. The application of oversized DCB at low pressure is effective and feasible for preventing late restenosis comparative to the standard technique of DCB.


Cellular Phone Use and Risk of Tumors: Systematic Review and Meta-Analysis.
We investigated whether cellular phone use was associated with increased risk of tumors using a meta-analysis of case-control studies. PubMed and EMBASE were searched from inception to July 2018. The primary outcome was the risk of tumors by cellular phone use, which was measured by pooling each odds ratio (OR) and its 95% confidence interval (CI). In a meta-analysis of 46 case-control studies, compared with never or rarely having used a cellular phone, regular use was not associated with tumor risk in the random-effects meta-analysis. However, in the subgroup meta-analysis by research group, there was a statistically significant positive association (harmful effect) in the Hardell et al. studies (OR, 1.15-95% CI, 1.00 to 1.33- n = 10), a statistically significant negative association (beneficial effect) in the INTERPHONE-related studies (case-control studies from 13 countries coordinated by the International Agency for Research on Cancer (IARC); (OR, 0.81-95% CI, 0.75 to 0.89-n = 9), and no statistically significant association in other research groups' studies. Further, cellular phone use with cumulative call time more than 1000 h statistically significantly increased the risk of tumors. This comprehensive meta-analysis of case-control studies found evidence that linked cellular phone use to increased tumor risk.


Clinical features suggestive of meningitis in children: a systematic review of prospective data.
Clinical diagnosis of pediatric meningitis is fundamental; therefore, familiarity with evidence underscoring clinical features suggestive of meningitis is important. To seek evidence supporting accuracy of clinical features of pediatric bacterial meningitis. A review of Medline, Embase, the Cumulative Index to Nursing and Allied Health Literature, Web of Science, and PubMed was conducted for all articles of relevance. Articles contained prospective data of clinical features in children with laboratory-confirmed bacterial meningitis and in comparison groups of those without it. Two authors independently assessed quality and extracted data to calculate accuracy data of clinical features. Of 14 145 references initially identified, 10 met our inclusion criteria. On history, a report of bulging fontanel (likelihood ratio [LR]: 8.00 [95% confidence interval (CI): 2.4-26]), neck stiffness (7.70 [3.2-19]), seizures (outside febrile-convulsion age range) (4.40 [3.0-6.4]), or reduced feeds (2.00 [1.2-3.4]) raised concern about the presence of meningitis. On examination, jaundice (LR: 5.90 [95% CI: 1.8-19]), being toxic or moribund (5.80 [3.0-11]), meningeal signs (4.50 [2.4-8.3]), neck stiffness (4.00 [2.6-6.3]), bulging fontanel (3.50 [2.0-6.0]), Kernig sign (3.50 [2.1-5.7]), tone up (3.20 [2.2-4.5]), fever of >40°C (2.90 [1.6-5.5]), and Brudzinski sign (2.50 [1.8-3.6]) independently raised the likelihood of meningitis. The absence of meningeal signs (LR: 0.41 [95% CI: 0.30-0.57]) and an abnormal cry (0.30 [0.16-0.57]) independently lowered the likelihood of meningitis. The absence of fever did not rule out meningitis (LR: 0.70 [95% CI: 0.53-0.92]). Evidence for several useful clinical features that influence the likelihood of pediatric meningitis exists. No isolated clinical feature is diagnostic, and the most accurate diagnostic combination is unclear.


Range-expanding populations of a globally introduced weed experience negative plant-soil feedbacks.
Biological invasions are fundamentally biogeographic processes that occur over large spatial scales. Interactions with soil microbes can have strong impacts on plant invasions, but how these interactions vary among areas where introduced species are highly invasive vs. naturalized is still unknown. In this study, we examined biogeographic variation in plant-soil microbe interactions of a globally invasive weed, Centaurea solstitialis (yellow starthistle). We addressed the following questions (1) Is Centaurea released from natural enemy pressure from soil microbes in introduced regions? and (2) Is variation in plant-soil feedbacks associated with variation in Centaurea's invasive success? We conducted greenhouse experiments using soils and seeds collected from native Eurasian populations and introduced populations spanning North and South America where Centaurea is highly invasive and noninvasive. Soil microbes had pervasive negative effects in all regions, although the magnitude of their effect varied among regions. These patterns were not unequivocally congruent with the enemy release hypothesis. Surprisingly, we also found that Centaurea generated strong negative feedbacks in regions where it is the most invasive, while it generated neutral plant-soil feedbacks where it is noninvasive. Recent studies have found reduced below-ground enemy attack and more positive plant-soil feedbacks in range-expanding plant populations, but we found increased negative effects of soil microbes in range-expanding Centaurea populations. While such negative feedbacks may limit the long-term persistence of invasive plants, such feedbacks may also contribute to the success of invasions, either by having disproportionately negative impacts on competing species, or by yielding relatively better growth in uncolonized areas that would encourage lateral spread. Enemy release from soil-borne pathogens is not sufficient to explain the success of this weed in such different regions. The biogeographic variation in soil-microbe effects indicates that different mechanisms may operate on this species in different regions, thus establishing geographic mosaics of species interactions that contribute to variation in invasion success.


In vivo melanoma imaging based on dynamic nuclear polarization enhancement in melanin pigment of living mice using in vivo dynamic nuclear polarization magnetic resonance imaging.
Melanin is a pigment that includes free radicals and is widely distributed in living animals. Malignant melanoma is one of the most progressive tumors in humans with increasing incidence worldwide, and has shown resistance to chemotherapy, resulting in high mortality at the metastatic stage. In general, melanoma involves the abnormal accumulation of melanin pigment produced by malignant melanocytes. Electron paramagnetic resonance (EPR) spectroscopy and imaging is a powerful technique to directly visualize melanomas using endogenous free radicals in the melanin pigment. Because melanin radicals have a large linewidth, the low spatial resolution of EPR imaging results in blurred images and a lack of anatomical information. Dynamic nuclear polarization (DNP)-MRI is a noninvasive imaging method to obtain the spatio-temporal information of free radicals with MRI anatomical resolution. Proton signals in tissues, including free radicals, can be dramatically enhanced by EPR irradiation at the resonance frequency of the free radical prior to applying the MRI pulse sequence. However, the DNP effects of free radicals in the pigment of living organisms is unclear. Therefore, if endogenous free radicals in melanin pigment could be utilized as a bio-probe for DNP-MRI, this will be an advantage for the specific enhancement of melanoma tissues and might allow the separate noninvasive visualization of melanoma tissues without the need for probe administration. Here, we report that biological melanin pigment induced a in vivo DNP effect by interacting with water molecules. In addition, we demonstrated in vivo melanoma imaging based on the DNP effects of endogenous free radicals in the melanin pigment of living mice.


Increased sialylation of surface glycopeptides of human trophoblast compared with fetal cells from the same conceptus.
The surface glycopeptides of human trophoblastic cells have been compared with those of fetal cells from the same embryos using double-labeling methods with isotopes of L-fucose and D-glucosamine. A faster eluting, neuraminidase-sensitive, fraction was observed on Sephadex chromatography of the trophoblast spectra when D-glucosamine was used as precursor. Labeling with fucose did not appear to result in any differences, thus suggesting that the glycopeptides characertistic of trophoblast contained glucosamine-derived metabolic products, including sialic acid, but excluding fucose. This increased sialylation is similar to, but not identical with, modifications observed in neoplastic cells, and on this basis it is postulated that two species of glycopeptides may be involved in atypical cellular behavior. The first contains sialic acid and other sugars excluding fucose, and is associated with localized cellular growth and invasion. The second contains both sialic acid and fucose and is characteristic of neoplastic cells.


Adjustable intragastric balloons: a 12-month pilot trial in endoscopic weight loss management.
Intragastric balloons are associated with (1) early period intolerance, (2) diminished effect within 3-4 months, and (3) bowel obstruction risk mandating removal at 6 months. The introduction of an adjustable balloon could improve comfort and offer greater efficacy. A migration prevention function, safely enabling prolonged implantation, could improve efficacy and weight maintenance post-extraction. The first implantations of an adjustable balloon with an attached migration prevention anchor are reported. The primary endpoint was the absence of bowel perforation, obstruction, or hemorrhage. Eighteen patients with mean BMI of 37.3 were implanted with the Spatz Adjustable Balloon system (ABS) for 12 months. Balloon volumes were adjusted for intolerance or weight loss plateau. Mean weight loss at 24 weeks was 15.6 kg with 26.4% EWL (percent of excess weight loss) and 24.4 kg with 48.8% EWL at 52 weeks. Sixteen adjustments were successfully performed. Six downward adjustments alleviated intolerance, yielding additional mean weight loss of 4.6 kg. Ten upward adjustments for weight loss plateau yielded a mean additional weight loss of 7 kg. Seven balloons were removed prematurely. Complications necessitating early removal included valve malfunction (1), gastritis (1), Mallory-Weiss tear (1), NSAID (2× dose/2 weeks) perforating ulcer (1), and balloon deflation (1). Two incidents of catheter shear from the chain: one passed uneventfully and one caused an esophageal laceration without perforation during extraction. The Spatz ABS has been successfully implanted in 18 patients. (1) Upward adjustments yielded additional weight loss. (2) Downward adjustments alleviated intolerance, with continued weight loss. (3) Preliminary 1-year implantation results are encouraging.


Instrumentation needs of research universities.
This article assesses the status of scientific instruments in major research universities and identifies factors that facilitate or impede their development, acquisition, use, and maintenance. Sixteen universities, six national and government laboratories, and nine commercial laboratories were visited; over 700 individuals were interviewed. Data on instrument acquisition and age were collected. Instrumentation was examined in physics, chemistry, biological sciences, earth sciences, and electrical engineering. The study found that the quality of university instrumentation has seriously deteriorated, due principally to a relative decrease in instrumentation funding, inflexibility within the project grant system, and insufficient support for maintenance.


Extracting dynamical equations from experimental data is NP hard.
The behavior of any physical system is governed by its underlying dynamical equations. Much of physics is concerned with discovering these dynamical equations and understanding their consequences. In this Letter, we show that, remarkably, identifying the underlying dynamical equation from any amount of experimental data, however precise, is a provably computationally hard problem (it is NP hard), both for classical and quantum mechanical systems. As a by-product of this work, we give complexity-theoretic answers to both the quantum and classical embedding problems, two long-standing open problems in mathematics (the classical problem, in particular, dating back over 70 years).


A novel concept for scaffold-free vessel tissue engineering: self-assembly of microtissue building blocks.
Current scientific attempts to generate in vitro tissue-engineered living blood vessels (TEBVs) show substantial limitations, thereby preventing routine clinical use. In the present report, we describe a novel biotechnology concept to create living small diameter TEBV based exclusively on microtissue self-assembly (living cellular re-aggregates). A novel bioreactor was designed to assemble microtissues in a vascular shape and apply pulsatile flow and circumferential mechanical stimulation. Microtissues composed of human artery-derived fibroblasts (HAFs) and endothelial cells (HUVECs) were accumulated and cultured for 7 and 14 days under pulsatile flow/mechanical stimulation or static culture conditions with a diameter of 3mm and a wall thickness of 1mm. The resulting vessels were analyzed by immunohistochemistry for extracellular matrix (ECM) and cell phenotype (von Willebrand factor, alpha-SMA, Ki67, VEGF). Self-assembled microtissues composed of fibroblasts displayed significantly accelerated ECM formation compared to monolayer cell sheets. Accumulation of vessel-like tissue occurred within 14 days under both, static and flow/mechanical stimulation conditions. A layered tissue formation was observed only in the dynamic group, as indicated by luminal aligned alpha-SMA positive fibroblasts. We could demonstrate that self-assembled cell-based microtissues can be used to generate small diameter TEBV. The significant enhancement of ECM expression and maturation, together with the pre-vascularization capacity makes this approach highly attractive in terms of generating functional small diameter TEBV devoid of any foreign material.


Inflammatory effects of gene transfer into the CNS with defective HSV-1 vectors.
The use of viral vectors which infect and express genes in post-mitotic neurons is a potential strategy for the treatment of disorders affecting the central nervous system (CNS). However, the inflammatory consequences of such strategies have yet to be systematically examined. Preparations of non-replicating defective herpes simplex virus type 1 (HSV-1) amplicon vectors containing the lacZ gene were obtained by standard methods and stereotaxically injected into the adult rat dentate gyrus (DG). The consequent gene expression and inflammatory effects following microinjection were investigated. beta-Galactosidase activity was detected in neurons of the DG from 24 h to at least 12 days after vector injection. A strong inflammatory response developed within 2 days, characterized by diffuse up-regulation of major histocompatibility complex (MHC) class I antigens and the activation of microglia. After 4 days the recruitment of MHC class II+ cells, activated T lymphocytes and macrophages was detected. These features persisted for at least 31 days. Of importance was the finding of beta-galactosidase activity in a bilateral group of neurons in the supramammillary nuclei (SMN) of the posterior hypothalamus, known to send afferent projections to the DG. The onset of inflammation at this secondary site was delayed, but its cellular characteristics resembled those found at the primary site of injection. Thus, the use of preparations of defective HSV-1 vectors for gene transfer in the CNS has immunological implications both at primary and secondary sites within the CNS.(ABSTRACT TRUNCATED AT 250 WORDS)


Insights on the persistence of pines (Pinus species) in the Late Cretaceous and their increasing dominance in the Anthropocene.
Although gymnosperms were nearly swept away by the rise of the angiosperms in the Late Cretaceous, conifers, and pines (Pinus species) in particular, survived and regained their dominance in some habitats. Diversification of pines into fire-avoiding (subgenus Haploxylon) and fire-adapted (subgenus Diploxylon) species occurred in response to abiotic and biotic factors in the Late Cretaceous such as competition with emerging angiosperms and changing fire regimes. Adaptations/traits that evolved in response to angiosperm-fuelled fire regimes and stressful environments in the Late Cretaceous were key to pine success and are also contributing to a new "pine rise" in some areas in the Anthropocene. Human-mediated activities exert both positive and negative impacts of range size and expansion and invasions of pines. Large-scale afforestation with pines, human-mediated changes to fire regimes, and other ecosystem processes are other contributing factors. We discuss traits that evolved in response to angiosperm-mediated fires and stressful environments in the Cretaceous and that continue to contribute to pine persistence and dominance and the numerous ways in which human activities favor pines.


Fluctuation in deep groundwater chemistry and microbial community and their impact on corrosion of stainless-steels.
Low and intermediate level radioactive waste produced during the operation and decommissioning of nuclear power plants is disposed of in an underground geological repository. The majority of metallic waste is made of various stainless-steels and carbon steel. Microbial communities and groundwater composition in deep bedrock at repository sites were believed to stay stable over time, allowing the prediction of evolution of the repository environment. However, a two-year survey of chemical components and microbial community composition within deep bedrock revealed changes in both. An in situ corrosion monitoring system was developed to monitor real-time corrosion rates of two stainless-steel grades (AISI 304 and 316) to study the evolution of corrosion, and correlation between environmental changes and corrosion rate. Surprisingly, higher corrosion rates of steel coupons were detected in the higher alloyed stainless-steel grade 316 compared to the lower alloyed grade 304. Pitting was the main corrosion form. Sulphate reducing bacteria and methanogenic archaea were enriched on surfaces of both types of steel coupons. These microbes likely have a role in the corrosion of stainless-steel in this environment. The changes in groundwater conditions and microbial communities within deep bedrock groundwater at this repository site may have implications for the nuclide release and transport of radioactive material and the long-term evolution and safety of this repository and continental repositories in general and thus needs to be thoroughly understood.


A massive outbreak of food poisoning--a reminder of the importance of proper toxic waste control.
Because of rapid urbanisation in South Africa, scavenging from waste disposal sites by poor communities poses an increasing health risk. Reject cough lozenges, some of which contained larger amounts of dextromethorphan than usual, were illegally removed from a disposal site and, after resale by informal traders, caused moderately severe symptoms of toxicity in 171/540 (24%) primary school pupils. Although dextromethorphan was implicated as a cause, contributing effects of other toxins could not be excluded. Bacteriological cultures and a pesticide screen were negative. Had more toxic substances been involved, the consequences would have been disastrous. This incident supports calls for an integrated national waste management policy and waste control act to govern the management and control of waste from generation to disposal. Such a policy is necessary to prevent potentially serious incidents in the future.


Quantitative Fundus Autofluorescence: Advanced Analysis Tools.
To use multimodal retinal images (including quantitative fundus autofluorescence [QAF]) for spectral-domain optical coherence tomography (SD-OCT)-based image registration and alignment. For each age decade of healthy adults, normative fine-grained QAF retinal maps are generated and advanced methods for QAF image analysis are applied. Multimodal retinal images were obtained from 103 healthy subjects (age 19-77 years; unremarkable retina/macula, age-appropriate clear optic media). Custom written FIJI plugins enabled: (1) determination of the fovea in SD-OCT and the edge of the optic disc in infrared (IR) images; (2) alignment and superimposition of multimodal retinal images based on foveal and optic disc position; (3) plotting of normative QAF retinal maps for each decade; and (4) comparison of individual retinas with normative retinas of different decades using newly introduced analysis patterns (QAF97, freehand tool). SD-OCT based image registration enables easy image registration, alignment, and analysis of different modalities (QAF, IR, and SD-OCT here reported). In QAF, intensities significantly increase with age with two major inclines between the third/fourth and seventh/eighth decades. With aging, the parafoveal area of maximum QAF intensity slightly shifts from temporal-superior to temporal. Compared with standard QAF analysis, refined QAF analysis patterns reveal a more detailed analysis of QAF, especially in the diseased retina. Age-related QAF normative retinal maps can be used to directly compare and classify individual's QAF intensities. Advanced QAF analysis tools will further help to interpret autofluorescence changes in normal aging and in the diseased retina in a multimodal imaging setting. Advanced methods for QAF analysis link basic findings with clinical observations in normal aging and in the diseased macula.


Design of a hydrophobic tripeptide that self-assembles into amphiphilic superstructures forming a hydrogel biomaterial.
We report the rational design of a heterochiral hydrophobic tripeptide self-assembling into amphiphilic d-superstructures that yield a self-supportive hydrogel at physiological pH. The material endures cell culture conditions and sustains fibroblast proliferation. Tripeptide superstructures are thoroughly analysed by several techniques.


Search for dinucleon decay into kaons in Super-Kamiokande.
A search for the dinucleon decay pp → K+ K+ has been performed using 91.6 kton·yr data from Super-Kamiokande-I. This decay provides a sensitive probe of the R-parity-violating parameter λ112''. A boosted decision tree analysis found no signal candidates in the data. The expected background was 0.28±0.19 atmospheric neutrino induced events and the estimated signal detection efficiency was 12.6%±3.2%. A lower limit of 1.7×10(32) years has been placed on the partial lifetime of the decay O16 → C14K+ K+ at 90% C.L. A corresponding upper limit of 7.8×10(-9) has been placed on the parameter λ112''.


Amplification of DNA from whole blood.
A method is described for the amplification by PCR of human chromosomal DNA sequences from whole blood samples. Various amounts of blood samples, with either EDTA, citrate, or heparin used as the anticoagulant, have been used to determine optimal PCR conditions for each type of sample. Up to 80% (vol/vol) of whole blood sample is tolerated in PCR with Taq polymerase. Amplification from whole blood requires the optimization of salt (K+ and Mg++) according to sample volume and type of anticoagulant used. Pretreatment of fresh blood samples to lyse the leukocytes is required for EDTA-treated blood samples and is beneficent in PCR with heparin- and citrate-treated blood samples to obtain maximal amplicon amounts. A satisfactory method of pretreating samples is freeze/thawing. In addition, EDTA-treated blood samples require a heat treatment before PCR for maximal amplicon synthesis. It appears that purification of the DNA is not necessary for any of the whole blood samples analyzed by PCR. Results of amplification reactions from unpurified hepatitis B virus (HBV) sequences of whole sera samples are shown also.


Contrasting patterns of Y chromosome variation in Ashkenazi Jewish and host non-Jewish European populations.
The molecular basis of more than 25 genetic diseases has been described in Ashkenazi Jewish populations. Most of these diseases are characterized by one or two major founder mutations that are present in the Ashkenazi population at elevated frequencies. One explanation for this preponderance of recessive diseases is accentuated genetic drift resulting from a series of dispersals to and within Europe, endogamy, and/or recent rapid population growth. However, a clear picture of the manner in which neutral genetic variation has been affected by such a demographic history has not yet emerged. We have examined a set of 32 binary markers (single nucleotide polymorphisms; SNPs) and 10 microsatellites on the non-recombining portion of the Y chromosome (NRY) to investigate the ways in which patterns of variation differ between Ashkenazi Jewish and their non-Jewish host populations in Europe. This set of SNPs defines a total of 20 NRY haplogroups in these populations, at least four of which are likely to have been part of the ancestral Ashkenazi gene pool in the Near East, and at least three of which may have introgressed to some degree into Ashkenazi populations after their dispersal to Europe. It is striking that whereas Ashkenazi populations are genetically more diverse at both the SNP and STR level compared with their European non-Jewish counterparts, they have greatly reduced within-haplogroup STR variability, especially in those founder haplogroups that migrated from the Near East. This contrasting pattern of diversity in Ashkenazi populations is evidence for a reduction in male effective population size, possibly resulting from a series of founder events and high rates of endogamy within Europe. This reduced effective population size may explain the high incidence of founder disease mutations despite overall high levels of NRY diversity.


NKTR-181: A Novel Mu-Opioid Analgesic with Inherently Low Abuse Potential.
The increasing availability of prescription opioid analgesics for the treatment of pain has been paralleled by an epidemic of opioid misuse, diversion, and overdose. The development of abuse-deterrent formulations (ADFs) of conventional opioids such as oxycodone and morphine represents an advance in the field and has had a positive but insufficient impact, as most opioids are still prescribed in highly abusable, non-ADF forms, and abusers can tamper with ADF medications to liberate the abusable opioid within. The abuse liability of mu-opioid agonists appears to be dependent on their rapid rate of entry into the central nervous system (CNS), whereas analgesic activity appears to be a function of CNS exposure alone, suggesting that a new opioid agonist with an inherently low rate of influx across the blood-brain barrier could mediate analgesia with low abuse liability, regardless of formulation or route of administration. NKTR-181 is a novel, long-acting, selective mu-opioid agonist with structural properties that reduce its rate of entry across the blood-brain barrier compared with traditional mu-opioid agonists. NKTR-181 demonstrated maximum analgesic activity comparable to that of oxycodone in hot-plate latency and acetic-acid writhing models. NKTR-181 was distinguishable from oxycodone by its reduced abuse potential in self-administration and progressive-ratio break point models, with behavioral effects similar to those of saline, as well as reduced CNS side effects as measured by the modified Irwin test. The in vitro and in vivo studies presented here demonstrate that NKTR-181 is the first selective mu-opioid agonist to combine analgesic efficacy and reduced abuse liability through the alteration of brain-entry kinetics.


Critical features for the recognition of biological motion.
Humans can perceive the motion of living beings from very impoverished stimuli like point-light displays. How the visual system achieves the robust generalization from normal to point-light stimuli remains an unresolved question. We present evidence on multiple levels demonstrating that this generalization might be accomplished by an extraction of simple mid-level optic flow features within coarse spatial arrangement, potentially exploiting relatively simple neural circuits: (1) A statistical analysis of the most informative mid-level features reveals that normal and point-light walkers share very similar dominant local optic flow features. (2) We devise a novel point-light stimulus (critical features stimulus) that contains these features, and which is perceived as a human walker even though it is inconsistent with the skeleton of the human body. (3) A neural model that extracts only these critical features accounts for substantial recognition rates for strongly degraded stimuli. We conclude that recognition of biological motion might be accomplished by detecting mid-level optic flow features with relatively coarse spatial localization. The computationally challenging reconstruction of precise position information from degraded stimuli might not be required.