5-hydroxymethylcytosine marks promoters in colon that resist DNA hypermethylation in cancer.
The discovery of cytosine hydroxymethylation (5hmC) as a mechanism that potentially controls DNA methylation changes typical of neoplasia prompted us to investigate its behaviour in colon cancer. 5hmC is globally reduced in proliferating cells such as colon tumours and the gut crypt progenitors, from which tumours can arise. Here, we show that colorectal tumours and cancer cells express Ten-Eleven-Translocation (TET) transcripts at levels similar to normal tissues. Genome-wide analyses show that promoters marked by 5hmC in normal tissue, and those identified as TET2 targets in colorectal cancer cells, are resistant to methylation gain in cancer. In vitro studies of TET2 in cancer cells confirm that these promoters are resistant to methylation gain independently of sustained TET2 expression. We also find that a considerable number of the methylation gain-resistant promoters marked by 5hmC in normal colon overlap with those that are marked with poised bivalent histone modifications in embryonic stem cells. Together our results indicate that promoters that acquire 5hmC upon normal colon differentiation are innately resistant to neoplastic hypermethylation by mechanisms that do not require high levels of 5hmC in tumours. Our study highlights the potential of cytosine modifications as biomarkers of cancerous cell proliferation.


[Psychological and psychophysiological characteristics of arterial hypertension patients].
A total of 120 essentially-hypertensive patients and 30 coronary patients were investigated clinically and by means of psychophysiologic and psychological tests. Different patterns of physiological response to psychoemotional stress were demonstrated in hypertensive and coronary patients. The markedness and duration of stress-induced hypertensive response was found to be related to psychosocial status of essentially-hypertensive patients. Hypertensive patients showing high anxiety and high interpersonal and intrapersonal conflict develop more marked blood pressure elevation under stress, a possible evidence of a greater contribution of psychosocial factors to the disease in these patients.


Leadership development plan. A CQI approach to management performance appraisals.
In 1993 managers at St. Mary's Hospital Medical Center established the attributes necessary to be a successful leader in St. Mary's continuous quality improvement culture. These leadership attributes formed the basis of a new performance appraisal system for managers. The medical center adopted its new performance appraisal system at the beginning of fiscal year 1994-1995. The objective of the plan is to develop St. Mary's managers' leadership skills. St. Mary's Leadership Development Plan is an ongoing cycle, with three phases. First, managers and administrative representatives jointly agree on objectives to discuss throughout the fiscal year. The objectives reflect the hospital planning and financial goals and objectives, department goals and objectives, and leadership growth opportunities. Each manager is then responsible for gathering feedback from subordinates on how well he or she is meeting the set objectives. Finally, each manager and administrative representative highlight accomplishments achieved during the fiscal year. St. Mary's decided to discontinue pay-for-performance salary increases beginning with the 1994-1995 fiscal year, coinciding with the initiation of the Leadership Development Plan. Manager's compensation is now a flat percentage increase granted to all managers.


Lateral tegmental field neurons sensitive to muscular contraction: a role in pressor reflexes?
The medullary lateral tegmental field (LTF) has a major role in sympathetic nerve discharge (SND) rhythmicity, but its role in pressor reflexes generated by hindlimb muscular contraction (MC) is unknown. Therefore, two sets of experiments were performed in 17 chloralose-urethane anesthetized cats. First, responses of single LTF neurons to MC induced by L7-S1 ventral root stimulation were examined. The majority (30 of 47) of LTF neurons increased firing during MC. Most LTF neurons had a basal discharge correlated with the 2-10 Hz rhythm of SND or the cardiac cycle and responded to increases in blood pressure. Only seven neurons were inhibited by MC, most having a respiratory rhythm. Second, pressor responses to MC and to caudal hypothalamic stimulation were examined before and after bilateral LTF microinjections of a synaptic blocker (CoCl2) as well as with lidocaine. Microinjection of CoCl2 or lidocaine significantly attenuating the dominant 2-10 Hz power coefficient of SND had no effect on the pressor responses to MC or caudal hypothalamic stimulation. Therefore, LTF may be important for basal rhythms in SND and may help synchronize SND during MC, but its contribution to basal rhythms is apparently not required for pressor reflexes evoked by hindlimb MC or hypothalamic stimulation.


Cell-specific expression and regulation of a glucokinase gene locus transgene.
Transgenic mice containing one or more extra copies of the entire glucokinase (GK) gene locus were generated and characterized. The GK transgene, an 83-kilobase pair mouse genomic DNA fragment containing both promoter regions, was expressed and regulated in a cell-specific manner, and rescued GK null lethality when crossed into mice bearing a targeted mutation of the endogenous GK gene. Livers from the transgenic mice had elevated GK mRNA, protein, and activity levels, compared with controls, and the transgene was regulated in liver by dietary manipulations. The amount of GK immunoreactivity in hepatocyte nuclei, where GK binds to the GK regulatory protein, was also increased. Pancreatic islets displayed increased GK immunoreactivity and NAD(P)H responses to glucose, but only when isolated and cultured in 20 mM glucose, as a result of the hypoglycemic phenotype of these mice (Niswender, K. D., Shiota, M., Postic, C., Cherrington, A. D., and Magnuson, M. A. (1997) J. Biol. Chem. 272, 22604-22609). Together, these results indicate that the region of the gene from -55 to +28 kilobase pairs (relative to the liver GK transcription start site) contains all the regulatory sequences necessary for expression of both GK isoforms, thereby placing an upper limit on the size of the GK gene locus.


Characterizing analgesic use during air medical transport of injured children.
Pain management is an important aspect of emergency care for children suffering traumatic injuries. The objectives of this study were to characterize analgesic administration to injured children during air medical transport, to describe factors associated with analgesic use, and to examine the effects of patient race on analgesia. We used electronic records for patients transported by a regional air medical transport agency. We retrospectively examined data from 2003-2012 for patients ≤ 15 years old suffering traumatic injuries. We used bivariable analyses to identify associations for multivariable logistic regression models to determine factors associated with our outcomes -documentation of pain score and analgesic administration. Of 5,057 patients, the median age was 8 (IQR 3-12) years. The majority of patients were male (66%, 95% CI 64-66%), were white non-Hispanic (83%, 95% CI 82-84%), and had no pain score documented (61%, 95% CI 60-62%). While only 15% of patients received analgesics overall, 70% with an initial pain score ≥ 5 received analgesics. In unadjusted models, non-white race was associated with lower odds of having a pain score documented (OR 0.52, 95% CI 0.44-0.62) and receiving analgesics (OR 0.64, 95% CI 0.50-0.82). After adjusting for confounders, there was no evidence of racial differences in pain management. Multivariable analysis revealed that younger age, lack of intravenous access, higher Glasgow Coma Scale, systolic blood pressure <100, transportation from the scene, initial pain score <5, and not having a pain score documented were associated with lower odds of receiving analgesics. Few pediatric patients had pain scores documented and fewer received analgesics during air medical transport for injuries. Racial differences in analgesia seen in unadjusted analyses did not persist after controlling for confounders. Resources, training, and appropriate pain management protocols should be made available to facilitate pain assessment in children as a strategy for increasing appropriate analgesic use during transport.


The P66Shc/mitochondrial permeability transition pore pathway determines neurodegeneration.
Mitochondrial-mediated oxidative stress and apoptosis play a crucial role in neurodegenerative disease and aging. Both mitochondrial permeability transition (PT) and swelling of mitochondria have been involved in neurodegeneration. Indeed, knockout mice for cyclophilin-D (Cyc-D), a key regulatory component of the PT pore (PTP) that triggers mitochondrial swelling, resulted to be protected in preclinical models of multiple sclerosis (MS), Parkinson's disease (PD), and amyotrophic lateral sclerosis (ALS). However, how neuronal stress is transduced into mitochondrial oxidative stress and swelling is unclear. Recently, the aging determinant p66Shc that generates H2O2 reacting with cytochrome c and induces oxidation of PTP and mitochondrial swelling was found to be involved in MS and ALS. To investigate the role of p66Shc/PTP pathway in neurodegeneration, we performed experimental autoimmune encephalomyelitis (EAE) experiments in p66Shc knockout mice (p66Shc-/-), knock out mice for cyclophilin-D (Cyc-D-/-), and p66Shc Cyc-D double knock out (p66Shc/Cyc-D-/-) mice. Results confirm that deletion of p66Shc protects from EAE without affecting immune response, whereas it is not epistatic to the Cyc-D mutation. These findings demonstrate that p66Shc contributes to EAE induced neuronal damage most likely through the opening of PTP suggesting that p66Shc/PTP pathway transduces neurodegenerative stresses.


Bupropion in the treatment of depression in Parkinson's disease.
A 78-year-old female with a nine-year history of depression was hospitalized due to worsening depression and symptoms associated with Parkinson's disease (PD). Her motor abilities improved on levodopa and the depression improved after a trial of bupropion, following unsuccessful treatment with other antidepressants. We found four reports on successful treatment of depression in PD with bupropion. However, no controlled double-blind studies have been conducted so clinicians should be cautious when administering bupropion in depression in PD.


A comparison between the Boussignac™ continuous positive airway pressure mask and the venturi mask in terms of improvement in the PaO2/F(I)O2 ratio in morbidly obese patients undergoing bariatric surgery: a randomized controlled trial.
This study compared the Boussignac™ continuous positive airway pressure (CPAP) mask with the venturi face mask in terms of the postoperative PaO2/F(I)O2 (PF) ratio in morbidly obese patients after bariatric surgery. Following hospital Research Ethics Board approval and written informed consent, morbidly obese (body mass index > 35 kg·m(-2)) patients undergoing bariatric surgery were recruited. The patients were anesthetized and laparoscopic Roux-en-Y gastric bypass was performed. Patients were assigned randomly to receive either the Boussignac (Boussignac Group) or the venturi face mask (Venturi Group) immediately after tracheal extubation. Patients were transported to the postanesthesia care unit, and the respective devices were applied for one hour. The PF ratio was recorded after tracheal intubation and at one hour and two hours post extubation. The percent forced expiratory volume (%FEV1) and the percent forced vital capacity (%FVC) were recorded preoperatively and at one hour and two hours post extubation. Independent Student's t tests were used for continuous variables, and the Chi square test was used for categorical variables. P < 0.05 was considered statistically significant. Eighty-one patients (Group Boussignac, n = 43; Group Venturi, n = 38) completed the study. Mean ages and body mass indices were similar in the two groups. At one hour post extubation, the PF ratio in the Boussignac Group was 361 (170) compared with 279 (91) in the Venturi Group (P = 0.007), and at two hours post extubation, the PF ratio in the Boussignac Group was 371 (162) compared with 323 (127) in the Venturi Group (P = 0.1). The postoperative %FEV(1) and %FVC were comparable in both groups at all time points. Compared with the venturi mask, the Boussignac CPAP mask improves the postoperative PF ratio in morbidly obese patients after bariatric surgery. The postoperative %FEV1 and %FVC are comparable for both groups.


Genetics of a de novo origin of undifferentiated multicellularity.
The evolution of multicellularity was a major transition in evolution and set the stage for unprecedented increases in complexity, especially in land plants and animals. Here, we explore the genetics underlying a de novo origin of multicellularity in a microbial evolution experiment carried out on the green alga Chlamydomonas reinhardtii. We show that large-scale changes in gene expression underlie the transition to a multicellular life cycle. Among these, changes to genes involved in cell cycle and reproductive processes were overrepresented, as were changes to C. reinhardtii-specific and volvocine-specific genes. These results suggest that the genetic basis for the experimental evolution of multicellularity in C. reinhardtii has both lineage-specific and shared features, and that the shared features have more in common with C. reinhardtii's relatives among the volvocine algae than with other multicellular green algae or land plants.


Comparison of global left ventricular function using 20 phases with 10-phase reconstructions in multidetector-row computed tomography.
To compare the measurement of global left-ventricular (LV) function parameters of 64-slice multidetector-row computed tomography (MDCT) between 20- and 10-reconstruction phases. Fifty five patients with suspected or known coronary artery disease underwent 64-slice MDCT. LV end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), and ejection fraction (EF) were measured from MDCT data sets using threshold-based volume segmentation and reconstruction at every 5% (20 phases) and 10% (10 phases) step through the R-R interval. These global functional parameters were compared to those obtained via two-dimensional transthoracic echocardiography (2D-TTE), considering the reference standard. The required time for CT data analysis was checked. Agreement for parameters of LV global function was determined using Pearson's correlation coefficient (r) and Bland-Altman analysis. LV volumes (EDV(-5%) 87.5 ± 17.1 ml, EDV(-10%) 87.7 ± 16.3 ml; ESV(-5%) 32.4 ± 10.6 ml, ESV(-10%) 31.9 ± 9.9 ml; SV(-5%) 55.1 ± 10.5 ml, SV(-10%) 55.8 ± 9.9 ml; mean ± SD) and EF (EF(-5%) 63.4 ± 6.2%, EF(-10%) 63.9 ± 5.8%) did not differ significantly between the 20- and 10 phase reconstructions, and evidenced good to excellent correlation (r = 0.786-0.896, all P < 0.001) with the 2D-TTE results. The mean required time for CT data analysis in the 20- and 10 phase reconstructions were 15.5 ± 4.0 and 7.3 ± 2.5 min. Within MDCT, using 10-phase image reconstruction is sufficient to evaluate LV volumes and EF, and is also more time-effective than 20-phase reconstruction.


LX-1031, a tryptophan 5-hydroxylase inhibitor, and its potential in chronic diarrhea associated with increased serotonin.
LX-1031 is an oral, small molecule tryptophan 5-hydroxylase (TPH) inhibitor that reduces serotonin (5-HT) synthesis peripherally. It has potential for illnesses characterized by excess 5-HT, such as diarrhea-predominant irritable bowel syndrome (IBS-D) and carcinoid diarrhea. In vitro, inhibition of TPH1 occurred in 10(-8) -10(-7) mol L(-1) range. In vivo in rodents, LX-1031 has no effect on brain 5-HT while dose-dependently reducing 5-HT, particularly in the small bowel. After oral LX1031 in humans, systemic exposure is very low, plasma concentrations are linear in dose range 250-750 mg q.i.d.; the median T(1/2) for elimination is ∼ 20 h, and repeat administration for 14 days doubles C(max) . In ascending-single-dose and multiple-dose (14 days) trials in healthy volunteers, LX-1031, 2-4 g day(-1) significantly reduced urinary 5-hydroxyindoleacetic acid (5-HIAA) starting by Day 5, and persisting over the 14 day exposure. There are no dose-limiting toxicities in healthy subjects or remarkable adverse effects in clinical trials to date. Over a 28-day treatment period, LX-1031 was associated with improved weekly global scores (2/4 weeks) and improved stool consistency with lower urinary 5-HIAA excretion. LX-1031 appears promising for chronic diarrhea associated with increased 5-HT expression including IBS-D. Optimal doses, efficacy and safety in IBS clinical trials need to be fully elucidated; low systemic exposure, selectivity for TPH1 over TPH2, and lack of effect on brain 5-HT in several species suggest that LX-1031 is unlikely to cause affective disorders.


Discovering shared dynamics in physiological signals: application to patient monitoring in ICU.
Modern clinical databases include time series of vital signs, which are often recorded continuously during a hospital stay. Over several days, these recordings may yield many thousands of samples. In this work, we explore the feasibility of characterizing the "state of health" of a patient using the physiological dynamics inferred from these time series. The ultimate objective is to assist clinicians in allocating resources to high-risk patients. We hypothesize that "similar" patients exhibit similar dynamics and the properties and duration of these states are indicative of health and disease. We use Bayesian nonparametric machine learning methods to discover shared dynamics in patients' blood pressure (BP) time series. Each such "dynamic" captures a distinct pattern of evolution of BP and is possibly recurrent within the same time series and shared across multiple patients. Next, we examine the utility of this low-dimensional representation of BP time series for predicting mortality in patients. Our results are based on an intensive care unit (ICU) cohort of 480 patients (with 16% mortality) and indicate that the dynamics of time series of vital signs can be an independent useful predictor of outcome in ICU.


The transition from rookie to genuine nurse: narratives from Swedish nurses 1 year after graduation.
This article describes nurses' experiences during their first year after graduation, based on the qualitative content analysis of eight interviews. The results show that the nurses experienced a transition from "being a rookie," including being accepted as a member of the team and respected as a colleague, to "becoming a genuine nurse," including the feeling of being able to shoulder responsibility, prioritize tasks, and convey confidence to patients. This transition seems to occur regardless of support, and it is important to be aware of nurses' level of maturity and to adjust the degree of responsibility given to them because they may not be comfortable revealing their limitations to others.


A multicenter evaluation of pandemic influenza A/H1N1 in hematopoietic stem cell transplant recipients.
Hematopoietic stem cell transplant (HSCT) recipients have increased morbidity from respiratory viral infections. Pandemic influenza A - A(H1N1)/pdm09 - in 2009-2010 was associated with increased severity of illness in patients with underlying co-morbidities including HSCT, but the factors that contribute to severe disease in HSCT patients are not well characterized. We conducted a multicenter review of microbiologically proven influenza A(H1N1)pdm09 in the HSCT population between April 2009 and April 2010 to determine factors that are associated with severe disease. We identified 37 adult patients (26 allogeneic and 11 autologous HSCT recipients). Median time from transplant to diagnosis was 411 days (range 4 days-14.9 years). Three cases were hospital acquired. Twenty-eight of 37 (75.7%) had confirmed A(H1N1)pdm09. Presumed viral lower respiratory tract infection was present in 12/37 (32.4%) patients. Antiviral therapy was given to 33/37 (89%) patients, primarily oseltamivir (n = 24) and oseltamivir before or after another antiviral (n = 8). Excluding those with nosocomial A(H1N1)pdm09, 18/34 (52.9%) were hospitalized and 6 (33%) required admission to an intensive care unit. Mortality within 30 and 60 days of symptom onset was 7/37 (18.9%) and 11/37 (29.7%), respectively. Factors associated with mortality included nosocomial acquisition (P = 0.023), receipt of mycophenolate mofetil (P = 0.001), or antilymphocyte antibody (P = 0.005) within the past 6 months, reduced-intensity conditioning (P = 0.027), and bacteremia (P = 0.021). A(H1N1)pdm09 infection was particularly severe in HSCT recipients, specifically among those receiving augmented immunosuppression for graft-versus-host disease. The high mortality of the nosocomial cases highlights the need for strict infection-control measures in hospitals during influenza outbreaks.


Modern simulation utilities for genetic analysis.
Statistical geneticists employ simulation to estimate the power of proposed studies, test new analysis tools, and evaluate properties of causal models. Although there are existing trait simulators, there is ample room for modernization. For example, most phenotype simulators are limited to Gaussian traits or traits transformable to normality, while ignoring qualitative traits and realistic, non-normal trait distributions. Also, modern computer languages, such as Julia, that accommodate parallelization and cloud-based computing are now mainstream but rarely used in older applications. To meet the challenges of contemporary big studies, it is important for geneticists to adopt new computational tools. We present TraitSimulation, an open-source Julia package that makes it trivial to quickly simulate phenotypes under a variety of genetic architectures. This package is integrated into our OpenMendel suite for easy downstream analyses. Julia was purpose-built for scientific programming and provides tremendous speed and memory efficiency, easy access to multi-CPU and GPU hardware, and to distributed and cloud-based parallelization. TraitSimulation is designed to encourage flexible trait simulation, including via the standard devices of applied statistics, generalized linear models (GLMs) and generalized linear mixed models (GLMMs). TraitSimulation also accommodates many study designs: unrelateds, sibships, pedigrees, or a mixture of all three. (Of course, for data with pedigrees or cryptic relationships, the simulation process must include the genetic dependencies among the individuals.) We consider an assortment of trait models and study designs to illustrate integrated simulation and analysis pipelines. Step-by-step instructions for these analyses are available in our electronic Jupyter notebooks on Github. These interactive notebooks are ideal for reproducible research. The TraitSimulation package has three main advantages. (1) It leverages the computational efficiency and ease of use of Julia to provide extremely fast, straightforward simulation of even the most complex genetic models, including GLMs and GLMMs. (2) It can be operated entirely within, but is not limited to, the integrated analysis pipeline of OpenMendel. And finally (3), by allowing a wider range of more realistic phenotype models, TraitSimulation brings power calculations and diagnostic tools closer to what investigators might see in real-world analyses.


[The blood bank as a consultation service].
This is a 6-month prospective clinical trial which evaluates the importance that blood bank may have as a consultation service for in-patient care. From January 31-July 31 1994, the blood bank physicians gave 108 consults which helped the consulting physician in an adequate diagnostic and therapeutic management of transfusion-related problems. The main reasons for consultation were coagulopathies, suspicion of refractoriness to platelet concentrates, and presence of antibodies against red cell antigens. The main underlying disorders were hematological, surgical, hepatic, renal and rheumatic diseases. We conclude that our blood bank as a consultation service was helpful in improving the transfusional management of our patients.


Respiratory effect of beta-blockers in people with asthma and cardiovascular disease: population-based nested case control study.
Cardiovascular disease (CVD) is a common comorbidity in people with asthma. However, safety concerns have caused heterogeneity in clinical guideline recommendations over the use of cardioselective beta-blockers in people with asthma and CVD, partly because risk in the general population has been poorly quantified. The aim of this study was to measure the risk of asthma exacerbations with beta-blockers prescribed to a general population with asthma and CVD. Linked data from the UK Clinical Practice Research Datalink was used to perform nested case-control studies among people with asthma and CVD matched on age, sex and calendar time. Adjusted incidence rate ratios (IRR) were calculated for the association between oral beta-blocker use and moderate asthma exacerbations (rescue oral steroids) or severe asthma exacerbations (hospitalisation or death) using conditional logistic regression. The cohort consisted of 35,502 people identified with active asthma and CVD, of which 14.1% and 1.2% were prescribed cardioselective and non-selective beta-blockers, respectively, during follow-up. Cardioselective beta-blocker use was not associated with a significantly increased risk of moderate or severe asthma exacerbations. Consistent results were obtained following sensitivity analyses and a self-controlled case series approach. In contrast, non-selective beta-blockers were associated with a significantly increased risk of moderate asthma exacerbations when initiated at low to moderate doses (IRR 5.16, 95% CI 1.83-14.54, P = 0.002), and both moderate and severe exacerbations when prescribed chronically at high dose (IRR 2.68, 95% CI 1.08-6.64, P = 0.033 and IRR 12.11, 95% CI 1.02-144.11, P = 0.048, respectively). Cardioselective beta-blockers prescribed to people with asthma and CVD were not associated with a significantly increased risk of moderate or severe asthma exacerbations and potentially could be used more widely when strongly indicated.


Photodynamic therapy of oral Candida infection in a mouse model.
Species of the fungal genus Candida, can cause oral candidiasis especially in immunosuppressed patients. Many studies have investigated the use of photodynamic therapy (PDT) to kill fungi in vitro, but this approach has seldom been reported in animal models of infection. This study investigated the effects of PDT on Candida albicans as biofilms grown in vitro and also in an immunosuppressed mouse model of oral candidiasis infection. We used a luciferase-expressing strain that allowed non-invasive monitoring of the infection by bioluminescence imaging. The phenothiazinium salts, methylene blue (MB) and new methylene blue (NMB) were used as photosensitizers (PS), combined or not with potassium iodide (KI), and red laser (660nm) at four different light doses (10J, 20J, 40J and 60J). The best in vitro log reduction of CFU/ml on biofilm grown cells was: MB plus KI with 40J (2.31 log; p<0.001); and NMB without KI with 60J (1.77 log; p<0.001). These conditions were chosen for treating the in vivo model of oral Candida infection. After 5days of treatment the disease was practically eradicated, especially using MB plus KI with 40J. This study suggests that KI can potentiate PDT of fungal infection using MB (but not NMB) and could be a promising new approach for the treatment of oral candidiasis.


College students' evaluations of alcohol consequences as positive and negative.
Alcohol expectancy, motivation, and consequences measures assume a known valence of 'positive' and 'negative' outcomes. However, different individuals may rate the same consequences of alcohol use as good or bad. The current study examines the extent to which: (a) college students rate researcher-defined positive consequences as good and researcher-defined negative consequences as bad, and (b) these evaluations predict alcohol use and problems after controlling for previous use. In longitudinal self-reports via web-surveys across the first three semesters of college, students (N=600; 54% women) reported their alcohol use and problems, experienced consequences, and evaluations of those consequences. Contrary to the generally-accepted valence of positive consequences, Fun/Social consequences were viewed as neutral or negative by 22% (having more fun) to 73% (relieving boredom) of participants. Over half of participants evaluated each of the Relaxation, Sex, and Image consequences items as neutral or negative. Consistent with the generally-accepted valence of negative consequences, Physical/Behavioral consequences were viewed by the majority as negative, although 11% (getting in trouble with police/authorities) to 34% (doing/saying something embarrassing) of students rated these consequences as neutral or positive. Independent of levels of previous drinking, more positive evaluations of Fun/Social consequences prospectively predicted frequency, quantity, and maximum drinks. Less negative evaluations of Physical/Behavioral consequences predicted more alcohol problems. There is variation in the evaluations of consequences among college students, and understanding characteristics of those who view consequences as positive or negative may have implications for future alcohol-related behaviors and problems.


Airborne infection with Bacillus anthracis--from mills to mail.
The lack of identified exposures in 2 of the 11 cases of bioterrorism-related inhalation anthrax in 2001 raised uncertainty about the infectious dose and transmission of Bacillus anthracis. We used the Wells-Riley mathematical model of airborne infection to estimate 1) the exposure concentrations in postal facilities where cases of inhalation anthrax occurred and 2) the risk for infection in various hypothetical scenarios of exposure to B. anthracis aerosolized from contaminated mail in residential settings. These models suggest that a small number of cases of inhalation anthrax can be expected when large numbers of persons are exposed to low concentrations of B. anthracis. The risk for inhalation anthrax is determined not only by bacillary virulence factors but also by infectious aerosol production and removal rates and by host factors.


A dual transcriptomic approach reveals contrasting patterns of differential gene expression during drought in arbuscular mycorrhizal fungus and carrot.
While arbuscular mycorrhizal (AM) fungi are known for providing host plants with improved drought tolerance, we know very little about the fungal response to drought in the context of the fungal-plant relationship. In this study, we evaluated the drought responses of the host and symbiont, using the fungus Rhizophagus irregularis with carrot as a plant model. Carrots (Daucus carota) inoculated with spores of R. irregularis DAOM 197198 grew in a greenhouse. During taproot development, carrots were exposed to a 10-day water restriction. Compared with well-watered conditions, drought caused diminished photosynthetic activity and reduced plant growth in carrot with and without AM fungi. Droughted carrots had lower root colonization. For R. irregularis, 93% of 826 differentially expressed genes (DEGs) were upregulated during drought, including phosphate transporters, several predicted transport proteins of potassium, and the aquaporin RiAQPF2. In contrast, 78% of 2,486 DEGs in AM carrot were downregulated during drought, including the symbiosis-specific genes FatM, RAM2, and STR, which are implicated in lipid transfer from the host to the fungus and were upregulated exclusively in AM carrot during well-watered conditions. Overall, this study provides insight into the drought response of an AM fungus in relation to its host; the expression of genes related to symbiosis and nutrient exchange were downregulated in carrot but upregulated in the fungus. This study reveals that carrot and R. irregularis exhibit contrast in their regulation of gene expression during drought, with carrot reducing its apparent investment in symbiosis and the fungus increasing its apparent symbiotic efforts.


Functional cerebral reorganization following motor sequence learning through mental practice with motor imagery.
The goal of the present study was to examine, via positron emission tomography, the functional changes associated with the learning of a sequence of foot movements through mental practice with motor imagery (MI). Following intensive MI training over several days, which led to a modest but significant improvement in performance, healthy subjects showed an increase in activity restricted to the medial aspect of the orbitofrontal cortex (OFC), and a decrease in the cerebellum. These main results are largely consistent with those found in a previous study of sequence learning performed in our laboratory after physical practice of the same task [NeuroImage 16 (2002) 142]. Further analyses showed a positive correlation between the blood flow increase in the OFC and the percentage of improvement on the foot sequence task. Moreover, the increased involvement of the medial OFC revealed a modality specific anatomo-functional organization, as imagination of the sequential task after MI practice activated a more posterior region than its execution. These results demonstrate that learning a sequential motor task through motor imagery practice produces cerebral functional changes similar to those observed after physical practice of the same task. Moreover, the findings are in accord with the hypothesis that mental practice with MI, at least initially, improves performance by acting on the preparation and anticipation of movements rather than on execution per se.


Cofilin reduces the mechanical properties of actin filaments: approach with coarse-grained methods.
An actin filament is an essential cytoskeleton protein in a cell. Various proteins bind to actin for cell functions such as migration, division, and shape control. ADF/cofilin is a protein that severs actin filaments and is related to their dynamics. Actin is known to have excellent mechanical properties. Binding cofilin reduces its mechanical properties, and is related to the severing process. In this research, we applied a coarse-grained molecular dynamics simulation (CGMD) method to obtain actin filaments and cofilin-bound actin (cofilactin) filaments. Using these two obtained models, we constructed an elastic network model-based structure and conducted a normal mode analysis. Based on the low-frequency normal modes of the filament structure, we applied the continuum beam theory to calculate the mechanical properties of the actin and cofilactin filaments. The CGMD method provided structurally accurate actin and cofilactin filaments in relation to the mechanical properties, which showed good agreement with the established experimental results.


Homocamptothecins: synthesis and antitumor activity of novel E-ring-modified camptothecin analogues.
Homocamptothecin (hCPT), a camptothecin (CPT) analogue with a seven membered beta-hydroxylactone which combines enhanced plasma stability and potent topoisomerase I (Topo I)-mediated activity, is an attractive template for the elaboration of new anticancer agents. Like CPT, hCPT carries an asymmetric tertiary alcohol and displays stereoselective inhibition of Topo I. The preparation and biological screening of racemic hCPT analogues are described. The 10 hCPTs tested were better Topo I inhibitors than CPT. Fluorinated hCPTs 23c, d,f,g were found to have potent cytotoxic activity on A427 and PC-3 tumor cell lines. Their cytotoxicity remained high on the K562adr and MCF7mdr cell lines, which overexpress a functionally active P-glycoprotein. Fluorinated hCPTs were more efficacious in vivo than CPT on HT-29 xenografts. In this model, a tumor growth delay of 25 days was reached with hCPT 23g at a daily dose of 0.32 mg/kg, compared to 4 days with CPT at 0.625 mg/kg. Thus difluorinated hCPT 23g warrants further investigation as a novel Topo I inhibitor with high cytotoxicity toward tumor cells and promising in vivo efficacy.


Effects of Diquafosol Ophthalmic Solution on Quality of Life in Dry Eye Assessed Using the Dry Eye-Related Quality-of-Life Score Questionnaire: Effectiveness in Patients While Reading and Using Visual Display Terminals.
Diquafosol ophthalmic solution improves objective findings in the ocular surface and subjective symptoms in patients with dry eye. The Dry Eye-Related Quality-of-Life Score (DEQS) questionnaire was developed to assess dry eye symptoms and their effects on quality of life. However, because little research using the DEQS has been reported, we evaluated the effects of diquafosol ophthalmic solution on ocular surface findings and quality of life using the DEQS in patients with dry eye. Sixty-three patients with dry eye were assigned to the control group (artificial tears) or diquafosol group. Both groups instilled 1 drop of the solution in both eyes 6 times daily and were evaluated after 2 weeks; the diquafosol group also was instructed to be examined at 1 and 3 months. We evaluated the subjective symptoms using the DEQS, fluorescein staining score, tear film breakup time (BUT), Schirmer testing, and lower tear meniscus height with anterior-segment optical coherence tomography. In the diquafosol group, the fluorescein staining score, BUT, tear meniscus height, and DEQS scores improved significantly compared with before treatment in contrast to the control group. Furthermore, in the diquafosol group, the staining score and BUT improved significantly compared with the control group. Analysis of each DEQS item indicated that diquafosol ophthalmic solution relieved foreign body sensation and problems when reading and using visual display terminals compared with the control group. Diquafosol ophthalmic solution was effective in patients with dry eye, especially those with foreign body sensation and problems when reading and using visual display terminals.


Expression of the mammalian c-fes protein in hematopoietic cells and identification of a distinct fes-related protein.
The avian c-fps and mammalian c-fes proto-oncogenes are cognate cellular sequences. Antiserum raised against the P140gag-fps transforming protein of Fujinami avian sarcoma virus specifically recognized a 92,000-Mr protein in human and mouse hematopoietic cells which was closely related in structure to Snyder-Theilen feline sarcoma virus P87gag-fes. This polypeptide was apparently the product of the human c-fes gene and was therefore designated p92c-fes. Human p92c-fes was associated with a tyrosine-specific protein kinase activity in vitro and was capable of both autophosphorylation and phosphorylation of enolase as an exogenous protein substrate. The synthesis of human and mouse p92c-fes was largely, though not entirely, confined to myeloid cells. p92c-fes was expressed to relatively high levels in a multipotential murine myeloid cell line, in more mature human and mouse granulocyte-macrophage progenitors, and in differentiated macrophage like cells as well as in the mononuclear fraction of normal and leukemic human peripheral blood. p92c-fes was not found in erythroid cells, with the exception of a human erythroleukemia line which retains the capacity to differentiate into macrophage like cells. These results suggest a normal role for the p92c-fes tyrosine kinase in hematopoiesis, particularly in granulocyte-macrophage differentiation. In addition, a distinct 94,000-Mr polypeptide, antigenically related to p92c-fes, was identified in a number of hematopoietic and nonhematopoietic human and mouse cells and was also found to be associated with a tyrosine-specific protein kinase activity.


Multistage decision-based heart sound delineation method for automated analysis of heart sounds and murmurs.
A robust multistage decision-based heart sound delineation (MDHSD) method is presented for automatically determining the boundaries and peaks of heart sounds (S1, S2, S3, and S4), systolic, and diastolic murmurs (early, mid, and late) and high-pitched sounds (HPSs) of the phonocardiogram (PCG) signal. The proposed MDHSD method consists of the Gaussian kernels based signal decomposition (GSDs) and multistage decision-based delineation (MDBD). The GSD algorithm first removes the low-frequency (LF) artefacts and then decomposes the filtered signal into two subsignals: the LF sound part (S1, S2, S3, and S4) and the high-frequency sound part (murmurs and HPSs). The MDBD algorithm consists of absolute envelope extraction, adaptive thresholding, and fiducial point determination. The accuracy and robustness of the proposed method is evaluated using various types of normal and pathological PCG signals. Results show that the method achieves an average sensitivity of 98.22%, positive predictivity of 97.46%, and overall accuracy of 95.78%. The method yields maximum average delineation errors of 4.52 and 4.14 ms for determining the start-point and end-point of sounds. The proposed multistage delineation algorithm is capable of improving the delineation accuracy under time-varying amplitudes of heart sounds and various types of murmurs. The proposed method has significant potential applications in heart sounds and murmurs classification systems.


2011 Marshall Urist Young Investigator Award: when to release patients to high-impact activities after hip resurfacing.
Surface replacement arthroplasties are commonly performed in young, active patients who desire return to high-impact activities including heavy manual labor and recreational sports. Femoral neck fracture is an arthroplasty-related complication unique to surface replacement arthroplasty. However, it is unclear regarding whether patients are at lower risk for fracture after a certain postoperative time. We therefore raised the following questions: (1) does stress shielding occur after surface replacement arthroplasty, and (2) when does bone mineral density return to normal so patients can return to high-impact activities without excessive risk of fracture? We prospectively enrolled 90 patients (96 hips) with either surface replacement arthroplasty or THA, and performed dual energy x-ray absorptiometry scans at 6 weeks, 6 months, 1 year, and 2 years. We analyzed bone density by Gruen zone in both groups, and six femoral neck zones in the patients who had surface replacement arthroplasties. We calculated 6-month, 1-year, and 2-year ratios for the change in bone density compared with baseline. Bone density was greater in patients who had surface replacement arthroplasties than for patients who had THAs at 6 months and 1 year in Gruen Zones 1, 2, 6, and 7, with the largest increase in femoral neck bone density on the tension side at 6 months in Zone L1. We saw no decrease in bone density in patients who had surface replacement arthroplasties in any Gruen zone at any time, and observed no decrease in bone density in female patients. Increased bone density at 6 months postoperatively in patients who had surface replacement arthroplasties provides evidence that clinically relevant stress shielding does not occur after surface replacement arthroplasty. Owing to the increased bone mineral density at 6 months, we believe patients who underwent surface replacement arthroplasties may to return to high-impact activities at that time without increased risk of fracture.


Motor symptoms and schizophrenia.
Classical schizophrenia literature reports motor symptoms as characteristic of the disorder. After the introduction of neuroleptic drugs, the existence of genuine motor disorders was challenged. Renewed interest arose as symptoms were found in never-medicated patients. Reports focused on abnormal involuntary movements, parkinsonism, neurological soft signs, catatonia, negative symptoms, or psychomotor slowing. Since these syndromes refer to different concepts, however, the definitions are not congruent and the symptoms overlap. The prevalence rates of motor symptoms in schizophrenia are surprisingly high, and recent studies indicate a possible pathobiology. In particular, the development and maturation of the human motor system appears to be closely linked to the emergence of motor symptoms observed in schizophrenia. Post-mortem and neuroimaging results demonstrated aberrant structure and function of premotor and motor cortices, basal ganglia, thalamus, and the connecting white matter tracts. Animal models have focused on aberrant neurotransmission and genetic contributions. Findings of localized abnormal oligodendrocyte function and myelination point to the special role of the white matter in schizophrenia, and recent studies specifically found an association between motor abnormalities and white matter structure in schizophrenia. This review of the literature supports the idea that motor symptoms are closely related to the neurodevelopmental disturbances of schizophrenia and a distinct syndromal dimension with its own pathophysiology.


'Someday it will be the norm': physician perspectives on the utility of genome sequencing for patient care in the MedSeq Project.
To describe practicing physicians' perceived clinical utility of genome sequencing. We conducted a mixed-methods analysis of data from 18 primary care physicians and cardiologists in a study of the clinical integration of whole-genome sequencing. Physicians underwent brief genomics continuing medical education before completing surveys and semi-structured interviews. Physicians described sequencing as currently lacking clinical utility because of its uncertain interpretation and limited impact on clinical decision-making, but they expressed the idea that its clinical integration was inevitable. Potential clinical uses for sequencing included complementing other clinical information, risk stratification, motivating patient behavior change and pharmacogenetics. Physicians given genomics continuing medical education use the language of both evidence-based and personalized medicine in describing the utility of genome-wide testing in patient care.


Pharmacokinetics, efficacy, and safety evaluation of docetaxel/hydroxypropyl-sulfobutyl-β-cyclodextrin inclusion complex.
Hydroxypropyl-sulfobutyl-β-cyclodextrin (HP-SBE-β-CD) inclusion complex was developed and used as a drug delivery system for DTX (DTX/HP-SBE-β-CD). The objective of the present study was to evaluate and compare the biological properties of DTX/HP-SBE-Β-CD with Taxotere®. The pharmacokinetics, biodistribution, antitumor efficacy in vivo and in vitro, and safety evaluation of DTX/HP-SBE-β-CD were studied. The most significant finding was that it was possible to prepare a Polysorbate-80-free inclusion complex for DTX. Studies based on pharmacokinetics, biodistribution, and antitumor efficacy indicated that DTX/HP-SBE-β-CD had similar pharmacokinetic properties and antitumor efficacy both in vitro and in vivo as Taxotere®. Fortunately, this new drug delivery system attenuated the side effects when used in vivo. As a consequence, DTX/HP-SBE-β-CD may be a promising alternative to Taxotere® for cancer chemotherapy treatment with reduced side effects. The therapeutic potential against a variety of human tumors and low toxicity demonstrated in a stringent study clearly warrant clinical investigation of DTX/HP-SBE-β-CD for possible use against human tumors.


Effective immobilization of BMP-2 mediated by polydopamine coating on biodegradable nanofibers for enhanced in vivo bone formation.
Although bone morphogenic proteins (BMPs) have been widely used for bone regeneration, the ideal delivery system with optimized dose and minimized side effects is still active area of research. In this study, we developed bone morphogenetic protein-2(BMP-2) immobilized poly(l-lactide) (PLLA) nanofibers inspired by polydopamine, which could be ultimately used as membranes for guided bone regeneration, and investigated their effect on guidance of in vitro cell behavior and in vivo bone formation. Surface chemical analysis of the nanofibers confirmed successful immobilization of BMP-2 mediated by polydopamine, and about 90% of BMP-2 was stably retained on the nanofiber surface for at least 28 days. The alkaline phosphatase activity and calcium mineralization of human mesenchymal stem cells (hMSCs) after 14 days of in vitro culture was significantly enhanced on nanofibers immobilized with BMP-2. More importantly, BMP-2 at a relatively small dose was highly active following implantation to the critical-sized defect in the cranium of mice; radiographic analysis demonstrated that 77.8 ± 11.7% of newly formed bone was filled within the defect for a BMP-2-immobilized groups at the concentration of 124 ± 9 ng/cm(2), as compared to 5.9 ± 1.0 and 34.1 ± 5.5% recovery, for a defect-only and a polydopamine-only group, respectively. Scanning and transmission electron microscopy of samples from the BMP-2 immobilized group showed fibroblasts and osteoblasts with nanofiber strands in the middle of regenerated bone tissue, revealing the importance of interaction between implanted nanofibers and the neighboring extracellular environment. Taken together, our data support that the presentation of BMP-2 on the surface of nanofibers as immobilized by utilizing polydopamine chemistry may be an effective method to direct bone growth at relatively low local concentration.


Synthesis and Single-Molecule Conductances of Neutral and Cationic Indenofluorene-Extended Tetrathiafulvalenes: Kondo Effect Molecules.
Development of molecules that can switch between redox states with paired and unpaired electrons is important for molecular electronics and spintronics. In this work, a selection of redox-active indenofluorene-extended tetrathiafulvalenes (IF-TTFs) with thioacetate end groups was prepared from a readily obtainable dibromo-functionalized IF-TTF building block using palladium-catalyzed cross-coupling reactions, such as the Suzuki reaction. The end groups served as electrode anchoring groups for single-molecule conductance studies, and the molecules were subjected to mechanically controlled break-junction measurements with gold contacts and to low-bias charge transport measurements in gated three-terminal electromigration junctions. The neutral molecules showed clear conductance signatures, and somewhat surprisingly, we found that a meta-meta anchoring configuration gave a higher conductance than a para-meta configuration. We explain this behavior by "through-space" coupling between the gold electrode and the phenyl on which the anchoring group is attached. Upon charging the molecule in a gated junction, we found reproducibly a Kondo effect (zero-bias conductance) attributed to a net spin. Ready generation of radical cations was supported by cyclic voltammetry measurements, revealing stepwise formation of radical cation and dication species in solution. The first oxidation event was accompanied by association reactions as the appearance of the first oxidation peak was strongly concentration dependent.


Varicella zoster virus infection after allogeneic hematopoietic cell transplantation in children using a relatively short duration of acyclovir prophylaxis: A retrospective study.
Although acyclovir prophylaxis against varicella zoster virus (VZV) infection for ≥1 year is recommended after allogeneic hematopoietic cell transplantation (HCT), the emergence of acyclovir-resistant viruses and adverse drug effects cannot be ignored. We investigated the cumulative incidence of VZV infection after allogeneic HCT in children receiving a shorter duration of acyclovir prophylaxis than recommended and evaluated the appropriateness of the short duration of acyclovir prophylaxis.Medical records of 217 children who received allogeneic HCT were retrospectively reviewed until a median of 25 months (range = 1-59 months) after HCT. Acyclovir prophylaxis was given for a median of 9 weeks (range = 3-24 weeks) after HCT.VZV infection was diagnosed in 33 (15.2%) children at a median time of 5 months (range = 2-41 months) after HCT. The 1-year and 2-year cumulative incidences of VZV infection after allogeneic HCT were 11.2% and 15.5%, respectively. These incidences were between the previously reported 1-year incidence of 25% to 30% in patients not receiving prophylaxis and 1-year incidence of 4% to 5% in patients receiving ≥1 year duration of prophylaxis. Male sex and older age were significantly associated with VZV infection after allogeneic HCT. Only 1 chickenpox patient experienced severe complications because of VZV infection, and there were no deaths attributable to VZV infection.In conclusion, a shorter duration of acyclovir prophylaxis may be appropriate for children receiving allogeneic HCT, based on the rare occurrence of severe complications because of VZV infection and the expected discomfort because of daily oral medication for a long time.


A comparison of treatment adherence in individuals with a first episode of psychosis and inpatients with psychosis.
In predicting treatment compliance in individuals with severe mental illness, research has focused on variables such as substance abuse, personality, history of child abuse, and symptomatology, although these relationships have not been investigated in great detail in individuals at the onset of mental illness. To better understand these correlates of treatment compliance, two samples were examined: a sample of 117 individuals presenting with a first episode of psychosis and a more chronic forensic sample of 65 participants recruited from a psychiatric hospital. These samples were investigated for service engagement in terms of violence history, substance abuse, symptom severity, psychopathic traits and history of childhood abuse. Linear regressions performed for the first episode sample revealed that childhood physical abuse was the strongest predictor of poor service engagement, followed by problems with alcohol, a history of physical violence, any history of violence and higher psychopathic traits. Linear regression revealed for the forensic group that a lower level of service engagement was most strongly predicted by a history of childhood abuse and a higher score on the Brief Psychiatric Rating Scale (BPRS). Results are presented in light of the existing literature and clinical implications are discussed.


Novel medium-term carcinogenesis model for lung squamous cell carcinoma induced by N-nitroso-tris-chloroethylurea in mice.
Targeted treatments for lung cancer based on pathological diagnoses are required to enhance therapeutic efficacy. There are few well-established animal models for lung squamous cell carcinoma although several highly reproducible mouse models for lung adenoma and adenocarcinoma are available. This study was carried out to establish a new lung squamous cell carcinoma mouse model. In the first experiment, female A/J mice were painted topically on back skin twice weekly with 75 μL 0.013 M N-nitroso-tris-chloroethylurea for 2, 4, and 8 weeks (n = 15-20 per group) as initiation of lung lesions, and surviving mice were killed at 18 weeks. In the second experiment, mice were treated as above for 4 weeks and killed at 6, 12, or 18 weeks (n = 3 per group). Lung lobes were subjected to histopathological, immunohistochemical, immunoblotting, and ultrastructural analyses. In the case of treatment for 2, 4, and 8 weeks, incidences of lung squamous cell carcinoma were 25, 54, and 71%, respectively. Cytokeratin 5/6 and epidermal growth factor receptor were clearly expressed in dysplasia and squamous cell carcinoma. Desmosomes and tonofilaments developed in the squamous cell carcinoma. Considering the carcinogenesis model, we conclude that 2 or 4 weeks of N-nitroso-tris-chloroethylurea treatment may be suitable for investigating new chemicals for promotional or suppressive effects on lung squamous cell carcinoma.


Levels of advertised unprotected vaginal and oral sex by independent indoor female sex workers in West Yorkshire, UK.
To assess the proportion of independent indoor female sex workers (FSW) in West Yorkshire, UK who advertise unprotected sex, and to investigate any association with cost, location and provision of anal sex. Data on whether independent indoor FSW (defined as those not advertising via an escort agency or through a parlour) advertised unprotected sexual services, along with demographic data, were collected from 462 advertisement profiles of FSW in West Yorkshire from the website http://www.adultwork.com. Independent t test and χ(2) statistics were used to test the association between advertised unprotected vaginal and oral sex, and FSW age, cost of services, location and whether they advertised anal sex. Unprotected vaginal sex was advertised by 8% of FSW, and unprotected oral sex by 74% of FSW. FSW advertising unprotected vaginal sex were more likely to live in Wakefield and Bradford than in Leeds, had significantly lower hourly rates, and were more likely to advertise anal sex. Advertised condom use for vaginal and oral sex by independent indoor FSW in West Yorkshire was significantly lower than reported rates of protected sex found in previous studies based in London and the south of England. The advertisement of unprotected vaginal sex is associated with factors such as lower hourly rates and the advertisement of higher risk anal sex, which may signify greater economic need. FSW offering unprotected sex therefore represent an at-risk target group for health promotion.


Experiences of teenage pregnancy among Xhosa families.
to explore and describe the experiences of teenage pregnancy among Xhosa families, and, depending on the results of the study, to recommend a strategy to assist midwives to enhance pregnancy outcomes. a qualitative, explorative, descriptive, phenomenological and contextual research design. In-depth face-to-face interviews were performed to collect data. 10 pregnant teenagers, eight mothers, two fathers, seven grandmothers and three grandfathers from the same families were interviewed independently and privately. pregnant teenagers experienced emotional turmoil as they strived to cope with their pregnancy, and experienced a change in their relationships with significant others due to expectations that were not met and role confusion which led to crisis. Parents experienced overwhelming emotions due to the unexpected pregnancy of their child, and loss of control as the pregnancy could not be reversed. Grandparents of pregnant teenagers experienced the pregnancy as a family disturbance, and acknowledged that healing should take place in the family. teenage pregnancy was experienced differently by different generations within the same family, but all the experiences culminated in anger that hampered the necessary parental support for the pregnant teenager. Lack of support during pregnancy can easily affected the well-being of the unborn child, as teenagers are not supervised and experience acute emotional stress. Intervention by a midwife could help to relieve the teenager's stress and optimise the pregnancy outcome.


Characterization and clinical application of human CD34+ stem/progenitor cell populations mobilized into the blood by granulocyte colony-stimulating factor.
A phase I study was performed to determine the safety and tolerability of injecting autologous CD34(+) cells into five patients with liver insufficiency. The study was based on the hypothesis that the CD34(+) cell population in granulocyte colony-stimulating factor (G-CSF)-mobilized blood contains a subpopulation of cells with the potential for regenerating damaged tissue. We separated a candidate CD34(+) stem cell population from the majority of the CD34(+) cells (99%) by adherence to tissue culture plastic. The adherent and nonadherent CD34(+) cells were distinct in morphology, immunophenotype, and gene expression profile. Reverse transcription-polymerase chain reaction-based gene expression analysis indicated that the adherent CD34(+) cells had the potential to express determinants consistent with liver, pancreas, heart, muscle, and nerve cell differentiation as well as hematopoiesis. Overall, the characteristics of the adherent CD34(+) cells identify them as a separate putative stem/progenitor cell population. In culture, they produced a population of cells exhibiting diverse morphologies and expressing genes corresponding to multiple tissue types. Encouraged by this evidence that the CD34(+) cell population contains cells with the potential to form hepatocyte-like cells, we gave G-CSF to five patients with liver insufficiency to mobilize their stem cells for collection by leukapheresis. Between 1 x 10(6) and 2 x 10(8) CD34(+) cells were injected into the portal vein (three patients) or hepatic artery (two patients). No complications or specific side effects related to the procedure were observed. Three of the five patients showed improvement in serum bilirubin and four of five in serum albumin. These observations warrant further clinical trials.


Obstructive sleep apnoea. Part I: Diagnosis, aetiology, and current treatment.
Obstructive sleep apnoea is a multi-factorial condition associated with high morbidity and mortality. Its prevalence is highest in middle-aged males with a predisposition to obesity. Certain facial types have been identified as being at risk. Cephalometric parameters have now been established to identify those patients who are anatomically compromised. Treatment modalities are dependent on the correct diagnosis of the site(s) of obstruction. Overnight polysomographic testing is the only definitive measure to quantify the presence and severity of obstructive sleep apnoea. Treatment options include behavioural, medical, surgical, and the use of oral appliances.


Versatile 3D-Printed Micro-Reference Electrodes for Aqueous and Non-Aqueous Solutions.
While numerous reference electrodes suitable for aqueous electrolytes exist, there is no well-defined standard for non-aqueous electrolytes. Furthermore, reference electrodes are often large and do not meet the size requirements for small cells. In this work, we present a simple method for fabricating stable 3D-printed micro-reference electrodes. The prints are made from polyvinylidene fluoride, which is chemically inert in strong acids, bases, and commonly used non-aqueous solvents. We chose six different reference systems based on Ag, Cu, Zn, and Na, including three aqueous and three non-aqueous systems to demonstrate the versatility of the approach. Subsequently, we conducted cyclic voltammetry experiments and measured the potential difference between the aqueous homemade reference electrodes and a commercial Ag/AgCl-electrode. For the non-aqueous reference electrodes, we chose the ferrocene redox couple as an internal standard. From these measurements, we deduced that this new class of micro-reference electrodes is leak-tight and shows a stable electrode potential.


Strong nonlinear terahertz response induced by Dirac surface states in Bi2Se3 topological insulator.
Electrons with a linear energy/momentum dispersion are called massless Dirac electrons and represent the low-energy excitations in exotic materials such as graphene and topological insulators. Dirac electrons are characterized by notable properties such as a high mobility, a tunable density and, in topological insulators, a protection against backscattering through the spin-momentum locking mechanism. All those properties make graphene and topological insulators appealing for plasmonics applications. However, Dirac electrons are expected to present also a strong nonlinear optical behaviour. This should mirror in phenomena such as electromagnetic-induced transparency and harmonic generation. Here we demonstrate that in Bi2Se3 topological insulator, an electromagnetic-induced transparency is achieved under the application of a strong terahertz electric field. This effect, concomitantly determined by harmonic generation and charge-mobility reduction, is exclusively related to the presence of Dirac electron at the surface of Bi2Se3, and opens the road towards tunable terahertz nonlinear optical devices based on topological insulator materials.


Operative Wound Complications Following Total Ankle Arthroplasty.
Wound complications following total ankle replacement (TAR) potentially lead to devastating consequences. The aim of this study was to compare the operative and demographic differences in patients with and without major wound problems which required operative management. We hypothesized that increased tourniquet and operative time would negatively influence wound healing. We identified a consecutive series of 762 primary TARs performed between December 1999 and April 2014 whose data were prospectively collected. We then identified the subset of patients who required a secondary surgery to treat major wound complications (ie, operative debridement, split-thickness skin grafting, and soft tissue reconstruction). All patients requiring a second surgery had operative wound debridement. We then compared the demographics, operative characteristics, and functional scores to see if any differences existed between patients with and without major wound complications. Clinical outcomes including secondary procedures and implant failure rates were recorded. Twenty-six patients (3.4%) had a total of 49 operative procedures to treat major wound issues. Eighteen patients had flaps and 14 had split-thickness skin grafts. The median time to operatively treating the wound was 1.9 (range: 0.5-12.5) months after the index TAR. The median follow-up time from the wound procedure was 12.7 (range: 1.2-170.8) months. Compared to the control group, patients with major wounds had a significantly longer mean surgery (214.8 vs 189.3 minutes, P = .041) time and trended toward a longer median tourniquet time (151 vs 141 minutes, P = .060). Patients without wound complications were more likely to have posttraumatic arthritis, whereas those with wound complications were more likely to have primary osteoarthritis ( P = .006). The control group trended toward having a higher mean BMI (29.5 vs 27.2, P = .056). There were 6 failures in the major wound complication cohort (23.1%), including 2 below the knee amputations. Ankle wounds that required operative management had high failure rates and some resulted in devastating outcomes. We did not find any increase in major wound complications in those with various risk factors as identified by other studies. Given our data, we recommend limiting operative time. While correcting hindfoot and midfoot alignment is important for improving patient functionality and survivorship of the implant, thought should be given to staging the TAR if multiple pathologies are to be addressed at the time of surgery to limit operative time. Level III, retrospective comparative series.


Laterally Extended Pelvic Resection for Gynaecological Malignancies: A Multicentric Experience with Out-of-the-Box Surgery.
To evaluate morbidity and oncological outcome in a multicentre series of women with gynaecological malignancies infiltrating pelvic side wall (PSW) that received laterally extended pelvic resection (LEPR). Patients operated between 2007 and 2017 at three institutions were included. LEPR was defined as an en bloc lateral resection of a pelvic tumour involving sidewall muscle, and/or bone, and/or major nerve, and/or major vascular structure. Postsurgical complications and survivals were evaluated. Sixty-three women with gynaecological tumours involving PSW were treated with LEPR. Five women underwent primary LEPR, whereas 58 (92%) patients needed LEPR because of recurrence. Twenty-four women (38%) received previous radiation therapy before the surgery. R0 resection was achieved in 54 patients (85.7%), whereas the pathologic margins were microscopically and macroscopically positive in 8 (12.7%) patients and 1 (1.6%) patient, respectively. There was one perioperative death, whereas major postoperative complications occurred in 17 patients (27.7%). Thirty (47.5%) women experienced recurrences: 24/54 (44.4%) were in the R0 group, and 6/9 (66.6%) were in the R1 group, with a median PFS of 15 months and 7 months, respectively (p = 0.024). In total, 11 of 54 (20.3%) patients died of disease in the R0 group and 5 of 9 (55.5%) in the R1 group; a median OS was not reached and was 32 months for R0 and R1 groups, respectively (p = 0.033). Involvement of the PSW should not prevent obtaining R0 resection. Although the LEPR is associated with considerable morbidity (≈ 30%), a long-term survival seems to be achieved in those women with complete resection.


Determinants of re-operation for bleeding in head and neck cancer surgery.
Post-operative bleeding in the head and neck area is potentially fatal. This 'real world' study sought to assess factors that increase the risk of re-operation for post-operative bleeding in head and neck cancer surgery. A total of 456 patients underwent surgery for head and neck cancer (591 operations). The primary endpoint was re-operation for bleeding. The rate of re-operation for bleeding was 5 per cent of all operations. Re-operation for bleeding was an independent risk factor for 30-day mortality (odds ratio = 5.27, p = 0.014). Risk factors for re-operation because of bleeding included excessive (more than 4000 ml) fluid administration (over 24 hours) (p < 0.001), heavy alcohol consumption (p = 0.014), pre-operative oncological treatment (p = 0.017), advanced disease stage (p = 0.020) and higher tumour (T) classification (p = 0.034). Operations with more excessive bleeding (700 ml or more) were associated with an increased risk (p = 0.001) of re-operation for post-operative bleeding. Moreover, the risk of re-operation was significantly higher in patients undergoing microvascular surgery compared to those who had no oncological treatment pre-operatively (18 vs 6 per cent, p = 0.001). The 30-day mortality risk increased over 5-fold in patients undergoing re-operation for bleeding.


Carbonic acid buffer species measured in real time with an intracellular microelectrode array.
Carbonic acid buffer anions, HCO3- and CO3(2-), play an instrumental role in a host of vital processes in animal cells and tissues. Yet study of carbonic acid buffer species is hampered because no means are available to simultaneously monitor them at a cellular level in a rapid and dynamic fashion. An ion-selective cocktail, previously reported to measure changes in bicarbonate activity (alpha HCO3-), was instead shown to be principally selective for alpha CO3(2-). Ion-selective micropipettes (ISMs) based on this exchanger and consisting of a 3:1:6 (volume) mixture of tri-n-octylpropylammonium chloride, 1-octanol, and trifluoroacetyl-p-butylbenzene showed no significant interference from bicarbonate, chloride, phosphate, ascorbate, lactate, glutamate, acetate, or hydroxyl ions at concentrations expected in vivo. Intracellular and triple-barrel ISMs, consisting of a CO3(2-) sensitive, pH-sensitive, and reference barrel, were fabricated. Skeletal muscle cells (n = 17) were penetrated in vivo and showed values of 74 +/- 7 mV for membrane potential, 6.94 +/- 0.09 pHi, and 11 +/- 5 microM intracellular alpha CO3(2-), from which intracellular alpha HCO3- of 25 +/- 10 mM and CO2 tension of 120 +/- 55 Torr were calculated. All ion measurements reached a new steady state in 9 +/- 2 s after cell penetration. Thus measurements of intracellular alpha CO3(2-) and pH and associated levels of alpha HCO3(2-) and CO2 tension can be determined in biological tissues and cells with a spatial and temporal resolution previously unattainable.


3D-printed, patient-specific DIEP flap templates for preoperative planning in breast reconstruction: a prospective case series.
Modern imaging technologies, such as computed tomographic angiography (CTA), can be useful for preoperative assessment in deep inferior epigastric artery perforator (DIEP) flap surgery. Planning perforator flap design can lead to improved surgical efficiency. However, current imaging modalities are limited by being displayed on a two-dimensional (2D) surface. In contrast, a 3D-printed model provides tactile feedback that facilitates superior understanding. Hence, we have 3D-printed patient-specific deep inferior epigastric artery perforator (DIEP) templates, in an affordable and convenient manner, for preoperative planning. Twenty consecutive patients undergoing 25 immediate or delayed post-mastectomy autologous breast reconstruction with DIEP or muscle-sparing transverse rectus abdominis (MS-TRAM) flaps are recruited prospectively. Using free, open-source softwares (3D Slicer, Autodesk MeshMixer, and Cura) and desktop 3D printers (Ultimaker 3E and Moment), we created a template based on a patient's abdominal wall anatomy from CTA, with holes and lines indicating the position of perforators, their intramuscular course and the DIEA pedicle. The mean age of patients was 52 [38-67]. There were 15 immediate and 10 delayed reconstructions. 3D printing time took mean 18 hours and 123.7 g of plastic filament, which calculates to a mean material cost of AUD 8.25. DIEP templates accurately identified the perforators and reduced intraoperative perforator identification by 7.29 minutes (P=0.02). However, the intramuscular dissection time was not affected (P=0.34). Surgeons found the template useful for preoperative marking (8.6/10) and planning (7.9/10), but not for intramuscular dissection (5.9/10). There were no immediate flap-related complications. Our 3D-printed, patient-specific DIEP template is accurate, significantly reduces intraoperative perforator identification time and, hence, may be a useful tool for preoperative planning in autologous breast reconstruction.


Role of HLA-DP Expression in Graft-Versus-Host Disease After Unrelated Donor Transplantation.
The main aim of this study was to evaluate the significance of HLA-DPB1 expression in acute graft-versus-host disease (GVHD) after hematopoietic cell transplantation (HCT) from HLA-A, -B, -C, -DRB1, -DQB1-matched and -mismatched unrelated donors. Between January 1, 2017, and January 10, 2019, we assessed 19,136 patients who received HCT from an HLA-A, -B, -C, -DRB1, -DQB1-matched or -mismatched unrelated donor performed in Australia, the European Union, Japan, North America, and the United Kingdom between 1988 and 2016. Among transplant recipients with one HLA-DPB1 mismatch, the patient's mismatched HLA-DPB1 allotype was defined as low or high expression. Multivariable regression models were used to assess risks of GVHD associated with high expression relative to low expression HLA-DPB1 mismatches. The effect of increasing numbers of HLA-DPB1 mismatches on clinical outcome was assessed in HLA-mismatched transplant recipients. In HLA-A, -B, -C, -DRB1,-DQB1-matched transplant recipients, donor mismatching against one high-expression patient HLA-DPB1 increased moderate (odds ratio [OR], 1.36; P = .001) and severe acute GVHD (OR, 1.32; P = .0016) relative to low-expression patient mismatches, regardless of the expression level of the donor's mismatched HLA-DPB1. Among transplant recipients with one HLA-A, -B, -C, -DRB1, or -DQB1 mismatch, the odds of acute GVHD increased with increasing numbers of HLA-DPB1 mismatches (OR, 1.23 for one; OR, 1.40 for two mismatches relative to zero mismatches for moderate GVHD; OR, 1.19 for one; OR, 1.40 for two mismatches relative to zero for severe GVHD), but not with the level of expression of the patient's mismatched HLA-DPB1 allotype. The level of expression of patient HLA-DPB1 mismatches informs the risk of GVHD after HLA-A, -B, -C, -DRB1, -DQB1-matched unrelated HCT, and the total number of HLA-DPB1 mismatches informs the risk of GVHD after HLA-mismatched unrelated HCT. Prospective consideration of HLA-DPB1 may help to lower GVHD risks after transplantation.


Development of an optimized tetracycline-inducible expression system to increase the accumulation of interleukin-10 in tobacco BY-2 suspension cells.
Plant cell suspension cultures can be used for the production of valuable pharmaceutical and industrial proteins. When the recombinant protein is secreted into the culture medium, restricting expression to a defined growth phase can improve both the quality and quantity of the recovered product by minimizing proteolytic activity. Temporal restriction is also useful for recombinant proteins whose constitutive expression affects cell growth and viability, such as viral interleukin-10 (vIL-10). We have developed a novel, tetracycline-inducible system suitable for tobacco BY-2 suspension cells which increases the yields of vIL-10. The new system is based on a binary vector that is easier to handle than conventional vectors, contains an enhanced inducible promoter and 5'-UTR to improve yields, and incorporates a constitutively-expressed visible marker gene to allow the rapid and straightforward selection of the most promising transformed clones. Stable transformation of BY-2 cells with this vector, without extensive optimization of the induction conditions, led to a 3.5 fold increase in vIL-10 levels compared to constitutive expression in the same host. We have developed an effective and straightforward molecular farming platform technology that improves both the quality and the quantity of recombinant proteins produced in plant cells, particularly those whose constitutive expression has a negative impact on plant growth and development. Although we tested the platform using vIL-10 produced in BY-2 cells, it can be applied to other host/product combinations and is also useful for basic research requiring strictly controlled transgene expression.


Direct curvature correction for noncontact imaging modalities applied to multispectral imaging.
Noncontact optical imaging of curved objects can result in strong artifacts due to the object's shape, leading to curvature biased intensity distributions. This artifact can mask variations due to the object's optical properties, and makes reconstruction of optical/physiological properties difficult. In this work we demonstrate a curvature correction method that removes this artifact and recovers the underlying data, without the necessity of measuring the object's shape. This method is applicable to many optical imaging modalities that suffer from shape-based intensity biases. By separating the spatially varying data (e.g., physiological changes) from the background signal (dc component), we show that the curvature can be extracted by either averaging or fitting the rows and columns of the images. Numerical simulations show that our method is equivalent to directly removing the curvature, when the object's shape is known, and accurately recovers the underlying data. Experiments on phantoms validate the numerical results and show that for a given image with 16.5% error due to curvature, the method reduces that error to 1.2%. Finally, diffuse multispectral images are acquired on forearms in vivo. We demonstrate the enhancement in image quality on intensity images, and consequently on reconstruction results of blood volume and oxygenation distributions.


An evaluation of the burden placed on the General Internal Medicine team at the Role 3 Hospital in Camp Bastion by UK Armed Forces personnel presenting with symptoms resulting from previously identified disease.
During previous deployments of the British Armed Forces, a significant proportion of aeromedical evacuations were accounted for with recurrent symptoms from a known disease that had often triggered occupational medical downgrading. Many servicemen and women had deployed inappropriately, and by doing so became a burden on the deployed medical facilities. Commanders performing systematic medical risk assessments prior to departure might have prevented these individuals from deploying. This study was designed to assess the avoidable burden from recurrent disease during the current Afghanistan operation. A cross-sectional study reviewing the hospital and computerised primary care medical records of consecutive patients admitted under the General Physicians to the Role 3 Hospital in Camp Bastion over 9 months from April 2011. The occupational medical grading, diagnosis, disposal and whether the disease was recurrent were recorded. Of 270 patients admitted, 14 (5.2%) were medically downgraded. The computerised records were unavailable for 31 (11.5%) patients. All those patients who were medically downgraded were graded 'Medically Limited Deployable'. In the downgraded group, only one patient presented with recurrent symptoms from their pre-existing condition (Crohn's disease). In the non-downgraded group, two patients presented with symptoms relating to their previous diagnoses. One presented with a second heat illness and should have been medically downgraded and not have been deployed, while the other patient had previously been investigated for recurrent syncope and was upgraded 6 months prior to deploying. All three patients underwent aeromedical evacuation but only two of these were considered to have been avoidable. The number of general medical admissions to the Role 3 Hospital due to a pre-existing disease is very low.


What does beta-bungarotoxin do at the neuromuscular junction?
beta-Bungarotoxin from the Taiwan banded krait, Bungarus multicinctus is a basic protein (pI=9.5), with a molecular weight of 21,800 consisting of two different polypeptide subunits. A phospholipase A(2) subunit named the A-chain and a non-phospholipase A(2) subunit named the B-chain, which is homologous to Kunitz protease inhibitors. The A-chain and the B-chain are covalently linked by one disulphide bridge. On mouse hemi-diaphragm nerve-muscle preparations, partially paralysed by lowering the external Ca(2+) concentration, beta-bungarotoxin classically produces triphasic changes in the contraction responses to indirect nerve stimulation. The initial transient inhibition of twitches (phase 1) is followed by a prolonged facilitatory phase (phase 2) and finally a blocking phase (phase 3). These changes in twitch tension are mimicked, to some extent, by similar changes to end plate potential amplitude and miniature end plate potential frequency. The first and second phases are phospholipase-independent and are thought to be due to the B-chain (a dendrotoxin mimetic) binding to or near to voltage-dependent potassium channels. The last phase (phase 3) is phospholipase dependent and is probably due to phospholipase A(2)-mediated destruction of membrane phospholipids in motor nerve terminals.


Population structure of spotted salamanders (Ambystoma maculatum) in a fragmented landscape.
Understanding the impacts of landscape-level processes on the population biology of amphibians is critical, especially for species inhabiting anthropogenically modified landscapes. Many pond-breeding amphibians are presumed to exist as metapopulations, but few studies demonstrate the extent and consequences of this metapopulation structure. Gene flow measures may facilitate the construction of more realistic models of population structure than direct measures of migration. This is especially true for species that are cryptic, such as many amphibians. We used eight polymorphic microsatellite loci to determine the genetic population structure of spotted salamanders (Ambystoma maculatum) breeding at 17 ponds in northeastern Ohio, a landscape fragmented by roads, agriculture, urban areas and the Cuyahoga River. Using a variety of analyses (Bayesian clustering, F-statistics, AMOVA) we generated a model of salamander population genetic structure. Our data revealed patterns of genetic connectivity that were not predicted by geographical distances between ponds (no isolation by distance). We also tested for a relationship between population structure and several indices of landscape resistance, but found no effect of potential barriers to dispersal on genetic connectivity. Strong overall connectivity among ponds, despite the hostile habitat matrix, may be facilitated by a network of riparian corridors associated with the Cuyahoga River; however, high gene flow in this system may indicate a general ability to disperse and colonize beyond particular corridors.


The immunological landscape in pancreatic ductal adenocarcinoma and overcoming resistance to immunotherapy.
Pancreatic ductal adenocarcinoma is associated with a poor prognosis and there are few treatment options. The development of immunotherapy in pancreatic ductal adenocarcinoma has been difficult, and immune checkpoint inhibitors are only effective in a very small subset of patients. Most obstacles for treatment have been related to intertumoural and intratumoural heterogeneity, the composition of tumour stroma, and crosstalk with cancer cells. Improved molecular characterisation of pancreatic ductal adenocarcinoma and a better understanding of its microenvironment have paved the way for novel immunotherapy strategies, including the identification of predictive biomarkers, the development of rational combinations to optimise effectiveness, and the targeting of new mechanisms. Future immunotherapy strategies should consider individual characteristics to move beyond the traditional immune targets and circumvent the resistance to therapies that have been developed so far.


Pharmacokinetics and pharmacodynamics of milrinone lactate in pediatric patients with septic shock.
The objectives of this study were to determine the pharmacokinetics of milrinone lactate in pediatric patients with septic shock and to determine whether a relationship exists between steady-state plasma milrinone concentrations and changes in hemodynamic variables. This was a randomized, double-blind, placebo-controlled, interventional study. In study phase 1 patients were randomized and underwent loading and infusion with milrinone lactate (50 microg/kg, then 0.5 microg/kg/min), and invasive hemodynamic values were determined. Steady-state was determined by obtaining plasma samples at 30, 15, and 0 minutes before the end of the milrinone infusion. Study phase 2 started when milrinone was discontinued by the patient care team. Steady-state was reaffirmed and plasma samples were obtained at 0.5, 1, 2, 4, 6, and 8 hours after the end of the infusion. The average plasma concentration at steady-state (Css avg) and total body clearance for phase 1 were 81.3+/-38.6 ng/ml (mean +/- SD) and 0.0106+/-0.0053 L/kg/min, respectively (n = 9). All but two patients underwent reloading with milrinone. In phase 2 Css avg and total body clearance were 65.8+/-42.1 ng/ml and 0.0110+/-0.0096 L/kg/min, respectively (n = 11). The average time of infusion was 51+/-21 hours. Eight patients were evaluated for phase 2 elimination. The mean elimination rate constant was 0.0091+/-0.0061 min(-1) (n = 8). The median half-life was 1.47 hours (range, 0.62 to 10.85 hours). All patients had creatinine clearances greater than 61 ml/min/1.73 m2. The volume of distribution at steady-state was 1.47+/-1.03 L/kg. No correlation existed between age and the elimination rate constant or the volume of distribution at steady-state. All patients achieved at least a 20% change in cardiac index and systemic vascular resistance index while maintaining a Css avg of 35 to 160 ng/ml. No adverse effects were noted. All patients achieved primary hemodynamic end points (cardiac index and systemic vascular resistance index) during the milrinone infusion. Loading doses of 75 microg/kg milrinone lactate and starting infusion rates of 0.75 to 1.0 microg/kg/min for patients with normal renal function should be used; the infusion rate should then be titrated to effect. We recommend that for every increase of 0.25 microg/kg/min, a 25 microg/kg bolus dose be given. Because the median half-life is 1.47 hours, immediate hemodynamic effects may not be seen unless appropriate loading doses and infusion adjustments are made.


[Electrocardiography and arterial blood gases monitoring during fiberoptic bronchoscopy].
Electrocardiography (ECG) and arterial blood gases (ABG) were taken to monitor 90 patients prior to, during and after bronchofiberscopy. PaO2 generally decreased during the procedure without supplemental oxygen. Patients who underwent bronchofiberscopy with PaO2 less then 9.31 kPa were 66.67%. The mean of the decrease of SaO2 was 5.79 +/- 7.74%. 83.33% of the patients developed sinus tachycardia. There were 26.67% of the patients showed abnormal P wave, ST-T wave, Q-T interval etc during the procedure. 93.33% of them were given oxygen via a nasal stuff during the procedure and had an increase of PaO2.


Prenatal lead exposure and childhood executive function and behavioral difficulties in project viva.
Lead is an established neurotoxicant and early life exposure to lead is associated with detrimental impacts on IQ and several neurobehavioral domains. Less is known, however, about effects of prenatal lead exposure below 5 μg/dL on executive function and on social, emotional and self-regulatory behaviors in childhood. To examine the association between prenatal lead exposure and childhood executive function and social, emotional and self-regulatory behaviors. We included 1006 mother-child pairs from the Project Viva prospective pre-birth cohort. We measured prenatal maternal lead in second-trimester erythrocytes. In mid-childhood (median 7.7 years), parents and teachers rated executive function related behaviors using the Behavior Rating Inventory of Executive Function (BRIEF) and behavioral difficulties using the Strengths and Difficulties Questionnaire (SDQ). We used multivariable linear regression models adjusted for maternal, paternal, and child characteristics and metal co-exposures. Mean maternal erythrocyte lead concentration was 1.2 μg/dL (interquartile range [IQR] 0.8-1.5 μg /dL), equivalent to approximately 0.4 μg/dL in whole blood. In adjusted models, associations with parent and teacher-rated scales were largely null, although effect estimates were consistently positive, suggesting worse scores with increasing lead levels. For an IQR increase in lead, BRIEF Global Executive Composite (GEC) was 0.73 (95% CI: -0.06, 1.52) points higher for parent-rated scores and 0.42 (95% CI: -0.39, 1.23) points higher for teacher-rated scores. Associations were strongest for parent-rated BRIEF plan/organize (β = 0.85; 95% CI: 0.12, 1.59) and shift (β = 0.88; 95% CI: 0.01, 1.75) subscales, as well as the SDQ emotional problems subscale (β = 0.18; 95% CI: 0.03, 0.33). In this cohort with lead levels commonly experienced by U.S. women, there were few statistically significant associations with childhood executive function and behavior. However, there was a trend of worse neurobehavioral scores with increasing prenatal lead concentrations, in particular for childhood emotional problems and capacity to plan/organize and shift. Our results highlight the importance of continuing efforts to eliminate lead exposure in the general population.


Effect of controlled and uncontrolled cooling rate on motility parameters of cryopreserved ram spermatozoa.
Ram spermatozoa are sensitive to extreme changes in temperature during the freeze-thaw process. The degree of damage depends on a combined effect of various factors including freezing temperature. The aim of this study was to determine the effects of two cooling method (controlled-rate and uncontrolled-rate) on pre-freezing and post-thaw sperm motility parameters. Ejaculates were collected using the artificial vagina from four Chal rams and three replicates of the ejaculates were diluted with a Tris-based extender and packed in 0.25 ml straws. Then, sample processed according to the two methods. Method 1: straws cooled from 37 to 5°C, at a liner rate of -0.3°C/min in a controlled-rate cooling machine (custom-built) and equilibrated at 5°C for 80 min, then the straws were frozen at rate of -0.3°C/min from 5°C to -10°C and -25°C/min from -10°C to -150°C and plunged into liquid nitrogen for storage. Method 2: straws were transferred to refrigerator and maintained at 5°C for 3 h, then the straws were frozen in liquid nitrogen vapor, 4 cm above the liquid nitrogen for 15 min and plunged into liquid nitrogen. Computer-assisted sperm motility analysis was used to analyze sperm motion characteristics. Controlled rate of freezing (Method 1) significantly improve the pre-freezing and post-thaw total and progressive motility compared to uncontrolled rate (Method 2). In specific kinetic parameters, Method 1 gives significantly higher value for VSL and VCL in comparison with Method 2. There are no significant differences between the two methods for VAP and LIN. In conclusion, controlled rate of cooling conferred better cryopreserving ability to ram spermatozoa compared to uncontrolled rate of cooling prior to programmable freezing.


The changing role of gesture in linguistic development: a developmental trajectory and a cross-cultural comparison between British and Finnish children.
We studied how gesture use changes with culture, age and increased spoken language competence. A picture-naming task was presented to British (N = 80) and Finnish (N = 41) typically developing children aged 2-5 years. British children were found to gesture more than Finnish children and, in both cultures, gesture production decreased after the age of two. Two-year-olds used more deictic than iconic gestures than older children, and gestured more before the onset of speech, rather than simultaneously or after speech. The British 3- and 5-year-olds gestured significantly more when naming praxic (manipulable) items than non-praxic items. Our results support the view that gesture serves a communicative and intrapersonal function, and the relative function may change with age. Speech and language therapists and psychologists observe the development of children's gestures and make predictions on the basis of their frequency and type. To prevent drawing erroneous conclusions about children's linguistic development, it is important to understand developmental and cultural variations in gesture use.


Isolation and chemical analyses of nonfermented fiber fractions of oat hulls and cottonseed hulls.
The purpose of this study was to isolate, using both in situ and in vivo methodology, the nonfermented fiber fraction of oat hulls (OH) and cottonseed hulls (CSH) and to compare the concentrations of alkali-labile phenolic monomers, nitrobenzene oxidizable phenolic monomers, and neutral monosaccharides, as well as the cross polarization/magic angle spinning (CP/MAS) carbon-13 (13C) nuclear magnetic resonance (NMR) spectra, of the nonfermented fraction with the original OH or CSH. The in situ isolation procedure involved a 30-h ruminal pretreatment and an 8-h acid:pepsin pretreatment followed by 1 to 7 additional days of incubation in the rumen. Fractions not fermented in vivo were isolated from duodena, ileal, and fecal material obtained from a site and extent of digestion trial in which these byproducts were fed to sheep at 80% of the diet (as-fed basis) and they represented the sole source of dietary fiber. Based on nonfermented fraction composition, both in situ and in vivo, all components analyzed were degraded to some extent. Also, all components present in original byproduct material were present in both the in situ and in vivo nonfermented fractions. Based on NMR analysis, cellulose crystallinity did not change during either long-term in situ or in vivo fermentation. However, CSH cellulose was more crystalline than that of OH. The ADL content of OH and CSH was 6.1% and 19.4%, respectively, and very little (15%) of the ADL disappeared during either in situ or in vivo fermentation. Much of the p-coumaric and ferulic acid of OH, associated with the cell wall matrix as lignin-carbohydrate and phenolic-carbohydrate complexes, was recovered in the fermented fractions. Data are interpreted to indicate that lignin encrustation and cellulose crystallinity are factors affecting CSH fermentation. Lignin encrustation and the presence of lignin-carbohydrate/phenolic-carbohydrate complexes are factors that inhibit OH fermentation.


Sulfur flows and biosolids processing: Using Material Flux Analysis (MFA) principles at wastewater treatment plants.
High flows of sulfur through wastewater treatment plants (WWTPs) may cause noxious gaseous emissions, corrosion of infrastructure, inhibit wastewater microbial communities, or contribute to acid rain if the biosolids or biogas is combusted. Yet, sulfur is an important agricultural nutrient and the direct application of biosolids to soils enables its beneficial re-use. Flows of sulfur throughout the biosolids processing of six WWTPs were investigated to identify how they were affected by biosolids processing configurations. The process of tracking sulfur flows through the sites also identified limitations in data availability and quality, highlighting future requirements for tracking substance flows. One site was investigated in more detail showing sulfur speciation throughout the plant and tracking sulfur flows in odour control systems in order to quantify outflows to air, land and ocean sinks. While the majority of sulfur from WWTPs is removed as sulfate in the secondary effluent, the sulfur content of biosolids is valuable as it can be directly returned to soils to combat the potential sulfur deficiencies. Biosolids processing configurations, which focus on maximising solids recovery, through high efficiency separation techniques in primary sedimentation tanks, thickeners and dewatering centrifuges retain more sulfur in the biosolids. However, variations in sulfur loads and concentrations entering the WWTPs affect sulfur recovery in the biosolids, suggesting industrial emitters, and chemical dosing of iron salts are responsible for differences in recovery between sites.