Efficacy of Electrical Baroreflex Activation Is Independent of Peripheral Chemoreceptor Modulation.
Arterial baroreflex activation through electrical carotid sinus stimulation has been developed for the treatment of resistant hypertension. Previous studies suggested that the peripheral chemoreflex is tonically active in hypertensive patients and may inhibit baroreflex responses. We hypothesized that peripheral chemoreflex activation attenuates baroreflex efficacy evoked by electrical carotid sinus stimulation. We screened 35 patients with an implanted electrical carotid sinus stimulator. Of those, 11 patients with consistent acute depressor response were selected (7 men/4 women, age: 67±8 years, body mass index: 31.6±5.2 kg/m2, 6±2 antihypertensive drug classes). We assessed responses to electrical baroreflex stimulation during normoxia, isocapnic hypoxia (SpO2: 79.0±1.5%), and hyperoxia (40% end-tidal O2 fraction) by measuring heart rate, blood pressure, ventilation, oxygen saturation, end-tidal CO2 and O2 fractions, and muscle sympathetic nerve activity. During normoxia, baroreflex activation reduced systolic blood pressure from 164±27 to 151±25 mm Hg (mean±SD, P<0.001), heart rate from 64±13 to 61±13 bpm (P=0.002), and muscle sympathetic nerve activity from 42±12 to 36±12 bursts/min (P=0.004). Hypoxia increased systolic blood pressure 8±12 mm Hg (P=0.057), heart rate 10±6 bpm (P<0.001), muscle sympathetic nerve activity 7±7 bursts/min (P=0.031), and ventilation 10±7 L/min (P=0.002). However, responses to electrical carotid sinus stimulation did not differ between hypoxic and hyperoxic conditions: systolic blood pressure: -15±7 versus -14±8 mm Hg (P=0.938), heart rate: -2±3 versus -2±2 bpm (P=0.701), and muscle sympathetic nerve activity: -6±4 versus -4±3 bursts/min (P=0.531). We conclude that moderate peripheral chemoreflex activation does not attenuate acute responses to electrical baroreflex activation therapy in patients with resistant hypertension. These patients provided insight into human baroreflex-chemoreflex interactions that could not be gained otherwise.


Incomplete stent apposition after implantation of paclitaxel-eluting stents or bare metal stents: insights from the randomized TAXUS II trial.
The clinical impact of late incomplete stent apposition (ISA) for drug-eluting stents is unknown. We sought to prospectively investigate the incidence and extent of ISA after the procedure and at 6-month follow-up of paclitaxel-eluting stents in comparison with bare metal stents (BMS) and survey the clinical significance of ISA over a period of 12 months. TAXUS II was a randomized, double-blind study with 536 patients in 2 consecutive cohorts comparing slow-release (SR; 131 patients) and moderate-release (MR; 135 patients) paclitaxel-eluting stents with BMS (270 patients). This intravascular ultrasound (IVUS) substudy included patients who underwent serial IVUS examination after the procedure and at 6 months (BMS, 240 patients; SR, 113; MR, 116). The qualitative and quantitative analyses of ISA were performed by an independent, blinded core laboratory. More than half of the instances of ISA observed after the procedure resolved at 6 months in all groups. No difference in the incidence of late-acquired ISA was observed among the 3 groups (BMS, 5.4%; SR, 8.0%; MR, 9.5%; P=0.306), with a similar ISA volume (BMS, 11.4 mm3; SR, 21.7 mm3; MR, 8.5 mm3; P=0.18). Late-acquired ISA was the result of an increase of vessel area without change in plaque behind the stent. Predictive factors of late-acquired ISA were lesion length, unstable angina, and absence of diabetes. No stent thrombosis occurred in the patients diagnosed with ISA over a period of 12 months. The incidence and extent of late-acquired ISA are comparable in paclitaxel-eluting stents and BMS. ISA is a pure IVUS finding without clinical repercussions.


Cognitions in Sleep: Lucid Dreaming as an Intervention for Nightmares in Patients With Posttraumatic Stress Disorder.
About 80% of posttraumatic stress disorder (PTSD) patients suffer from nightmares or dysphoric dreams that cause major distress and impact nighttime or daytime functioning. Lucid dreaming (LD) is a learnable and effective strategy to cope with nightmares and has positive effects on other sleep variables. In LDs, the dreamer is aware of the dreaming state and able to control the dream content. The aim of this study is to evaluate the effectiveness of lucid dreaming therapy (LDT) in patients suffering from PTSD. We suggest that learning a technique that enables the affected subjects to regulate the occurrence and content of nightmares autonomously increases the chance of coping with the complex symptoms of PTSD and can reduce suffering. Sleep quality (PSQI, Pittsburgh Sleep Quality Index), daytime sleepiness (ESS, Epworth Sleepiness Scale), quality of life (MQLI, Multicultural Quality of Life Index), psychological distress (SCL-90-R, Symptom Checklist 90-Revised), distress caused by traumatic events (IE-S, Impact of Events Scale), anxiety (SAS, Self-Rating Anxiety Scale), depression (SDS, Self-Rating Depression Scale), and nightmare severity were assessed in a self-rating questionnaire before and after the intervention. LDT had no effect on the investigated sleep variables. No correlation between reduction of nightmare severity and changes in PTSD-profile (IE-S) was found. Nevertheless, levels of anxiety and depression decreased significantly in the course of therapy. LDT could provide an alternate or complementary treatment option for nightmares in PTSD, specifically for symptoms of anxiety and depression.


Cost Effectiveness of Fish Skin Grafts Versus Standard of Care on Wound Healing of Chronic Diabetic Foot Ulcers: A Retrospective Comparative Cohort Study.
Health care policy decision makers seek the highest quality products at the lowest cost for their patients. Cost-benefit analysis is a helpful tool and can be used together with other sources of information to ensure the most efficient use of medical resources. The objective of this retrospective comparative cohort study is to evaluate the cost effectiveness of fish skin therapy compared with standard of care (SOC) on chronic diabetic foot ulcers (DFUs). Retrospective patient data collected in a single wound care setting from 2014 to 2017 were included. In total, 59 DFUs treated with fish skin were used to calculate transition probabilities for a Markov model in which a hypothetical patient cohort treated with fish skin was compared with an identical hypothetical patient cohort treated with SOC. Cost was from the perspective of the payer, and the time horizon was set at 1 year. The model indicated that fish skin treatment could result in lower costs ($11 210 vs. $15 075 per wound), more wounds healing (83.2% vs. 63.4%), fewer amputations (4.6% vs. 6.9%), and a higher quality of life (0.676 vs. 0.605 quality-adjusted life year [QALY]) than the SOC. A probabilistic sensitivity analysis, based on a Monte Carlo simulation, indicated that the fish skin treatment (on DFUs) would be 93.6% likely to be cost effective for a willingness to pay at $100 000 per QALY and 71.4% likely to be cheaper than SOC. Including fish skin grafts in the SOC for DFU treatment has the potential to reduce costs while improving patient outcomes.


The Tn3 beta-lactamase gene acts as a hotspot for meiotic recombination in yeast.
Although genetic distances are often assumed to be proportional to physical distances, chromosomal regions with unusually high (hotspots) or low (coldspots) levels of meiotic recombination have been described in a number of genetic systems. In general, the DNA sequences responsible for these effects have not been determined. We report that the 5' region of the beta-lactamase (ampR) gene of the bacterial transposon Tn3 is a hotspot for meiotic recombination when inserted into the chromosomes of the yeast Saccharomyces cerevisiae. When these sequences are homozygous, both crossing over and gene conversion are locally stimulated. The 5' end of the beta-lactamase gene is about 100-fold "hotter" for crossovers than an average yeast DNA sequence.


Application of 3D digital image correlation to track displacements and strains of canvas paintings exposed to relative humidity changes.
This paper introduces a methodology for tracking displacements in canvas paintings exposed to relative humidity changes. Displacements are measured by means of the 3D digital image correlation method that is followed by a postprocessing of displacement data, which allows the separation of local displacements from global displacement maps. The applicability of this methodology is tested on measurements of a model painting on canvas with introduced defects causing local inhomogeneity. The method allows the evaluation of conservation methods used for repairing canvas supports.


The effectiveness of peer health coaching in improving glycemic control among low-income patients with diabetes: protocol for a randomized controlled trial.
Although self-management support improves diabetes outcomes, it is not consistently provided in health care settings strained for time and resources. One proposed solution to personnel and funding shortages is to utilize peer coaches, patients trained to provide diabetes education and support to other patients. Coaches share similar experiences about living with diabetes and are able to reach patients within and beyond the health care setting. Given the limited body of evidence that demonstrates peer coaching significantly improves chronic disease care, this present study examines the impact of peer coaching delivered in a primary care setting on diabetes outcomes. The aim of this multicenter, randomized control trial is to evaluate the effectiveness of utilizing peer coaches to improve clinical outcomes and self-management skills in low-income patients with poorly controlled diabetes. A total of 400 patients from six primary health centers based in San Francisco that serve primarily low-income populations will be randomized to receive peer coaching (n = 200) or usual care (n = 200) over 6 months. Patients in the peer coach group receive coaching from patients with diabetes who are trained and mentored as peer coaches. The primary outcome is change in HbA1c. Secondary outcomes include change in: systolic blood pressure, body mass index (BMI), LDL cholesterol, diabetes self-care activities, medication adherence, diabetes-related quality of life, diabetes self-efficacy, and depression. Clinical values (HbA1c, LDL cholesterol and blood pressure) and self-reported diabetes self-efficacy and self-care activities are measured at baseline and after 6 months for patients and coaches. Peer coaches are also assessed at 12 months. Patients with diabetes, who are trained as peer health coaches, are uniquely poised to provide diabetes self management support and education to patients. This study is designed to investigate the impact of peer health coaching in patients with poorly controlled diabetes. Additionally, we will assess disease outcomes in patients with well controlled diabetes who are trained and work as peer health coaches. ClinicalTrials.gov identifier: NCT01040806.


Novel App-Based Portable Spirometer for the Early Detection of COPD.
Chronic obstructive pulmonary disease (COPD) is preventable and treatable. However, many patients remain undiagnosed and untreated due to the underutilization or unavailability of spirometers. Accordingly, we used Spirobank Smart, an app-based spirometer, for facilitating the early detection of COPD in outpatient clinics. This prospective study recruited individuals who were at risk of COPD (i.e., with age of ≥40 years, ≥10 pack-years of smoking, and at least one respiratory symptoms) but had no previous COPD diagnosis. Eligible participants were examined with Spirobank Smart and then underwent confirmatory spirometry (performed using a diagnostic spirometer), regardless of their Spirobank Smart test results. COPD was defined and confirmed using the postbronchodilator forced expiratory volume in 1 s/forced vital capacity values of <0.70 as measured by confirmatory spirometry. A total of 767 participants were enrolled and examined using Spirobank Smart; 370 participants (94.3% men, mean age of 60.9 years and mean 42.6 pack-years of smoking) underwent confirmatory spirometry. Confirmatory spirometry identified COPD in 103 participants (27.8%). At the optimal cutoff point of 0.74 that was determined using Spirobank Smart for COPD diagnosis, the area under the receiver operating characteristic was 0.903 (95% confidence interval (CI) = 0.860-0.947). Multivariate logistic regression revealed that participants who have an FEV1/FVC ratio of <74% that was determined using Spirobank Smart (odds ratio (OR) = 58.58, 95% CI = 27.29-125.75) and old age (OR = 3.23, 95% CI = 1.04-10.07 for 60 ≤ age < 65; OR = 5.82, 95% CI = 2.22-15.27 for age ≥ 65) had a higher risk of COPD. The Spirobank Smart is a simple and adequate tool for early COPD detection in outpatient clinics. Early diagnosis and appropriate therapy based on GOLD guidelines can positively influence respiratory symptoms and quality of life.


Calculations of flow resistance in the juxtacanalicular meshwork.
The structure of the juxtacanalicular meshwork (JCM) was analyzed morphometrically, and the resulting data were used to calculate the resistance to flow through this tissue. Two models of the JCM were presented and compared. In the first (Model A), aqueous humor was assumed to flow via open channels within a solid framework, while, in the second (Model B), these open spaces were assumed to be filled with extracellular matrix gel. An expression giving the resistance of such a gel as a function of gel concentration was presented and tested on corneal and scleral stroma. Morphometry of normal and glaucomatous human eyes showed that Model A underpredicted the resistance of the JCM by factors of 10-100, suggesting that a GAG or proteoglycan gel may control the flow resistance of this tissue. This was supported by Model B, which showed that measured bulk concentrations of GAGs were consistent with gel concentrations needed to account for the estimated resistance of the JCM in vivo. Some limitations and implications of Model B were discussed.


Cortical thickness and surface area in neonates at high risk for schizophrenia.
Schizophrenia is a neurodevelopmental disorder associated with subtle abnormal cortical thickness and cortical surface area. However, it is unclear whether these abnormalities exist in neonates associated with genetic risk for schizophrenia. To this end, this preliminary study was conducted to identify possible abnormalities of cortical thickness and surface area in the high-genetic-risk neonates. Structural magnetic resonance images were acquired from offspring of mothers (N = 21) who had schizophrenia (N = 12) or schizoaffective disorder (N = 9), and also matched healthy neonates of mothers who were free of psychiatric illness (N = 26). Neonatal cortical surfaces were reconstructed and parcellated as regions of interest (ROIs), and cortical thickness for each vertex was computed as the shortest distance between the inner and outer surfaces. Comparisons were made for the average cortical thickness and total surface area in each of 68 cortical ROIs. After false discovery rate (FDR) correction, it was found that the female high-genetic-risk neonates had significantly thinner cortical thickness in the right lateral occipital cortex than the female control neonates. Before FDR correction, the high-genetic-risk neonates had significantly thinner cortex in the left transverse temporal gyrus, left banks of superior temporal sulcus, left lingual gyrus, right paracentral cortex, right posterior cingulate cortex, right temporal pole, and right lateral occipital cortex, compared with the control neonates. Before FDR correction, in comparison with control neonates, male high-risk neonates had significantly thicker cortex in the left frontal pole, left cuneus cortex, and left lateral occipital cortex; while female high-risk neonates had significantly thinner cortex in the bilateral paracentral, bilateral lateral occipital, left transverse temporal, left pars opercularis, right cuneus, and right posterior cingulate cortices. The high-risk neonates also had significantly smaller cortical surface area in the right pars triangularis (before FDR correction), compared with control neonates. This preliminary study provides the first evidence that early development of cortical thickness and surface area might be abnormal in the neonates at genetic risk for schizophrenia.


Pre-induced Lac Operon Effect on Non Specific Sugars: Pre-culture Effect is Dependent on Strength of Induction, Exponential Phase and Substrate Concentration.
The source and history of the cell plays an important role in influencing the phenotypic properties of the organism in a particular environmental condition. Pre-induced lac operon provides benefit on lactose environment. During metabolism lactose is broken down into glucose and galactose. The fate of cells with pre-induced lac operon on glucose and galactose milieu is not known. The influence of nutritional status of the medium, level of pre-induction and growth phase on pre-culture effect is not investigated. Effect of pre-induced lac operon on non specific sugars along with the factors that influence this effect was enumerated in the present study. Results of this present study indicate that pre-induced lac operon provide benefit in terms of growth on galactose milieu. This study also suggests that Pre induced lac operon effect depends on the (i) strength of induction in the pre-culture, (ii) nutritional content of the environment and (iii) exponential growth phase of the organism. The above study will help in the better characterization of the pre culture effect. It will also help in the better understanding of the relation between gene expression and growth physiology.


Identification of a novel protein, PDIP38, that interacts with the p50 subunit of DNA polymerase delta and proliferating cell nuclear antigen.
The yeast two-hybrid screening method was used to identify novel proteins that associate with human DNA polymerase delta (pol delta). Two baits were used in this study. These were the large (p125) and small (p50) subunits of the core pol delta heterodimer. p50 was the only positive isolated with p125 as the bait. Two novel protein partners, named PDIP38 and PDIP46, were identified from the p50 screen. In this study, the interaction of PDIP38 with pol delta was further characterized. PDIP38 encodes a protein of 368 amino acids whose C terminus is conserved with the bacterial APAG protein and with the F box A protein. It was found that PDIP38 also interacts with proliferating cell nuclear antigen (PCNA). The ability of PDIP38 to interact with both the p50 subunit of pol delta and with PCNA was confirmed by pull-down assays using glutathione S-transferase (GST)-PDIP38 fusion proteins. The PCNA-PDIP38 interaction was also demonstrated by PCNA overlay experiments. The association of PDIP38 with pol delta was shown to occur in calf thymus tissue and mammalian cell extracts by GST-PDIP38 pull-down and coimmunoprecipitation experiments. PDIP38 was associated with pol delta isolated by immunoaffinity chromatography. The association of PDIP38 with pol delta could also be demonstrated by native gel electrophoresis.


Auto-oxidation products of epigallocatechin gallate activate TRPA1 and TRPV1 in sensory neurons.
The sensation of astringency is elicited by catechins and their polymers in wine and tea. It has been considered that catechins in green tea are unstable and auto-oxidized to induce more astringent taste. Here, we examined how mammalian transient receptor potential V1 (TRPV1) and TRPA1, which are nociceptive sensors, are activated by green tea catechins during the auto-oxidation process. Neither TRPV1 nor TRPA1 could be activated by any of the freshly prepared catechin. When one of the major catechin, epigallocatechin gallate (EGCG), was preincubated for 3h in Hank's balanced salt solution, it significantly activated both TRP channels expressed in HEK293 cells. Even after incubation, other catechins showed much less effects. Results suggest that only oxidative products of EGCG activate both TRPV1 and TRPA1. Dorsal root ganglion (DRG) sensory neurons were also activated by the incubated EGCG through TRPV1 and TRPA1 channels. Liquid chromatography-mass spectrometry revealed that theasinensins A and D are formed during incubation of EGCG. We found that purified theasinensin A activates both TRPV1 and TRPA1, and that it stimulates DRG neurons through TRPV1 and TRPA1 channels. Results suggested a possibility that TRPV1 and TRPA1 channels are involved in the sense of astringent taste of green tea.


B vitamins, homocysteine, and neurocognitive function in the elderly.
Evidence of the importance of the B vitamins folic acid, vitamin B-12, and vitamin B-6 for the well-being and normal function of the brain derives from data showing neurologic and psychologic dysfunction in vitamin deficiency states and in cases of congenital defects of one-carbon metabolism. The status of these vitamins is frequently inadequate in the elderly and recent studies have shown associations between loss of cognitive function or Alzheimer disease and inadequate B vitamin status. The question that arises is whether these B vitamin inadequacies contribute to such brain malfunctions or result from aging and disease. From a theoretical standpoint, these inadequacies could give rise to impairment of methylation reactions that are crucial to the health of brain tissue. In addition or perhaps instead, these inadequacies could result in hyperhomocysteinemia, a recently identified risk factor for occlusive vascular disease, stroke, and thrombosis, any of which may result in brain ischemia. Advances in the understanding of this putative relation between inadequate vitamin status and loss of cognitive function in the elderly are likely to be slow and may depend on the outcomes of both prospective studies and longitudinal studies in which nutritional intervention is provided before cognitive decline occurs.


Disrupted coupling of gating charge displacement to Na+ current activation for DIIS4 mutations in hypokalemic periodic paralysis.
Missense mutations at arginine residues in the S4 voltage-sensor domains of NaV1.4 are an established cause of hypokalemic periodic paralysis, an inherited disorder of skeletal muscle involving recurrent episodes of weakness in conjunction with low serum K(+). Expression studies in oocytes have revealed anomalous, hyperpolarization-activated gating pore currents in mutant channels. This aberrant gating pore conductance creates a small inward current at the resting potential that is thought to contribute to susceptibility to depolarization in low K(+) during attacks of weakness. A critical component of this hypothesis is the magnitude of the gating pore conductance relative to other conductances that are active at the resting potential in mammalian muscle: large enough to favor episodes of paradoxical depolarization in low K(+), yet not so large as to permanently depolarize the fiber. To improve the estimate of the specific conductance for the gating pore in affected muscle, we sequentially measured Na(+) current through the channel pore, gating pore current, and gating charge displacement in oocytes expressing R669H, R672G, or wild-type NaV1.4 channels. The relative conductance of the gating pore to that of the pore domain pathway for Na(+) was 0.03%, which implies a specific conductance in muscle from heterozygous patients of ∼ 10 µS/cm(2) or 1% of the total resting conductance. Unexpectedly, our data also revealed a substantial decoupling between gating charge displacement and peak Na(+) current for both R669H and R672G mutant channels. This decoupling predicts a reduced Na(+) current density in affected muscle, consistent with the observations that the maximal dV/dt and peak amplitude of the action potential are reduced in fibers from patients with R672G and in a knock-in mouse model of R669H. The defective coupling between gating charge displacement and channel activation identifies a previously unappreciated mechanism that contributes to the reduced excitability of affected fibers seen with these mutations and possibly with other R/X mutations of S4 of NaV, CaV, and KV channels associated with human disease.


Mucosal vaccines: where do we stand?
Mucosal vaccinology is a relatively young but rapidly expanding discipline. At present the vast majority of vaccines are administered by injection, including vaccines that protect against mucosally acquired pathogens such as influenza virus and human papilloma virus. However, mucosal immune responses are most efficiently induced by the administration of vaccines onto mucosal surfaces. The small number of currently licensed mucosal vaccines have reduced the burden of disease and mortality caused by enteric pathogens including rotavirus, V. cholerae and S. typhi, or those that spread to affect distal organs such as poliovirus. Expanding knowledge about the special features of the mucosal immune system promises to accelerate development of mucosal vaccines that could contribute significantly to protection against pathogens that colonize or invade via mucosal surfaces including HIV, Shigella, ETEC, Campylobacter jejuni, Helicobacter pylori and many others.


Optogenetic Downregulation of Protein Levels to Control Programmed Cell Death in Mammalian Cells with a Dual Blue-Light Switch.
Optogenetic approaches facilitate the study of signaling and metabolic pathways in animal cell systems. In the past 10 years, a plethora of light-regulated switches for the targeted control over the induction of gene expression, subcellular localization of proteins, membrane receptor activity, and other cellular processes have been developed and successfully implemented. However, only a few tools have been engineered toward the quantitative and spatiotemporally resolved downregulation of proteins. Here we present a protocol for reversible and rapid blue light-induced reduction of protein levels in mammalian cells. By implementing a dual-regulated optogenetic switch (Blue-OFF), both repression of gene expression and degradation of the target protein are triggered simultaneously. We apply this system for the blue light-mediated control of programmed cell death. HEK293T cells are transfected with the proapoptotic proteins PUMA and BID integrated into the Blue-OFF system. Overexpression of these proteins leads to programmed cell death, which can be prevented by irradiation with blue light. This experimental approach is very straightforward, requires just simple hardware, and therefore can be easily implemented in state-of-the-art equipped mammalian cell culture labs. The system can be used for targeted cell signaling studies and biotechnological applications.


Variance heterogeneity analysis for detection of potentially interacting genetic loci: method and its limitations.
Presence of interaction between a genotype and certain factor in determination of a trait's value, it is expected that the trait's variance is increased in the group of subjects having this genotype. Thus, test of heterogeneity of variances can be used as a test to screen for potentially interacting single-nucleotide polymorphisms (SNPs). In this work, we evaluated statistical properties of variance heterogeneity analysis in respect to the detection of potentially interacting SNPs in a case when an interaction variable is unknown. Through simulations, we investigated type I error for Bartlett's test, Bartlett's test with prior rank transformation of a trait to normality, and Levene's test for different genetic models. Additionally, we derived an analytical expression for power estimation. We showed that Bartlett's test has acceptable type I error in the case of trait following a normal distribution, whereas Levene's test kept nominal Type I error under all scenarios investigated. For the power of variance homogeneity test, we showed (as opposed to the power of direct test which uses information about known interacting factor) that, given the same interaction effect, the power can vary widely depending on the non-estimable direct effect of the unobserved interacting variable. Thus, for a given interaction effect, only very wide limits of power of the variance homogeneity test can be estimated. Also we applied Levene's approach to test genome-wide homogeneity of variances of the C-reactive protein in the Rotterdam Study population (n = 5959). In this analysis, we replicate previous results of Pare and colleagues (2010) for the SNP rs12753193 (n = 21,799). Screening for differences in variances among genotypes of a SNP is a promising approach as a number of biologically interesting models may lead to the heterogeneity of variances. However, it should be kept in mind that the absence of variance heterogeneity for a SNP can not be interpreted as the absence of involvement of the SNP in the interaction network.


Plasma melatonin rhythms in young and older humans during sleep, sleep deprivation, and wake.
To determine the effects of sleep and sleep deprivation on plasma melatonin concentrations in humans and whether these effects are age-dependent. At least 2 weeks of regular at-home, sleep/wake schedule followed by 3 baseline days in the laboratory and at least one constant routine (sleep deprivation). General Clinical Research Center (GCRC), Brigham and Women's Hospital, Boston, MA. In Study 1, one group (<10 lux when awake) of 19 young men (18-30 y) plus a second group (<2 lux when awake) of 15 young men (20-28 y) and 10 young women (19-27 y); in Study 2, 90 young men (18-30 y), 18 older women (65-81 y), and 11 older men (64-75 y). All participants were in good health, as determined by medical and psychological screening. One to three constant routines with interspersed inversion of the sleep/wake cycle in those with multiple constant routines. Examination of plasma melatonin concentrations and core body temperature. Study 1. There was a small, but significant effect of sleep deprivation of up to 50 hours on melatonin concentrations (increase of 9.81 +/- 3.73%, P <0.05, compared to normally timed melatonin). There was also an effect of circadian phase angle with the prior sleep episode, such that if melatonin onset occurred <8 hours after wake time, the amplitude was significantly lower (22.4% +/- 4.79%, P <0.001). Study 2. In comparing melatonin concentrations during sleep to the same hours during constant wakefulness, in young men, melatonin amplitude was 6.7% +/- 2.1% higher(P <0.001) during the sleep episode. In older men, melatonin amplitude was 37.0% +/- 12.5% lower (P <0.05) during the sleep episode and in older women, melatonin amplitude was non-significantly 10.9% +/- 8.38% lower (P = 0.13) during the sleep episode. Both sleep and sleep deprivation likely influence melatonin amplitude, and the effect of sleep on melatonin appears to be age dependent.


Circulating angiogenic cells can be derived from cryopreserved peripheral blood mononuclear cells.
Cell transplantation for regenerative medicine has become an appealing therapeutic method; however, stem and progenitor cells are not always freshly available. Cryopreservation offers a way to freeze cells as they are generated, for storage and transport until required for therapy. This study was performed to assess the feasibility of cryopreserving peripheral blood mononuclear cells (PBMCs) for the subsequent in vitro generation of their derived therapeutic population, circulating angiogenic cells (CACs). PBMCs were isolated from healthy human donors. Freshly isolated cells were either analyzed immediately or cryopreserved in media containing 6% plasma serum and 5% dimethyl sulfoxide. PBMCs were thawed after being frozen for 1 (early thaw) or 28 (late thaw) days and analyzed, or cultured for 4 days to generate CACs. Analysis of the cells consisted of flow cytometry for viability and phenotype, as well as functional assays for their adhesion and migration potential, cytokine secretion, and in vivo angiogenic potential. The viability of PBMCs and CACs as well as their adhesion and migration properties did not differ greatly after cryopreservation. Phenotypic changes did occur in PBMCs and to a lesser extent in CACs after freezing; however the potent CD34(+)VEGFR2(+)CD133(+) population remained unaffected. The derived CACs, while exhibiting changes in inflammatory cytokine secretion, showed no changes in the secretion of important regenerative and chemotactic cytokines, nor in their ability to restore perfusion in ischemic muscle. Overall, it appears that changes do occur in cryopreserved PBMCs and their generated CACs; however, the CD34(+)VEGFR2(+)CD133(+) progenitor population, the secretion of pro-vasculogenic factors, and the in vivo angiogenic potential of CACs remain unaffected by cryopreservation.


Nutrient intakes during diets including unkilned and large amounts of oats in celiac disease.
We have shown earlier that consumption of moderate amount of oats improve intakes of vitamin B(1), fiber, magnesium and iron in celiac patients using gluten-free diet (GFD). The objective of this study was to clarify the effect of high amount of both kilned and unkilned oats on food and nutrient intakes in celiac patients in remission. Kilning as an industrial heating process is performed to preserve the main properties of oats and to lengthen its useableness. Kilning may, however, change the protein structure of oats and therefore influence on the intake of nutrients. The study group consisted of 13 men and 18 women with celiac disease in remission. The patients who were earlier using moderate amount of oats as part of their GFD were randomized to consume kilned or unkilned oats. After 6 months, the patients changed the treatment groups. The goal of daily intake of oats was 100 g. Food records and frequency questionnaire were used to follow nutrient intakes. Type of oats did not affect the amount of oats used. In the group using kilned oats, the intake of vitamin B1 and magnesium and in the group of unkilned oats that of magnesium and zinc increased significantly during the first 6 months (P<or=0.05). :Large amounts of oats, both kilned and unkilned in GFD, can increase intakes of nutrients in celiac patients in remission. Oats improve the nutritional value of GFD.


Plasmatic extracellular vesicle microRNAs in malignant pleural mesothelioma and asbestos-exposed subjects suggest a 2-miRNA signature as potential biomarker of disease.
Malignant Pleural Mesothelioma (MPM) is an aggressive cancer mainly caused by asbestos exposure and refractory to current therapies. Specific diagnostic markers for early MPM diagnosis are needed. Changes in miRNA expression have been implicated in several diseases and cancers, including MPM. We examined if a specific miRNA signature in plasmatic extracellular vesicles (EV) may help to discriminate between malignant pleural mesothelioma patients (MPM) and subjects with Past Asbestos Exposure (PAE). We investigated 23 MPM patients and 19 cancer-free subjects with past asbestos exposure (PAE). We screened 754 miRNAs in plasmatic EVs by OpenArray and found 55 differential miRNAs using logistic regression models adjusted for age, sex, BMI, and smoking. Among the top-20 differential miRNAs chosen for validation by Real time PCR, 16 were confirmed. Using receiver operating characteristic (ROC) curve analysis, the most discriminating miRNA combination was given by miR-103a-3p + miR-30e-3p, which generated an AUC of 0.942 (95% CI 0.87-1.00), with a sensitivity of 95.5% and a specificity of 80.0%. Using multivariate Cox regression analysis including gender, age, BMI and smoking we found a Hazard Ratio for miR-103a-3p above the median of 0.37 (95%CI 0.13-1.13) and of 0.51 (95%CI 0.17-1.52) for miR-30e-3p. This study suggests EV-associated miR-103a-3p and miR-30e-3p are able to discriminate MPM from PAE subjects. Larger and prospective studies are needed to confirm these two-miRNA signature alone or in combination with other biomarkers as diagnostic tools for MPM.


Postoperative outcome after modified unilateral-approach microendoscopic midline decompression for degenerative spinal stenosis.
The object of this study was to assess the feasibility and efficacy of a novel, minimally invasive spinal surgery technique to correct degenerative lumbar spinal stenosis involving a modified unilateral-approach microendoscopic midline decompression. In this prospective study, 41 patients with lumbar stenosis were randomly assigned to undergo either a novel, median-approach microendoscopic laminectomy (20 patients) or a conventional laminectomy (21 patients). Spinal anteroposterior diameter, cross-sectional area, lateral recess distance, spinal stability, postoperative back pain, functional outcomes, and muscle trauma were evaluated. Follow-up ranged from 16 to 24 months, with a mean of 17.8 months for the novel procedure group and 18.6 months for the conventional laminectomy group. Compared with patients in the conventional laminectomy group, patients who received the novel procedure had a reduced mean duration of hospital stay, a lower mean creatine phosphokinase muscular-type isoenzyme level, a lower visual analog scale score for back pain at 1-year follow-up, and a faster recovery rate. These patients also had less mean blood loss compared with the conventionally treated group. Satisfactory neurological decompression and symptom relief were achieved in 90% of these patients. There was no significant clinical difference compared with the conventional laminectomy group's results. There was no evidence of spinal instability in any patient, and no patient required a follow-up conventional laminectomy. This novel procedure provides effective spinal decompression. Although this method requires more operating time than a conventional method, it requires only minimal muscle trauma and spinal stability maintenance, and allows for early mobilization. This shortens the hospital stay, reduces postoperative back pain, and leads to satisfactory neurological and functional outcomes. Moreover, with the midline approach, decompression was accomplished without compromising the facet joints, even with a narrow width of lamina.


When cues collide: use of stress and statistical cues to word boundaries by 7- to 9-month-old infants.
Prior research suggests that stress cues are particularly important for English-hearing infants' detection of word boundaries. It is unclear, though, how infants learn to attend to stress as a cue to word segmentation. This series of experiments was designed to explore infants' attention to conflicting cues at different ages. Experiment 1 replicated previous findings: When stress and statistical cues indicated different word boundaries, 9-month-old infants used syllable stress as a cue to segmentation while ignoring statistical cues. However, in Experiment 2, 7-month-old infants attended more to statistical cues than to stress cues. These results raise the possibility that infants use their statistical learning abilities to locate words in speech and use those words to discover the regular pattern of stress cues in English. Infants at different ages may deploy different segmentation strategies as a function of their current linguistic experience.


Military chaplaincy in Sweden: A contemporary perspective.
Pastors serving as military chaplains have been an integral part of the Swedish Armed Forces for hundreds of years. This close relationship has continued even after the Church of Sweden was formally separated from the state in 2000. Yet contemporary research drawing from experiences of Swedish military chaplains during their pastoral and spiritual assignments within a military context is virtually non-existent. This research explores the contemporary situation for military chaplaincy in Sweden through the lenses of proportional accessibility for military personnel, availability to civilian congregations, increasing cultural and ethnic diversity, religious embeddedness in military culture, theological tension, and personal identity development among chaplains. It furthermore suggests that military chaplains navigate a complex situation: they are expected to serve professional service members, many of whom have war zone experiences, in addition to shepherding increasing volumes of conscripts now that conscription has been reinstated, all within a new interreligious paradigm.


GmcA is a putative glucose-methanol-choline oxidoreductase required for the induction of asexual development in Aspergillus nidulans.
Aspergillus nidulans asexual differentiation is induced by Upstream Developmental Activators (UDAs) that include the bZIP-type Transcription Factor (TF) FlbB. A 2D-PAGE/MS-MS-coupled screen for proteins differentially expressed in the presence and absence of FlbB identified 18 candidates. Most candidates belong to GO term classes involved in osmotic and/or oxidative stress response. Among these, we focused on GmcA, a putative glucose-methanol-choline oxidoreductase which is upregulated in a ΔflbB background. GmcA is not required for growth since no differences were detected in the radial extension upon deletion of gmcA. However, its activity is required to induce conidiation under specific culture conditions. A ΔgmcA strain conidiates profusely under acid conditions but displays a characteristic fluffy aconidial phenotype in alkaline medium. The absence of asexual development in a ΔgmcA strain can be suppressed, on one hand, using high concentrations of non-fermentable carbon sources like glycerol, and on the other hand, when the cMyb-type UDA TF flbD is overexpressed. Overall, the results obtained in this work support a role for GmcA at early stages of conidiophore initiation.


Air pollution, smoking, and plasma homocysteine.
Mild hyperhomocysteinemia is independently associated with an increased risk of cardiovascular disease. Air pollution exposure induces short-term inflammatory changes that may determine hyperhomocysteinemia, particularly in the presence of a preexisting proinflammatory status such as that found in cigarette smokers. We examined the relation of air pollution levels with fasting and postmethionine-load total homocysteine (tHcy) in 1,213 normal subjects from Lombardia, Italy. We obtained hourly concentrations of particulate matter < 10 mum in aerodynamic diameter (PM(10)) and gaseous pollutants (carbon monoxide, nitrogen dioxide, sulfur dioxide(,) ozone) from 53 monitoring sites covering the study area. We applied generalized additive models to compute standardized regression coefficients controlled for age, sex, body mass index, smoking, alcohol, hormone use, temperature, day of the year, and long-term trends. The estimated difference in tHcy associated with an interquartile increase in average PM(10) concentrations in the 24 hr before the study was nonsignificant [0.4%; 95% confidence interval (CI), -2.4 to 3.3 for fasting; and 1.1%, 95% CI, -1.5 to 3.7 for postmethionine-load tHcy]. In smokers, 24-hr PM(10) levels were associated with 6.3% (95% CI, 1.3 to 11.6; p < 0.05) and 4.9% (95% CI, 0.5 to 9.6; p < 0.05) increases in fasting and postmethionine-load tHcy, respectively, but no association was seen in nonsmokers (p-interaction = 0.005 for fasting and 0.039 for postmethionine-load tHcy). Average 24-hr O(3) concentrations were associated with significant differences in fasting tHcy (6.7%; 95% CI, 0.9 to 12.8; p < 0.05), but no consistent associations were found when postmethionine-load tHcy and/or 7-day average O(3) concentrations were considered. Air particles may interact with cigarette smoking and increase plasma homocysteine in healthy subjects.


Explorative Study on Isolation and Characterization of a Microviridae G4 Bacteriophage, EMCL318, against Multi-Drug-resistant Escherichia coli 15-318.
Bacteriophages screened and isolated from sewage water samples exhibited antibacterial activities against multi-drug-resistant Escherichia coli strains. Five different water samples from Canadian habitats such as Kamloops Wastewater Treatment Center, Domtar, the Pacific Ocean, Bisaro Anima Cave, and alkali ponds, were used in this study. Four Enterobacteriaceae strains including one non-resistant and three clinical multi-drug Escherichia coli strains (E. coli 15-102, E. coli 15-124, and E. coli 15-318) were selected as target bacteria to screen for the bacteriophages from these collected water samples. Seeded agar assay technique was implemented for the screening. It was found that only sewage water sample exhibited a significant number of plaques count with the E. coli 15-318 (1.82 × 10² plaques/plate) cells in comparison to E. coli non-resistant strain K12 (8 plaques/plate). The phage did not produce plaques in the E. coli 15-124 and E. coli 15-102 strains. The bacteriophage, designated EMCL318, was isolated, purified, characterized, and identified to belong to the G4 species of the Family Microviridae, GenBank accession number MG563770. This is an explorative study conducted in order to reveal the viruses as alternative potentials to fight against emerging and existing multi-drug-resistant infectious diseases.


Genome-wide quantitative assessment of variation in DNA methylation patterns.
Genomic DNA methylation contributes substantively to transcriptional regulations that underlie mammalian development and cellular differentiation. Much effort has been made to decipher the molecular mechanisms governing the establishment and maintenance of DNA methylation patterns. However, little is known about genome-wide variation of DNA methylation patterns. In this study, we introduced the concept of methylation entropy, a measure of the randomness of DNA methylation patterns in a cell population, and exploited it to assess the variability in DNA methylation patterns of Alu repeats and promoters. A few interesting observations were made: (i) within a cell population, methylation entropy varies among genomic loci; (ii) among cell populations, the methylation entropies of most genomic loci remain constant; (iii) compared to normal tissue controls, some tumors exhibit greater methylation entropies; (iv) Alu elements with high methylation entropy are associated with high GC content but depletion of CpG dinucleotides and (v) Alu elements in the intronic regions or far from CpG islands are associated with low methylation entropy. We further identified 12 putative allelic-specific methylated genomic loci, including four Alu elements and eight promoters. Lastly, using subcloned normal fibroblast cells, we demonstrated the highly variable methylation patterns are resulted from low fidelity of DNA methylation inheritance.


[The factors influencing carboxyhemoglobin kinetics in acid induced lung injury].
Anesthetized dogs were ventilated with 1% carbon monoxide (CO) in air for 10 minutes to produce CO poisoning. They were then ventilated with room air (n = 5) or pure oxygen (n = 5) for 3 hours. They served as controls. Acute lung injury was produced by intratracheal injection of 0.1 N HCl (2 ml/kg) 30 minutes before CO poisoning in another group of dogs. Arterial blood gas and COHb were monitored before and after CO poisoning. Pharmacokinetic analysis was used to find the half-time of COHb elimination (T1/2). Pulmonary shunt was measured before CO poisoning in the injured dogs. Half-time of COHb elimination was prolonged in lung injury dogs resuscitated on air as compared with the control group (275 +/- 28 vs. 203 +/- 24 min, P less than 0.05). Oxygen accelerated COHb excretion in both injured and control animals. No significant correlation was found between T1/2 and Qs/Qt in the injured dogs ventilated with air or oxygen. A negative linear relationship was found between T1/2 of COHb elimination and the pre-CO-poisoning PaO2 (r = -0.98, P less than 0.05) in the injured dogs treated with oxygen. The data suggest that hypoxemia may represent the underlying mechanism of altered COHb kinetics in acute inhalation lung injury.


Treatment of metastatic melanoma B16F10 by the flavonoids tangeretin, rutin, and diosmin.
Melanoma is one of the most frequently metastasizing malignant neoplasias. This study examines an experimental model of pulmonary metastasis and the B16F10 cell subline, highly metastatic in the lung. Antimetastatic effects of the flavonoids tangeretin, rutin, and diosmin were analyzed, and at the same time an analysis of the metastatic activity of ethanol was performed, considered to be necessary because it is used as a vehicle for administering the flavonoids. Lentini's model, which complements the macroscopic evaluation of nodule numbers by using a stereoscopic microscope and image analysis at the microscopic level, was used. The greatest reduction in the number of metastatic nodules (52%) was obtained with diosmin; similarly, the percentages of implantation, growth index, and invasion index (79.40, 67.44, and 45.23%, respectively), were all compared with those of the ethanol group, considered to be an effective control group. Rutin- and tangeretin-treated groups also showed reductions of the same index compared with the ethanol group. It would seem that structural factors would better explain these results and the antimetastatic activity of each flavonoid and the respective metabolites.


Automated 3D Segmentation of Intraretinal Surfaces in SD-OCT Volumes in Normal and Diabetic Mice.
To describe an adaptation of an existing graph-theoretic method (initially developed for human optical coherence tomography [OCT] images) for the three-dimensional (3D) automated segmentation of 10 intraretinal surfaces in mice scans, and assess the accuracy of the method and the reproducibility of thickness measurements. Ten intraretinal surfaces were segmented in repeat spectral domain (SD)-OCT volumetric images acquired from normal (n = 8) and diabetic (n = 10) mice. The accuracy of the method was assessed by computing the border position errors of the automated segmentation with respect to manual tracings obtained from two experts. The reproducibility was statistically assessed for four retinal layers within eight predefined regions using the mean and SD of the differences in retinal thickness measured in the repeat scans, the coefficient of variation (CV) and the intraclass correlation coefficients (ICC; with 95% confidence intervals [CIs]). The overall mean unsigned border position error for the 10 surfaces computed over 97 B-scans (10 scans, 10 normal mice) was 3.16 ± 0.91 μm. The overall mean differences in retinal thicknesses computed from the normal and diabetic mice were 1.86 ± 0.95 and 2.15 ± 0.86 μm, respectively. The CV of the retinal thicknesses for all the measured layers ranged from 1.04% to 5%. The ICCs for the total retinal thickness in the normal and diabetic mice were 0.78 [0.10, 0.92] and 0.83 [0.31, 0.96], respectively. The presented method (publicly available as part of the Iowa Reference Algorithms) has acceptable accuracy and reproducibility and is expected to be useful in the quantitative study of intraretinal layers in mice. The presented method, initially developed for human OCT, has been adapted for mice, with the potential to be adapted for other animals as well. Quantitative in vivo assessment of the retina in mice allows changes to be measured longitudinally, decreasing the need for them.


Flexible solid-state supercapacitors based on three-dimensional graphene hydrogel films.
Flexible solid-state supercapacitors are of considerable interest as mobile power supply for future flexible electronics. Graphene or carbon nanotubes based thin films have been used to fabricate flexible solid-state supercapacitors with high gravimetric specific capacitances (80-200 F/g), but usually with a rather low overall or areal specific capacitance (3-50 mF/cm(2)) due to the ultrasmall electrode thickness (typically a few micrometers) and ultralow mass loading, which is not desirable for practical applications. Here we report the exploration of a three-dimensional (3D) graphene hydrogel for the fabrication of high-performance solid-state flexible supercapacitors. With a highly interconnected 3D network structure, graphene hydrogel exhibits exceptional electrical conductivity and mechanical robustness to make it an excellent material for flexible energy storage devices. Our studies demonstrate that flexible supercapacitors with a 120 μm thick graphene hydrogel thin film can exhibit excellent capacitive characteristics, including a high gravimetric specific capacitance of 186 F/g (up to 196 F/g for a 42 μm thick electrode), an unprecedented areal specific capacitance of 372 mF/cm(2) (up to 402 mF/cm(2) for a 185 μm thick electrode), low leakage current (10.6 μA), excellent cycling stability, and extraordinary mechanical flexibility. This study demonstrates the exciting potential of 3D graphene macrostructures for high-performance flexible energy storage devices.


Moral Punishment in Everyday Life.
The present research investigated event-related, contextual, demographic, and dispositional predictors of the desire to punish perpetrators of immoral deeds in daily life, as well as connections among the desire to punish, moral emotions, and momentary well-being. The desire to punish was reliably predicted by linear gradients of social closeness to both the perpetrator (negative relationship) and the victim (positive relationship). Older rather than younger adults, conservatives rather than people with other political orientations, and individuals high rather than low in moral identity desired to punish perpetrators more harshly. The desire to punish was related to state anger, disgust, and embarrassment, and these were linked to lower momentary well-being. However, the negative effect of these emotions on well-being was partially compensated by a positive indirect pathway via heightened feelings of moral self-worth. Implications of the present field data for moral punishment research and the connection between morality and well-being are discussed.


Quantum chemical insights into the dependence of porphyrin basicity on the meso-aryl substituents: thermodynamics, buckling, reaction sites and molecular flexibility.
The chemical and sensing properties of porphyrins are frequently tuned via the introduction of peripheral substituents. In the context of the exceptionally fast second protonation step in the case of 5,10,15,20-tetraphenylporphyrin (TPP), as compared to porphin and 5,10,15,20-tetramesitylporphyrin (TMesP), we investigated the macrocycle-substituent interactions of these three porphyrin derivatives in detail. Using quantum chemical thermodynamics calculations, the analysis of geometric structures, torsional profiles, electrostatic potential distributions, and particularly the analysis of molecular flexibilities via ab initio molecular dynamics simulations, we obtained a comprehensive picture of the reactivities of the studied porphyrins and how these are influenced by the meso-substituents. As compared to porphin and TMesP the second protonation of TPP is energetically more favorable and is particularly energetically comparable to its first protonation, instead of being significantly less favorable like in the case of porphyrin and TMesP. Additionally, the second TPP protonation is facilitated by an interplay between out-of-plane (oop) distortion of the protonation site and a pronounced electrostatic binding spot at the protonation site. Furthermore, the second protonation is particularly facilitated in the case of TPP by the large oop-flexibility of the diprotonated species as unraveled by ab initio molecular dynamics simulations.


How can research on plants contribute to promoting human health?
One of the most pressing challenges for the next 50 years is to reduce the impact of chronic disease. Unhealthy eating is an increasing problem and underlies much of the increase in mortality from chronic diseases that is occurring worldwide. Diets rich in plant-based foods are strongly associated with reduced risks of major chronic diseases, but the constituents in plants that promote health have proved difficult to identify with certainty. This, in turn, has confounded the precision of dietary recommendations. Plant biochemistry can make significant contributions to human health through the identification and measurement of the many metabolites in plant-based foods, particularly those known to promote health (phytonutrients). Plant genetics and metabolic engineering can be used to make foods that differ only in their content of specific phytonutrients. Such foods offer research tools that can provide significant insight into which metabolites promote health and how they work. Plant science can reduce some of the complexity of the diet-health relationship, and through building multidisciplinary interactions with researchers in nutrition and the pathology of chronic diseases, plant scientists can contribute novel insight into which foods reduce the risk of chronic disease and how these foods work to impact human health.


Parallel, minimally-invasive implantation of ultra-flexible neural electrode arrays.
Implanted microelectrodes provide a unique means to directly interface with the nervous system but have been limited by the lack of stable functionality. There is growing evidence suggesting that substantially reducing the mechanical rigidity of neural electrodes promotes tissue compatibility and improves their recording stability in both the short- and long-term. However, the miniaturized dimensions and ultraflexibility desired for mitigating tissue responses preclude the probe's self-supported penetration into the brain tissue. Here we demonstrate the high-throughput implantation of multi-shank ultraflexible neural electrode arrays with surgical footprints as small as 200 µm2 in a mouse model. This is achieved by using arrays of tungsten microwires as shuttle devices, and bio-dissolvable adhesive polyethylene glycol (PEG) to temporarily attach a shank onto each microwire. We show the ability to simultaneously deliver electrode arrays in designed patterns, to adjust the implantation locations of the shanks by need, to target different brain structures, and to control the surgical injury by reducing the microwire diameters to cellular scale. These results provide a facile implantation method to apply ultraflexible neural probes in scalable neural recording.


Award for Distinguished Professional Contributions to Institutional Practice.
This award is intended to recognize outstanding practitioners in psychology. Nominations are considered for psychologists working in a wide variety of institutional practice settings (e.g., schools, military, state hospital, Department of Veterans Affairs). Services provided to diverse client groups or patient populations, including but not limited to children/ adolescents/adults/older adults, urban/rural/ frontier populations, minority populations, and persons with serious mental illness, are considered. Contributions are judged distinguished by virtue of peer recognition, advancement of the public's recognition of psychology as a profession, relevant professional association honors, or other meritorious accomplishments denoting excellence as a practitioner, including improvement of institutional service delivery systems or development of psychologically informed public policy. The 2020 recipients of the APA Professional Contributions Awards were selected by the 2019 Board of Professional Affairs (BPA). (PsycInfo Database Record (c) 2020 APA, all rights reserved).


An In-Depth View of the Porcine Trabecular Meshwork Proteome.
The house swine (Sus scrofa domestica Linnaeus 1758) is an important model organism regarding the study of neurodegenerative diseases, especially ocular neuropathies such as glaucoma. This is due to the high comparability of the porcine and human eye regarding anatomy and molecular features. In the pathogenesis of glaucoma, the trabecular meshwork (TM) forms a key ocular component in terms of intraocular pressure (IOP) elevation. Thereby, functional TM abnormalities are correlated with distinct proteomic alterations. However, a detailed analysis of the TM proteome has not been realized so far. Since the porcine eye has high potential as a model system to study ocular diseases such as glaucoma, the present study focuses on the in-depth analysis of the porcine TM proteome. By use of a bottom-up (BU) mass spectrometric (MS) platform utilizing electrospray ionization liquid chromatography tandem MS (LC-ESI-MS/MS) considering database-dependent and peptide de novo sequencing, more than 3000 TM proteins were documented with high confidence (FDR < 1%). A distinct number of proteins with neuronal association were revealed. To the best to our knowledge, many of these protein species have not been reported for TM tissue before such as reelin, centlein and high abundant neuroblast differentiation-associated protein AHNAK (AHNAK). Thereby, AHNAK might play a superordinate role in the TM regarding proposed tissue involvement in barrier function. Also, a high number of secretory proteins could be identified. The generated TM proteomic landscape underlines a multifunctional character of the TM beyond representing a simple drainage system. Finally, the protein catalogue of the porcine TM provides an in-depth view of the TM molecular landscape and will serve as an important reference map in terms of glaucoma research utilizing porcine animal models, porcine TM tissues and/or cultured TM cells.


Sublethal mechanisms of Pb and Zn toxicity to the purple sea urchin (Strongylocentrotus purpuratus) during early development.
In order to understand sublethal mechanisms of lead (Pb) and zinc (Zn) toxicity, developing sea urchins were exposed continuously from 3h post-fertilization (eggs) to 96 h (pluteus larvae) to 55 (±2.4) μgPb/L or 117 (±11)μgZn/L, representing ~ 70% of the EC50 for normal 72 h development. Growth, unidirectional Ca uptake rates, whole body ion concentrations (Na, K, Ca, Mg), Ca(2+) ATPase activity, and metal bioaccumulation were monitored every 12h over this period. Pb exhibited marked bioaccumulation whereas Zn was well-regulated, and both metals had little effect on growth, measured as larval dry weight, or on Na, K, or Mg concentrations. Unidirectional Ca uptake rates (measured by (45)Ca incorporation) were severely inhibited by both metals, resulting in lower levels of whole body Ca accumulation. The greatest disruption occurred at gastrulation. Ca(2+) ATPase activity was also significantly inhibited by Zn but not by Pb. Interestingly, embryos exposed to Pb showed some capacity for recovery, as Ca(2+)ATPase activities increased, Ca uptake rates returned to normal intermittently, and whole body Ca levels were restored to control values by 72-96 h of development. This did not occur with Zn exposure. Both Pb and Zn rendered their toxic effects through disruption of Ca homeostasis, though likely through different proximate mechanisms. We recommend studying the toxicity of these contaminants periodically throughout development as an effective way to detect sublethal effects, which may not be displayed at the traditional toxicity test endpoint of 72 h.


Biologic Significance of Magnetic Resonance Imaging Invisibility in Localized Prostate Cancer.
Multiparametric magnetic resonance imaging (mpMRI) is used widely for prostate cancer (PCa) evaluation. Approximately 35% of aggressive tumors, however, are not visible on mpMRI. We sought to identify the molecular alterations associated with mpMRI-invisible tumors and determine whether mpMRI visibility is associated with PCa prognosis. Discovery and validation cohorts included patients who underwent mpMRI before radical prostatectomy and were found to harbor both mpMRI-visible (Prostate Imaging and Reporting Data System 3 to 5) and -invisible (Prostate Imaging and Reporting Data System 1 or 2) foci on surgical pathology. Next-generation sequencing was performed to determine differential gene expression between mpMRI-visible and -invisible foci. A genetic signature for tumor mpMRI visibility was derived in the discovery cohort and assessed in an independent validation cohort. Its association with long-term oncologic outcomes was evaluated in a separate testing cohort. The discovery cohort included 10 patients with 26 distinct PCa foci on surgical pathology, of which 12 (46%) were visible and 14 (54%) were invisible on preoperative mpMRI. Next-generation sequencing detected prioritized genetic mutations in 14 (54%) tumor foci (n = 8 mpMRI visible, n = 6 mpMRI invisible). A nine-gene signature (composed largely of cell organization/structure genes) associated with mpMRI visibility was derived (area under the curve = 0.89), and the signature predicted MRI visibility with 75% sensitivity and 100% specificity (area under the curve = 0.88) in the validation cohort. In the testing cohort (n = 375, median follow-up 8 years) there was no significant difference in biochemical recurrence, distant metastasis, or cancer-specific mortality in patients with predicted mpMRI-visible versus -invisible tumors (all P > .05). Compared with mpMRI-invisible disease, mpMRI-visible tumors are associated with underexpression of cellular organization genes. mpMRI visibility does not seem to be predictive of long-term cancer outcomes, highlighting the need for biopsy strategies that detect mpMRI-invisible tumors.


Oxytocin, a new determinant of bone mineral density in post-menopausal women: analysis of the OPUS cohort.
Oxytocin (OT), a neurohypophysial hormone regulated by estrogen and leptin, may play a role in bone metabolism in humans as suggested by animal studies. This study assessed the relationship between OT and bone status in a large population of postmenopausal women. Subjects were included in the Osteoporosis and Ultrasound study, a 6-year prospective study in a population-based cohort. Final visit data were used for this cross-sectional study. OT, leptin, and estradiol serum levels were measured in 1097 postmenopausal women and compared with bone mineral density (BMD), fractures, and the bone turnover markers (BTMs) procollagen type 1 N-terminal propeptide, bone alkaline phosphatase, and C-telopeptide of type 1 collagen. The median age was 70.8 years, 16% were osteoporotic, 48% were osteopenic, and 29% had at least one fracture. The OT serum level was related to spine (r = +0.12, P = .0002) and total hip BMD (r = +0.21, P < .0001) and with BTM (procollagen type 1 N-terminal propeptide: r = -0.13, P < .0001, bone alkaline phosphatase: r = -0.07, P = .02, C-telopeptide of type 1 collagen: r = -0.18, P < .0001). The relationship of OT with BMD was independent of BTM. After adjustment for confounding factors, the correlation between OT serum level and BMD remains significant at the hip in women with unmeasurable estradiol or leptin above the median value. There was no significant relationship between OT serum levels and fractures. High OT levels are associated with high BMD, especially at the hip in women with low estradiol or high leptin serum levels. The mechanism may be explained by the effect of OT on bone turnover.


Using Clinical Data Standards to Measure Quality: A New Approach.
Value-based payment for care requires the consistent, objective calculation of care quality. Previous initiatives to calculate ambulatory quality measures have relied on billing data or individual electronic health records (EHRs) to calculate and report performance. New methods for quality measure calculation promoted by federal regulations allow qualified clinical data registries to report quality outcomes based on data aggregated across facilities and EHRs using interoperability standards. This research evaluates the use of clinical document interchange standards as the basis for quality measurement. Using data on 1,100 patients from 11 ambulatory care facilities and 5 different EHRs, challenges to quality measurement are identified and addressed for 17 certified quality measures. Iterative solutions were identified for 14 measures that improved patient inclusion and measure calculation accuracy. Findings validate this approach to improving measure accuracy while maintaining measure certification. Organizations that report care quality should be aware of how identified issues affect quality measure selection and calculation. Quality measure authors should consider increasing real-world validation and the consistency of measure logic in respect to issues identified in this research.


PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility.
We previously demonstrated that PACRG plays a role in regulating dynein-driven microtubule sliding in motile cilia. To expand our understanding of the role of PACRG in ciliary assembly and motility, we used a combination of functional and structural studies, including newly identified Chlamydomonas pacrg mutants. Using cryo-electron tomography we show that PACRG and FAP20 form the inner junction between the A- and B-tubule along the length of all nine ciliary doublet microtubules. The lack of PACRG and FAP20 also results in reduced assembly of inner-arm dynein IDA b and the beak-MIP structures. In addition, our functional studies reveal that loss of PACRG and/or FAP20 causes severe cell motility defects and reduced in vitro microtubule sliding velocities. Interestingly, the addition of exogenous PACRG and/or FAP20 protein to isolated mutant axonemes restores microtubule sliding velocities, but not ciliary beating. Taken together, these studies show that PACRG and FAP20 comprise the inner junction bridge that serves as a hub for both directly modulating dynein-driven microtubule sliding, as well as for the assembly of additional ciliary components that play essential roles in generating coordinated ciliary beating.


Dexamethasone for adult patients with a symptomatic chronic subdural haematoma (Dex-CSDH) trial: study protocol for a randomised controlled trial.
Chronic subdural haematoma (CSDH) is a common neurosurgical condition, typically treated with surgical drainage of the haematoma. However, surgery is associated with mortality and morbidity, including up to 20% recurrence of the CSDH. Steroids, such as dexamethasone, have been identified as a potential therapy for reducing recurrence risk in surgically treated CSDHs. They have also been used as a conservative treatment option, thereby avoiding surgery altogether. The hypothesis of the Dex-CSDH trial is that a two-week course of dexamethasone in symptomatic patients with CSDH will lead to better functional outcome at six months. This is anticipated to occur through reduced number of hospital admissions and surgical interventions. Dex-CSDH is a UK multi-centre, double-blind randomised controlled trial of dexamethasone versus placebo for symptomatic adult patients diagnosed with CSDH. A sample size of 750 patients has been determined, including an initial internal pilot phase of 100 patients to confirm recruitment feasibility. Patients must be recruited within 72 h of admission to a neurosurgical unit and exclusions include patients already on steroids or with steroid contraindications, patients who have a cerebrospinal fluid shunt and those with a history of psychosis. The decision regarding surgical intervention will be made by the clinical team and patients can be included in the trial regardless of whether operative treatment is planned or has been performed. The primary outcome measure is the modified Rankin Scale (mRS) at six months. Secondary outcomes include the number of CSDH-related surgical interventions during follow-up, length of hospital stay, mRS at three months, EQ-5D at three and six months, adverse events, mortality and a health-economic analysis. This multi-centre trial will provide high-quality evidence as to the effectiveness of dexamethasone in the treatment of CSDH. This has implications for patient morbidity and mortality as well as a potential economic impact on the overall health service burden from this condition. ISRCTN, ISRCTN80782810 . Registered on 7 November 2014. EudraCT, 2014-004948-35 . Registered on 20 March 2015. Dex-CSDH trial protocol version 3, 27 Apr 2017. This protocol was developed in accordance with the SPIRIT checklist. Available as a separate document on request.


Increasing prominence of implantology research: a chronological trend analysis of 100 top-cited articles in periodontal journals.
To identify 100 top-cited articles published in periodontal journals and analyse the research trends by using citation analysis. 100 top-cited articles published in periodontal journals were retrieved by searching the database of the ISI Web of Science and Journal Citation reports. For each article, the following principal bibliometric parameters: authorship, geographic and institute origin, manuscript type, study design, scope of study, and citation count of each time period were analysed from 1965 to 2015. The identified 100 top-cited articles were retrieved from five periodontal journals and citation counts were recorded between 262 and 1,693 times. For the institute of origin, the most productive institute, in terms of the number of 100 top-cited articles published, was the University of Gothenburg (Sweden) (n = 19), followed by the Forsyth Dental Center (USA) (n = 15). Most manuscripts were original research (n = 74), and the inflammatory periodontal disease (n = 59) was the most frequent topic studied. Interestingly, the trend of increase average citation reached significance for implantology (β = 26.75, P = 0.003) and systemic interactions (β = 29.83, P = 0.005), but not for inflammatory disease (β = -10.30, P = 0.248) and tissue regeneration (β = 9.04, P = 0.081). By using multivariable linear regression in a generalised linear model, suitable published journal (Journal of Clinical Periodontology), geographic regions (Europe), more intense international collaboration, adequate manuscript type (review article) and study design (systematic review) could be attributed to escalating average citation counts in implantology (all P < 0.05). However, for systemic interactions, only geographic region and study design were significantly associated with the increasing citation trend. These principal bibliometric characteristics revealed escalated trends in average citation count in implantology throughout time.


High plant diversity is needed to maintain ecosystem services.
Biodiversity is rapidly declining worldwide, and there is consensus that this can decrease ecosystem functioning and services. It remains unclear, though, whether few or many of the species in an ecosystem are needed to sustain the provisioning of ecosystem services. It has been hypothesized that most species would promote ecosystem services if many times, places, functions and environmental changes were considered; however, no previous study has considered all of these factors together. Here we show that 84% of the 147 grassland plant species studied in 17 biodiversity experiments promoted ecosystem functioning at least once. Different species promoted ecosystem functioning during different years, at different places, for different functions and under different environmental change scenarios. Furthermore, the species needed to provide one function during multiple years were not the same as those needed to provide multiple functions within one year. Our results indicate that even more species will be needed to maintain ecosystem functioning and services than previously suggested by studies that have either (1) considered only the number of species needed to promote one function under one set of environmental conditions, or (2) separately considered the importance of biodiversity for providing ecosystem functioning across multiple years, places, functions or environmental change scenarios. Therefore, although species may appear functionally redundant when one function is considered under one set of environmental conditions, many species are needed to maintain multiple functions at multiple times and places in a changing world.


Force-deformation response of the lumbar spine: a sagittal plane model of posteroanterior manipulation and mobilization.
To develop a mathematical model capable of describing the static and dynamic motion response of the lumbar spine to posteroanterior forces. Static, impulsive and oscillatory forces with varying thrust angles and offsets were applied to the model, and the resulting motion responses were compared to experimental data published for spinal mobilization and manipulation of prone-lying subjects. Methods are sought to improve understanding of the dynamic force-induced displacement response of the lumbar spine during spinal mobilization and manipulation treatment. The thorax, pelvis and five lumbar vertebrae were represented as seven rigid structures and eight flexible joint structures. Flexible joint structures were modeled using spring and damper elements with three displacement degrees-of-freedom (posterior-anterior and axial displacement, and flexion-extension rotation). The resulting 21 degrees-of-freedom lumped parameter model was solved in modal space. The fundamental natural frequency of vibration was 5.24 Hz. Simulations performed using 100 N static and dynamic posteroanterior forces applied to the L3 vertebrae indicated that peak L3 segmental displacements were up to 2.40 mm (impulsive) and 8.23 mm (oscillatory at 2 Hz). Appreciable axial displacements (0.41 mm) and flexion-extension rotations (1.49 degrees ) were also observed for oscillatory forces at L3. The posteroanterior motion response of the lumbar vertebrae was relatively insensitive to both the thrust force angle and thrust force offset, but axial displacements and flexion-extension rotations showed a large change (2-fold or greater) for thrust angles greater than -5 degrees (caudal) in comparison to vertical thrusts. Intersegmental motion responses for static, impulsive and oscillatory loads were more comparable than their segmental counterparts. The model predicts lumbar segmental and inter-segmental motion responses to manipulative forces that are otherwise difficult to obtain experimentally. This study assists clinicians to understand the biomechanics of posteroanterior forces applied to the lumbar spine of prone-lying subjects. Of particular clinical relevance is the finding that greater spinal mobility is possible by targeting specific load-time histories.


Developing a practical algorithm for a pediatric emergency department's response to radiological dispersal device events.
In the aftermath of the detonation of a radiological dispersal device (RDD), or "dirty bomb," a large influx of children would be expected to present to the emergency department, including many patients not directly affected by the event who present with concerns regarding radiation exposure. Our objective was to develop an algorithm for efficiently and effectively triaging and appropriately treating children based on the likelihood of their having been contaminated or exposed. The hospital's disaster preparedness committee with the help of disaster planning experts engaged in an iterative process to develop a triage questionnaire and patient flow algorithm for a pediatric hospital following an RDD event. The questionnaire and algorithm were tested using hypothetical patients to ensure that they resulted in appropriate triage and treatment for the full range of anticipated patient presentations and were then tested in 2 live drills to evaluate their performance in real time. The triage questionnaire reduced triage times and accurately sorted children into groups based on the type of intervention they required. Nonmedical personnel were able to administer the triage questionnaire effectively with minimal training, relieving professional staff. The patient flow algorithm and supporting materials provided direction to staff about how to appropriately treat patients once they had been triaged. In the event of the detonation of an RDD, the triage questionnaire and patient flow algorithm presented would enable pediatric hospitals to direct limited resources to children requiring intervention due to injury, contamination, or exposure.


Optimizing DyNaChron instrument for assessing chronic nasal dysfunction symptoms by Rasch analysis.
The DyNaChron (Dysfonctionnement Nasal Chronique) questionnaire is a self-reporting 78-item instrument assessing six symptoms and their consequences of chronic nasal dysfunction. Patients complete items of a symptom domain only when it is present but in case the patient presents several or all symptoms, its length can limit its use. Here, we aimed to optimize, or shorten, the DyNaChron for clinical use. A total of 640 patients in 14 rhinology outpatient clinics all over France completed the original DyNaChron questionnaire before the first rhinologic clinic and 15 days later. The optimization process involved Rasch analysis and then qualitative content analyses. Rasch analysis flagged items with a floor/ceiling effect or with important differential item functioning and an expert committee decided whether to retain the flagged items on the basis of clinical importance and statistical characteristics. The psychometric properties of the optimized version were studied according to classical test theory and Rasch analysis. Rasch analysis revealed 4 items with underfit, 6 with an extreme score, 2 that were highly locally dependent and 16 with differential item functioning which 5 of these 16 items were retained after content analysis. In total, 19 flagged items were removed. Factorial analysis confirmed the preservation of the initial instrument structure in the optimized scale; psychometrics properties and scale calibration were the same as or better than the original version. The shortened DyNaChron optimizes the quality of assessment by deleting redundant items and reduces the burden on respondents; the structure is preserved and the psychometrics properties are improved.


Effective suckling in relation to naked maternal-infant body contact in the first hour of life: an observation study.
Best practice guidelines to promote breastfeeding suggest that (i) mothers hold their babies in naked body contact immediately after birth, (ii) babies remain undisturbed for at least one hour and (iii) breastfeeding assistance be offered during this period. Few studies have closely observed the implementation of these guidelines in practice. We sought to evaluate these practices on suckling achievement within the first hour after birth. Observations of seventy-eight mother-baby dyads recorded newborn feeding behaviours, the help received by mothers and birthing room practices each minute, for sixty minutes. Duration of naked body contact between mothers and their newborn babies varied widely from 1 to 60 minutes, as did commencement of suckling (range = 10 to 60 minutes). Naked maternal-infant body contact immediately after birth, uninterrupted for at least thirty minutes did not predict effective suckling within the first hour of birth. Newborns were four times more likely to sustain deep rhythmical suckling when their chin made contact with their mother's breast as they approached the nipple (OR 3.8; CI 1.03 - 14) and if their mothers had given birth previously (OR 6.7; CI 1.35 - 33). Infants who had any naso-oropharyngeal suctioning administered at birth were six times less likely to suckle effectively (OR .176; CI .04 - .9). Effective suckling within the first hour of life was associated with a collection of practices including infants positioned so their chin can instinctively nudge the underside of their mother's breast as they approach to grasp the nipple and attach to suckle. The best type of assistance provided in the birthing room that enables newborns to sustain an effective latch was paying attention to newborn feeding behaviours and not administering naso-oropharyngeal suction routinely.


Hippocampal pyramidal cell disarray in schizophrenia as a bilateral phenomenon.
In a continuing quantitative study of hippocampal cell orientation in schizophrenic subjects and nonschizophrenic control subjects, a pattern of right hemispheric cell disorganization in schizophrenic subjects, as robust as that previously described in the left hemisphere, was shown. The study was based on tissue from 11 schizophrenic and seven approximately age-matched control subjects from a Veterans Administration Medical Center. Using a new measuring technique that proved complementary and superior to the method formerly used, we determined the axes of orientation for approximately 10,800 neurons. A significant difference in cell disorganization was found between schizophrenic and control subjects. From separate analyses of the interface zones among the various hippocampal cell sectors, this difference was found to be significant at the interfaces between cornu ammonis zones 1 and 2 and zones 2 and 3. Analysis of the pattern of disorganization numbers suggests a bimodal distribution in which the cell orientation values of one group of schizophrenic subjects overlap those of the nonschizophrenic control group, while the cell orientation values of a second group significantly exceed those of the control population. Although some recent studies have suggested that schizophrenia may be related exclusively to the left hemisphere, the results of this study indicate that schizophrenia-related structural changes involve the right hemisphere as well. Thus, schizophrenia may be a bilateral rather than unilateral disorder.


Lack of recent condom use among detained adolescent males: a multilevel investigation.
To investigate multiple levels of influence with respect to the lack of recent condom use among a high risk sample of adolescent males recruited from short term detention facilities. A cross sectional survey of 231 adolescent males serving, predominantly, short term detention sentences. Assessments were conducted using audiocomputer assisted self interviewing. Condom use during the most recent sexual event was assessed as well as 20 potential correlates of not using condoms. Correlates were assessed within five levels of causation: personal, relational, peer affiliation, family, and societal. Nine correlates achieved bivariate significance (p<0.05). Of these, the personal level correlates were particularly important in a multivariate model. The motivation subscale from the Condom Barriers Scale was the strongest multivariate correlate of recent condom use. Adolescents scoring below the median were about 3.4 times more likely to report lack of recent condom use (p=0.0006). Adolescents indicating they had ever caused a pregnancy were about 2.5 times more likely to report lack of condom use (p=0.02). Finally, those reporting their peers did not use condoms were about twice as likely to report lack of use (p=0.048). Upon investigating multiple levels of potential influence on condom use, the multivariate findings suggest that personal level factors may be the most important determinant of non-use among adolescent males in short term detention facilities. Although structural changes may be needed to influence some forms of safer sex behaviour, direct intervention with adolescent males may be justified to favourably alter determinants of condom use.


A Mouse Model of Post-Stroke Pneumonia Induced by Intra-Tracheal Inoculation with Streptococcus pneumoniae.
Stroke-induced immunodeficiency increases the risk of infectious complications, which adversely affects neurological outcome. Among those, pneumonia affects as many as one third of stroke patients and is the main contributor to mortality in the post-acute phase of stroke. Experimental findings on post-stroke susceptibility to spontaneous pneumonia in mice are contradictory. Here, we established a mouse model inducing standardized bacterial pneumonia and characterized the impaired pulmonary cellular and humoral immune responses after experimental stroke. Bacterial pneumonia was induced by intra-tracheal inoculation with Streptococcus pneumoniae at different time points after transient middle cerebral artery occlusion (MCAO). Bacterial counts in lungs and blood, histological changes, and cytokine production in the lungs were assessed. Furthermore, we investigated the effect of pneumonia on stroke outcome. Intra-tracheal inoculation resulted in reproducible pneumonia and bacteraemia, and demonstrated post-stroke susceptibility to streptococcal pneumonia developing with a delay of at least 24 h after MCAO. Higher bacterial counts in mice infected 3 days after stroke induction correlated with reduced neutrophil and macrophage infiltration in the lungs and lower levels of pro-inflammatory cytokines in the broncho-alveolar lavage compared to sham-operated animals. Pneumonia increased mortality without affecting brain-infiltrating leukocytes. In this standardized mouse model of post-stroke pneumonia, we describe attenuated leukocyte infiltration and cytokine production in response to bacterial infection in the lungs that has a profound effect on outcome.


Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAHenu2 mouse model of PKU.
Phenylketonuria (PKU) is a genetic disease characterized by the inability to convert dietary phenylalanine to tyrosine by phenylalanine hydroxylase. Given the importance of gut microbes in digestion, a genetically engineered microbe could potentially degrade some ingested phenylalanine from the diet prior to absorption. To test this, a phenylalanine lyase gene from Anabaena variabilis (AvPAL) was codon-optimized and cloned into a shuttle vector for expression in Lactobacillus reuteri 100-23C (pHENOMMenal). Functional expression of AvPAL was determined in vitro, and subsequently tested in vivo in homozygous PAHenu2 (PKU model) mice. Initial trials of two PAHenu2 homozygous (PKU) mice defined conditions for freeze-drying and delivery of bacteria. Animals showed reduced blood phe within three to four days of treatment with pHENOMMenal probiotic, and blood phe concentrations remained significantly reduced (P < 0.0005) compared to untreated controls during the course of experiments. Although pHENOMMenal probiotic could be cultured from fecal samples at four months post treatment, it could no longer be cultivated from feces at eight months post treatment, indicating eventual loss of the microbe from the gut. Preliminary screens during experimentation found no immune response to AvPAL. Collectively these studies provide data for the use of a genetically engineered probiotic as a potential treatment for PKU.


Silver enhanced TiO2 thin films: photocatalytic characterization using aqueous solutions of tris(hydroxymethyl)aminomethane.
The photocatalytic activity in aqueous solutions of TiO2 and Ag enhanced TiO2 sol-gel produced films was characterized using tris(hydroxymethyl)aminomethane (Tris) under black light (365 nm) and the observed differences in efficiency were further investigated by O2 adsorption studies using the same probe. Hydrogen abstracting species, such as hydroxyl radicals formed upon photocatalysis, are able to abstract hydrogen from Tris. This reaction leads to the formation of formaldehyde which was detected and quantified through a modified version of the Hantzsch reaction. It was found that the Ag enhanced TiO2 film increased the apparent quantum yield from 7% to 12%, partly as a result of a Schottky barrier formation at the metal-semiconductor interface and partly as the sensitizing effect of Ag nanoparticles extends the visible light absorption, which through electron transfer processes enable an efficient charge separation in the TiO2 by attracting acceptor species more efficiently than pure TiO2. The O2 adsorption studies in this paper showed that the Ag enhanced TiO2 film has a stronger adsorption affinity than pure TiO2 towards O2, which make the reduction of O2 more efficient with a subsequent enhanced electron-hole lifetime. It was also found that the Ag enhanced TiO2 film had a poorer adsorption affinity for Tris than the pure TiO2 film, which is a consequence of fewer available surface adsorption sites due to the Ag coverage at 64% which agrees well with the obtained adsorption equilibrium constants (K(LH(TiO2)) = 615 M(-1) and K(LH(Ag-TiO2)) = 320 M(-1)).


Characterization of adenosine A1 receptors in intact DDT1 MF-2 smooth muscle cells.
Adenosine receptors in the smooth muscle cell line DDT1 MF-2 were studied by radioligand binding using the A1 receptor-selective antagonist [3H]8-cyclopentyl-1,3-dipropylxanthine [( 3H]DPCPX) as the ligand. Binding characteristics were similar in intact cells and in membranes (KD value of approximately 1 nM). The maximum binding amounted to 183 fmol/10(6) intact cells or 344 fmol/mg of membranes. To characterize the receptor, competition experiments were performed by inhibiting [3H]DPCPX binding with several adenosine agonists and antagonists. Adenosine receptor antagonists appeared to bind to a single class of binding site, both in membranes and intact cells. The order of potency was DPCPX = CGS 15943A greater than 8-cyclopentyl-1,3-dimethylxanthine greater than 8-(p-sulfophenyl)-theophylline greater than 3-isobutyl-1-methylxanthine greater than theophylline. Competition curves with adenosine agonists in membranes were best described by a two-site rather than a one-site model. At equilibrium in intact cells, only a single site was detected at both 4 degrees and 25 degrees. However, short term incubations (1-4 min) at 25 degrees showed biphasic binding curves in intact cells. The equilibrium KD values for intact cells were similar to the low affinity KD values in membranes (KL). The order of potency was N6-cyclopentyladenosine greater than or equal to (-)-(R)-N6-phenylisopropyladenosine[(R)-PIA] greater than or equal to N6-cyclohexyl adenosine greater than 5'-N-ethylcarboxamidoadenosine (NECA) greater than 2-chloroadenosine greater than adenosine (intact cells only) greater than 2-phenylaminoadenosine (CV 1808). Treatment of cells with pertussis toxin ADP-ribosylated GTP-binding proteins and eliminated the high affinity agonist binding in membranes but did not affect binding to intact cells. The addition of GTP (100 microM) also shifted the competition curves from bi- to monophasic curves in membranes. Adenosine receptor agonists inhibited the formation of cAMP induced by isoprenaline (IC50 for (R)-PIA, 0.4 nM). This inhibition could be prevented with adenosine receptor antagonists. Pretreatment with pertussis toxin also reversed these effects and actually revealed functional A2 receptors, as shown by the formation of cAMP induced by NECA. In conclusion, the equilibrium binding of A1 receptor agonists to intact smooth muscle cells is similar to the low affinity binding observed in membranes. In addition, it is suggested that agonists may transiently convert the A1 receptor from a "resting" low affinity state to a high affinity state coupled to a GTP-binding protein.(ABSTRACT TRUNCATED AT 400 WORDS)


Surprising similarities in structure comparison.
Examination of a protein's structural 'neighbors' can reveal distant evolutionary relationships that are otherwise undetectable, and perhaps suggest unsuspected functional properties. In the past, such analyses have often required specialized software and computer skills, but new structural comparison methods, developed in the past two years, increasingly offer this opportunity to structural and molecular biologists in general. These methods are based on similarity-search algorithms that are fast enough to have effectively removed the computer-time limitation for structure-structure search and alignment, and have made it possible for several groups to conduct systematic comparisons of all publicly available structures, and offer this information via the World Wide Web. Furthermore, and perhaps surprisingly given the difficulty of the structure-comparison problem, these groups seem to have converged on quite similar approaches with respect to both fast search algorithms and the identification of statistically significant similarities.


Sialic acids in the brain: gangliosides and polysialic acid in nervous system development, stability, disease, and regeneration.
Every cell in nature carries a rich surface coat of glycans, its glycocalyx, which constitutes the cell's interface with its environment. In eukaryotes, the glycocalyx is composed of glycolipids, glycoproteins, and proteoglycans, the compositions of which vary among different tissues and cell types. Many of the linear and branched glycans on cell surface glycoproteins and glycolipids of vertebrates are terminated with sialic acids, nine-carbon sugars with a carboxylic acid, a glycerol side-chain, and an N-acyl group that, along with their display at the outmost end of cell surface glycans, provide for varied molecular interactions. Among their functions, sialic acids regulate cell-cell interactions, modulate the activities of their glycoprotein and glycolipid scaffolds as well as other cell surface molecules, and are receptors for pathogens and toxins. In the brain, two families of sialoglycans are of particular interest: gangliosides and polysialic acid. Gangliosides, sialylated glycosphingolipids, are the most abundant sialoglycans of nerve cells. Mouse genetic studies and human disorders of ganglioside metabolism implicate gangliosides in axon-myelin interactions, axon stability, axon regeneration, and the modulation of nerve cell excitability. Polysialic acid is a unique homopolymer that reaches >90 sialic acid residues attached to select glycoproteins, especially the neural cell adhesion molecule in the brain. Molecular, cellular, and genetic studies implicate polysialic acid in the control of cell-cell and cell-matrix interactions, intermolecular interactions at cell surfaces, and interactions with other molecules in the cellular environment. Polysialic acid is essential for appropriate brain development, and polymorphisms in the human genes responsible for polysialic acid biosynthesis are associated with psychiatric disorders including schizophrenia, autism, and bipolar disorder. Polysialic acid also appears to play a role in adult brain plasticity, including regeneration. Together, vertebrate brain sialoglycans are key regulatory components that contribute to proper development, maintenance, and health of the nervous system.