Multiple Forms of Nicotinamide Adenine Dinucleotide Glycohydrolase from Bull Semen

Nicotinamide adenine dinucleotide glycohydrolase (NADase) of bull semen is an extracellular, soluble enzyme capable of catalyzing the hydrolysis of the nicotinamide N-ribosidic linkage of NAD+. In search for a rapid, simple procedure for the preparation of this enzyme, it was found that Matrex Gel Red A (Amicon) column packing served very well as an affinity gel for NADase. The enzyme was applied to the Matrex Red A column (21 cm x 1.5 cm) in 10 mM sodium phosphate buffer, pH 6.0. Surprisingly, two fractions of NADase activity were obtained when the column was eluted with a sodium chloride linear gradient from Oto 2.5 M. The two forms, NADase I and NADase II, were eluted at the approximate salt concentrations of 0.25 Mand 1.20 M, respectively. Investigation of these two forms was continued. Gel filtration chromatography was performed to determine the approximate molecular weights of 55,000 to 60,000 daltons and the Stokes\u27 radii of 31.8 Å for NADase I and 0 31.5 Å for NADase II. Kinetic studies of the active substances showed differences in their abilities to catalyze the hydrolysis of NAD+. NADase I was irnmediately active in hydrolyzing the substrate, whereas NADase II possessed a characteristic lag phase before catalyzing the reaction. The apparent Michealis constant, Km, for NADase I and for the maximum velocity of the m NADase II reaction was determined to be 1.0 x 10-4 M, which is the same value determined for the homogeneous bull semen NADase. Further studies of the dual forms of the enzyme were made by observing the ability to catalyze the base exchange reaction with 3-acetylpyridine and NAD+ to form free nicotinamide and 3-acetylpyridine adenine inucleotide. Earlier investigation showed that bull semen NADase was unable to catalyze the base exchange. However, both NADase I and NADase II were found to cause the base exchange as the other mammalian NADase\u27s. Thin layer chromatography was used to provide visual evidence ·of the existence of the exchange products

The Influence of the Early Retirement Process on Satisfaction with Early Retirement and Psychological Well-Being

The present study explores the influence of the early retirement process on adjustment to early retirement, taking into account the roles of individual characteristics and social context in this process. We proposed a systematic model integrating perceived ability to continue working, organizational pressures toward early retirement and group norms about early retirement as antecedents of the early retirement process and subsequent satisfaction with early retirement and psychological well-being. In addition, we examined the moderating role of the voluntariness of the early retirement transition in the proposed model. Our hypotheses were tested using a sample of 213 early retirees. We found that while high organizational pressures were related to lower retirement age, low perceived ability to continue working and group norms favorable to early retirement were related to higher levels of early retirement intentions. Furthermore, group norms favorable to early retirement and low perceived ability to continue working predicted higher satisfaction with early retirement, both directly and indirectly, through early retirement intentions. Finally, satisfaction with early retirement was related to psychological well-being. However, when the sample was divided into subgroups of voluntary and involuntary early retirees, two different relationships were observed in each group. For instance, organizational pressures positively predicted satisfaction with early retirement in involuntary early retirees, whereas this relationship was negative in voluntary early retirees. In addition, income was positively related to retirement age in involuntary early retirees and negatively in voluntarily early retired

A Likelihood-Based Approach for Computing the Operating Characteristics of the Standard Phase I Clinical Trial Design

In phase I clinical trials, the standard ‘3+3’ design has passed the test of time and survived various sample size adjustments, or other dose-escalation dynamics. The objective of this study is to provide a probabilistic support for analyzing the heuristic performance of the ‘3+3’ design. Our likelihood method is based on the evidential paradigm that uses the likelihood ratio to measure the strength of statistical evidence for one simple hypothesis over the other. We compute the operating characteristics and compare the behavior of the standard algorithm under different hypotheses, levels of evidence, and true (or best guessed) toxicity rates. Given observed toxicities per dose level, the likelihood-ratio is evaluated according to a certain k threshold (level of evidence). Under an assumed true toxicity scenario the following statistical characteristics are computed and compared: i) probability of weak evidence, ii) probability of favoring H1 under H1(analogous to 1-α), iii) probability of favoring H2 under H2 (analogous to 1-β). This likelihood method allows consistent inferences to be made and evidence to be quantified regardless of cohort size. Moreover, this approach can be extended and used in phase I designs for identifying the highest acceptably safe dose and is akin to the sequential probability ratio test

Brief Report: Factors Associated with Asthma Management Self-Efficacy Among 7th and 8th Grade Students

Objective Examine correlates of asthma self-management among 12,154 adolescents with physician-diagnosed asthma. Methods All 7th and 8th grade students in North Carolina completed a survey to assess asthma prevalence and self-management behaviors among those with asthma. Results Adolescents who were allowed to carry their inhaled medication at school, shown how to use a peak flow meter, and had access to more asthma care resources were more confident that they could prevent an asthma exacerbation. Adolescents who were allowed to carry their inhaled medication at school and who had a private doctor were more confident that they could control their symptoms. Adolescents taking anti-inflammatory medicine were less confident that they could prevent an exacerbation and control their symptoms. Conclusions Various indicators of autonomy and control were associated with greater self-efficacy for managing asthma. Adolescents who require anti-inflammatory medicines would benefit from additional intervention efforts to improve their asthma management self-efficacy

Do hypoxia/normoxia culturing conditions change the neuroregulatory profile of Wharton Jelly mesenchymal stem cells secretome?

Introduction: The use of human umbilical cord Wharton Jelly-derived mesenchymal stem cells (hWJ-MSCs) has been considered a new potential source for future safe applications in regenerative medicine. Indeed, the application of hWJ-MSCs into different animal models of disease, including those from the central nervous system, has shown remarkable therapeutic benefits mostly associated with their secretome. Conventionally, hWJ-MSCs are cultured and characterized under normoxic conditions (21 % oxygen tension), although the oxygen levels within tissues are typically much lower (hypoxic) than these standard culture conditions. Therefore, oxygen tension represents an important environmental factor that may affect the performance of mesenchymal stem cells in vivo. However, the impact of hypoxic conditions on distinct mesenchymal stem cell characteristics, such as the secretome, still remains unclear. Methods: In the present study, we have examined the effects of normoxic (21 % O2) and hypoxic (5 % O2) conditions on the hWJ-MSC secretome. Subsequently, we address the impact of the distinct secretome in the neuronal cell survival and differentiation of human neural progenitor cells. Results: The present data indicate that the hWJ-MSC secretome collected from normoxic and hypoxic conditions displayed similar effects in supporting neuronal differentiation of human neural progenitor cells in vitro. However, proteomic analysis revealed that the use of hypoxic preconditioning led to the upregulation of several proteins within the hWJ-MSC secretome. Conclusions: Our results suggest that the optimization of parameters such as hypoxia may lead to the development of strategies that enhance the therapeutic effects of the secretome for future regenerative medicine studies and applications. © 2015 Teixeira et al.Portuguese Foundation for Science and Technology (FCT) (Ciência 2007 program and IF Development Grant (AJS); and pre-doctoral fellowships to FGT (SFRH/69637/ 2010) and SIA (SFRH/BD/81495/2011); Canada Research Chairs (LAB) and a SSE Postdoctoral Fellowship (KMP); The National Mass Spectrometry Network (RNEM) (REDE/1506/REM/2005); co-funded by Programa Operacional Regional do Norte (ON.2 – O Novo Norte), ao abrigo do Quadro de Referência Estratégico Nacional (QREN), através do Fundo Europeu de Desenvolvimento Regional (FEDER).info:eu-repo/semantics/publishedVersio

Recanalization and Angiographic Reperfusion Are Both Associated with a Favorable Clinical Outcome in the IMS III Trial.

BACKGROUND: Prompt revascularization is the main goal of acute ischemic stroke treatment. We examined which revascularization scale - reperfusion (modified Treatment in Cerebral Infarctions, mTICI) or recanalization (Arterial Occlusive Lesion, AOL) - better predicted the clinical outcome in ischemic stroke participants treated with endovascular therapy (EVT). Additionally, we determined the optimal thresholds for the predictive accuracy of each scale. METHODS: We included participants from the Interventional Management of Stroke (IMS) III trial with complete occlusion in the internal carotid artery terminus or proximal middle cerebral artery (M1 or M2) who completed EVT within 7 h of symptom onset. The abilities of the AOL and mTICI scales to predict a favorable outcome (defined as a modified Rankin Scale score of 0-2 at 3 months) were compared by receiver operating characteristic analyses. The maximal sensitivity and specificity for each revascularization scale were established. RESULTS: Among 240 participants who met the study inclusion criteria, 79 (33%) achieved a favorable outcome. Higher scores of mTICI and AOL increased the likelihood of a favorable outcome (2.7% with mTICI 0 vs. 83.3% with mTICI 3, and 3.0% with AOL 0 vs. 43% with AOL 3). The accuracy of mTICI reperfusion and AOL recanalization for a favorable outcome prediction was similar, with optimal thresholds of mTICI 2b/3 and AOL 3, respectively. CONCLUSION: Reperfusion (mTICI) and recanalization (AOL) predicted a favorable clinical outcome with comparable accuracy in ischemic stroke participants treated with EVT. Optimal revascularization goals to maximize clinical outcome (modified Rankin Scale score of 0-2) consisted of complete recanalization (AOL 3) and reperfusion of at least 50% of the arterial tree of the symptomatic artery (mTICI 2b/3) in the IMS III trial setting

Meta-analysis of microarray studies reveals a novel hematopoietic progenitor cell signature and demonstrates feasibility of inter-platform data integration

Microarray-based studies of global gene expression (GE) have resulted in a large amount of data that can be mined for further insights into disease and physiology. Meta-analysis of these data is hampered by technical limitations due to many different platforms, gene annotations and probes used in different studies. We tested the feasibility of conducting a meta-analysis of GE studies to determine a transcriptional signature of hematopoietic progenitor and stem cells. Data from studies that used normal bone marrow-derived hematopoietic progenitors was integrated using both RefSeq and UniGene identifiers. We observed that in spite of variability introduced by experimental conditions and different microarray platforms, our meta-analytical approach can distinguish biologically distinct normal tissues by clustering them based on their cell of origin. When studied in terms of disease states, GE studies of leukemias and myelodysplasia progenitors tend to cluster with normal progenitors and remain distinct from other normal tissues, further validating the discriminatory power of this meta-analysis. Furthermore, analysis of 57 normal hematopoietic stem and progenitor cell GE samples was used to determine a gene expression signature characteristic of these cells. Genes that were most uniformly expressed in progenitors and at the same time differentially expressed when compared to other normal tissues were found to be involved in important biological processes such as cell cycle regulation and hematopoiesis. Validation studies using a different microarray platform demonstrated the enrichment of several genes such as SMARCE, Septin 6 and others not previously implicated in hematopoiesis. Most interestingly, alpha-integrin, the only common stemness gene discovered in a recent comparative murine analysis (Science 302(5644):393) was also enriched in our dataset, demonstrating the usefulness of this analytical approach

Provider Demonstration and Assessment of Child Device Technique During Pediatric Asthma Visits

The purposes of this study were to (a) describe the extent to which children use metered dose inhalers, turbuhalers, diskuses, and peak flow meters correctly, and (b) investigate how often providers assess and demonstrate use of metered dose inhalers, turbuinhalers, diskuses, and peak flow meters during pediatric asthma visits

Communication During Pediatric Asthma Visits and Self-Reported Asthma Medication Adherence

Our objectives were to examine how certain aspects of provider-patient communication recommended by national asthma guidelines (ie, provider asking for child and caregiver input into the asthma treatment plan) were associated with child asthma medication adherence 1 month after an audio-taped medical visit