bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Resveratrol in Patients with Type 2 Diabetes Mellitus on Skeletal Muscle SIRT1 Expression and Energy Expenditure
Authors: Goh Kian Peng , Lee Hwei Yee , Lau Dawn Pingxi , Supaat Wilma , Chan Yiong Huak , Koh Angela Fang Yung
Published: April 2014

Objectives:

The primary aims of the study were to examine the effect of resveratrol on skeletal muscle SIRT1 expression and energy expenditure in subjects with Type 2 diabetes mellitus (T2DM).

Background:

Animal and in vivo studies indicate that resveratrol increases SIRT1 expression that stimulates PGC1α activity. Subsequent upregulation of AMPK and GLUT4 expression are associated with improved insulin sensitivity in peripheral tissues.

Methods:

Ten subjects with T2DM were randomized in a double-blind fashion to receive 3g resveratrol or placebo daily for 12 weeks. Secondary outcomes include measures of AMPK, p-AMPK and GLUT4 expression levels, energy expenditure, physical activity levels, distribution of abdominal adipose tissue and skeletal muscle fiber type composition, body weight, HbA1c, plasma lipid subfraction, adiponectin levels, and insulin sensitivity.

Results:

There was a significant increase in both SIRT1 expression (2.01 vs. 0.86 arbitrary units [AU], p = .016) and p-AMPK to AMPK expression ratio (2.04 vs. 0.79 AU, p = .032) in the resveratrol group compared with the placebo group. Although the percentage of absolute change (8.6 vs. –13.9%, p = .033) and percentage of predicted resting metabolic rate (RMR; 7.8 vs. –13.9%, p = .013) were increased following resveratrol, there was a significant reduction in average daily activity (–38 vs. 43.2%, p = .028) and step counts (–39.5 vs. 11.8%, p = .047) when compared with placebo.

Conclusions:

In patients with T2DM, treatment with resveratrol regulates energy expenditure through increased skeletal muscle SIRT1 and AMPK expression. These findings indicate that resveratrol may have beneficial exercise-mimetic effects in patients with T2DM.
Effect of High-Intensity Intermittent Exercise on Postprandial Plasma Triacylglycerol in Sedentary Young Women
Authors: Tan Martin , Fat Rachel Chan Moy , Boutcher Yati N. , Boutcher Stephen H.
Published: April 2014

High-intensity intermittent exercise (HIIE) such as the 30-s Wingate test attenuates postprandial triacylglycerol (TG), however, the ability of shorter versions of HIIE to reduce postprandial TG is undetermined. Thus, the effect of 8-s sprinting bouts of HIIE on blood TG levels of 12 females after consumption of a high-fat meal (HFM) was examined. Twelve young, sedentary women (BMI 25.1 ± 2.3 kg/m2; age 21.3 ± 2.1 years) completed a maximal oxygen uptake test and then on different days underwent either an exercise or a no-exercise postprandial TG condition. Both conditions involved consuming a HFM after a 12-hr fast. The HFM, in milkshake form provided 4170 kJ (993 Kcal) of energy and 98 g fat. Order was counter-balanced. In the exercise condition participants completed 20-min of HIIE cycling consisting of repeated bouts of 8 s sprint cycling (100–115 rpm) and 12 s of active rest (easy pedaling) 14 hr before consuming the HFM. Blood samples were collected hourly after the HFM for 4 hr. Total postprandial TG was 13% lower, p = .004, in the exercise (5.84 ± 1.08 mmol L−1 4 h−1) compared with the no-exercise condition (6.71 ± 1.63 mmol L−1 4 h−1). In conclusion, HIIE significantly attenuated postprandial TG in sedentary young women.
The Effect of Feeding during Recovery from Aerobic Exercise on Skeletal Muscle Intracellular Signaling
Authors: Reidy Paul T. , Konopka Adam R. , Hinkley J. Matthew , Suer Miranda K. , Harber Matthew P.
Published: April 2014

We previously reported an increase in skeletal muscle protein synthesis during fasted and fed recovery from nonexhaustive aerobic exercise (Harber et al., 2010). The current study examined skeletal muscle intracellular signaling in the same subjects to further investigate mechanisms of skeletal muscle protein metabolism with and without feeding following aerobic exercise. Eight males (VO2peak: 52 ± 2 ml−1.kg−1.min−1) performed 60-min of cycle ergometry at 72 ± 1% VO2peak on two occasions in a counter-balanced design. Exercise trials differed only in the postexercise nutritional intervention: EX-FED (5kcal, 0.83g carbohydrate, 0.37g protein, 0.03g fat per kg body weight) and EX-FAST (noncaloric, isovolumic placebo) ingested immediately and one hour after exercise. Muscle biopsies were obtained from the vastus lateralis at rest (on a separate day) and two hours postexercise to assess intracellular signaling via western blotting of p70S6K1, eEF2, 4EBP1, AMPKα and p38 MAPK. p70S6K1 phosphorylation was elevated (p < .05) in EX-FED relative to REST and EX-FAST. eEF2, 4EBP1, AMPKα and p38 MAPK signaling were unaltered at 2h after exercise independent of feeding status when expressed as the ratio of phosphorylated to total protein normalized to actin. These data demonstrate that feeding after a nonexhaustive bout of aerobic exercise stimulates skeletal muscle p70S6K1 intracellular signaling favorable for promoting protein synthesis which may, as recent literature has suggested, better prepare the muscle for subsequent exercise bouts. These data provide further support into the role of feeding on mechanisms regulating muscle protein metabolism during recovery from aerobic exercise.
Validation of the Fitbit Wireless Activity Tracker for Prediction of Energy Expenditure
Authors: Sasaki Jeffer Eidi , Hickey Amanda , Mavilia Marianna , Tedesco Jacquelynne , John Dinesh , Keadle Sarah Kozey , Freedson Patty S.
Published: April 2014

Objective:

The purpose of this study was to examine the accuracy of the Fitbit wireless activity tracker in assessing energy expenditure (EE) for different activities.

Methods:

Twenty participants (10 males, 10 females) wore the Fitbit Classic wireless activity tracker on the hip and the Oxycon Mobile portable metabolic system (criterion). Participants performed walking and running trials on a treadmill and a simulated free-living activity routine. Paired t tests were used to test for differences between estimated (Fitbit) and criterion (Oxycon) kcals for each of the activities.

Results:

Mean bias for estimated energy expenditure for all activities was −4.5 ± 1.0 kcals/6 min (95% limits of agreement: −25.2 to 15.8 kcals/6 min). The Fitbit significantly underestimated EE for cycling, laundry, raking, treadmill (TM) 3 mph at 5% grade, ascent/descent stairs, and TM 4 mph at 5% grade, and significantly overestimated EE for carrying groceries. Energy expenditure estimated by the Fitbit was not significantly different than EE calculated from the Oxycon Mobile for 9 activities.

Conclusion:

The Fitbit worn on the hip significantly underestimates EE of activities. The variability in underestimation of EE for the different activities may be problematic for weight loss management applications since accurate EE estimates are important for tracking/monitoring energy deficit.
The Quality of School Physical Activity Policies Within Maryland and Virginia
Authors: Smith Erin M. , Wilburn Grace , Estabrooks Paul A.
Published: April 2014

Background:

Since the adoption of the Healthy, Hunger-Free Kids Act of 2010, many researchers have examined changes in the school nutrition environment; however, far less research has focused on the evaluation of physical activity (PA) policies within public schools.

Methods:

School district wellness policies (n = 144) of Virginia and Maryland were coded using a previously validated audit tool with a scale of 0 (weakest, least comprehensive) to 1 (strongest, most comprehensive).

Results:

Mean policy strength was weak (.20 ± .15), and, on average, policies were moderately comprehensive (.40 ± .22). The strongest (.73 ± .44) and most comprehensive (.79 ± .40) policy subgroup addressed daily recess in elementary schools. Virginia had significantly higher scores in 9 policy groups, while Maryland had higher significant policy scores in the 2 following groups: (1) the strength and comprehensiveness of a written physical education (PE) curriculum for each grade level (Ps < .05) and (2) the strength and comprehensiveness of addressing the use of PE waivers (Ps < .05).

Conclusions:

PA wellness policies in Maryland and Virginia are extremely weak and only moderately comprehensive; it is unlikely that these policies will significantly influence school-based PA.
Parental Practices Encouraging and Discouraging Physical Activity in Hong Kong Chinese Preschoolers
Authors: Suen Yi-nam , Cerin Ester , Wu Sin-lung
Published: April 2014

Background:

Regular participation in physical activity (PA) can help reduce the risk of overweight/obesity. Parental practices related to PA are modifiable determinants of preschoolers’ PA that are still not well understood, especially in non-Western cultures. This qualitative explorative study aimed to identify parental practices encouraging or discouraging PA in Hong Kong preschoolers.

Methods:

Nominal Group Technique (NGT) sessions (n = 45; 6 to 9/group), complemented by a focus group (n = 6) and individual interviews (n = 12), were conducted with primary caregivers (mainly parents) of Hong Kong preschoolers to investigate what parents do to encourage (4 groups) and discourage (2 groups) PA in children. The groups were stratified by low and high neighborhood socioeconomic status. Results: Participants generated 21 and 16 items describing practices encouraging and discouraging preschoolers’ PA, respectively. Parental provision of instrumental, motivational, and conditional support were thought to encourage child’s PA, while parental safety concerns, focus on academic achievement, lack of time and resources, and promotion of sedentary behaviors were thought to discourage child’s PA.

Conclusions:

Several parental practices that were deemed to encourage or discourage Hong Kong preschoolers’ PA were identified. These can assist with development of a culturally sensitive scale of PA parenting practices and inform future quantitative research.
Which School- and Home-Based Factors in Elementary School–Age Children Predict Physical Activity and Sedentary Behavior in Secondary School–Age Children? A Prospective Cohort Study
Authors: Rutten Cindy , Boen Filip , Seghers Jan
Published: April 2014

Background:

To examine which school- and home-based factors at age 11 to 12 (6th grade, elementary school) predict physical activity (PA) and sedentary behavior (SB) at age 13 to 14 (8th grade, secondary school).

Methods:

Data at both time points were collected from 472 children (mean age baseline = 10.97, SD age = 0.41) and their parents. Children and parents completed self-reported questionnaires. Children’s height and weight were measured and children wore a pedometer for 7 days.

Results:

Hierarchical regression analyses revealed that predominantly boys (β = −.11), children with higher levels of pedometer-determined PA (β = .44) and more parental logistic support (β = .11) at age 11 to 12 displayed higher levels of pedometer-determined PA at age 13 to 14 (R2 = 39.1%). Similar results emerged for self-reported moderate-to-vigorous PA (R2 = 36.7%). Finally, lower levels of screen-based SB at age 13 to 14 (R2 = 32.5%) were most strongly related to lower levels of screen-based SB (β = .41), a medium/high socioeconomic status (β = −.18), and higher levels of parental PA explicit modeling (β = −.18) at age 11 to 12.

Conclusions:

Children’s PA/SB and the supportive role of parents at age 11 to 12 are strong predictors of PA and screen-based SB at age 13 to 14.
Prevalence of Physically Active and Sedentary Adolescents in 10 Eastern Mediterranean Countries and its Relation With Age, Sex, and Body Mass Index
Authors: Subhi Lyutha K. Al , Bose Shekar , Ani Maraim F. Al
Published: April 2014

Background:

A cross-country profile of physical activity and sedentary behavior is lacking within Eastern Mediterranean region (EMR) counties. The objectives were to examine prevalence of physical activity and sedentary behavior among adolescents of 10 EMR countries, and to describe potential differences in the 2 factors by sex, age, and BMI.

Methods:

A total of 23,562 adolescents were included from 10 EMR counties based on completeness of data (physical activity, sedentary behavior, age, sex, weight and height) from the Global school-based student health survey (GSHS).

Results:

Overall prevalence of physical activity (19%) is low and sedentary behavior is high (29%), with significant differences among counties. Oman had the highest (26%) and Egypt had the lowest (9%) prevalence of active students. Prevalence of sedentary behavior was the highest in United Arab Emirates (40%) and lowest in Pakistan (8%). Physical activity was lower and sedentary behavior was higher among female adolescents. A linear trend was observed between BMI and both physical activity and sedentary behavior; a similar pattern was seen with age.

Conclusions:

There is a need for interventions to increase the prevalence of adolescents meeting physical activity recommendations in the 10 countries. More investigation is required to understand the cultural context of sex and BMI influence on activity patterns.
Effects of Leisure-Time and Occupational Physical Activity on Total Mortality Risk in NHANES III According to Sex, Ethnicity, Central Obesity, and Age
Authors: Richard Aline , Martin Brian , Wanner Miriam , Eichholzer Monika , Rohrmann Sabine
Published: April 2014

Background:

Associations of physical activity with all-cause mortality seem to be quite strong, but little is known about potential effect modifiers as sex, race/ethnicity, age, and obesity.

Methods:

Data of the Third National Health and Nutrition Examination Survey (NHANES III), conducted 1988−1994 with mortality follow-up until 2006, were used to compare mortality risk between different levels of leisure-time physical activity (LTPA) and occupational physical activity (OPA). Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs).

Results:

LTPA (n = 15,307) was inversely associated with all-cause mortality (HR 0.75, 95% CI 0.64−0.88 for regular vs. no LTPA). There was a statistically significant interaction with age (P = .03), with participants over 60 years of age benefitting more from regular or irregular LTPA. OPA was positively associated with all-cause mortality (HR 1.25, 95% CI 0.85−1.84 for high vs. low OPA), particularly among Mexican-Americans (HR 2.28, 95% CI 1.23−4.22); statistically significant interactions were observed for obesity and gender.

Conclusions:

LTPA clearly predicts all-cause mortality. However, associations between OPA and all-cause mortality are unclear and need further research with special regard to ethnic differences.
The Association Between Time Spent in Vigorous Physical Activity and Dietary Patterns in Adolescents: A Cross-Sectional Study
Authors: Durksen Anita , Downs Shauna , Mollard Rebecca , Forbes Laura , Ball Geoff D.C. , McGavock Jon
Published: April 2014

Background:

Physical activity interventions targeting weight status have yielded mixed results. This variability may be attributed to compensatory changes in dietary patterns after increasing physical activity (PA) levels. Therefore, we sought to determine whether dietary patterns varied with time spent in vigorous-intensity PA in youth.

Methods:

Cross-sectional analysis of 330 youth enrolled in a school-based prospective cohort in central Alberta. Physical activity was assessed with waist mounted accelerometers (Actical) worn for 7 days. Main outcomes included consumption of unhealthy foods and the unhealthy food index obtained from a validated web-based 24-hour dietary recall instrument. Secondary outcomes included macronutrient intake, food group (Canada’s Food Guide to Healthy Eating) intake, and diet quality.

Results:

Compared with youth participating in < 7 min/ day of vigorous physical activity, those achieving ≥ 7 min/day displayed no change in healthy or unhealthy food consumption. However, linear regression suggests a modest association between diet quality and vigorous-intensity PA.

Conclusion:

These data demonstrate that in this cohort of Canadian youth, time spent being physically active is associated with healthier dietary patterns and not with increased consumption of unhealthy foods.
Childhood Adversities and Socioeconomic Position as Predictors of Leisure-Time Physical Inactivity in Early Adulthood
Authors: Kestilä Laura , Mäki-Opas Tomi , Kunst Anton E , Borodulin Katja , Rahkonen Ossi , Prättälä Ritva
Published: April 2014

Background:

Limited knowledge exists on how childhood social, health-related and economic circumstances predict adult physical inactivity. Our aim was a) to examine how various childhood adversities and living conditions predict leisure-time physical inactivity in early adulthood and b) to find out whether these associations are mediated through the respondent’s own education.

Methods:

Young adults aged 18−29 were used from the Health 2000 Study of the Finnish. The cross-sectional data were based on interviews and questionnaires including retrospective information on childhood circumstances. The analyses were carried out on 68% of the original sample (N = 1894). The outcome measure was leisure-time physical inactivity.

Results:

Only a few of the 11 childhood adversities were related with physical activity in early adulthood. Having been bullied at school was associated with physical inactivity independently of the other childhood circumstances and the respondent’s own education. Low parental education predicted leisure-time physical inactivity in men and the association was mediated by the respondent´s own education. Respondents with only primary or vocational education were more likely to be physically inactive during leisure-time compared with those with secondary or higher education.

Conclusions:

There is some evidence that few specific childhood adversities, especially bullying at school, have long-lasting effects on physical activity levels.
Physical Activity Counseling by Primary Care Physicians: Attitudes, Knowledge, Implementation, and Perceived Success
Authors: Diehl Katharina , Mayer Manfred , Mayer Frank , Görig Tatiana , Bock Christina , Herr Raphael M. , Schneider Sven
Published: April 2014

Background:

In physical activity (PA) counseling, primary care physicians (PCPs) play a key role because they are in regular contact with large sections of the population and are important contact people in all health-related issues. However, little is known about their attitudes, knowledge, and perceived success, as well as about factors associated with the implementation of PA counseling.

Methods:

We collected data from 4074 PCPs including information on physician and practice characteristics, attitudes toward cardiovascular disease (CVD) prevention, and measures used during routine practice to prevent CVD. Here, we followed widely the established 5 A’s strategy (Assess, Advise, Agree, Assist, Arrange).

Results:

The majority (87.2%) of PCPs rated their own level of competence in PA counseling as ‘high,’ while 52.3% rated their own capability to motivate patients to increase PA as ‘not good.’ Nine of ten PCPs routinely provided at least 1 measure of the modified 5 A’s strategy, while 9.5% routinely used all 5 intervention strategies.

Conclusions:

The positive attitude toward PA counseling among PCPs should be supported by other stakeholders in the field of prevention and health promotion. An example would be the reimbursement of health counseling services by compulsory health insurance, which would enable PCPs to invest more time in individualized health promotion.
Active Transportation to and on Campus is Associated With Objectively Measured Fitness Outcomes Among College Students
Authors: Bopp Melissa , Bopp Christopher , Schuchert Megan
Published: April 2014

Background:

Active transportation (AT) has been associated with positive health outcomes, yet limited research has addressed this with college students, a population at-risk for inactivity. The purpose of this study was to examine the relationship between AT behavior and objectively measured fitness outcomes.

Methods:

A volunteer, convenience sample (n = 299) of college students from a large northeastern university completed a survey about their AT habits to and on campus and psychosocial constructs related to AT and participated in a laboratory-based fitness assessment (cardiovascular endurance, muscular strength and endurance, flexibility, body composition).Off-campus students were dichotomized as nonactive (0−1 AT trips/day) or active travelers (> 1 AT trips/day) to campus; t-tests compared nonactive and active travelers for psychosocial and fitness variables.

Results:

Students were 56.3% male, 79.2% non-Hispanic White, and primarily living off-campus (87%). Most students (n = 177, 59.2%) reported active travel between classes. Off-campus students were primarily active travelers (76.1%). Active travelers to campus had greater cardiovascular fitness (P = .005), were more flexible (P = .006) and had lower systolic blood pressure (P = .05) compared with nonactive travelers.

Conclusion:

This study documents a relationship between AT behavior and objectively measured fitness among college students and provides a rationale for targeting this behavior as a method for improving health outcomes.
Affective Response to Exercise and Preferred Exercise Intensity Among Adolescents
Authors: Schneider Margaret , Schmalbach Priel
Published: April 2014

Background:

Little information exists as to the exercise intensity that adolescents enjoy and whether identifiable subgroups of adolescents will choose higher-intensity exercise.

Methods:

Healthy adolescents (N = 74; mean age = 11.09 years) completed a cardiorespiratory fitness test, a moderate-intensity exercise task, and an exercise task at an intensity that felt “good.” Heart rate (HR), work rate (WR), and ratings of perceived exertion (RPE) were assessed every 3 minutes.

Results:

During the “feels good” task, adolescents exercised at a HR recognized as beneficial for cardiovascular health (mean HR = 66% to 72% of HR at VO2peak). Adolescents who experienced a positive affective shift during the moderate-intensity task engaged in higher-intensity exercise during the feels-good task as compared with those whose affective response to moderate-intensity exercise was neutral or negative (76% of peak HR vs. 70% of peak HR, P < .01).There was no difference between groups in RPE.

Conclusions:

Adolescents tend to select an exercise intensity associated with fitness benefits when afforded the opportunity to choose an intensity that feels good. An identified subgroup engaged in higher-intensity exercise without a commensurate perception of working harder. Encouraging adolescents to exercise at an intensity that feels good may increase future exercise without sacrificing fitness.
Comment on “A Complete Skull from Dmanisi, Georgia, and the Evolutionary Biology of Early
Homo
”
Authors: Schwartz Jeffrey H. , Tattersall Ian , Chi Zhang
Published: April 2014

Lordkipanidze
et al
. (Research Article, 18 October 2013, p. 326) conclude, from gross morphological comparisons and geometric-morphometric analysis of general shape, that the five hominid crania from Dmanisi in Georgia represent a single regional variant of
Homo erectus
. However, dental, mandibular, and cranial morphologies all suggest taxic diversity and, in particular, validate the previously named
H. georgicus
.
Detection of the Gravitational Lens Magnifying a Type Ia Supernova
Authors: Quimby Robert M. , Oguri Masamune , More Anupreeta , More Surhud , Moriya Takashi J. , Werner Marcus C. , Tanaka Masayuki , Folatelli Gaston , Bersten Melina C. , Maeda Keiichi , Nomoto Ken’ichi
Published: April 2014

Magnified Flare-Up

The rise and fall of the luminosity of a supernova detected in 2010 was typical for its class, but its apparent brightness was 30 times greater than similar events.

Quimby
et al.

(p.
396
) compared spectra from the time of peak brightness and after the supernova faded, from which they concluded that something was interfering with our line of sight to the supernova. A previously unknown foreground galaxy turned out to be acting as a lens, bending and magnifying the light from the supernova. Potentially, spacetime warping like this could allow direct testing of cosmic expansion.
The 30-nm Fiber Redux
Authors: Travers Andrew
Published: April 2014

Do alternating stacking modes of nucleosomes underlie the compaction of chromatin?

[Also see Research Article by

Song
et al.

]
Climate Outsider Finds Missing Global Warming
Authors: Kintisch Eli
Published: April 2014

New analyses show that major climate data sets have overstated the recent slowdown in global warming.
At NIH, Two Strikes Policy Is Out
Authors: Kaiser Jocelyn
Published: April 2014

Responding to complaints, the National Institutes of Health will no longer limit the number of times a researcher can resubmit a rejected grant proposal.
Potential Alzheimer's Drug Spurs Protein Recycling
Authors: Garber Ken
Published: April 2014

A research team has identified a potential new drug lead for Alzheimer's disease: a small molecule that stabilizes the components of retromers, molecular complexes that act like recycling bins in cells.
Tanks for the Batteries
Authors: Service Robert F.
Published: April 2014

The need to store energy from wind, solar, and other renewable energy sources could spark a revival of a dormant battery technology.
After the Deluge
Authors: Normile Dennis
Published: April 2014

Hard data and survivor interviews are helping researchers unravel the science of Supertyphoon Haiyan and its storm surge.
The Cyclone Addict
Authors: Normile Dennis
Published: April 2014

Storm chaser Josh Morgerman witnessed Supertyphoon Haiyan as it hit Tacloban, and he went back months later to document the storm's track and impact.
Aspirin and the Risk of Colorectal Cancer in Relation to the Expression of 15-Hydroxyprostaglandin Dehydrogenase (
HPGD
)
Authors: Fink Stephen P. , Yamauchi Mai , Nishihara Reiko , Jung Seungyoun , Kuchiba Aya , Wu Kana , Cho Eunyoung , Giovannucci Edward , Fuchs Charles S. , Ogino Shuji , Markowitz Sanford D. , Chan Andrew T.
Published: April 2014

Aspirin use is associated with a lower risk of colorectal cancer arising in association with high expression of
15-PGDH
(
HPGD
) in normal colon mucosa.
Educational Games for Malaria Diagnosis
Authors: Ozcan Aydogan
Published: April 2014

On World Malaria Day, BioGames will release its malaria cell image library to the public as an educational training module and expert scoring system.
Putting the Brakes on Inflammation to Control Sepsis
Authors: Round June L.
Published: April 2014

During sepsis, VEGF-C is induced by blood-borne bacterial products and binds to its receptor, down-regulating inflammation.
Human Antibody Responses to the Polyclonal Dryvax Vaccine for Smallpox Prevention Can Be Distinguished from Responses to the Monoclonal Replacement Vaccine ACAM2000
Authors: Pugh Christine , Keasey Sarah , Korman Lawrence , Pittman Phillip R. , Ulrich Robert G.
Published: April 2014

Dryvax (Wyeth Laboratories, Inc., Marietta, PA) is representative of the vaccinia virus preparations that were previously used for preventing smallpox. While Dryvax was highly effective, the national supply stocks were depleted, and there were manufacturing concerns regarding sterility and the clonal heterogeneity of the vaccine. ACAM2000 (Acambis, Inc./Sanofi-Pasteur Biologics Co., Cambridge, MA), a single-plaque-purified vaccinia virus derivative of Dryvax, recently replaced the polyclonal smallpox vaccine for use in the United States. A substantial amount of sequence heterogeneity exists within the polyclonal proteome of Dryvax, including proteins that are missing from ACAM2000. Reasoning that a detailed comparison of antibody responses to the polyclonal and monoclonal vaccines may be useful for identifying unique properties of each antibody response, we utilized a protein microarray comprised of approximately 94% of the vaccinia poxvirus proteome (245 proteins) to measure protein-specific antibody responses of 71 individuals receiving a single vaccination with ACAM2000 or Dryvax. We observed robust antibody responses to 21 poxvirus proteins in vaccinated individuals, including 11 proteins that distinguished Dryvax responses from ACAM2000. Analysis of protein sequences from Dryvax clones revealed amino acid level differences in these 11 antigenic proteins and suggested that sequence variation and clonal heterogeneity may contribute to the observed differences between Dryvax and ACAM2000 antibody responses.
Validation of a Microbead-Based Format for Spoligotyping of Legionella pneumophila
Authors: Gomgnimbou Michel Kiréopori , Ginevra Christophe , Peron-Cane Caroline , Versapuech Margaux , Refrégier Guislaine , Jacotin Nathalie , Sola Christophe , Jarraud Sophie
Published: April 2014

A 42-plex clustered regularly interspaced short palindromic repeat (CRISPR)-based typing technique (spoligotyping) was recently developed at the French National Reference Center for
Legionella
. It allows the subtyping of the
Legionella pneumophila
sequence type 1/Paris pulsotype. In this report, we present the transfer of the membrane-based spoligotyping technique to a microbead-based multiplexed format. This microbead-based high-throughput assay uses devices such as Luminex 200 or the recently launched Magpix system (Luminex Corp., Austin, TX). We designated this new technique LP-SPOL (for

L.

p

neumophila
spol
igotyping). We used two sets of samples previously subtyped by the membrane-based spoligotyping method to set up and validate the transfer on the two microbead-based systems. The first set of isolates (
n
= 56) represented the whole diversity of the CRISPR patterns known to date. These isolates were used for transfer setup (determination of spacer cutoffs for both devices). The second set of isolates (
n
= 245) was used to validate the transfer to the two microbead-based systems. The results obtained by the Luminex 200 system were 100% concordant with those obtained by the Magpix system for the 2 sets of isolates. In total, 10 discrepant results were observed when comparing the membrane-based method to the microbead-based method. These discrepancies were further resolved by repeating either the membrane-based or the microbead-based assay. This new assay is expected to play an emerging role for surveillance of
L. pneumophila
, starting with one of the most frequent genotypes, the sequence type 1/Paris pulsotype. However, the generalization of this typing method to all
L. pneumophila
strains is not feasible, since not all
L. pneumophila
strains contain CRISPRs.
Comparison of ESwab with Traditional Swabs for Detection of Methicillin-Resistant Staphylococcus aureus Using Two Different Walk-Away Commercial Real-Time PCR Methods
Authors: Silbert Suzane , Kubasek Carly , Uy Dominic , Widen Raymond
Published: April 2014

The ESwab system (Copan Diagnostics) was evaluated as a nasopharyngeal specimen collection device to be used for methicillin-resistant
Staphylococcus aureus
(MRSA) detection by the GeneXpert and BD Max MRSA assays. Different MRSA strains and dilutions of each strain were tested in triplicate. ESwabs proved to be a suitable collection system for the two assays tested.
Molecular Test Based on Isothermal Helicase-Dependent Amplification for Detection of the Clostridium difficile Toxin A Gene
Authors: Eckert Catherine , Holscher Eleonore , Petit Amandine , Lalande Valérie , Barbut Frédéric
Published: April 2014

The AmpliVue
Clostridium difficile
assay and a glutamate dehydrogenase (GDH)-
illumi
gene algorithm were evaluated using 308 diarrheal stool specimens of patients suspected of having
C. difficile
infection. Compared to the enriched toxigenic culture method, the sensitivities, specificities, and positive and negative predictive values of the AmpliVue
C. difficile
assay and the GDH-
illumi
gene-based algorithm were 91.7% (95% confidence interval [CI], 76.4 to 97.8), 100% (95% CI, 98.3 to 100), 100% (95% CI, 87 to 100), and 98.9% (95% CI, 96.6 to 99.7), respectively.
Bruker Biotyper Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry System for Identification of Nocardia, Rhodococcus, Kocuria, Gordonia, Tsukamurella, and Listeria Species
Authors: Hsueh Po-Ren , Lee Tai-Fen , Du Shin-Hei , Teng Shih-Hua , Liao Chun-Hsing , Sheng Wang-Hui , Teng Lee-Jene
Published: April 2014

We evaluated whether the Bruker Biotyper matrix-associated laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) system provides accurate species-level identifications of 147 isolates of aerobically growing Gram-positive rods (GPRs). The bacterial isolates included
Nocardia
(
n
= 74),
Listeria
(
n
= 39),
Kocuria
(
n
= 15),
Rhodococcus
(
n
= 10),
Gordonia
(
n
= 7), and
Tsukamurella
(
n
= 2) species, which had all been identified by conventional methods, molecular methods, or both. In total, 89.7% of
Listeria monocytogenes
, 80% of
Rhodococcus
species, 26.7% of
Kocuria
species, and 14.9% of
Nocardia
species (
n
= 11, all
N. nova
and
N. otitidiscaviarum
) were correctly identified to the species level (score values, ≥2.0). A clustering analysis of spectra generated by the Bruker Biotyper identified six clusters of
Nocardia
species, i.e., cluster 1 (
N. cyriacigeorgica
), cluster 2 (
N. brasiliensis
), cluster 3 (
N. farcinica
), cluster 4 (
N. puris
), cluster 5 (
N. asiatica
), and cluster 6 (
N. beijingensis
), based on the six peaks generated by ClinProTools with the genetic algorithm, i.e.,
m/z
2,774.477 (cluster 1),
m/z
5,389.792 (cluster 2),
m/z
6,505.720 (cluster 3),
m/z
5,428.795 (cluster 4),
m/z
6,525.326 (cluster 5), and
m/z
16,085.216 (cluster 6). Two clusters of
L. monocytogenes
spectra were also found according to the five peaks, i.e.,
m/z
5,594.85,
m/z
6,184.39, and
m/z
11,187.31, for cluster 1 (serotype 1/2a) and
m/z
5,601.21 and
m/z
11,199.33 for cluster 2 (serotypes 1/2b and 4b). The Bruker Biotyper system was unable to accurately identify
Nocardia
(except for
N. nova
and
N. otitidiscaviarum
),
Tsukamurella
, or
Gordonia
species. Continuous expansion of the MALDI-TOF MS databases to include more GPRs is necessary.
Determination of Disk Diffusion and MIC Quality Control Guidelines for GSK2140944, a Novel Bacterial Type II Topoisomerase Inhibitor Antimicrobial Agent
Authors: Ross James E. , Scangarella-Oman Nicole E. , Flamm Robert K. , Jones Ronald N.
Published: April 2014

GSK2140944 is a novel bacterial type II topoisomerase inhibitor in development for the treatment of conventional and biothreat pathogens, including Gram-positive pathogens and methicillin-resistant
Staphylococcus aureus
. This quality control study was performed to establish ranges for selected control strains:
S. aureus
ATCC 29213 and ATCC 25923,
Escherichia coli
ATCC 25922,
Haemophilus influenzae
ATCC 49247, and
Streptococcus pneumoniae
ATCC 49619. The control ranges will be crucial for the accurate evaluation of GSK2140944 potency as it progresses through clinical trial development.
Retrospective Study of Etiologic Agents Associated with Nonsuppurative Meningoencephalitis in Stranded Cetaceans in the Canary Islands
Authors: Sierra Eva , Sánchez Susan , Saliki Jeremiah T. , Blas-Machado Uriel , Arbelo Manuel , Zucca Daniele , Fernández Antonio
Published: April 2014

Nineteen natural cases of etiologically undetermined encephalitides in free-ranging cetaceans were studied retrospectively. Histological examination of the brains revealed variable degrees of nonsuppurative encephalitis or meningoencephalitis, characterized predominantly by perivascular lymphohistiocytic infiltrates. A PCR assay was used on brain and other available tissues to detect the presence of morbillivirus, herpesvirus, West Nile virus,
Toxoplasma gondii
, and
Brucella
spp. In addition, immunohistochemical (IHC) staining was performed on selected tissues to determine the presence of morbilliviral antigens. Six animals (5 striped dolphins and 1 common dolphin) showed IHC and/or molecular evidence of morbilliviral antigens and/or genomes, mainly in brain tissue. Conventional nested PCR detected herpesviral DNA in brain tissue samples from two striped dolphins. There was no evidence of West Nile virus,
T. gondii
, or
Brucella
spp. in any of the brain tissue samples examined. The information presented here increases the number of confirmed morbillivirus-positive cases within the Canarian archipelago from two previously reported cases to eight. Furthermore, a new nested-PCR method for the detection of morbillivirus is described here. Regarding herpesvirus, the phylogenetic analysis performed in the current study provides valuable information about a possible pathogenic branch of cetacean alphaherpesviruses that might be responsible for some fatal cases worldwide.
Ezrin Is a Component of the HIV-1 Virological Presynapse and Contributes to the Inhibition of Cell-Cell Fusion
Authors: Roy Nathan H. , Lambelé Marie , Chan Jany , Symeonides Menelaos , Thali Markus
Published: April 2014

During cell-to-cell transmission of HIV-1, viral and cellular proteins transiently accumulate at the contact zone between infected (producer) and uninfected (target) cells, forming the virological synapse. Rearrangements of the cytoskeleton in producer and target cells are required for proper targeting of viral and cellular components during synapse formation, yet little is known about how these processes are regulated, particularly within the producer cell. Since ezrin-radixin-moesin (ERM) proteins connect F-actin with integral and peripheral membrane proteins, are incorporated into virions, and interact with cellular components of the virological presynapse, we hypothesized that they play roles during the late stage of HIV-1 replication. Here we document that phosphorylated (i.e., active) ezrin specifically accumulates at the HIV-1 presynapse in T cell lines and primary CD4
+
lymphocytes. To investigate whether ezrin supports virus transmission, we sought to ablate ezrin expression in producer cells. While cells did not tolerate a complete knockdown of ezrin, even a modest reduction of ezrin expression (∼50%) in HIV-1-producing cells led to the release of particles with impaired infectivity. Further, when cocultured with uninfected target cells, ezrin-knockdown producer cells displayed reduced accumulation of the tetraspanin CD81 at the synapse and fused more readily with target cells, thus forming syncytia. Such an outcome likely is not optimal for virus dissemination, as evidenced by the fact that,
in vivo
, only relatively few infected cells form syncytia. Thus, ezrin likely helps secure efficient virus spread not only by enhancing virion infectivity but also by preventing excessive membrane fusion at the virological synapse.

IMPORTANCE
While viruses, in principal, can propagate through successions of syncytia, HIV-1-infected cells in the majority of cases do not fuse with potential target cells during viral transmission. This mode of spread is coresponsible for key features of HIV-1 pathogenesis, including killing of bystander cells and establishment of latently infected T lymphocytes. Here we identify the ERM protein family member ezrin as a cellular factor that contributes to the inhibition of cell-cell fusion and thus to suppressing excessive syncytium formation. Our analyses further suggest that ezrin, which connects integral membrane proteins with actin, functions in concert with CD81, a member of the tetraspanin family of proteins. Additional evidence, documented here and elsewhere, suggests that ezrin and CD81 cooperate to prevent cytoskeleton rearrangements that need to take place during the fusion of cellular membranes.
Phosphorylation of Herpes Simplex Virus 1 dUTPase Upregulated Viral dUTPase Activity To Compensate for Low Cellular dUTPase Activity for Efficient Viral Replication
Authors: Kato Akihisa , Hirohata Yoshitaka , Arii Jun , Kawaguchi Yasushi
Published: April 2014

We recently reported that herpes simplex virus 1 (HSV-1) protein kinase Us3 phosphorylated viral dUTPase (vdUTPase) at serine 187 (Ser-187) to upregulate its enzymatic activity, which promoted HSV-1 replication in human neuroblastoma SK-N-SH cells but not in human carcinoma HEp-2 cells. In the present study, we showed that endogenous cellular dUTPase activity in SK-N-SH cells was significantly lower than that in HEp-2 cells and that overexpression of cellular dUTPase in SK-N-SH cells increased the replication of an HSV-1 mutant with an alanine substitution for Ser-187 (S187A) in vdUTPase to the wild-type level. In addition, we showed that knockdown of cellular dUTPase in HEp-2 cells significantly reduced replication of the mutant vdUTPase (S187A) virus but not that of wild-type HSV-1. Furthermore, the replacement of Ser-187 in vdUTPase with aspartic acid, which mimics constitutive phosphorylation, and overexpression of cellular dUTPase restored viral replication to the wild-type level in cellular dUTPase knockdown HEp-2 cells. These results indicated that sufficient dUTPase activity was required for efficient HSV-1 replication and supported the hypothesis that Us3 phosphorylation of vdUTPase Ser-187 upregulated vdUTPase activity in host cells with low cellular dUTPase activity to produce efficient viral replication.virus.

IMPORTANCE
It has long been assumed that dUTPase activity is important for replication of viruses encoding a dUTPase and that the viral dUTPase (vdUTPase) activity was needed if host cell dUTPase activity was not sufficient for efficient viral replication. In the present study, we showed that the S187A mutation in HSV-1 vdUTPase, which impaired its enzymatic activity, reduced viral replication in SK-N-SH cells, which have low endogenous cellular dUTPase activity, and that overexpression of cellular dUTPase restored viral replication to the wild-type level. We also showed that knockdown of cellular dUTPase in HEp-2 cells, which have higher dUTPase activity than do SK-N-SH cells, reduced replication of HSV-1 with the vdUTPase mutation but had no effect on wild-type virus replication. This is the first report, to our knowledge, directly showing that dUTPase activity is critical for efficient viral replication and that vdUTPase compensates for low host cell dUTPase activity to produce efficient viral replication.
IRF7-Dependent Type I Interferon Production Induces Lethal Immune-Mediated Disease in STAT1 Knockout Mice Infected with Lymphocytic Choriomeningitis Virus
Authors: Li Wen , Hofer Markus J. , Jung So Ri , Lim Sue-Ling , Campbell Iain L.
Published: April 2014

Following systemic infection with lymphocytic choriomeningitis virus (LCMV), STAT1 knockout (KO) mice but not wild-type, STAT2 KO, IRF9 KO, or IFNAR KO mice develop lethal disease perpetrated by CD4+T cells. IRF7 is a key transcriptional activator of type I IFN (IFN-I) during LCMV infection. Here, the role of IRF7 in the lethal host response to LCMV infection in STAT1 KO mice was examined. In contrast to STAT1 KO mice, STAT1/IRF7 double KO (DKO) mice survived LCMV infection with a reduced immune pathology in key organs, such as the liver and spleen. However, similar to STAT1 KO mice, STAT1/IRF7 DKO mice failed to control LCMV replication and spread. LCMV infection in STAT1 KO mice was associated with a significant elevation in the levels of a number of cytokines in serum, including IFN-Is, but this was largely absent in STAT1/IRF7 DKO mice, which had a modest increase in the levels of gamma interferon and CCL2 only. Since IRF7 is known to be a key transcriptional regulator of IFN-I gene expression, the possible role of IFN-I in lethal disease was examined further. STAT1/IFNAR DKO mice, in contrast to STAT1 KO mice, all survived infection with LCMV and exhibited little tissue immune pathology. Additionally, STAT1 KO mice that were deficient for either of the two IFN-I signaling molecules, STAT2 or IRF9, also survived LCMV infection. We conclude that the lethal immune-mediated disease resulting from LCMV infection in STAT1 KO mice is (i) dependent on IRF7-induced IFN-I production and (ii) driven by noncanonical IFN-I signaling via STAT2 and IRF9.

IMPORTANCEHere we report on the basis for the novel, fatal immune-mediated disease of STAT1 KO mice infected with LCMV. Our findings show that, surprisingly, the pathogenesis of this disease is dependent on IRF7-mediated type I interferon production. Moreover, our study identifies noncanonical type I interferon signaling via STAT2 and IRF9 to be essential for the type I IFN-driven fatal disease in LCMV-infected STAT1 KO mice. These results further highlight the significance of noncanonical type I IFN signaling in the pathogenesis of host-mediated injury following viral infection.
Statistical Linkage Analysis of Substitutions in Patient-Derived Sequences of Genotype 1a Hepatitis C Virus Nonstructural Protein 3 Exposes Targets for Immunogen Design
Authors: Quadeer Ahmed A. , Louie Raymond H. Y. , Shekhar Karthik , Chakraborty Arup K. , Hsing I-Ming , McKay Matthew R.
Published: April 2014

Chronic hepatitis C virus (HCV) infection is one of the leading causes of liver failure and liver cancer, affecting around 3% of the world's population. The extreme sequence variability of the virus resulting from error-prone replication has thwarted the discovery of a universal prophylactic vaccine. It is known that vigorous and multispecific cellular immune responses, involving both helper CD4
+
and cytotoxic CD8
+
T cells, are associated with the spontaneous clearance of acute HCV infection. Escape mutations in viral epitopes can, however, abrogate protective T-cell responses, leading to viral persistence and associated pathologies. Despite the propensity of the virus to mutate, there might still exist substitutions that incur a fitness cost. In this paper, we identify groups of coevolving residues within HCV nonstructural protein 3 (NS3) by analyzing diverse sequences of this protein using ideas from random matrix theory and associated methods. Our analyses indicate that one of these groups comprises a large percentage of residues for which HCV appears to resist multiple simultaneous substitutions. Targeting multiple residues in this group through vaccine-induced immune responses should either lead to viral recognition or elicit escape substitutions that compromise viral fitness. Our predictions are supported by published clinical data, which suggested that immune genotypes associated with spontaneous clearance of HCV preferentially recognized and targeted this vulnerable group of residues. Moreover, mapping the sites of this group onto the available protein structure provided insight into its functional significance. An epitope-based immunogen is proposed as an alternative to the NS3 epitopes in the peptide-based vaccine IC41.

IMPORTANCE
Despite much experimental work on HCV, a thorough statistical study of the HCV sequences for the purpose of immunogen design was missing in the literature. Such a study is vital to identify epistatic couplings among residues that can provide useful insights for designing a potent vaccine. In this work, ideas from random matrix theory were applied to characterize the statistics of substitutions within the diverse publicly available sequences of the genotype 1a HCV NS3 protein, leading to a group of sites for which HCV appears to resist simultaneous substitutions possibly due to deleterious effect on viral fitness. Our analysis leads to completely novel immunogen designs for HCV. In addition, the NS3 epitopes used in the recently proposed peptide-based vaccine IC41 were analyzed in the context of our framework. Our analysis predicts that alternative NS3 epitopes may be worth exploring as they might be more efficacious.