bluuebunny/crossref_metadata_2025 huggingface dataset:
A tool for selecting patients with a high probability of sustained virological response to peginterferon alfa‐2a (40kD)/ribavirin
Authors: Ferenci Peter , Aires Rodrigo , Ancuta Ioan , Arohnson Andrew , Cheinquer Hugo , Delic Dragan , Gschwantler Michael , Larrey Dominique , Tallarico Ludovico , Schmitz Manuela , Tatsch Fernando , Ouzan Denis
Published: December 2013

Background & Aims

Pretreatment identification of patients likely to achieve a sustained virological response (SVR) with peginterferon alfa‐2a/ribavirin would be useful for individualizing treatment choices. The aim of this analysis was to devise a simple scoring system to identify patients with high probability of achieving an SVR with peginterferon alfa‐2a/ribavirin.

Methods

Using data from 2109 Caucasian treatment‐naive hepatitis C virus (HCV) genotype 1 mono‐infected patients from the PROPHESYS cohorts, the relationship between favourable baseline characteristics and SVR was explored using generalized additive model analysis, and a scoring system was devised to predict SVR.

Results

Points were assigned for: age (years) (≤35: 2; >35, ≤45: 1; >45: 0); body mass index (kg/m2) (≤20: 2; >20, ≤22: 1; >22: 0); HCV RNA (IU/ml) (≤100 000: 3; >100 000–400 000: 2; >400 000–800 000: 1; >800 000: 0); platelets (>150 ×109/l: 1; ≤150 ×109/l: 0); alanine aminotransferase [×upper limit of normal (ULN)] (>3: 1; ≤3: 0); serum aspartate aminotransferase (×ULN) (≤1: 1; >1: 0). 1029, 698 and 382 patients had scores of 0–2, 3–4 and ≥5, respectively, among whom SVR rates were 35.0, 54.9 and 76.7%. SVR in patients with scores ≥5 and undetectable HCV RNA by week 4 was 86.7%. The score was tested against two databases of patients who received peginterferon alfa‐2a/ribavirin in other clinical trials; similar high SVR rates in patients with scores ≥5 were reported.

Conclusions

The scoring system can reliably identify treatment‐naive HCV genotype 1 mono‐infected Caucasian patients who have a high probability of achieving an SVR with peginterferon alfa‐2a/ribavirin and will be particularly useful where protease inhibitors are not readily available.
Expression of toll‐like receptors 1–5 but not TLR 6–10 is elevated in livers of patients with non‐alcoholic fatty liver disease
Authors: Kanuri Giridhar , Ladurner Ruth , Skibovskaya Julia , Spruss Astrid , Königsrainer Alfred , Bischoff Stephan C. , Bergheim Ina
Published: December 2013

Background & Aims

Animal models of non‐alcoholic fatty liver disease (NAFLD) suggest that an increased translocation of bacterial endotoxins, leading to an activation of toll‐like receptor‐dependent signalling cascades (TLRs) and increased formation of reactive oxygen species, may add to development of insulin resistance and induction of plasminogen activator inhibitor‐1 (PAI‐1) in the liver. If similar mechanisms are also involved in the development of NAFLD in humans remains to be determined.

Methods

Toll‐like receptor (1–10), myeloid differentiation primary response gene (MyD88), interferon regulatory transcription factor 3 (IRF‐3) and insulin receptor substrate 1 (IRS‐1) mRNA expression was determined in liver samples of 11 patients with NAFLD and 11 controls. Hepatic PA1‐1 and 4‐hydroxynonenal protein adducts (4‐HNE) levels were determined by immunohistochemistry.

Results

Hepatic TLR 1–5 mRNAs expression was significantly higher in livers of NAFLD patients than in controls, whereas expression of TLR 6–10 mRNAs did not differ between groups. Expression of MyD88 but not IRF‐3 was also significantly higher in livers of NAFLD patients than in controls. These alterations were associated with significantly higher levels of 4‐HNE and PAI‐1 protein levels in livers of NAFLD patients than in controls, whereas IRS‐1 mRNA expression was ~80% lower in livers of NAFLD patients than in controls.

Conclusions

Taken together, these findings add further weight to the hypothesis that alterations at the level of intestine and intestinal barrier function may be critical in the development of NAFLD in humans.
Perspectives on Language as a Source of Social Markers
Authors: Roberts Gareth
Published: December 2013

Because linguistic forms vary between groups of speakers, language serves as a source of social markers, allowing people to distinguish between those who do and do not belong to the same social group. This review surveys interdisciplinary perspectives on four issues concerning social markers: (a) the role of inter‐ and intra‐individual variation, (b) the purpose of social markers, (c) language as an especially good source of social markers and (d) social marking as a source of new dialects.
Low‐Ir IAB irons from Morasko and other locations in central Europe: One fall, possibly distinct from IAB‐MG
Authors: Pilski A. S. , Wasson J. T. , Muszyński A. , Kryza R. , Karwowski Ł. , Nowak M.
Published: December 2013

Differences in texture and discovery location prompted us to analyze 16 irons from Morasko; one from Seeläsgen, known to have a similar composition; and a new mass found at Jankowo Dolne. These were analyzed in duplicate by instrumental neutron‐activation analysis (INAA). The results show that all 18 samples have very similar compositions, distinct from all other IAB irons except Burgavli; we conclude that they are all from a single shower. Eight of the samples were from regions with large amounts of cohenite (but were largely free of inclusions) and six were from samples with very little cohenite; we could find no resolvable difference in composition between these sets, a fact that suggests that the C contents of the metal phases were similar in the two areas. Although Morasko has been classified into the IAB main group (IAB‐MG), its Ir plots well outside the main group field on an Ir‐Au diagram. We considered the possibility that the low Ir reflected contamination by a melt from a IAB region that ponded and experienced fractional crystallization; however, because Morasko has Pt, W, and Ga values that are the same as the highest values in IAB‐MG, we rejected this model. We therefore conclude that Morasko formed from a different melt than the IAB‐MG irons; the Morasko melt was produced by impact heating, but one or more of the main Ir carriers did not melt, leaving much of the Ir in the unmelted residue. Copper is the only element that shows resolvable differences among Morasko samples. Most (13 of 18) samples have 149 ± 4 μg g−1 Cu, but three have 213 ± 10 μg g−1; we interpret this to mean that the low‐Cu samples have equilibrated with a Cu‐rich phase, whereas there was none of the latter phase within a few diffusion lengths of the samples with high Cu contents.
81Kr‐Kr cosmic ray exposure ages of individual chondrules from Allegan
Authors: Strashnov I. , Gilmour J. D.
Published: December 2013

81Kr‐Kr cosmic ray exposure (CRE) ages of individual chondrules (6–10 mg) and adjacent matrix samples (5–10 mg) from the Allegan H5 chondrite have been measured using a new highly sensitive resonance ionization mass spectrometer. No conclusive evidence of variations among the CRE ages of individual chondrules or between chondrules and matrix has been observed—average CRE ages of 5.90 ± 0.42 Ma (81Kr‐78Kr) and 5.04 ± 0.37 Ma (81Kr‐80+82Kr) are identical within error to those determined for the matrix (7.42 ± 1.27 Myr, 81Kr‐80+82Kr) and agree well with the literature value for bulk Allegan. If any accumulation of cosmogenic krypton in the early solar system took place, either it was below our detection limit in these samples (<100 atoms), or any such gas was lost during parent body metamorphism. However, this demonstration that useful 81Kr‐Kr ages can be obtained from few milligram samples of chondritic material has clear relevance to the analysis of samples returned by planned missions to asteroids and to the search for a signature of pre‐exposure in other, less processed meteorites.
I‐Xe ages of Campo del Cielo silicates as a record of the complex early history of theIABparent body
Authors: Pravdivtseva O. , Meshik A. , Hohenberg C. M. , Kurat G.
Published: December 2013

Using in situ laser analyses of a polished thin section from theIABiron meteorite Campo del Cielo, we identified two silicate grains rich in radiogenic129*Xe, Cr‐diopside, and oligoclase, excavated them from the metal, and irradiated them with thermal neutrons for I‐Xe dating. The release profiles of129*Xe and128*Xe are consistent with these silicates being diopside and oligoclase, with activation energies, estimated using Arrhenius plots, of ∼201 and ∼171 kcal mole−1, respectively. The 4556.4 ± 0.4 Ma absolute I‐Xe age of the more refractory diopside isyoungerthan the 4558.0 ± 0.7 Ma I‐Xe age of the less refractory oligoclase. We suggest that separate impact events at different locations and depths on a porous initial chondriticIABparent body led to the removal of the melt and recrystallization of diopside and oligoclase at the times reflected by their respective I‐Xe ages. The diopside and oligoclase grains were later brought into the studied inclusion by a larger scale catastrophic collision that caused breakup and reassembly of the debris, but did not reset the I‐Xe ages dating the first events. The metal melt most probably was <1250 °C when it surrounded studied silicate grains. This reassembly could not have occurred earlier than the I‐Xe closure in diopside at 4556.4 ± 0.4 Ma.
Dehydroascorbate: a possible surveillance molecule of oxidative stress and programmed cell death in the green alga Chlamydomonas reinhardtii
Authors: Murik Omer , Elboher Ahinoam , Kaplan Aaron
Published: December 2013

Chlamydomonas reinhardtii tolerates relatively high H2O2 levels that induce an array of antioxidant activities. However, rather than rendering the cells more resistant to oxidative stress, the cells become far more sensitive to an additional H2O2 dose.

If H2O2 is provided 1.5–9 h after an initial dose, it induces programmed cell death (PCD) in the wild‐type, but not in the dum1 mutant impaired in the mitochondrial respiratory complex III. This mutant does not exhibit a secondary oxidative burst 4–5 h after the inducing H2O2, nor does it activate metacaspase‐1 after the second H2O2 treatment.

The intracellular dehydroascorbate level, a product of ascorbate peroxidase, increases under conditions leading to PCD. The addition of dehydroascorbate induces PCD in the wild‐type and dum1 cultures, but higher levels are required in dum1 cells, where it is metabolized faster. The application of dehydroascorbate induces the expression of metacaspase‐2, which is much stronger than the expression of metacaspase‐1.

The presence or absence of oxidative stress, in addition to the rise in internal dehydroascorbate, may determine which metacaspase is activated during Chlamydomonas PCD. Cell death is strongly affected by the timing of H2O2 or dehydroascorbate admission to synchronously grown cultures, suggesting that the cell cycle phase may distinguish cells that perish from those that do not.
Photoacclimation Responses of the Brown MacroalgaSargassum Cymosumto the Combined Influence ofUVRadiation and Salinity: Cytochemical and Ultrastructural Organization and Photosynthetic Performance
Authors: Polo Luz K. , de L. Felix Marthiellen R. , Kreusch Marianne , Pereira Debora T. , Costa Giulia B. , Simioni Carmen , Ouriques Luciane C. , Chow Fungyi , Ramlov Fernanda , Maraschin Marcelo , Bouzon Zenilda L. , Schmidt Éder C.
Published: December 2013

The photoacclimation responses of the brown macroalgaSargassum cymosumwere studied to determine its cytochemical and ultrastructural organization, as well as photosynthetic pigments and performance.S. cymosumwas cultivated in three salinities (30, 35 and 40 psu) under four irradiation treatments:PAR‐only,PAR + UVA,PAR + UVBandPAR + UVA + UVB. Plants were exposed toPARat 70 μmol photons m−2 s−1,PAR + UVBat 0.35 W m−2andPAR +UVAat 0.70 W m−2for 3 h per day during 7 daysin vitro. Growth rate was not significantly affected by any type of radiation or salinity. The amount of pigments inS. cymosumwas significantly influenced by the interaction of salinity and radiation treatments. Compared withPAR‐only,UVRtreatments modified the kinetics patterns of the photosynthesis/irradiance curve. After exposure toUVR,S. cymosumincreased cell wall thickness and the presence of phenolic compounds. The number of mitochondria increased, whereas the number of chloroplasts showed few changes. AlthoughS. cymosumshowed insensitivity to changes in salinity, it can be concluded that samples treated under four irradiation regimes showed structural changes, which were more evident, but not severe, underPAR + UVBtreatment.
Fusarium crown rot under continuous cropping of susceptible and partially resistant wheat in microcosms at elevated CO2
Authors: Khudhair M. , Melloy P. , Lorenz D. J. , Obanor F. , Aitken E. , Datta S. , Luck J. , Fitzgerald G. , Chakraborty S.
Published: December 2013

This study examines the CO2‐mediated influence of plant resistance on crown rot dynamics under continuous cropping of partially resistant wheat line 249 and the susceptible cultivar Tamaroi. Disease incidence, severity, deoxynivalenol and Fusarium biomass were assessed after each cycle in microcosms established at ambient and 700 mg kg−1 CO2 using soil and stubble of these wheat lines from a field experiment with free to air CO2 enrichment. Monoconidial isolates from wheat stubble were collected initially, and after five cropping cycles, to compare the frequency and aggressiveness of Fusarium species in the two populations. Aggressiveness was measured using a high‐throughput seedling bioassay. At elevated CO2, the higher initial incidence in Tamaroi increased with cropping cycles, but incidence in 249 remained unchanged. Incidence at ambient CO2 did not change for either line. Elevated CO2 induced partial resistance in Tamaroi, but not in 249. Increased Fusarium biomass in wheat tissue at elevated CO2 matched raised deoxynivalenol of the stem base in both lines. After five cycles of continuous wheat cropping, aggressiveness increased in pathogenic F. culmorum and F. pseudograminearum by 110%, but decreased in weakly pathogenic F. equiseti and F. oxysporum by 50%. CO2 and host resistance interactively influenced species frequency, and the highly aggressive F. pseudograminearum became dominant on Tamaroi irrespective of CO2 concentration, while its frequency declined on 249. This study shows that induced resistance at elevated CO2 will not reduce crown rot severity, or impede the selection and enrichment of Fusarium populations with increased aggressiveness.
How to Improve the Domain of Starting Points for Steffensen's Method
Authors: Ezquerro J. A. , Hernández‐Verón M. A.
Published: December 2013

We analyze the semilocal convergence of Steffensen's method, using a novel technique, which is based on recurrence relations, for solving systems of nonlinear equations. This technique allows analyzing the convergence of Steffensen's method to solutions of equations, where the function involved can be both differentiable and nondifferentiable. Moreover, this technique also allows enlarging the domain of starting points for Steffensen's method from certain predictions with the simplified Steffensen method.
The structure analysis of the Norway spruce stands after schematic thinning in the 1st protected zone of water-supply storage
Authors: Bátor M. , Danková L.
Published: December 2013

The water-supply storages are important resources of drinking water in some regions of Slovakia, especially in surrounding of Banská Bystrica. Málinec is one of the three storages in this region. In protected zone of this storage are planted Norway spruce (hereinafter „spruce“) stands monocultures because of ensuring required water quality in storage. There are 24 years old spruce stands without admixture of deciduous trees in the upper layer. Understory is formed by blackthorn (PRP 1). On this locality was carried out geometric (schematic) thinning what is not recommended in protected zones because of worse influencing of soil permeability, humus decomposition and open forest edge creating. There was identified one older permanent research plot (PRP) and established one new PRP where was performed own measurements. Into the PRPs there are transekts and remaining area. Acreage of PRP is 0,234 and 0,315 ha. On the whole area were measured tree diameter and assessed tree classes (5 degrees–Polanský 1955). Moreover, on transekts were measured tree and also crown height. Based of measured variables were calculated stand characteristics like slenderness ratio, crown ratio, growing stock and basal area. If it is possible, these were compared with older results. The aim of this work is evaluation of spruce stands structure and their stability after schematic thinning. This kind of silvicultural treatment is absolutely unfit in stands with specific (water-protested) function (Slodičák et al. 2010). These stands are from the perspective of static stability too unfit–basal area and abundance of trees is nearly as in yield tables (Kolektív 1992) but growing stock is much higher and a lot of trees is too slim. The slenderness ratio is from 75 to 90 %. But the crown lenght is quite good with crown ratio 52–64 %. In these ages of stands are crowns reduced too much. Suitable individual thinning has been in these stands neglected (growing stock is higher than normal). There is assumed reduced stand lifetime after schematic thinning. Current stand structure is due to the young age inappropriate.
Development of cambioxylophagous insect fauna on blue spruce after chemical thinning
Authors: Kula E. , Kajfosz R. , Polívka J.
Published: December 2013

Chemical thinning performed in stands of blue spruce (Picea pungens) in air-polluted areas of the Ore Mountains does not pose a significant threat of sub-bark pest outbreaks. Application of the herbicide Roundup in autumn and in spring caused dieback of the treated trees where mainly Hylurgops palliatus became active and completed its development. Pests Ips amitinus and Pityogenes chalcographus died mostly at larval stage, despite their high abundance. Cryphalus abietis completed its development only on thicker branches and was not able to occupy tops of trees due to fast dieback of phloem. Quality of phloem was strongly affected by the speed of defoliation.
Structural diversity and regeneration patterns of the Norway spruce (Picea abies (L.) Karst) dominated virgin forest in NNR Zadná Poľana
Authors: Saniga M. , Balanda Miroslav , Pittner J. , Kucbel S. , Jaloviar P.
Published: December 2013

The paper analyzes the structural diversity of the spruce dominated old-growth forest in National Nature Reserve (NNR) Zadná Poľana on the series of three permanent research plots (PRP) with the area of 0.5 ha each. The height and diameter diversity as well as the spatial distribution were expressed using standard structural indices (Füldner’s TH and TD index, Gini index, Clark-Evans index). Obtained results confirmed the low structural diversity regardless of the developmental stage of investigated forest. The extensive ungulate browsing retards the height growth of the subsequent tree generation and leads toward to the structural homogenization of the future forest stand.
Autonomic, locomotor and cardiac abnormalities in a mouse model of muscular dystrophy: targeting the renin–angiotensin system
Authors: Sabharwal Rasna , Chapleau Mark W.
Published: December 2013

New Findings

What is the topic of this review?

This symposium report summarizes autonomic, cardiac and skeletal muscle abnormalities in sarcoglycan‐δ‐deficient mice (Sgcd−/−), a mouse model of limb girdle muscular dystrophy, with emphasis on the roles of autonomic dysregulation and activation of the renin–angiotensin system at a young age.

What advances does it highlight?

The contributions of the autonomic nervous system and the renin–angiotensin system to the pathogenesis of muscular dystrophy are highlighted. Results demonstrate that autonomic dysregulation precedes and predicts later development of cardiac dysfunction in Sgcd−/− mice and that treatment of young Sgcd−/− mice with the angiotensin type 1 receptor antagonist losartan or with angiotensin‐(1–7) abrogates the autonomic dysregulation, attenuates skeletal muscle pathology and increases spontaneous locomotor activity.

Muscular dystrophies are a heterogeneous group of genetic muscle diseases characterized by muscle weakness and atrophy. Mutations in sarcoglycans and other subunits of the dystrophin–glycoprotein complex cause muscular dystrophy and dilated cardiomyopathy in animals and humans. Aberrant autonomic signalling is recognized in a variety of neuromuscular disorders. We hypothesized that activation of the renin–angiotensin system contributes to skeletal muscle and autonomic dysfunction in mice deficient in the sarcoglycan‐δ (Sgcd) gene at a young age and that this early autonomic dysfunction contributes to the later development of left ventricular (LV) dysfunction and increased mortality. We demonstrated that young Sgcd−/− mice exhibit histopathological features of skeletal muscle dystrophy, decreased locomotor activity and severe autonomic dysregulation, but normal LV function. Autonomic regulation continued to deteriorate in Sgcd−/− mice with age and was accompanied by LV dysfunction and dilated cardiomyopathy at older ages. Autonomic dysregulation at a young age predicted later development of LV dysfunction and higher mortality in Sgcd−/− mice. Treatment of Sgcd−/− mice with the angiotensin type 1 receptor blocker losartan for 8–9 weeks, beginning at 3 weeks of age, decreased fibrosis and oxidative stress in skeletal muscle, increased locomotor activity and prevented autonomic dysfunction. Chronic infusion of the counter‐regulatory peptide angiotensin‐(1–7) resulted in similar protection. We conclude that activation of the renin–angiotensin system, at a young age, contributes to skeletal muscle and autonomic dysfunction in muscular dystrophy. We speculate that the latter is mediated via abnormal sensory nerve and/or cytokine signalling from dystrophic skeletal muscle to the brain and contributes to age‐related LV dysfunction, dilated cardiomyopathy, arrhythmias and premature death. Therefore, correcting the early autonomic dysregulation and renin–angiotensin system activation may provide a novel therapeutic approach in muscular dystrophy.
Elevated levels of branched‐chain amino acids have little effect on pancreatic islet cells, but l‐arginine impairs function through activation of the endoplasmic reticulum stress response
Authors: Mullooly Niamh , Vernon Wendy , Smith David M. , Newsholme Philip
Published: December 2013

New Findings

What is the central question of this study?

Recent studies have demonstrated strong correlations between circulating branched‐chain amino acid (AA) levels and insulin resistance, a predictor of susceptibility to type 2 diabetes. This study investigated how elevations in various AA concentrations over a chronic incubation period affect rat pancreatic islet cell viability and insulin secretion.

What is the main finding and its importance?

We report that chronic exposure to specific AAs provoked islet cell toxic effects, particularly the insulinotropic AA l‐arginine. Dose‐dependent decreases in insulin secretion and cell proliferation were observed in addition to activation of the cell endoplasmic reticulum stress response. These results show, for the first time, that elevated l‐arginine levels may impact negatively on islet function, providing further insights into β‐cell dysfunction in type 2 diabetes.

Recent metabolic profiling studies have identified a correlation between branched‐chain amino acid levels, insulin resistance associated with prediabetes and susceptibility to type 2 diabetes. Glucose and lipids in chronic excess have been reported to induce toxic effects in pancreatic β‐cells, but the effect of elevated amino acid concentrations on primary islet cell function has not been investigated to date. The aim of this study was to investigate the effect of chronic exposure to various amino acids on islet cell function in vitro. Isolated rat islets were incubated over periods of 48 h with a range of concentrations of individual amino acids (0.1 μm to 10 mm). After 48 h, islets were assessed for glucose‐dependent insulin secretion capacity, proliferation or islet cell apoptosis. We report that elevated levels of branched‐chain amino acids have little effect on pancreatic islet cell function or viability; however, increased levels of the amino acid l‐arginine were found to be β‐cell toxic, causing a dose‐dependent decrease in insulin secretion accompanied by a decrease in islet cell proliferation and an increase in islet cell apoptosis. These effects were not due to l‐arginine‐dependent increases in production of nitric oxide but arose through elicitation of the islet cell endoplasmic reticulum stress response. This novel finding indicates, for the first time, that the l‐arginine concentration in vitro may impact negatively on islet cell function, thus indicating further complexity in relationship to in vivo susceptibility of β‐cells to nutrient‐induced dysfunction.
Efficacy of Using Physical Activity Mentors to Increase the Daily Steps of Older Adults in the Primary Care Setting: A Pilot Study
Authors: Croteau Karen A. , Suresh Vijiayurani , Farnham Elanna
Published: December 2013

The purpose of this pilot study was to determine if using physical activity (PA) mentors has any additional impact on daily steps of older adults participating in the Maine in Motion (MIM) program in the primary care setting. Participants were randomly assigned to a MIM-only group (n = 14) or a MIM+ mentor group (n = 14). The MIM intervention lasted 6 months with follow-up at 12 months. Average age of participants was 64 ± 8.8 years and most participants had multiple chronic illnesses. At baseline, mean body mass index (BMI) was 32.2 ± 5.1 and average daily steps were 4,236 ± 2,266. Repeated-measures ANOVA revealed significant main effects for steps, F(2.324, 59.104) = 4.168, p = .015, but no main effects for group, F(1, 25) = 2.988, p = .096, or time-by-group interaction, F(2.324, 59.104) = 0.905, p = .151. All participants significantly increased daily steps over the course of the intervention, with MIM+ participants maintaining increases at follow-up. No significant findings were found for BMI.
Barriers to Physical Activity and Restorative Care for Residents in Long-Term Care: A Review of the Literature
Authors: Benjamin Kathleen , Edwards Nancy , Ploeg Jenny , Legault Frances
Published: December 2013

Despite the benefits of physical activity, residents living in long-term care (LTC) are relatively sedentary. Designing successful physical activity and restorative care programs requires a good understanding of implementation barriers. A database search (2002–2013) yielded seven studies (nine articles) that met our inclusion criteria. We also reviewed 31 randomized controlled trials (RCTs) to determine if the authors explicitly discussed the barriers encountered while implementing their interventions. Eleven RCTs (13 articles) included a discussion of the barriers. Hence, a total of 18 studies (22 articles) were included in this review. Barriers occurred at resident (e.g., health status), environmental (e.g., lack of space for physical activity), and organizational (e.g., staffing and funding constraints) levels. These barriers intersect to adversely affect the physical activity of older people living in LTC. Future studies targeting physical activity interventions for residents living in LTC are needed to address these multiple levels of influence.
How Do Older Adults Respond to Active Australia Physical Activity Questions? Lessons From Cognitive Interviews
Authors: Heesch Kristiann C. , van Uffelen Jannique , Brown Wendy J.
Published: December 2013

The aim of this study was to examine older adults’ understanding and interpretation of a validated questionnaire for physical activity surveillance, the Active Australia Survey (AAS). To address this aim, cognitive interviewing techniques were used during face-to-face semistructured interviews with 44 adults age 65–89 years. Qualitative data analysis revealed that participants were confused with questionnaire phrasing, misunderstood the scope of activities to include in answers, and misunderstood the time frame of activities to report. They also struggled to accurately estimate the frequency and duration of their activities. Our findings suggest that AAS questions may be interpreted differently by older adults than intended by survey developers. Findings also suggest that older adults use a range of methods for calculating PA frequency and duration. The issues revealed in this study may be useful for adapting AAS for use in older community-dwelling adults.
Normative Functional Fitness Standards and Trends of Portuguese Older Adults: Cross-Cultural Comparisons
Authors: Marques Elisa A. , Baptista Fátima , Santos Rute , Vale Susana , Santos Diana A. , Silva Analiza M. , Mota Jorge , Sardinha Luís B.
Published: December 2013

This cross-sectional study was designed to develop normative functional fitness standards for the Portuguese older adults, to analyze age and gender patterns of decline, to compare the fitness level of Portuguese older adults with that of older adults in other countries, and to evaluate the fitness level of Portuguese older adults relative to recently published criterion fitness standards associated with maintaining physical independence. A sample of 4,712 independent-living older adults, age 65–103 yr, was evaluated using the Senior Fitness Test battery. Age-group normative fitness scores are reported for the 10th, 25th, 50th, 75th, and 90th percentiles. Results indicate that both women and men experience age-related losses in all components of functional fitness, with their rate of decline being greater than that observed in other populations, a trend which may cause Portuguese older adults to be at greater risk for loss of independence in later years. These newly established normative standards make it possible to assess individual fitness level and provide a basis for implementing population-wide health strategies to counteract early loss of independence.
Association of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) Elements with Specific Serotypes and Virulence Potential of Shiga Toxin-Producing Escherichia coli
Authors: Toro Magaly , Cao Guojie , Ju Wenting , Allard Marc , Barrangou Rodolphe , Zhao Shaohua , Brown Eric , Meng Jianghong
Published: December 2013

Shiga toxin-producing
Escherichia coli
(STEC) strains (
n
= 194) representing 43 serotypes and
E. coli
K-12 were examined for clustered regularly interspaced short palindromic repeat (CRISPR) arrays to study genetic relatedness among STEC serotypes. A subset of the strains (
n
= 81) was further analyzed for subtype I-E
cas
and virulence genes to determine a possible association of CRISPR elements with potential virulence. Four types of CRISPR arrays were identified. CRISPR1 and CRISPR2 were present in all strains tested; 1 strain also had both CRISPR3 and CRISPR4, whereas 193 strains displayed a short, combined array, CRISPR3-4. A total of 3,353 spacers were identified, representing 528 distinct spacers. The average length of a spacer was 32 bp. Approximately one-half of the spacers (54%) were unique and found mostly in strains of less common serotypes. Overall, CRISPR spacer contents correlated well with STEC serotypes, and identical arrays were shared between strains with the same H type (O26:H11, O103:H11, and O111:H11). There was no association identified between the presence of subtype I-E
cas
and virulence genes, but the total number of spacers had a negative correlation with potential pathogenicity (
P
< 0.05). Fewer spacers were found in strains that had a greater probability of causing outbreaks and disease than in those with lower virulence potential (
P
< 0.05). The relationship between the CRISPR-
cas
system and potential virulence needs to be determined on a broader scale, and the biological link will need to be established.
The Presence of Conjugative Plasmid pLS20 Affects Global Transcription of Its Bacillus subtilis Host and Confers Beneficial Stress Resistance to Cells
Authors: Rösch Thomas C. , Golman Wladislaw , Hucklesby Laura , Gonzalez-Pastor Jose E. , Graumann Peter L.
Published: December 2013

Conjugation activity of plasmid pLS20 from
Bacillus subtilis
subsp.
natto
is induced when cells are diluted into fresh medium and diminishes as cells enter into stationary-phase growth. Transcriptional profiling shows that during mid-exponential growth, more than 5% of the host genes are affected in the presence of the plasmid, in contrast to the minor changes seen in freshly diluted and stationary-phase cells. Changes occurred in many metabolic pathways, although pLS20 does not confer any detectable burden on its host cell, as well as in membrane and cell wall-associated processes, in the large motility operon, and in several other cellular processes. In agreement with these changes, we found considerable alterations in motility and enzyme activity and increased resistance against several different forms of stress in cells containing the plasmid, revealing that the presence of pLS20 has a broad impact on the physiology of its host cell and increases its stress resistance in multiple aspects. Additionally, we found that the lack of chromosomal gene
yueB
, known to encode a phage receptor protein, which is upregulated in cells containing pLS20, strongly reduced conjugation efficiency, revealing that pLS20 not only increases fitness of its host but also employs host proteins for efficient transfer into a new cell.
Ubiquity of Insect-Derived Nitrogen Transfer to Plants by Endophytic Insect-Pathogenic Fungi: an Additional Branch of the Soil Nitrogen Cycle
Authors: Behie Scott W. , Bidochka Michael J.
Published: December 2013

The study of symbiotic nitrogen transfer in soil has largely focused on nitrogen-fixing bacteria. Vascular plants can lose a substantial amount of their nitrogen through insect herbivory. Previously, we showed that plants were able to reacquire nitrogen from insects through a partnership with the endophytic, insect-pathogenic fungus
Metarhizium robertsii
. That is, the endophytic capability and insect pathogenicity of
M. robertsii
are coupled so that the fungus acts as a conduit to provide insect-derived nitrogen to plant hosts. Here, we assess the ubiquity of this nitrogen transfer in five
Metarhizium
species representing those with broad (
M. robertsii
,
M. brunneum
, and
M. guizhouense
) and narrower insect host ranges (
M. acridum
and
M. flavoviride
), as well as the insect-pathogenic fungi
Beauveria bassiana
and
Lecanicillium lecanii
. Insects were injected with
15
N-labeled nitrogen, and we tracked the incorporation of
15
N into two dicots, haricot bean (
Phaseolus vulgaris
) and soybean (
Glycine max
), and two monocots, switchgrass (
Panicum virgatum
) and wheat (
Triticum aestivum
), in the presence of these fungi in soil microcosms. All
Metarhizium
species and
B. bassiana
but not
L. lecanii
showed the capacity to transfer nitrogen to plants, although to various degrees. Endophytic association by these fungi increased overall plant productivity. We also showed that in the field, where microbial competition is potentially high,
M. robertsii
was able to transfer insect-derived nitrogen to plants.
Metarhizium
spp. and
B. bassiana
have a worldwide distribution with high soil abundance and may play an important role in the ecological cycling of insect nitrogen back to plant communities.
Survival and Transmission of Potato Virus Y, Pepino Mosaic Virus, and Potato Spindle Tuber Viroid in Water
Authors: Mehle N. , Gutiérrez-Aguirre I. , Prezelj N. , Delić D. , Vidic U. , Ravnikar M.
Published: December 2013

Hydroponic systems and intensive irrigation are used widely in horticulture and thus have the potential for rapid spread of water-transmissible plant pathogens. Numerous plant viruses have been reported to occur in aqueous environments, although information on their survival and transmission is minimal, due mainly to the lack of effective detection methods and to the complexity of the required transmission experiments. We have assessed the role of water as a source of plant infection using three mechanically transmissible plant pathogens that constitute a serious threat to tomato and potato production: pepino mosaic virus (PepMV), potato virus Y (PVY), and potato spindle tuber viroid (PSTVd). PepMV remains infectious in water at 20 ± 4°C for up to 3 weeks, PVY (NTN strain) for up to 1 week, and PSTVd for up to 7 weeks. Experiments using a hydroponic system show that PepMV (Ch2 genotype) and PVY (NTN strain) can be released from plant roots into the nutrient solution and can infect healthy plants through their roots, ultimately spreading to the green parts, where they can be detected after a few months. In addition, tubers developed on plants grown in substrate watered with PSTVd-infested water were confirmed to be the source of viroid infection. Our data indicate that although well-known pathways of virus spread are more rapid than water-mediated infection, like insect or mechanical transmission through leaves, water is a route that provides a significant bridge for rapid virus/viroid spread. Consequently, water should be taken into account in future epidemiology and risk assessment studies.
Analysis of the Transcriptional Regulator GlpR, Promoter Elements, and Posttranscriptional Processing Involved in Fructose-Induced Activation of the Phosphoenolpyruvate-Dependent Sugar Phosphotransferase System in Haloferax mediterranei
Authors: Cai Lei , Cai Shuangfeng , Zhao Dahe , Wu Jinhua , Wang Lei , Liu Xiaoqing , Li Ming , Hou Jing , Zhou Jian , Liu Jingfang , Han Jing , Xiang Hua
Published: December 2013

Among all known archaeal strains, the phosphoenolpyruvate-dependent phosphotransferase system (PTS) for fructose utilization is used primarily by haloarchaea, which thrive in hypersaline environments, whereas the molecular details of the regulation of the archaeal PTS under fructose induction remain unclear. In this study, we present a comprehensive examination of the regulatory mechanism of the fructose PTS in the haloarchaeon
Haloferax mediterranei
. With gene knockout and complementation, microarray analysis, and chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR), we revealed that GlpR is the indispensable activator, which specifically binds to the PTS promoter (P

PTS

) during fructose induction. Further promoter-scanning mutation indicated that three sites located upstream of the
H. mediterranei
P

PTS

, which are conserved in most haloarchaeal P

PTS

s, are involved in this induction. Interestingly, two PTS transcripts (named T
8
and T
17
) with different lengths of 5′ untranslated region (UTR) were observed, and promoter or 5′ UTR swap experiments indicated that the shorter 5′ UTR was most likely generated from the longer one. Notably, the translation efficiency of the transcript with this shorter 5′ UTR was significantly higher and the ratio of T
8
(with the shorter 5′ UTR) to T
17
increased during fructose induction, implying that a posttranscriptional mechanism is also involved in PTS activation. With these insights into the molecular regulation of the haloarchaeal PTS, we have proposed a working model for haloarchaea in response to environmental fructose.
Cross-Institute Evaluations of Inhibitor-Resistant PCR Reagents for Direct Testing of Aerosol and Blood Samples Containing Biological Warfare Agent DNA
Authors: Minogue Timothy D. , Rachwal Phillip A. , Trombley Hall Adrienne , Koehler Jeffery W. , Weller Simon A.
Published: December 2013

Rapid pathogen detection is crucial for the timely introduction of therapeutics. Two groups (one in the United Kingdom and one in the United States) independently evaluated inhibitor-resistant PCR reagents for the direct testing of substrates. In the United Kingdom, a multiplexed
Bacillus anthracis
(target) and
Bacillus subtilis
(internal-control) PCR was used to evaluate 4 reagents against 5 PCR inhibitors and down-selected the TaqMan Fast Virus 1-Step master mix (Life Technologies Inc.). In the United States, four real-time PCR assays (targeting
B. anthracis
,
Brucella melitensis
, Venezuelan equine encephalitis virus [VEEV], and
Orthopoxvirus
spp.) were used to evaluate 5 reagents (plus the Fast Virus master mix) against buffer, blood, and soil samples and down-selected the KAPA Blood Direct master mix (KAPA Biosystems Inc.) with added Platinum
Taq
(Life Technologies). The down-selected reagents underwent further testing. In the United Kingdom experiments, both reagents were tested against seven contrived aerosol collector samples containing
B. anthracis
Ames DNA and
B. subtilis
spores from a commercial formulation (BioBall). In PCR assays with reaction mixtures containing 40% crude sample, an airfield-collected sample induced inhibition of the
B. subtilis
PCR with the KAPA reagent and complete failure of both PCRs with the Fast Virus reagent. However, both reagents allowed successful PCR for all other samples—which inhibited PCRs with a non-inhibitor-resistant reagent. In the United States, a cross-assay limit-of-detection (LoD) study in blood was conducted. The KAPA Blood Direct reagent allowed the detection of agent DNA (by four PCRs) at higher concentrations of blood in the reaction mixture (2.5%) than the Fast Virus reagent (0.5%), although LoDs differed between assays and reagent combinations. Across both groups, the KAPA Blood Direct reagent was determined to be the optimal reagent for inhibition relief in PCR.
Functional Analysis of Environmental DNA-Derived Microviridins Provides New Insights into the Diversity of the Tricyclic Peptide Family
Authors: Gatte-Picchi Douglas , Weiz Annika , Ishida Keishi , Hertweck Christian , Dittmann Elke
Published: December 2013

Microviridins represent a unique family of ribosomally synthesized cage-like depsipeptides from cyanobacteria with potent protease-inhibitory activities. The natural diversity of these peptides is largely unexplored. Here, we describe two methodologies that were developed to functionally characterize cryptic microviridin gene clusters from metagenomic DNA. Environmental samples were collected and enriched from cyanobacterial freshwater blooms of different geographical origins containing predominantly
Microcystis
sp. Microviridins were produced either directly from fosmid clones or after insertion of environmental DNA-derived gene cassettes into a minimal expression platform in
Escherichia coli
. Three novel microviridin variants were isolated and tested against different serine-type proteases. The comparison of the bioactivity profiles of the new congeners allows deduction of further structure-function relationships for microviridins. Moreover, this study provides new insights into microviridin processing and gene cluster organization.
Regio- and Stereospecific Hydroxylation of Various Steroids at the 16α Position of the D Ring by the Streptomyces griseus Cytochrome P450 CYP154C3
Authors: Makino Takuya , Katsuyama Yohei , Otomatsu Toshihiko , Misawa Norihiko , Ohnishi Yasuo
Published: December 2013

Cytochrome P450 monooxygenases (P450s), which constitute a superfamily of heme-containing proteins, catalyze the direct oxidation of a variety of compounds in a regio- and stereospecific manner; therefore, they are promising catalysts for use in the oxyfunctionalization of chemicals. In the course of our comprehensive substrate screening for all 27 putative P450s encoded by the
Streptomyces griseus
genome, we found that
Escherichia coli
cells producing an
S. griseus
P450 (CYP154C3), which was fused C terminally with the P450 reductase domain (RED) of a self-sufficient P450 from
Rhodococcus
sp., could transform various steroids (testosterone, progesterone, Δ
4
-androstene-3,17-dione, adrenosterone, 1,4-androstadiene-3,17-dione, dehydroepiandrosterone, 4-pregnane-3,11,20-trione, and deoxycorticosterone) into their 16α-hydroxy derivatives as determined by nuclear magnetic resonance and high-resolution mass spectrometry analyses. The purified CYP154C3, which was not fused with RED, also catalyzed the regio- and stereospecific hydroxylation of these steroids at the same position with the aid of ferredoxin and ferredoxin reductase from spinach. The apparent equilibrium dissociation constant (

K
d

) values of the binding between CYP154C3 and these steroids were less than 8 μM as determined by the heme spectral change, indicating that CYP154C3 strongly binds to these steroids. Furthermore, kinetic parameters of the CYP154C3-catalyzed hydroxylation of Δ
4
-androstene-3,17-dione were determined (

K
m

, 31.9 ± 9.1 μM;
k
cat
, 181 ± 4.5 s
−1
). We concluded that CYP154C3 is a steroid D-ring 16α-specific hydroxylase which has considerable potential for industrial applications. This is the first detailed enzymatic characterization of a P450 enzyme that has a steroid D-ring 16α-specific hydroxylation activity.
Deregulation of Feedback Inhibition of Phosphoenolpyruvate Carboxylase for Improved Lysine Production in Corynebacterium glutamicum
Authors: Chen Zhen , Bommareddy Rajesh Reddy , Frank Doinita , Rappert Sugima , Zeng An-Ping
Published: December 2013

Allosteric regulation of phosphoenolpyruvate carboxylase (PEPC) controls the metabolic flux distribution of anaplerotic pathways. In this study, the feedback inhibition of
Corynebacterium glutamicum
PEPC was rationally deregulated, and its effect on metabolic flux redistribution was evaluated. Based on rational protein design, six PEPC mutants were designed, and all of them showed significantly reduced sensitivity toward aspartate and malate inhibition. Introducing one of the point mutations (N917G) into the
ppc
gene, encoding PEPC of the lysine-producing strain
C. glutamicum
LC298, resulted in ∼37% improved lysine production.
In vitro
enzyme assays and
13
C-based metabolic flux analysis showed ca. 20 and 30% increases in the PEPC activity and corresponding flux, respectively, in the mutant strain. Higher demand for NADPH in the mutant strain increased the flux toward pentose phosphate pathway, which increased the supply of NADPH for enhanced lysine production. The present study highlights the importance of allosteric regulation on the flux control of central metabolism. The strategy described here can also be implemented to improve other oxaloacetate-derived products.
Discovery of Novel Rickettsiella spp. in Ixodid Ticks from Western Canada
Authors: Anstead Clare A. , Chilton Neil B.
Published: December 2013

The genomic DNA from four species of ixodid ticks in western Canada was tested for the presence of
Rickettsiella
by PCR analyses targeting the 16S rRNA gene. Eighty-eight percent of the
Ixodes angustus
(
n
= 270), 43% of the
I. sculptus
(
n
= 61), and 4% of the
I. kingi
(
n
= 93) individuals examined were PCR positive for
Rickettsiella
, whereas there was no evidence for the presence of
Rickettsiella
in
Dermacentor andersoni
(
n
= 45). Three different single-strand conformation polymorphism profiles of the 16S rRNA gene were detected among amplicons derived from
Rickettsiella
-positive ticks, each corresponding to a different sequence type. Furthermore, each sequence type was associated with a different tick species. Phylogenetic analyses of sequence data of the 16S rRNA gene and three other genes (
rpsA
,
gidA
, and
sucB
) revealed that all three sequence types were placed in a clade that contained species and pathotypes of the genus
Rickettsiella
. The bacterium in
I. kingi
represented the sister taxon to the
Rickettsiella
in
I. sculptus
, and both formed a clade with
Rickettsiella
grylli
from crickets (
Gryllus bimaculatus
) and “
R. ixodidis
” from
I. woodi
. In contrast, the
Rickettsiella
in
I. angustus
was not a member of this clade but was placed external to the clade comprising the pathotypes of
R. popilliae
. The results indicate the existence of at least two new species of
Rickettsiella
: one in
I. angustus
and another in
I. kingi
and
I. sculptus
. However, the
Rickettsiella
strains in
I. kingi
and
I. sculptus
may also represent different species because each had unique sequences for all four genes.