The aim of the current study was to evaluate the effects of and adherence to an active video game promotion intervention on anthropometrics, sedentary screen time and consumption of sugar-sweetened beverages and snacks among non-active video gaming adolescents who primarily were of healthy weight.We assigned 270 gaming (i.e. ≥2 hours/week non-active video game time) adolescents randomly to an intervention group (n = 140) (receiving active video games and encouragement to play) or a waiting-list control group (n = 130). BMI-SDS (SDS = adjusted for mean standard deviation score), waist circumference-SDS, hip circumference and sum of skinfolds were measured at baseline, at four and ten months follow-up (primary outcomes). Sedentary screen time, physical activity, consumption of sugar-sweetened beverages and snacks, and process measures (not at baseline) were assessed with self-reports at baseline, one, four and ten months follow-up. Multi-level-intention to treat-regression analyses were conducted.The control group decreased significantly more than the intervention group on BMI-SDS , and sum of skinfolds  . The intervention group had a significantly higher decrease in self-reported non-active video game time  and total sedentary screen time  than the control group . The process evaluation showed that 14% of the adolescents played the Move video games every week ≥1 hour/week during the whole intervention period.The active video game intervention did not result in lower values on anthropometrics in a group of ‘excessive’ non-active video gamers (mean ~ 14 hours/week) who primarily were of healthy weight compared to a control group throughout a ten-month-period. Even some effects in the unexpected direction were found, with the control group showing lower BMI-SDS and skin folds than the intervention group. The intervention did result in less self-reported sedentary screen time, although these results are likely biased by social desirability.NTR3228Dutch Trial Register  Overweight and insufficient physical activity in youth are major public health concerns because of their associations with multiple chronic diseases –4. IndepA common and popular activity among adolescents is playing video games –14. VideIn addition to increasing physical activity and decreasing sedentary behavior, active video gaming might have an additional effect on the prevention of overweight. Compared to non-active video gaming, active video gaming might provide fewer opportunities for snacking while gaming. In contrast active video gaming might also lead to increased intakes of snacks and beverages following the activity. The first studies that compared energy intake during active and non-active video gaming reported no differences ,24. HoweThus far, there are a few large-scale randomized controlled trials (RCTs) that have evaluated the effects of active video games on body weight in adolescents . MoreoveThe current trial aimed to evaluate whether providing active video games to non-active video gaming adolescents could contribute to the prevention of excessive weight gain. The adolescents were provided with the newest active video games to optimize engagement and an additional controller to facilitate the playing of the video games in multi-players modes. Furthermore, active video gaming was actively encouraged as a replacement for non-active video gaming.a lower body mass index adjusted for mean standard deviation score (BMI-SDS), a smaller waist circumference-SDS, a smaller hip circumference and a lower skin fold thicknessless self-reported time engaged in sedentary screen activitieslower self-reported intake of sugar-sweetened beverages and snacks.We tested the hypotheses that four and ten months after randomization, the adolescents in the active video gaming intervention group would exhibit the following differences compared with the adolescents in the control group:adherence to the interventionappreciation of the Move video games and the interventiongame contextpotential adverse effects (occurrence of injuries due to playing the Move video games)activity replacement intention to continue playing the Move video games.To inform future active gaming studies and to gain additional more insight into the findings, a comprehensive process evaluation was conducted in the intervention group, to examine the following issues:The intervention was evaluated in a randomized controlled trial in adolescents aged 12- to 17-year-old adolescents. The study protocol has been described in detail in a separate publication . The adohttp://www.trialregister.nl/trialreg/admin/rctview.asp?TC=3228).The study was approved by the Medical Ethics Committee of the VU Medical Centre. The trial is registered in the Dutch Trial Register with number NTR3228 .The adolescent had access to a PlayStation 3 at home.The family did not have a Move upgrade for the PlayStation 3.The adolescent lived in the same home as the participating family members at least 4 days per week .At least one other family member (parent or sibling aged 8–18 years old) was willing to participate in the study .The current study focused primarily on adolescents aged 12–17 years. Additionally, the family members of the adolescent  completed questionnaires, but these data were not included in the current analyses. The inclusion criteria were as follows:The recruitment of the adolescents occurred in four cities in the Netherlands; i.e., Amsterdam, Amersfoort, Leiden and Breda. Detailed information about the recruitment is described in Simons et al. . Adoleschttp://nl.playstation.com/ps3/games/.The adolescents assigned to the intervention group received a PlayStation Move upgrade package to play the active video games on a PlayStation 3 console in their homes. The PlayStation Move uses a handheld motion controller wand, a motion-capture PlayStation Eye camera that tracks the player’s position and inertial sensors in the wand that detect its motion. Thus, every movement of the player is mimicked on-screen in the game. The following active video games were provided during the intervention: Sport Champions, Move Fitness, Start the Party and Medieval Moves, Dance Star Party and Sorcery. A detailed description of these Move video games can be found at: Although, there are multiple active video game systems we chose for the PlayStation Move because the PlayStation appeared to be the most frequently owned video game console among 12–16 year old adolescents [We included three elements to support continuing active video game play: 1) because variation in video games is important ,33,34 thAdolescents in the control group were asked to continue their normal gaming behavior. They received PlayStation Move starter packs at the end of the study as an incentive for their participation. Further, they received a small gift  as an incentive after participation at each measure moment.The participants started in three waves for which baseline measurements were collected in January/February 2012, March 2012, and June 2012. The participants completed online questionnaires at baseline and at one, four and ten months of follow-up. Anthropometric measurements were collected at baseline and after four and ten months. Anthropometric measurements were conducted by trained research assistants according to a standardized protocol and occurred on pre-scheduled days at a central and attractive location . Adolescents who were unable to attend to central measurement location on the specified day were measured at home.0 but were not blinded atT4m and T10m. The data analyses were not conducted in a blinded manner.It was not possible to keep the participants blinded to the treatment allocation because the intervention group received an active video game upgrade package, and the control group did not. The participants and research assistants were blinded to group assignment at T0, T4m, and T10m. BMI (kg/m2) was calculated by dividing the weight (kg) by the height squared (m2). Next, the BMI-SDS was determined using the data from the fourth Dutch growth study among children in 1997 as a reference [All measurements are described in detail in Simons et al. 2014 . In shoreference . Regardi1m, T4m, and T10m. Furthermore, the adolescents in the intervention group were asked to provide daily reports on their use of the Move video games over the entire ten-month period on a calendar. All of these self-reported measures are described in detail in Simons et al. [The questionnaires assessed demographics  and country of birth (to define ethnicity according the definition of Statistics Netherlands ). Furthes et al. . We prov0, T1m, T4m, and T10m assessed the total hours per week that were spent on active and non-active video games by asking about gaming frequencies and durations separately for week and weekend days.Questionnaires administered at TTo assess physical activity, we used the validated (correlation with CSA: r = 0.48–0.78) Flemish Physical Activity Computerized Questionnaire (FPACQ)  and focuTo assess sedentary screen time, we used the questions about computer time and TV time from the FPACQ . Both thConsumption of sugar-sweetened beverages was assessed based on the methods of Van der Horst et al. , which iThe consumption of snacks was assessed using the appropriate items from a validated questionnaire ,40. SnacAt all measurements time points, we assessed the game consoles and game applications that the adolescents had access to in their homes to determine whether any participants in the control group purchased any active video game devices.1m, T4m and T10m. Findings from focus groups [A comprehensive process evaluation was conducted in the intervention group at Ts groups  and a sus groups  providedIn addition to the questionnaires, the adolescents in the intervention group reported the time that they spent playing the Move video games on a daily basis over the entire 10-month period in a calendar format. The adolescents were asked to report the type of Move video game and the number of hours and minutes spent playing the Move video games .The analyses were performed according the pre-defined analyses plan described in Simons et al. . First, Additionally, a group * time interaction was added to estimate the time-specific intervention effects. In the results section, only the fully adjusted models (model 2) are reported in the text. Model 1 is included in the tables as background information. Second, analyses were performed according to the per-protocol principle. Adherence to the protocol was separately defined for each follow-up measurement, based on the reported Move video game time in the questionnaire. The intervention group adolescents who played the Move video games at least one hour per week were allocated a score of 1 for adherence, and the intervention group adolescents who played less than one hour per week and the control group adolescents were scored 0. Again, two models were constructed for each outcome measure (with the exception of active video gaming) and were adjusted for the same variables as in the main analysis. Finally, descriptive statistics were used for the process evaluation measures and the use of the Move video games over the ten-month-intervention period based on the Move video game calendar.Multiple imputation with chained equations and analyses of the imputed data were performed using STATA. Other statistical analyses were performed using IBM SPSS Statistics version 21 with a statistical significance level of p<0.05 for detecting main intervention effects.P = 0.19), sex (Fisher’s exact P = 0.51), educational level (Fisher’s exact P = 0.392) or ethnicity (Fisher’s exact P = 0.620) from the adolescents with complete anthropometric data (n = 240). Further, the adolescents who missed one or more questionnaire measurement (n = 42) did not differ in age (T(268): -1.235; P = 0.218), sex (Fisher’s exact P = 1.0), educational level (Fisher’s exact P = 0.064) or ethnicity (Fisher’s exact P = 0.074) from the adolescents with complete questionnaire data (n = 228).In total, 270 adolescents completed the anthropometric baseline measures and were randomly allocated to the intervention or control group. Of these 270 randomized adolescents, 260 participated in at least one of the anthropometric follow-up measurements and were included in the main analyses of the primary outcomes (anthropometrics). Two hundred sixty-two adolescents completed the baseline and at least one follow-up questionnaire and were included in the main analyses based on the questionnaire data . Missing4m, and T10m for the intervention and control group and the results of the main multilevel analyses. In the fully adjusted analyses (model 2), we observed significant intervention effects in the unintended direction at T10m and overall for BMI-SDS and the sum of the skin folds. Regarding hip circumference, we found a time x group interaction effect (P = 0.015); there was no significant difference at T4m, but at T10m, hip circumference was significantly higher in the intervention group than in the control group. We observed no significant intervention effects on waist circumference-SDS in the fully adjusted models. The sensitivity analyses on the imputed data produced very similar results (not reported here).The per-protocol analyses revealed no significant overall effects of adherence to the protocol  on BMI-SDS : -0.029 to 0.126; Model 2: β = -0.05, 95% CI: -0.186 to 0.084), waist circumference-SDS , or hip circumference  and a significant effect in the unintended direction on the sum of the skinfolds .1m, T4m, and T10m for the intervention and control groups and the results of the multilevel analyses. Based on the fully adjusted models (model 2), the intervention group was more likely to play active video games for at least one hour per week. Regarding active video gaming we observed a time x group interaction effect (P = 0.015) in which the effect diminished over time  on non-active gaming , total sedentary screen time (Model 1: Exp (β)  = 0.94, 95% CI: 0.87 to 1.03; Model 2: Exp (β) = 0.96, 95% CI: 0.88 to 1.05), physical activity , consumption of sugar-sweetened beverages , or consumption of snacks .1m, slightly more than half of the adolescents in the intervention group reported playing the Move video games for at least one hour per week, and after ten months, this proportion was approximately one-third. Across the entire intervention period, 14% of the adolescents reported playing the Move video games every week for at least one hour per week at all the time points.The mean enjoyment score was moderately positive and exhibited a decreasing trend over the intervention period. The participants thought that the Move video games were easy to handle based on the Ease of Use scores. Furthermore, the participants thought that they were moderately competent in playing the Move video games. The most-often played and most enjoyed Move video games were DanceStar Party and Start the Party: Save the World. The least played Move video game was Sports Champions. Most of the adolescents were satisfied with the number of Move video games provided. Further, approximately 1/4 of the adolescents bought extra Move video games themselves . Further, the majority of the adolescents perceived playing the Move video games to be a moderate-intense activity. Regarding game context, the majority of the adolescents played the Move video games alone, and most placed the PlayStation Move in the living room or in their own bedroom.10m, 1/5 of the intervention group reported having experienced an injury (the most frequently mentioned injuries were bruises or strained muscles/tendons) while playing the Move video games. The majority of the adolescents reported that the Move video games primarily replaced non-active video games. Finally, nearly half of the adolescents intended to continue playing with the Move video games.At T10m one additional adolescent in the control group reported having a PlayStation Move application.One adolescent in the control group reported having a PlayStation Move at baseline but not during the follow-up measurements. At TThe aim of the current study was to evaluate whether providing active video games could prevent unnecessary increases in anthropometrics among gaming adolescents who were primarily of healthy weight. The findings regarding the anthropometrics revealed that the active video game intervention had no effects in favor of the intervention group and even some effects in the unexpected direction were observed. The intervention group remained relatively stable regarding most of the anthropometric outcomes over time, and the control group improved somewhat in terms of their anthropometric outcomes. Furthermore, some beneficial effects on self-reported non-active gaming and total sedentary screen time  were found in favor of the intervention group. We found that the intervention had no effects, either positive or negative, on the levels of self-reported physical activity or the intakes of sugar-sweetened beverages and snacks. Compliance with the intervention was very low, although the adolescents in the intervention group were significantly more likely to play the active video games for at least one hour per week than the adolescents in the control group (OR = 49). It is important to acknowledge that the ORs for active gaming  should not be interpreted as risk ratios due to the high proportion of the sample who reported that they engaged in active gaming for at least one hour per week (or consumed >1400 ml/week sugar-sweetened beverages) at each time point.Unexpectedly, the control group exhibited improvements in BMI-SDS, while the intervention group generally remained stable. The process evaluation did not provide any reason to believe that the adolescents in the control group bought Move packages themselves and began playing (more) active video games. Further, no evidence was found that indicated meaningful differences in life style behaviors between the intervention and control group. Although the intervention aimed to enable and motivate the participants to engage in active video gaming, only 14% of the participants in the intervention group managed to play for at least one hour per week throughout the intervention period. The dose of active video game play might thus have been insufficient to induce differences in the anthropometrics between the intervention and the control group. A second reason for the lack of an effect on the BMI-SDS of the intervention group might be that our study focused on a general group of gaming adolescents who primarily were of healthy weight. In accordance with our study’s purpose,–i.e., to evaluate the role of active gaming in the primary prevention of overweight and obesity in youth,- we therefore did not select a high-risk group such as adolescents who were already overweight or obese. Earlier studies that focused on overweight children, and thus more on the ‘treatment’ of overweight, have reported beneficial effects on BMI and/or body composition ,27,29. TOther differences between our study and earlier studies that did find beneficial effects were included the targeting younger children (8–14 years) ,29, the These potential reasons for the lack of effects in the intervention group do not however explain the observation that the control group exhibited anthropometric improvements while the intervention group remained stable. Moreover, it is remarkable that the self-reported sedentary screen time of the intervention group was less than that of the control group, which might be related to social desirability bias see  below.During recruitment, we communicated that all participants would receive the Move video games (either at the start of the study or after ten months). Therefore, at the end of the study, the control group also received the active video games. If we had known the study results in advance, we would not have provided the active video games to the control group. Nevertheless, based on the intervention effects we observed , we do not believe that providing the active video games to the control group would have been harmful.The limitations of the current study are the self-reported secondary outcome measures and their susceptibility to social desirability and recall biases. Furthermore, the active video game questionnaires and calendars were not checked for validity. The intervention group might have provided more socially desirable answers than the control group because, as part of the intervention, the participants in the intervention group were asked to replace non-active gaming with active gaming. Unfortunately, we could not use objective data to monitor physical activity and game behavior. Video game play data are stored in the game consoles, but due to privacy regulations these data could not be obtained.The measured anthropometrics are clear strengths of the present study. In our analyses, we used BMI-SDS scores that were based on an external reference  as recoThus far, the evidence relating to the use of active video games as a weight management tool for overweight or obese children is mixed, and evidence for active gaming as a means to prevent excessive weight gain in normal-weight children is still lacking after the present study. Given the popularity of video gaming among youth, further high-quality research is needed regarding on how, when and among whom active video gaming can be effectively applied to promote health and prevent weight gain. Based on findings from earlier studies on the treatment of overweight and obesity, the integrating of active video games into broader health promotion programs seems most promising. Therefore, future studies should examine this issue further and focus on, for example, how to combine and target multiple settings, such as the home, school, and neighborhood, and examine the effectiveness of such integral multi-setting programs in the long-term. When evaluating active video game interventions, we recommend to use objective measurements (e.g. accelerometry) in combination with self-reports for assessing physical activity and sedentary behavior. Regarding assessing video gaming behavior, preferably the data that are electronically registered in the consoles should be used because this process would avoid participant burden and overcome the problems of under or over reporting and the providing of socially desirable answers. Therefore, it is important that the game industry make the stored game data available for research purposes. Also we recommend the use of objective measures of physical activity (e.g. accelerometry) in combination with self-report.The process evaluation results indicated that there is a need for a new generation of active video games that are better able to compete with non-active games in terms of fun, attractiveness, persuasiveness and sustainability. Thus far, the majority of studies have focused on commercially available active games for consoles such as the Xbox 360 Kinect, PlayStation 3 Move and Nintendo Wii. In addition to these console-based active video games, one could also think of more mobile games on platforms such as smartphones or tablets. These platforms are suitable for the utilization of using technologies such as the Global Positioning System and augmented reality, which could increase the possibilities for physical activity in and beyond the game. Furthermore, the focus of research should not be limited to only active video games that are developed by the commercial entertainment game industry , whose main goal is to sell as many devices and video games as possible. Rather, future studies should also focus on active video games, such as the active game Olympus , that arDespite the finding that self-reported sedentary screen time was reduced, the current study suggests that providing active video games to a group of excessive non-active video gamers in their home environment is not effective and might even be counter-effective in altering anthropometrics. In conclusion, the present active video game intervention is not a suitable tool for the prevention of excessive weight gain among gaming adolescents.S1 CONSORT Checklist(DOC)Click here for additional data file.S1 Protocol(PDF)Click here for additional data file.S1 Data File(SAV)Click here for additional data file.S2 Data File(SAV)Click here for additional data file.


Due to the abundance of poorly modeled or incorrectly identified Mg2+ ions, the set of all sites was comprehensively validated and filtered to identify a benchmark dataset of 15 334 ‘reliable’ RNA-bound Mg2+ sites. The normalized frequencies by which specific RNA atoms coordinate Mg2+ were derived for both the inner and outer coordination spheres. A hierarchical classification system of Mg2+ sites in RNA structures was designed and applied to the benchmark dataset, yielding a set of 41 types of inner-sphere and 95 types of outer-sphere coordinating patterns. This classification system has also been applied to describe six previously reported Mg2+-binding motifs and detect them in new RNA structures. Investigation of the most populous site types resulted in the identification of seven novel Mg2+-binding motifs, and all RNA structures in the PDB were screened for the presence of these motifs.The ubiquitous presence of magnesium ions in RNA has long been recognized as a key factor governing RNA folding, and is crucial for many diverse functions of RNA molecules. In this work, Mg Metal ions are indispensable for proper RNA folding, stability and function in various biological processes . The pos2+ is generally accepted as the most important ion for RNA stabilization  for analysis.A potential pitfall in classifying and surveying Mgructures ,17 and Mxception . Mg2+ haps alone . Moreove2+ binding by RNA was performed with the following five objectives: (i) validate Mg2+ binding sites in RNA crystal structures deposited to the PDB and discard poorly modeled or misidentified sites; (ii) statistically analyze preferences of nucleotides and their individual atoms for Mg2+ coordination; (iii) craft and apply a hierarchical Mg2+ site classification system which takes into account both inner- and outer-sphere ligands; (iv) employ the classification system for describing and detecting Mg2+-binding motifs in RNA structures and (v) discover new Mg2+-binding motifs within populous site types.In the present study, a systematic analysis of MgTerminology, abbreviations and definitions used in this manuscript are provided as Supplementary Lexicon 1.2+ ion. This dataset of all Mg2+ sites is herein referred to as the ‘full dataset.’ Mg2+ sites were analyzed using the NEIGHBORHOOD database  Oph, phosphate oxygen (OP1/OP2); (ii) Or, oxygens in ribose (O2′/O4′) or oxygens bridging phosphate and ribose (O3′/O5′); (iii) Ob, nucleobase oxygen and (iv) Nb, nucleobase nitrogen.The atoms from common ribonucleotides that can potentially coordinate Mgdideal) for Mg2+−O (2.08 Å) and Mg2+−N (2.20 Å) bonds were defined as the mean distances observed in the Cambridge Structural Database . This second step identified additional nearby oxygen and nitrogen atoms which could potentially complete the octahedral geometry of the inner sphere. Three additional rules were applied in this second step to preclude chemically unfavorable interactions: (i) a second oxygen from the same phosphate group was not accepted since a phosphate group cannot form a bidentate interaction with Mg2+  thThe coordination number (CN) of a magnesium ion was defined as the number of inner-sphere ligand atoms identified by the procedure outlined above.Probe program , Rout (ribose) and Bout (nucleobase).For annotation of outer-sphere interactions, each individual RNA moiety  was counted only once, and only if it did not form inner-sphere interactions with a given Mgv, QsQ and eQ were used to quantitatively identify the ‘quality’ of Mg2+sites (in addition to other criteria). The value of each parameter has a maximum value of 1, with a lower value indicating poorer reliability and a higher value indicating better reliability.Three customized parameters vQ measures the agreement of the bond valence summation .ligand i . A tablesQ measures the geometrical symmetry of the ligands distribution around the Mg2+ required for octahedral geometry by calculating the amplitude of the vector sum of the bond valence vectors vi (sQ is defined as ctors vi . The sumeQ measures the agreement of the isotropic atomic displacement parameter (B-factor) of the Mg2+ (mB) and its occupancy (mO) compared to those of all atoms in its environment . The environmental B-factor (eB) and occupancy (eO) were calculated as the valence weighted sum of those parameters for all non-hydrogen atoms within 4 Å of the Mg2+ divided by the overall valence  were calculated in a similar manner as has been reported previously (Fatom(X) was calculated by the formula p(Mg-X) is defined as the percentage of Mg-X interactions out of the total number of interactions for all coordinating atoms in the benchmark dataset. p(X) is defined as the percentage of atoms of type X out of all coordinating atoms in the full dataset. In other words, Fatom reflects the frequency that a certain type of atom is observed to coordinate Mg2+, normalized by the frequency of that type of atom in the full dataset. Hence, if Fatom(X) > 1 and thus p(Mg-X) > p(X), atoms of type X interact with Mg2+ with relatively high frequency. Conversely, if Fatom(X) < 1 and thus p(Mg-X) < p(X), Mg-X interactions occur with relatively low frequency.Normalized interaction frequencies of atoms (eviously . For an 2+ sites coordinated more than one phosphate group (Oph) in the inner sphere were differentiated by geometrical arrangement as either cis- or trans- isoforms (two or four Oph) or fac- or mer- isoforms (three Oph). The trans- or mer- isoforms were defined by the presence of two Oph atoms opposite to each other in a trans- conformation, defined as a ligand-Mg-ligand angle larger than 135º. In the absence of such pair of opposite Oph ligands, the site was defined as cis- (two Oph) or fac- (three Oph).Mg2+ sites from 1036 structures, consisting of 95406 sites from 494 ribosome structures and 3854 sites from 542 non-ribosome structures. Most sites in the full dataset are from low-resolution structures; only 2.5% of the sites (2508 sites) are from structures determined at a resolution better than 2.4 Å  in the range of 0–3  complete inner sphere (CN = 4–6) compose around 30% of all sites in ribosome and around 50% of all sites in non-ribosome structures. A small number of sites  had CN>6. Manual inspection of the sites with CN>6 revealed modeling errors with severe clashes between the inner-sphere ligands and/or unlikely bidentate coordination by phosphate . This trend reverses at resolution worse than 3.7 Å, and well-coordinated sites become more abundant in worse resolution structures, likely due to the common practice of using a restrained hexahydrated Mg2+ during refinement instead of a single Mg2+ at lower resolutions with very poor electron density maps  the site is RNA-bound (through inner and/or outer sphere); (ii) the site CN = 4–6; (iii) all three validation parameters are higher or equal to the threshold values  as determined in the preliminary research described in the supplementary data ; (iv) sites should not be coordinated by nucleobase nitrogen other than an endocyclic nitrogen with a lone electron pair in the plane of the aromatic ring (–N = ) in the inner sphere.The benchmark dataset, which excludes highly questionable Mg2+ binding sites (489 structures), which constitutes only 15% of the full dataset. The size of the benchmark dataset is significantly smaller than the size of the full dataset, mostly due to the abundance of sites with a highly incomplete Mg2+ inner sphere (CN = 0–3) in the full dataset . The inner-sphere Mg2+−O (non-water) and Mg2+−N distance distributions show that more often than not, sites in the full dataset were modeled with these distances much longer than ideal values. Many of the sites in the full dataset that are observed with interaction distances longer than 2.4 Å might have been incorrectly identified as Mg2+ instead of water oxygen atoms, potassium ions or sodium ions, which have ideal bond distances to oxygen of 2.9 Å, 2.8 Å and 2.5 Å, respectively , which measures the frequency a particular kind of atom served as a ligand normalized by the frequency of that atom type in RNA structures overall. The most commonly observed inner-sphere ligand atom type is oxygen from phosphate (Oph)  from nucleobase (Ob). The Fatom value of the most frequently observed Ob ligand atom (U-O4) is just two times smaller than the value for Oph atoms, even though U-O4 lacks the ability to significantly compensate for the positive charge of Mg2+. Not all Ob atoms exhibited similar normalized frequency to serve as ligands for Mg2+ binding. The Fatom values for Ob atoms adjacent to the ribose bond (C-O2 and U-O2) were up to 290 times lower than that for Ob atoms located opposite to the sugar edge (G-O6 and U-O4)  are rarely found in Mg2+ inner sphere.The frequencies of RNA atoms to coordinate Mg) Figure . The reab) are the only other RNA atoms to serve as inner-sphere ligands, but generally they were observed in the inner sphere less frequently , only two (G-N7 and A-N7) have an inner-sphere Fatom value higher than one. All other –N= atoms were almost never observed to coordinate Mg2+ in the inner sphere.Besides oxygen atoms, nitrogen atoms from nucleobases  are also among the most frequent outer-sphere atoms of Mg2+  are much greater than those for Ob atoms adjacent to the ribose bond (C-O2 and U-O2). In spite of the similar high frequencies of certain atoms to coordinate Mg2+ both through the outer and inner sphere, several types of atoms rarely found in the inner sphere were found to form outer-sphere hydrogen bonds quite frequently . A more extensive discussion of the differences in atomic Mg2+ coordination frequencies is presented in Supplementary Text 2.Though outer-sphere hydrogen bonds are chemically different from the inner-sphere coordination bonds, the most frequent inner-sphere atoms . The inner-sphere Fatom value for U-O4 is higher than that for A-N7, which makes uracil the second most frequently observed nucleobase. As a net result, the frequency for nucleobases to coordinate Mg2+ in the inner sphere is (in descending order) G > U > A > C. In the outer sphere, the guanine moiety is also the most frequently observed nucleobase, followed by adenine and uracil, exhibiting a slightly different trend (G > A > U > C) than is observed for the inner sphere.Guanine is the most frequently observed nucleobase to coordinate Mg2+ interactions in the benchmark dataset  sites with non-RNA, non-water atoms in the inner sphere; (iii) sites with only RNA atoms as non-water ligands in the inner sphere (RNA-inner) and (iv) sites with only water molecules in the inner sphere and at least one RNA moiety in the outer sphere (RNA-outer).All Mger Table : (i) sithttp://www.csgid.org/metalnas/  and (ii) comprise only a small fraction of sites in the benchmark dataset , Oph is most abundant , while the total number of other inner-sphere atoms (Or|Ob|Nb) was chosen to be the second criterion. The maximum number of RNA atoms serving as inner-sphere ligands is 4 per site  if more than one Oph is present. For the most populated branch , the number of outer-sphere phosphates was also taken as an additional criterion to subdivide sites.The RNA-inner class is the most populous class, representing 63% of the benchmark dataset Table . Of the e Figure , so combFatom values for nucleobase and ribose atoms . The only exception to this trend is the 2Nb type in the #Oph = 0, #(Or|Ob|Nb) = 2 subclass, which contains a disproportionately high population (158 sites). All other types with more than one Or|Ob|Nb inner-sphere ligand, regardless of subclass, contain 28 sites or less. Both fac- and mer- conformations have similar abundances for sites with #Oph = 3, while the cis- conformation is more frequently observed than the trans- conformation for all #Oph = 2 subclasses. Eighty-six percent of the sites in the subclass #Oph = 2, #(Or|Ob|Nb) = 0 are in the cis- conformation. The cis-2Oph conformation is even more predominant (283 out of 286 sites) in the sites in which non-Oph inner-sphere ligands are also present  = 1,2).The general trend seen in the inner-sphere interactions as was done for the inner sphere, individual outer-sphere moieties were used to subdivide RNA-outer sites. We believe that the number of moieties represents the uniqueness of a given structural arrangement of RNA, while the actual number of interactions (hydrogen bonds) of the moiety with inner-sphere water molecules just slightly contributes to the energy of binding and may easily vary between very similar sites. Similar to RNA-inner site classification, the number of outer-sphere phosphate moieties #Pout was chosen as the first criterion and the total number of ribose and nucleobase moieties #(Rout/Bout) was used as the second criterion to subdivide the RNA-outer class, resulting in 39 subclasses. The specific combinations of ribose (#Rout) and nucleobase (#Bout) moieties in the outer sphere produced a list of 95 types of RNA-outer sites (Supplementary Table S4).The RNA-outer class represents a significant fraction (33%) of all sites in the benchmark dataset Table . The outout with only two nucleobase moieties in its outer sphere (555 sites). This site type is often found in RNA helices, as base stacking results in very close positions of two consecutive bases coordinating one Mg2+. Site types containing only phosphate moieties are less populous yet still abundant, represented by Pout (94 sites), 2Pout (191 sites), 3Pout (207 sites) and 4Pout (174 sites). The rarity of the ribose moiety in the outer sphere  and outer-sphere phosphate moieties . The N7 atoms serving as ligands in motif are usually found in two guanine bases, but sometimes an adenine-guanine pair was found. Two single-nucleotide and guanine-specific ‘macrochelate’ motifs were identified with G-N7 serving as a ligand in either inner sphere or outer sphere, and ‘macrochelated’ with an outer-sphere phosphate moiety  sites were accepted in order to exclude those Mg2+ sites which are bound only to protein in RNA–protein complexes or do not have any clear connection to RNA. The Mg2+ validation procedure was largely based on the similarity of each site to the rigid octahedral arrangement of inner-sphere ligands expected for Mg2+ and the characteristically short Mg2+-ligand distances. Harnessing those intrinsic properties for Mg2+ identification required that all sites have a relatively complete coordination sphere; therefore only sites with CN = 4–6 were accepted. The three validation parameters used to select sites for the benchmark dataset evaluated Mg2+-ligand distances (vQ), the symmetry of the inner-sphere ligands arrangement (sQ) and the agreement of Mg2+ B-factors with the surrounding atoms (eQ). Using the combination of multiple parameters as filtering criteria effectively removed the majority of poorly modeled Mg2+ sites. However, a few Mg2+ sites with chemically infeasible interactions were still not caught by the criteria, for example, those with amino nitrogen atoms in the inner sphere. To exclude those cases, we introduced an additional criterion for sites with nitrogen atoms from nucleobases in the inner sphere: namely, accepting only those sites which have an endocyclic nitrogen with a lone electron pair in the plane of the aromatic ring (–N=) since only this type of nitrogen from nucleobase can feasibly coordinate Mg2+ (Supplementary Text 2).Since the scope of this study was Mg2+ binding sites in RNA are absent from the benchmark dataset because the electron density for Mg2+ was not observed, or density was present but was incorrectly modeled as another metal ion or as a water molecule. Other sites might have had a true magnesium ion, but an incompletely modeled inner sphere resulted in some of the validation parameters being below the benchmark dataset thresholds. In the latter case, the inner sphere can often be completed by placing additional water molecules during crystallographic re-refinement of problematic structures. In this way, the benchmark dataset can be extended by revealing additional ‘trustworthy’ Mg2+ sites, but would require manual inspection of each site and is beyond the scope of the current study. Nevertheless, our algorithms enabled extraction of many existing trustworthy Mg2+ sites which can be used for further statistical and classification studies. For example, the benchmark dataset is particularly valuable to be used as a training dataset for tools to predict the positions of Mg2+ ions in both experimental and theoretical models of RNA structures and/or increase the accuracy of Mg2+ prediction. Thus far, we have implemented the benchmark dataset developed in this work as one of the alternative reference datasets in MetalionRNA predictor at http://metalionrna.genesilico.pl/  or incorrect (2.18 Å) Mg2+-water distances used in some refinement programs by default .  Around 200 strongly restrained Mg2+ sites at another incorrect (1.83 Å) Mg2+-water distance were also identified  in a 40S ribosomal subunit  (2+−O (non-water) and Mg2+-N distances, even in sites that satisfy the validation criteria, suggests that those interactions are loosely restrained during crystallographic refinement .The reliability of a MgM, 2XZN) . On the 2+-ligand distance restraints is essential for correct modeling of a Mg2+ inner sphere, especially of low-resolution structures. However, improperly strict or weak restraints may result in misinterpretation of experimental data, and thus in turn impair subsequent research. Given the prevalence of poorly modeled Mg2+ sites in RNA structures and improper use of crystallographic restraints (either too strict or too loose), we propose that the crystallographic refinement software should include easy-to-use restraints to enforce correct CN and geometrical arrangement of ligands around metal ions.The proper use of Mg2+ binding sites. Redundant structures  were not excluded. It is beneficial to include sites that satisfy validation criteria from all structures in the analysis because different structures of the same macromolecule may carry different sets of reliable Mg2+ sites due to differences in diffraction data quality, refinement strategies, crystallization conditions, bound ligands and/or macromolecule conformation. Moreover, it is essential to preserve as much variety of the different coordination patterns that may be present for otherwise equivalent Mg2+ sites observed in different PDB deposits of homologous RNA molecules.The main objective in the construction of the benchmark dataset was ensuring the reliability of the Mg2+ are minimally affected by dataset redundancy, because the frequencies of individual interactions are normalized by the frequency of atoms or nucleotides in the dataset. Dataset redundancy is beneficial for the classification because it ensures that the architecture of each reasonable Mg2+ site was accounted for. The numbers of instances of each validated Mg2+ motif do indeed contain redundant sites, but each motif was manually confirmed to represent a variety of sites in non-redundant structures.Even though a certain level of redundancy was observed in the benchmark dataset, all further analyses were carefully designed to control for the presence of redundancy. The statistics used for estimation of the preferences of atoms and nucleotides to bind MgFatom values produced by statistical analysis of the benchmark dataset indicate the frequency at which certain types of RNA atoms served as ligands for Mg2+ in the inner or outer coordination sphere, and are normalized so that values for different atoms can be directly compared to one another. These values are consistent with the steric accessibility and chemical properties of each atom type (Supplementary Text 2), and we propose that these frequencies are reasonable estimates of the ‘propensity’ or ‘preference’ of these RNA atom types to coordinate Mg2+. However, we cannot formally rule out the possibility of sampling bias . For example, the analyzed set was necessarily limited to RNA molecules which form diffraction-quality crystals and, as mentioned above, may exclude sites that were incompletely or incorrectly modeled. The values of Fatom (and the preferences they imply) can be used as prior knowledge for predicting ‘probable’ versus ‘improbable’ Mg2+ sites in both computational modeling and crystallographic refinement.The 2+ is believed to be neutralization of the negative charge of phosphate moieties in RNA  present in the site type. The name of each site type is unique and contains enough information to infer the subclasses at broader levels of classification.The classification system is based on a simple dendrogram-like hierarchy of Mg2+-binding motifs, which  is sufficient to identify new instances of these motifs  and the server that describes them may be used by researchers to determine the extent of uniqueness of a particular coordinating pattern, allowing them to highlight candidate Mg2+ sites for specific consideration.We believe our classification system has potential to detect unique structural sites responsible for specific functional roles. Mg2+ motifs, i.e. some specific coordinating patterns will prove to be sufficiently populated. Moreover, increasing the number of sites could allow a detailed classification of more complicated sites with additional metal or non-RNA ligand in the Mg2+ coordination sphere. The growth of structural data will also permit the systematic investigation of other metals less commonly observed in RNA.With the availability of more data in the future, the method used may provide evidence of new validated Mg2+ sites in the benchmark dataset can be accessed via URL http://www.csgid.org/metalnas/. The main page of the server lists all site types with schematic drawings and the number of sites found for each site type. Detailed information for each site type may be accessed, which includes an image of a representative site and a list of all PDB entries containing a site of that type with chain and residue ids. Each Mg2+ site may be visualized in Jmol (The classification of Mg in Jmol . Users c in Jmol .Supplementary Data are available at NAR online.


On Friday, April 1, 2016, the United Nations (UN) General Assembly proclaimed 2016 to 2025 as a decade of action on nutrition.Lancet published the first series on maternal and child nutrition, which subsequently triggered the formation of the Scaling Up Nutrition (SUN) Movement.,Excited as I am about how all the chips are falling in place, a key question remains on my mind: Do we have the key ingredients to deliver the goods, come 2025 or 2030? My mind goes back to 2008 when the ,Six years ago, when the SUN Movement was established, many of us in the nutrition community were caught up in the fever of the moment. We became excitedly optimistic that we were finally going to make a major dent in the malnutrition situation in the developing world, and especially in the so-called “high-burden” countries. Although there are signs of significant progress being made through the movement, the “2015 Global Nutrition Report” points to a rather slow rate of change as well as persistant gaps, including insufficient financing and suboptimal country-level capacity needed to address malnutrition at sufficient scale.The UN General Assembly proclamation clearly identifies the framework within which the malnutrition situation could be addressed in the next 10 years. However, in the particular case of Africa, I would like to share a few thoughts on what I consider to be missing in the soup, based on past experience. First is that Africans must be at the forefront of the efforts to address malnutrition in Africa. There is a lot of investment already in nutrition in Africa. What is missing is leadership by Africans to make the investment work for the African situation. Much more progress would have been achieved already if African governments had sufficiently prioritized nutrition and made available sustained and adequate financial commitment from their treasuries to address malnutrition. This is especially needed when too many of the countries that are off-course in meeting global nutrition targets are in Africa. Such commitments should be married to a strong process of ensuring accountability for achievement of realistic but sufficiently ambitious targets to address malnutrition in all its forms. Further, African nutrition scientists must take leadership of the evidence economy and generate policy options for decision makers. As the saying goes, “It is never too late to do the right thing.” The time has come to stop having more of the usual situation in which the Global North is driving the efforts to address malnutrition, with Africans playing catch-up.Africans must be at the forefront of efforts to address malnutrition in Africa.,,Second, we have spent much time on repeating research to understand the determinants and characteristics of malnutrition; in some cases, the research agenda is driven from outside Africa and not designed to answer questions relevant to the African.Third, our governments must be pushed, through grassroots civic movements such as the country SUN civil society networks and traditional and religious leadership, to invest more in nutrition than they are currently doing. The projected economic growth patterns in resource-poor countries will be depleted quickly without governments investing to consolidate the gains that could be made.Governments must invest more in nutrition.Finally, more effort should be invested in promoting collaboration across various stakeholders within the African region. Cross-country experience and best-practice sharing can enhance faster progress and reduce costs of program implementation. Within countries, more collaboration across agencies and sectors  is warranted.Collaboration across countries can enhance faster progress and reduce program implementation costs.


Burkholderia pseuodmallei) present in soil and water. Here, we evaluated public awareness of melioidosis using a combination of population-based questionnaire, a public engagement campaign to obtain video clips made by the public, and viewpoints on these video clips as potential educational tools about the disease and its prevention.Melioidosis causes more than 1,000 deaths in Thailand each year. Infection occurs via inoculation, ingestion or inhalation of the causative organism . Focus group interviews were used to evaluate their perceptions of the video clips.Of 4,203 Thais who completed our study questionnaire, 74% had never heard of melioidosis, and 19% had heard of the disease but had no further knowledge. Most participants in all focus group sessions felt that video clips were beneficial and could positively influence them to increase adherence to recommended preventive behaviours, including drinking boiled water and wearing protective gear if in contact with soil or environmental water. Participants suggested that video clips should be presented in the local dialect with simple words rather than medical terms, in a serious manner, with a doctor as the one presenting the facts, and having detailed pictures of each recommended prevention method.In summary, public awareness of melioidosis in Thailand is very low, and video clips could serve as a useful medium to educate people and promote disease prevention.World Melioidosis Congress 2013, Bangkok, Thailand, 18–20 September 2013 (abstract OS VII-04). Burkholderia pseudomallei, which is present in soil and water in a defined geographic distribution . HoNearly all participants (>99%) reported that they had heard or knew of common infectious diseases, including HIV/AIDS (99.5%), tuberculosis (99.1%), malaria (98.6%), leptospirosis (99.5%), dengue (99.5%), influenza (99.6%), and bird flu (99.3%). However, for melioidosis, 3,177 (74%) participants reported that they had never heard of the disease, with 783 (19%) responding that they had heard of this but had no further knowledge about melioidosis, and only 303 (7%) responding that they knew about melioidosis . A higheFor risk factors of acquiring infectious diseases in general, most participants (>80%) thought that the risk increased with exposure to soil (85.2%), exposure to water (85.1%), being diabetic (81.1%), smoking (85.7%), drinking (85.3%), outdoor exposure to dust (89.6%), and needle sharing (96.9%). However, less than 80% of the participants thought that the risk of acquiring infectious diseases increased by drinking tap water (67.6%), or having outdoor exposure to rain (68.3%) .For preventive measures, more than 95% of participants thought that infectious diseases in general could be prevented by wearing protective gear during exposure to soil (97.9%) or environmental water (97.4%), by cleaning after exposure to soil or water (97.8%), eating cooked food (97.8%), drinking boiled water (96.8%), cleaning open wounds with disinfectants (96.8%), and eliminating mosquito larvae (96.0%). However, many participants wrongly thought that applying herbal remedies (80.4%) or soil (30.1%) directly to open wounds could prevent infectious diseases .A total of 71 diabetic patients without a history of melioidosis participated in the ten focus groups. Overall, 21 (30%) were male and 50 (70%) were female. The median age of participants was 52 years . Sixty-eight participants (96%) had never heard of melioidosis, and 3 (4%) had heard of the disease but had no further knowledge. The topics of the focus groups were defined a priori and included (1) general viewpoints about video clips, (2) potential use of video clips to raise public awareness of melioidosis, (3) potential use of video clips to provide education about melioidosis prevention, and (4) responses and obstacles to the recommended preventive guidelines.In general, participants felt that using video clips was a good idea and suggested that these should be shown to other diabetic patients in Thailand. All participants valued the information provided by the video clips.“We should show these video clips to other patients waiting downstairs to see the doctor” “This (the video clip) is good. It can be used to inform (my) friends and everyone in the country (about the disease).” In most sessions, participants suggested that they could understand and relate to the video clips better when these were spoken in the local dialect, and in simple words without the use of medical terms.“If you want Isan (northeast) people to understand, a local dialect (Isan) should be used so that we can understand.” “(The video clip) should use simple words for Isan people. It should not use medical terms as they are not understandable.” Most participants felt that the video clips should not be narrated in a lighthearted fashion, such as in a cartoon clip or through an entertaining story. However, some participants liked the cartoon clip and found that it was easy to understand.“This disease is dangerous but they (the presenters) are making it (the video clip) look entertaining. So, it might make the audience think that it (the disease) is not as dangerous as it is.” “Isan (northeast) people may not understand cartoons, think that it (the disease) is a funny matter, and may not pay attention to it.” “The cartoon clip is attractive. I understand (the disease)” The other common general viewpoint was that doctors should deliver the key messages as rural people put their trust in them.“Doctors must be the ones who speak. Most local people believe in doctors” Most participants had never heard of the disease name, and found that the video clips were very useful in informing them about this.“Is it rheumatoid disease? (I have) heard of only rheumatoid.” “The doctor has not said anything about this disease yet. I have never heard of it” “This is a scary disease. I just know of it now” Participants stated that some video clips provided good education about the prevention of melioidosis.“This video clip makes us picture clearly why we should wear (rubber boots)” “I like the video clips. They are informative. They show examples (of recommended preventive behaviours).” .Participants also stated that they would follow recommendations for prevention after watching the video clips, including the difficult or time-consuming recommendations such as wearing boots while working in the rice fields and boiling water before drinking.“It (the video clip) provides recommendations. We will try to follow what they recommended” “(After watching the video clips), I will try to wear rubber boots” “If it (wearing boots) is needed, I will do it” “This video clip is good about boiling water. It makes people know that water coming out of the tap is not clean. Now I will boil the water as I know that it (boiled water) will not contain germs” Most participants thought that the biggest obstacle to following preventive guidelines was to wear protective gear during exposure to water or soil, since wearing rubber boots made it difficult to walk in muddy paddy fields, and wearing rubber gloves made it difficult to plant rice. The problem of becoming hot while wearing rubber gear was also raised in most sessions.“It is impossible for farmers to wear boots while working in the rice paddies. The mud sucks the boots. If it is just for walking on the road, we can wear boots. However, when working in rice paddies, only walking bare foot is possible. We cannot work while wearing boots.” “Boots might be possible, but rubber gloves are impossible” “It is impossible to wear rubber gloves. It’s hot.” We found that many participants drank water without treatment, even though they knew that drinking boiled water was better, explaining that they do not have time to boil water.“I know (that I should drink boiled water), but I do not do it.” “I don’t boil (water). (I) work in the rice fields, (and I) can’t boil water in time. After working, I am tired. When I get back, I drink water immediately. (I) can’t boil water in time.” Our study has shown that public awareness of melioidosis in Thailand is very low, indicating a need to increase knowledge about its prevention. Video clips were responded to positively, suggesting that they could serve as a useful medium to educate and promote prevention of melioidosis. The results of our focus group study suggested that video clips should be presented in the local dialect using simple words rather than medical terms, in a serious manner, with a doctor presenting the facts, and having detailed pictures of each recommended prevention method. These findings will contribute towards the future development of educational tools about melioidosis in Thailand.The finding that 74% of adults in Thailand have not heard of melioidosis is not surprising. To our knowledge, information about melioidosis is rarely given to the public by the mass media, including television, newspapers and radio stations. Basic information about melioidosis and its prevention is also not taught in schools in Thailand. By contrast, public awareness of other common infections including HIV/AIDS, tuberculosis, malaria, leptospirosis, dengue and influenza, which are taught in schools and frequently mentioned by the national media, is very high. The result is consistent with our clinical observations that many melioidosis patients have not heard about the disease when they first present to hospital with this infection , and witB. pseudomallei [B. pseudomallei. Our previous study also showed that the application of herbal remedies or an organic substance to open wounds was associated with developing melioidosis [As people are not aware of the disease, it is not surprising that they do not know about how to prevent the disease. Finding that many people think that drinking tap water does not increase the risk of acquiring infectious diseases need urgent consideration by policy makers. Our previous environmental study showed that 10% of village tap water samples were contaminated with domallei , and thadomallei . Misundeioidosis . Policy Changing behaviour requires a systematic approach. In circumstances where awareness and knowledge are low, increasing the levels of these is fundamental for people to change their routine patterns of behaviour . Our stuhttp://youtu.be/l1VxcZTw-BU). This suggests that there are still people watching the video clip after the competition, and highlights the benefit of the YouTube as a platform for providing educational videos to the general public.The use of YouTube for sharing of educational videos via the Internet has been increasingly used and objectively evaluated –23. It iA limitation of this study is that questionnaire participants had relatively high educational levels compared with the general population in Thailand. The questionnaires were distributed by Suan Dusit Poll, and study samples may have over-represented people living in the center of each district, and under-represented people living in more remote, rural areas. It is possible that male gender, and people with a low level of education, were more likely to decline the invitation to complete the questionnaire. Nonetheless, this sampling bias suggests that awareness and knowledge might be even lower in the general population than our results suggest. Using competitions and targeting schools and universities to generate video clips, we found that many used medical jargon in video clips without considering their target population who spoke local dialects and preferred simple language. However, a few video clips fulfilled these criteria and could be used to educate target populations. These could be made to video compact disk (VCD) and distributed to every primary care unit (PCU) in Thailand, which has ready access to televisions and CD players that can be used to show video clips to the target population. Other educational tools for rural populations such as pamphlets and group education should also be considered for future interventions.Our findings suggest that policy makers together with program planners in education, health and workplaces should urgently consider raising public awareness and knowledge of melioidosis and its prevention, and systematically develop interventions to increase preventive behaviours based on careful, theoretically-informed analyses of the target behaviour in the context of the target populations.S1 Text(DOCX)Click here for additional data file.S1 Table(XLS)Click here for additional data file.


Parkinson’s disease (PD) is one of the most common movement disorders in adults, often cannot be adequately controlled with medical treatment, and thereby treated with the neurosurgical procedures. Unilateral pallidotomy, making a lesion in the globus pallidus pars interna (GPi), was the predominant surgical technique until early 1900s. Thereafter, deep brain stimulation (DBS) of the subthalamic nucleus (STN) has become a mainstream surgical technique for managing not only the cardinal dopaminergic features of PD, but also levodopa-induced motor fluctuations and dyskinesia.Recent review studies, however, reported that unilateral pallidotomy also has long-term effect on both cardinal symptoms  and motor complications . Currently, transcranial magnetic resonance guided focused ultrasound (MRgFUS), without necessity of opening the cranium, has been developed as a minimal invasive surgical technique, generating precise and focal thermal lesion in the brain compared to previous radiofrequency thermal lesion.The authors underwent world first MRgFUS pallidotomy to confirm its efficacy as well as potential side effects. We hereby demonstrated the beneficial effects of MRgFUS pallidotomy in PD patient for improving levodopa induced dyskinesia as well as motor symptoms.


This study evaluates whether the daily practice of an exercise routine might protect from endocrine pancreas damage in cola drinking rats.ad libitum, water (W) or cola beverage (C), and b) physical activity, sedentary (S) or treadmill running (R). Accordingly, 4 groups were studied: WS (water sedentary), WR (water runner), CS (cola sedentary) and CR (cola runner). Body weight, nutritional data, plasma levels of glucose, creatinine, total cholesterol and cholesterol fractions, and triglycerides (enzymocolorimetry), and systolic blood pressure (plethysmography) were measured. After 6 months, euthanasia was performed . Pancreatic tissue was immediately excised and conventionally processed for morphometrical and immunohistochemical determinations.Forty-eight Wistar rats were randomly assigned to 4 groups depending on a) beverage consumption 4 μm2 vs 2.58 104 μm2, p<0.0001) and median β-cell mass , and increased median α/β ratio . In water drinking rats (WR vs WS), running reduced median α-cell mass  and α/β ratio . Differently, in cola drinking rats (CR vs CS), running partially restored median islet area , increased median β-cell mass  and reduced median α/β ratio .The effects of running and chronic cola drinking on pancreas morphology showed interaction (p<0.001) rather than simple summation. Cola drinking (CS vs WS) reduced median pancreatic islet area (-30%, 1.8 10This study is likely the first reporting experimental evidence of the beneficial effect of exercise on pancreatic morphology in cola-drinking rats. Presently, the increase of nearly 50% in β cells mass by running in cola drinking rats is by far the most relevant finding. Moderate running, advisably indicated in cola consumers and patients at risk of diabetes, finds here experimental support. Metabolic syndrome (MetS) is a cluster of conditions: hypertension, hyperglycemia, overweight and dyslipidemia that pose a high risk of cardiovascular disease, stroke and diabetes. Occidental urban societies have high prevalence of cardiovascular disease, diabetes and obesity –4. The sThe number of diabetic patients with obesity has increased due to life-style changes which favor sedentarism and unhealthy nutritional habits ,9. FortuIn diabetic patients, aerobic exercise has been found not only to reduce hypertriglyceridemia and hyperglycemia but to contribute to weight-loss as well ,12. In rSo far, no experimental research has been conducted in order to evaluate the effects of combining an exercise routine with a nutritional factor on endocrine pancreas. Actually, even the individual effects of exercise on endocrine pancreas have been scarcely addressed not to mention that alpha cells have not been evaluated either.Nearly for ten years, we have been studying the variety of physiological and morphological alterations found after chronic ingestion of cola beverages in both genetically modified mice and normal rats. We found that chronic cola drinking accelerated aortic atherosclerosis and enlarged atherosclerotic lesions in atherosclerotic mice ,21. In hRecently we reported that 6 months of cola-drinking induce MetS-like features: hyperglycemia, hypertriglyceridemia, mild overweight and insulin resistance, and affects endocrine pancreas morphology in rats –24.Hence, consistently with our line of research on experimental metabolic syndrome mimicking life-style conditions concerning with a human nutritional habit as chronic cola-drinking, we conceived a model combining the systematic practice of an aerobic exercise with cola-drinking, a combination frequently observed these days in human life-style.This study evaluates whether an aerobic exercise program may help to ameliorate metabolic disorder and preserve normal morphology in pancreatic islets of cola-drinking rats.The experiment was conducted in accordance with the recommendations of the Weatherall report, "The use of non-human primates in research." The committee of Ethics in Animal Research of the Instituto de Investigaciones Cardiológicas (ININCA) and the Institutional Animal Care and Use Committee (IACUC) of the Faculty of Medicine of the University of Buenos Aires, namely the CICUAL  approved the study.In regard to our previous publications ,25, diff™, Argentina). In turn, each group was split into two subgroups depending on physical activity: S (sedentary) or R (runner). Accordingly, 4 groups were studied (n = 12 each): WS, WR, CS and CR for 6 months. Animal care followed the ‘Guide for the Care and Use of Laboratory Animals’ .Forty-eight adult male Wistar rats were randomly assigned to two groups depending on beverage consumption “ad libitum”: water (W) or cola  was measured by tail cuff plethysmography, and biochemical determinations were performed in blood collected from the tail vein after 4-hour fasting. Commercially available kits for enzymatic colorimetric assays  were used to measure concentration of: glucose (hexokinase/glucose-6-phosphate dehydrogenase reaction), creatinine , total cholesterol (enzymatic colorimetric test), HDL-c (selective accelerating detergent), LDL-c (homogeneous enzymatic colorimetric assay) and triglycerides (glycerophosphate oxidase). The atherogenic index in plasma was calculated as AIP = log (TG/HDL-c) .™) [The aerobic exercise program consisted in treadmill running on a 10° inclined plane for 30 minutes daily, 5 times per week, at a maximal speed of 20 m/minute (treadmill Columbus™) .After 6 months, rats were euthanized by subtotal exsanguination  under anesthesia . At autopsy, pancreas were removed, weighed, fixed in phosphate-buffered 10% formalin solution and processed for histology.Eight 3 μm sections of tissue blocks were stained with hematoxylin-eosin (H-E). For immunohistochemistry, the traditional streptavidin-biotin-peroxidase complex technique was used. A blocking solution  was used to block non-specific protein binding by incubation for 30 min at 37°C and 90 min at room temperature, in phosphate-buffered saline solution (PBS) at pH = 7.4. Overnight incubation at 4°C with primary monoclonal mouse antibodies against insulin (dilution 1/3000 in blocking solution) and glucagon (dilution 1/30000 in blocking solution) (Sigma-Aldrich) allowed labeling of β- and α-cells, followed by incubation with biotinilated goat anti mouse secondary antibody (diluted 1/500 in PBS) 2h at room temperature. Finally, streptavidin (1/1000 in PBS) was added for 1h at room temperature, followed by addition of chromogen DAB 3-5min. Control sections were incubated with non-immune normal mouse serum.4 μm2 at 40 X objective lens. Alpha- and β-cell fractional area was calculated as immunopositive-to-total islet area ratio for glucagon and insulin respectively and it was expressed as percentage. Alpha- and β-cell mass was estimated as the product of the relative cross-sectional area of β cells per total tissue and the weight of the pancreas [Islet cross-sectional area was estimated counting the number of points hitting an islet (point-counting method) in ≥ 30 islets/pancreas. An orthogonal grid with 300 test points projected onto the fields of view representing an area of 6.7 10pancreas . Images For variation of a given factor which is expressed in %, e.g. from 2% to 3%, stating that there is “a 1% increase” is ambiguous for it might be interpreted in two ways: a). the difference is 1% unit b). the change represents 1% of the initial or reference value. In the above example: the absolute change is 1% unit (3–2) while the relative change is 50%  were expressed as mean±SD, submitted to MANOVA and ANOVA, and compared using Bonferroni’s test. Morphology data (non- parametric variables) were expressed as median and interquartile range (IQR) to indicate central tendency and spread of data distribution respectively, submitted to Kruskal-Wallis test and between-group multiple comparisons were performed (Dunn’s test). Statistical significance was conventionally set at p≤0.05. SPSSCola-drinking (CS vs WS respectively) reduced food consumption -19% (p<0.05), increased liquid intake 37% (p<0.05) and calorie supply 15% (p<0.05) , and indper se effects of each treatment.Statistical analysis confirmed the existence of between-treatment interaction (p<0.001) for cola-drinking and running practice on pancreas morphology, masking their respective individual effects. Then, separate post-hoc analyses were performed in order to identify 4 μm2, IQR 1.71–2.11 vs 2.58 104 μm2, IQR 2.12–3.26; p<0.0001), median β-cell mass by 43% , median β-cell fractional area by 24% , and increased median α-cell fractional area by 62%  and median α/β ratio by 49%  (Cola drinking (CS vs WS) reduced median pancreatic islet area by 30% (1.8 10< 0.001) .In water drinking rats (WR vs WS), running reduced median α-cell mass by 48% , median α-cell fractional area by 27%  and median α/β ratio by 56%  .Differently, in cola drinking rats (CR vs CS), running caused a 15% restoration of median islet area , increased median β-cell mass by 47%  and median β-cell fractional area by 17% , and reduced median α-cell fractional area by 25%  and median α/β ratio by 6%  . Change per se (WR vs WS) showed no major changes on immunolabeling. However, in cola-drinking rats, running increased insulin immunopositive area and islet size (CR vs CS). In all cases, typical cells arrangement was found  in CR compared with CS suggesting that individual differences might be actually larger for higher islet area values. From a physiological viewpoint, a far more important finding was the increase in β-cell population induced by running in cola-drinking rats. The increase of nearly 50% in β cells mass by running in cola drinking rats is by far the most relevant finding in this study.Interestingly, rather than simple summation, running and cola-drinking showed mutual interaction concerning with the effects on endocrine pancreas morphology. Running ameliorated some of the detrimental effects of cola-drinking on pancreatic islet morphology. The median islet area was significantly higher in CR compared with CS. Regardless the respective medians differed by 0.22 10Physiologically dynamic interaction between cola-drinking and running practice interaction, strongly supported by statistical interaction as well, might unfold in the critical contingency of systemic and local inflammatory and oxidative conditions associated with hypertriglyceridemia ,36. HypeDifferences in either behavioral traits or vegetative signs of catecholaminergic stimulation such as increased heart rate or blood pressure, were not observed in relation with drinking fluid in the present study. Particularly, no signs of psychomotor stimulation were observed in cola drinking rats in agreement with a previous report . In thatIt is unlikely that behavioral changes might have gone undetected in our study. Laboratory technicians and personnel staff responsible for animal handling and care, bear years-long practical experience in behavioral studies, have received specialized professional training and have developed a keen eye on behavioral traits. Besides, some of the authors of this study have published behavioral findings , 40.Changes in β-cell mass were overall the highest contributors to the variation in islet area as confirmed by significant β-cell mass to islet area correlation across groups. This is inferable considering that the insulin-producing β-cells are the most abundant cell phenotype residing in the islets of Langerhans, and any change in β-cell mass is likely to yield a corresponding change in islet area.Genetic deficiency of glucagon receptor prevents β-cell loss in experimental diabetes induced in glucagon knock-out mice . Beta-ce™ , Coca cola™  and Pepsi cola™  [™ in our studies. In Celec et al report, daily drinking volume consumed by rats in all cola groups was up to three times higher than the water intake in the control group [Cola drinking for 6 months, induced insulin resistance, hyperglycemia and β-cell loss with increase in α-cell fractional area , affecte10 mg/L) , 43. In ol group . In the ol group . HoweverActually, the pivotal difference between our studies, including the present one, and Celec et al reports ,43 appeaContrasting with the effects of exercise on the cardiovascular system, experimental evidence of the effects of exercise training on pancreas is scarce and has been obtained in rats submitted to genetical, surgical or pharmacological manipulation, not to mention that α-cells were not evaluated in those protocols. Alternatively, we present a model combining a nutritional factor (cola drinking) with the practice of an exercise routine, a frequent combination in daily human life.This study is likely the first reporting experimental evidence of a beneficial effect of exercise on pancreatic morphology in cola-drinking rats by partially restoring β-cell population. An interesting physiological running-cola drinking interaction was observed. Moderate running advisably indicated in cola consumers and patients at risk of diabetes is supported by present findings.Awareness that a healthy life style including the regular practice of moderate exercise and a balanced diet according to individual needs seems to be definitely the best prevention ever.


To address these competitive requirements, the use of error-correcting codes is advised. Current barcoding systems are mostly built from short random error-correcting codes, a feature that strongly limits their multiplexing accuracy and experimental scalability. To overcome these problems on sequencing systems impaired by mismatch errors, the alternative use of binary BCH and pseudo-quaternary Hamming codes has been proposed. However, these codes either fail to provide a fine-scale with regard to size of barcodes (BCH) or have intrinsic poor error correcting abilities (Hamming). Here, the design of barcodes from shortened binary BCH codes and quaternary Low Density Parity Check (LDPC) codes is introduced. Simulation results show that although accurate barcoding systems of high multiplexing capacity can be obtained with any of these codes, using quaternary LDPC codes may be particularly advantageous due to the lower rates of read losses and undetected sample misidentification errors. Even at mismatch error rates of 10 Molecular barcoding provides the opportunity to multiplex next-generation sequencing  capacityN are first screened to ensure the satisfiability of chemistry constraints imposed by the target sequencing technology, e.g., barcodes designed for pyrosequencing platforms must avoid homopolymer regions. Candidate barcodes are then screened to ensure a minimum pairwise distance dmin that guarantees the unambiguous correction of dmin and the number M of legal barcodes must be accepted  was considered.Barcoding systems built from binary BCH, binary Golay, quaternary LDPC and BY barcodes were evaluated using a Quaternary Symmetric Channel (QSC) model . Under tollowing , 55, psN was limited to 25 bases. For each barcoding system of size N built with an error correcting code of size n, a wide range of error correction and multiplexing abilities were evaluated. This was accomplished by varying parameter t of binary BCH codes and parameter m of quaternary LDPC codes. For BCH barcodes, binary BCH codes of size n ∈ {15, 31, 63} and shortened versions of them were considered. For LDPC barcodes, quaternary LDPC codes of size n ≥ 16 were considered. LDPC codes of size n < 16 were disregarded due to difficulties in satisfying the mandatory LDPC sparse constraint. In addition, BY barcodes of size N ∈ {7, 8, 15} and Golay barcodes of size N = 12 were considered. For the sake of completeness, random barcodes reported in [N ∈ {8, 9, 10} and minimum edit (Levenshtein) distance dL ∈ {3, 5, 7} were also considered. Recalling that the Hamming distance is an upper bound of the edit distance, random barcodes were further screened to determine their minimum Hamming distance dH. Similar values were observed, i.e., dH ∈ {3, 5, 7} so that the correction of at least t ∈ {1, 2, 3} mismatch sequencing errors can be guaranteed.For practical purposes, orted in  of size M, their barcoding rate B, their probabilities pe of barcode identification errors and their probabilities pu of undetected multiplexation errors. For each N, M was defined as the maximum number of barcodes which were compatible with the given sequencing chemistry. Similarly, B was defined as the actual fraction of informative quads per barcode, i.e., B is close as possible to Barcoding systems were evaluated through their multiplexing capacity QSC type that generates mismatch sequencing errors with a probability ps. For a given set of sequencing chemistry constraints, the M and B accomplished by BCH and LDPC barcodes of size N will depend on the desired pe and pu for the given ps. With main focus on boosting experimental scalability without compromising multiplexation accuracy, BCH and LDPC barcodes of size N ≤ 25 able to fulfill the operational constraint M > ≥ 24 and pu ≤ 10−8 at ps = 10−2 were identified. For BCH barcodes, simulation results showed that the desired operational constraint could be only satisfied by shortening binary BCH codes of size n = 63. For LDPC barcodes, the desired operational constraint could be only satisfied by LDPC barcodes of size N ≥ 19.Let us consider an ideal sequencing channel of the N = 21 can be used to multiplex up to M = 86 samples with pe ≈ 10−5 and pu ≈ 10−8. By letting N to increase up to 25, one additional satisfactory configuration with pu ≈ 10−8 can be identified at N = 22 with M = 384 and pe ≈ 10−5. Note that for pu ≈ 0, pe essentially bounds the probability of read losses. Taking into account that the number of Illumina reads per flow cell currently ranges from 25 × 106 to 300 × 109, we may be interested in further pe reductions.As shown in pe ≈ 10−6, shortened BCH barcodes of size N at least 24 are required. BCH barcodes of size N = 24 can be used to multiplex up to M = 73 samples with pe ≈ 10−6 and pu ≈ 10−9. By letting N = 25, one additional satisfactory configuration with pu ≈ 10−9, M = 295 and pe ≈ 10−6 can be obtained. Details about the pe performance of BCH barcodes beyond ps = 0.01 are shown in Simulation results showed that to accomplish N = 19 can be used to multiplex up to M = 65 samples with pe ≈ 10−5 and pu ≈ 10−9. By letting N to increase up to 25, three additional satisfactory configurations with pu ≈ 10−9 can be identified at N = 21, 23, 24 with M = 210, 648, 1911 and pe ≈ 10−6. To further reduce pe in one order, LDPC barcodes of size N ≥ 23 are required. LDPC barcodes of size N = 23 can be used to multiplex up to 56 samples with pe ≈ 10−7 and pu ≈ 10−9. By letting N = 25, one additional satisfactory configuration with pu ≈ 10−9, M = 118 and pe ≈ 10−7 can be obtained. Details about the pe performance of LDPC barcodes beyond ps = 0.01 are shown in As shown in N = 7, 8 ) per iteration, being m = n − k the number of redundant symbols of the regular non-binary LDPC in GF(q) and j the number of non-zero entries per column of the LDPC matrix [q = 16 [N = 19 allowing the multiplexation of up to M = 65 samples using a quaternary regular  LDPC code with j = 3, the recovery of 12M identities in an Illumina MiSeq platform with a 177 MIPS processor using a maximum of 50 iterative decoding steps would take less than an hour.In this paper, LDPC barcodes demultiplexing was performed with the iterative decoding algorithm used for decoding non-binary LDPC codes for magnetic recording applications . Hence, C matrix , 61. Thi [q = 16 , is amen [q = 16 . For exanon-binary LDPC codes. Specifically, concatenated watermark codes [non-binary LDPC code and an inner sparse code, to which a watermark sequence has been added, could be used. Concatenated watermark codes rely on the ability of the inner decoder to transform insertion/deletion errors into mismatch errors and on the ability of the outer non-binary LDPC decoder to correct them. Regarding DNA barcoding applications of concatenated watermark codes, we expect samples identities are first mapped to hexadecimal strings, that these strings are LDPC encoded with a short hexadecimal LDPC code already optimized for transmissions over a quaternary symmetric channel, that hexadecimal LDPC codeword symbols are mapped to the quaternary sequencing layer with a non-linear quaternary sparse code and that resulting sparse quaternary sequences are finally perturbed with the addition of a well-known quaternary pseudorandom sequence defined as the pilot watermark signal. After this non-trivial processing of samples identities is performed, candidate barcode sequences able to deal with mismatch and indel sequencing errors could be obtained. We are currently putting together these pieces, looking forward to the design of concatenated watermark barcodes for the third generation Single-Molecule Real-Time (SMRT) long-read sequencing technology [Along this paper we have restricted our attention to sequencing errors of the mismatch type. Readers might reasonable argue that challenging sequencing errors are those dominated by insertions and deletions. We note, however, that these errors might be also tackled with short rk codes  built frchnology , for whichnology . We thuschnology  sequenciq = 4u and u ≥ 1. Let Γ be a bipartite graph with n left nodes called message nodes and m < n right nodes called check nodes. Let us map one-to-one the n message nodes of Γ to the n coordinates of q − ary codewords c = , ci ∈ {0, 1, 2, …,q−1}, i = 1,…,n. Provided the sum of neighboring positions for all check nodes among neighboring message nodes is zero in GF(q), Γ defines a q-ary linear code of size n able to carry k = n − m informative symbols. Thus, the code structure can be dissected into m component parity subcodes. In addition, if Γ is sparse, i.e., each message node is constrained by j < < m check nodes and each check node constraints v < < n message nodes, the code turns to be an LDPC code in GF(q). Finally, if n ⋅ j = m ⋅ v holds, a regular LDPC code is obtained.Let us start with a brief revision of LDPC codes in GF(q), ci and their noisy observations ri, i = 1, …, n. Rectangles are used to represent parity constraints over codeword symbols. Edges are put between codeword symbols and parity constraints. Legal LDPC codewords must fulfill the complete set of m parity constraints. On regular LDPC codes, any codeword symbol participates in exactly j ≥ 3 parity constraints. Rectangles are also used to represent probability functions pi = P(ri = a∣ci = b) modelling the transmission channel, i = 1…n. For a QSC model, pi = 1−p and a ≠ b holds. Iterative decoding provides estimates p and ri, i = 1,…,n , si ∈ {0, 1, 2, 3}, i = 1,…,k, by means of c = s ⋅ G.The construction of barcodes from quaternary LDPC codes is straightforward once Γ is given. Formally, Γ is described by the so-called parity check matrix D score to discriminate between prospective good and bad short quaternary LDPC codes, 20 sets of 50 random quaternary LDPC matrices were built from binary PEG templates with m rows, n columns and j = 3 non-zero entries per column. For each set, the best D-scoring code was selected and its barcoding performance assessed at ps = 10−2. The Spearman correlation coefficient S2 was then used to evaluate the correlation between D scores and the empirical log10pe accomplished by selected LDPC codes. Moderate negative associations were observed suggesting that the D score was indeed useful for the identification of prospective good short quaternary LDPC codes for barcoding purposes. For example, the observed correlation at ps = 10−2 for LDPC barcodes of size N = 24 carrying k = 5 informative quads was S2 = −0.52, p-value < 0.05. Practically, prospective good short quaternary LDPC codes were selected from sets of 1000 random instances. For each selected H, the corresponding generator matrix G with k rows and n columns was computed using the constraint G ⋅ Ht = 0. This was accomplished by means of an adaptation of I.V. Kozintsev software [H matrices in GF(q) using Gaussian elimination.To shed light into the ability of the software  for the QSC model. Corrupted BCH barcodes were first deinterleaved, mapped to the binary domain using inverse mapping tables and decoded with the implementation of the Berlekamp-Massey decoding algorithm in [GF(2) were then mapped to GF(4) to recover original barcode sequences. A similar procedure was used to recover corrupted Golay, LDPC and BY barcodes. Golay barcodes were first deinterleaved, mapped to the binary domain and then decoded with the implementation of the arithmetic decoding algorithm in [ps of a base mismatch used as input to the QSC channel model. BY barcodes were first deinterleaved and then decoded as indicated in [Simulation experiments were performed to analyze the robustness of BCH, Golay, LDPC, BY and Random barcodes over the rithm in . Decodedrithm in . LDPC barithm in  and implrithm in . The LDPcated in . Finallycated in  with exppe of barcode identification error was then estimated by Montecarlo simulation. For this purpose, T = 100 random samples comprising C = 107 barcode sequences were used. Samples were obtained by performing sampling with replacement over the sets of valid barcode sequences obtained after the Barcrawl filtering stage. For each sample Si, the proportion pe,i of barcodes identification errors and sample variances i = 1, …, T. At the end, the pooled sample mean pe estimations and thus, 95% confidence intervals for the case pu of undetected multiplexation errors.The probability ere used . A base g10sepp¯e. A similM and the barcoding rate B are fundamental properties of any coding-based barcoding system. They follow from counting all candidate barcode sequences that are compatible with a predefined set of DNA manipulation constraints. For random barcodes of size N, 4N candidate barcode sequences must be first individually screened to remove those with undesirable composition patterns. Concerning their posterior use for error correction purposes, remaining sequences must be then globally analyzed to ensure a predefined minimum distance. Hence, for random barcodes, the estimation of M and B scales exponentially with N both in time and memory. On the other hand, the minimum distance is a built-in property of candidate barcode sequences when systematic error correcting codes are used and thus, only the presence of undesirable composition patterns must be controlled. For barcodes of size N built from linear codes of size n able to carry k informative symbols in GF(q), just qk sequences must be individually screened, k < < n. For modest k and q settings, e.g., k < 8 for q = 4, exact estimation of M and B can be accomplished. For larger k settings, a Montecarlo approach is required.The multiplexing capacity


Neither the CNAQ nor the SNAQ have a high level of validity or reliability in this elderly population and are therefore not recommended for use in the ambulatory rehabilitation setting. Further work is necessary to assess the validity and reliability of other malnutrition screening tools to establish their usefulness in this population.Malnutrition is common in older adults and often goes unrecognised and untreated. Australian evidence-based guidelines for the management of malnutrition indicate that only the Mini Nutritional Assessment short form (MNA-sf) and Rapid Screen are recommended for use as malnutrition screening tools in the rehabilitation setting. The aim of this secondary analysis was to assess the validity and reliability of two malnutrition screening tools, validated in other adult sub-groups, in a rehabilitation population aged ≥60 years. The Council on Nutrition Appetite Questionnaire (CNAQ) and the Simplified Nutritional Appetite Questionnaire (SNAQ), were completed by 185 ambulatory rehabilitation patients  and results compared to the full MNA as a reference technique. Prevalence of risk of malnutrition was 63% according to the MNA. For identification of risk of malnutrition the CNAQ had sensitivity of 54%, specificity 81%, positive predictive value 83% and negative predictive value 51%, compared to 28%, 94%, 89% and 44%, respectively, using SNAQ. Assessment of reliability indicated significant slight to fair agreement between MNA with CNAQ (κ = 0.309,  Malnutrition, used here to refer to under nutrition, is widespread in older adults across all settings . It is wMalnutrition is costly to individuals and the hospital system alike, in terms of negative health impacts such as increased length of stay, delayed wound healing, and increased risk of infection . In addiEffective prevention and treatment of malnutrition relies on accurate diagnosis. Nutrition screening identifies individuals who are at risk of becoming malnourished. There is good evidence to support routine nutrition screening in all settings using a valid tool which is appropriate to the population in which it is to be applied . NumerouAs indicated previously there are many other validated malnutrition screening tools in existence and many have been shown to effectively identify the risk of malnutrition in a variety of other population groups. Two of those tools, the Council on Nutrition Appetite Questionnaire (CNAQ) and the Simplified Nutritional Appetite Questionnaire (SNAQ), which were developed in community dwelling adults and long term care residents , have beThus, this study aimed to test the validity and reliability of the SNAQ and the CNAQ to effectively detect risk of malnutrition in a sample of older adults participating in ambulatory rehabilitation.A secondary analysis was performed on baseline data collected as part of a randomized controlled trial (RCT) conducted at three public hospitals in southern Adelaide, Australia between June 2005 and June 2006. The RCT compared the effects of a hospital-based day rehabilitation program and a home-based rehabilitation program, and the methods used in the trial were previously described in detail . All parStudy participants completed two self-reported malnutrition screening tools; the CNAQ and the SNAQ  7]. The. The7]. per se. All other participants were classified as well-nourished (score 24–30).The nutritional status of study participants was assessed by a trained research dietitian using the Mini Nutritional Assessment (MNA) , an 18-ip < 0.05. After checking the distribution for normality sample characteristics were described by median (interquartile range (IQR)) for continuous variables and frequencies (%) for categorical variables. Contingency tables were used to determine sensitivity, specificity, positive predictive values (PPV) and negative predictive values (NPV) of both the CNAQ and the SNAQ, in comparison to the MNA. Kappa measure of agreement was used to assess consistency between the CNAQ and SNAQ, and the MNA and results categorised according to the classification of Landis and Koch [Data collected as part of this study were analysed using SPSS version 17.0  and the and Koch .From 225 ambulatory rehabilitation participants who were randomized in the primary RCT, 185 were ≥60 years and had data available for inclusion in this secondary analysis. The sample was 48% male and had median (IQR) age of 78  years. According to the MNA 63% of study participants were at risk of malnutrition. Sensitivity and specificity and PPV and NPV of CNAQ and SNAQ, compared to MNA, were calculated from the contingency tables displayed in When compared to the MNA, the CNAQ demonstrated sensitivity of 54%, specificity of 81%, PPV of 83% and NPV of 51%, compared to 28%, 94%, 89% and 44%, respectively, using the SNAQ.p < 0.001) and κ = 0.176 (p < 0.001), respectively.Assessment of reliability between the MNA and CNAQ and SNAQ resulted in statistically significant slight agreements with κ = 0.309  and SNAQ (44%) indicate that, among those who were identified as well-nourished, approximately half were in fact at risk of malnutrition, with similar consequences to the low levels of sensitivity already discussed.A recent study conducted in Malaysia attempted to validate the CNAQ and the SNAQ against the Subjective Global Assessment in hospitalized older adults and outpatients . The resReliability of both tools in comparison to the MNA, although statistically significant, was below acceptable levels. While the CNAQ demonstrated fair agreement (κ = 0.309) the SNAQ showed only slight agreement κ = 0.176) . These l6 . TheseThe characteristics of a good malnutrition screening tool are that it is quick, simple and easy to use . Both thThere are some strengths and limitations of the current study that should be acknowledged. The participants in the original RCT ranged in age from 18 years to 94 years however only those aged ≥60 years were included in this secondary analysis. Of those in the primary study, 13 (6%) were aged less than 50 years, and 82% of participants were aged 60 years or over with a median age of 76 years. This reflects the patient profile reported recently in South Australia , and NewBased on the results of this secondary analysis, the CNAQ and SNAQ do not have a high level of validity or reliability for the identification of malnutrition risk and are therefore not recommended for use in older ambulatory rehabilitation patients. There is however scope in the future to determine validity and reliability of other existing malnutrition screening tools to establish their usefulness in this population.