The vestibular system is responsible for a wide range of postural and oculomotor functions and maintains an internal, updated representation of the position and movement of the head in space. In this study, we assessed whether unilateral vestibular loss affects external space representation. Patients with Menière's disease and healthy participants were instructed to point to memorized targets in near  and far  spaces in the absence or presence of a visual background. These individuals were also required to estimate their body pointing direction. Menière's disease patients were tested before unilateral vestibular neurotomy and during the recovery period (one week and one month after the operation), and healthy participants were tested at similar times. Unilateral vestibular loss impaired the representation of both the external space and the body pointing direction: in the dark, the configuration of perceived targets was shifted toward the lesioned side and compressed toward the contralesioned hemifield, with higher pointing error in the near space. Performance varied according to the time elapsed after neurotomy: deficits were stronger during the early stages, while gradual compensation occurred subsequently. These findings provide the first demonstration of the critical role of vestibular signals in the representation of external space and of body pointing direction in the early stages after unilateral vestibular loss. The vestibular system is involved in a wide range of functions, from postural and oculomotor reflexes to spatial cognition. Vestibular organs  measure head angular and linear accelerations, respectively, for the coding and representation of head movements in space. Projections to ocular and spinal motoneurons mediate oculomotor and postural reflexes involved in stabilizing and orienting the gaze, head and body in space Asymmetry in vestibular signals in patients with unilateral peripheral vestibular loss induces asymmetrical postural, locomotor, oculomotor and perceptive responses The function of the vestibular system in spatial cognition is just being recognized, and the consequences of its loss at a higher level are important to acknowledge. The properties of the extrapersonal space have long been considered to be independent of vestibular system properties. In the present study, we determined whether symmetrical vestibular signals are necessary for the accurate representation of the external space. In healthy subjects, vestibular signals are used to estimate the target location in space during self-motion Pointing to memorized targets involves two distinct mental spatial representations: egocentric (or body-centered) and allocentric (or environment-centered) e.g. . In the Other contextual factors, such as the distance between the participant and the target, might differentially influence the representation of the target location. Indeed, a distinction between near  and far  space coding has been reported in brain-damaged patients Relatively few attempts have been made to quantify the consequences of vestibular loss in body cognition. Indeed, the body schema is modified after vestibular loss, resulting in an altered representation of the width of hemi-bodies Previous studies have shown that spatial impairments are maximal during the early stages following vestibular asymmetry The experiments were performed in 13 unilateral vestibular-defective (UVD) patients suffering from Menière's disease , when they had not received antivertigo medications, and they did not report attacks of vertigo during the preceding week. The patients' performance was recorded at two postoperative times throughout the recovery process: one week after UVN (D+7), where all patients were able to stand straight and motionless, and one month after UVN (D+30). The intervals between the three experimental sessions were the same for the healthy participants.The participants were seated, and their head was immobilized on a head-and-chin rest. The heights of the seat and resting system were independently adjusted until the participants felt that they were in a comfortable position, with their head aligned with their trunk and their midline aligned with the center of the experimental space. The participants held a laser pointer with both hands, which rested on a small platform located at chest level. Pointing to the target location only required rotating the wrists while keeping the hands on the platform. Each subject wore home-made glasses that limited the visual field to approximately 60 degrees horizontal and 40 degrees vertical, so that hand movement was not visible. The experimental room was surrounded with black curtains, and the roof and floor were painted black.To distinguish the consequences of vestibular loss in both the peripersonal and extrapersonal spaces, the target location was assessed in two bi-dimensional horizontal spaces, the “near horizontal plane” and the “far horizontal plane” . The tarThe horizontal near space  corresponded to the gripping space. The horizontal far space (145 cm wide by 108 cm deep) was located on the ground 180 cm away from the participant. The vertical or fronto-parallel space (145 cm wide by 108 cm high) was located 310 cm in front of the participant and centered on the line of sight. A panel with red laser LEDs for the projection of target stimuli (0.6 cm diameter dots) was mounted perpendicular to each space and managed using a computer program .The spatial arrangement of the targets was designed to prevent the participants from building a cognitive representation that could influence the accuracy of the pointing responses. Thus, the targets were located in directions and at distances that varied with respect to the participant's midline projection , bottom.The participants were required to point to a visual target that disappeared prior to movement. Each target was presented for 2 sec. When the target disappeared, the laser pointer was switched on and the participant was instructed to point to the memorized location immediately, as directly and as accurately as possible. No time constraint was imposed. After reaching the estimated target location, the participants were instructed to say “ok” and remain stationary. One experimenter used a fixed camera to take a snapshot of the response. Subsequently, the pointer was automatically switched off and moved to the starting location for the next trial.The pointing tasks were performed in total darkness and in light using a structured background to provide visual references (two concentric rectangles formed with 0.8-cm-large white stripes on a black backdrop), without particular depth cues. The pointing tasks were performed in blocks of balanced ordered sessions. Three blocks were performed in the dark (one block for each space) and two blocks were performed in the light . Each block was composed of 24 trials with four trials per target. The targets were randomly presented in each block, and no identical targets were presented in successive trials. A computer program displayed the targets one by one, and each response location was recorded for subsequent analysis. A five-minute resting period was provided between each block presentation. To become familiar with the tasks, the participants completed a few trials before starting data acquisition. The participants received no feedback during or after the experimental session.For calibration purposes, the target LED was activated and its location was automatically recorded before each session. To analyze the responses, each target location was compared with the participant's response for that target by computing the barycenter of the laser spot, applying the calibration corrections and computing the corresponding distances along the two axes. The barycenter computation was used to compensate for the unevenness of the target LED illumination spot and the small distortion of the spot resulting from the slight tremor of the participant's hands when holding the pointer. The resolution of the recording system was calculated considering the overall dimensions of the experimental field, the number of pixel rows and columns and the distance between the laser pointer and the experimental field. The resolution ranged from 0.1 to 0.2 degrees according to the target location.The data (in millimeters) were transformed into degrees computed from the participants' eyes to obtain the direct comparison of the performance between the different spaces. Pointing to the right of the real target location  was indicated with positive values, whereas negative values indicated a leftward pointing error . The x- (left-right) and y- (fore-aft) errors were analyzed using a mixed-design analysis of Variance (ANOVA) with Group  as the between-participant factor, and Session (n = 3), Condition  and Space  as within-participant factors. For the representation of body pointing direction task (see below), mixed ANOVAs with Group and Session were performed. The significance level for all means was less than or equal to 0.05.To determine whether the pointing error was associated with changes in representation of body pointing direction, the visual SSA was also recorded for each participant during each session. The SSA was assessed using a laser line  projected on the horizontal far plane and in the fore-aft direction with the rotation centered on the participant's midline. The participants were instructed to imagine the laser line starting from their navel and extending straight ahead from the trunk and to adjust the orientation of the pointer so that the two extremities were positioned on this virtual line. The experimenter placed the starting position of the line either to the left or to the right  of the participant's midline projection. The participants were subsequently instructed to align the laser straight ahead using a pair of pushbuttons held in each hand. The orientation of the laser line relative to straight ahead was automatically measured using a motorized system. The mean SSA was expressed in degrees and calculated by averaging the four consecutive trials of each participant. For each patient, the positive and negative signs referred to ipsilesional and contralesional SSA deviations, respectively; for the healthy participants, the positive and negative signs referred to rightward and leftward deviations, respectively.The potential consequences of eye movements on the representation of the target location and SSA were evaluated. After unilateral vestibular loss, perceptual changes might be associated with a spontaneous nystagmus or an altered resting location of the eyes through vestibular imbalance. Spontaneous nystagmus induces a drift in visual perception. However, this disturbance has been primarily revealed in darkness and in the absence of visual fixation. In the present study, only a few patients reported a drift of the target for the last trials when fatigue was established. In these cases, the trials were eliminated.Eye movements were recorded using videonystagmography. A CCD camera, illuminated with infrared LEDs, was fixed on the goggles in front of the non-dominant eye . Because eye movements were recorded while the participants monocularly viewed the targets, this task was performed after the pointing task. In both near and far spaces, the same targets as those of the pointing task were randomly presented one by one. The participants were required to fix each target. Eye movements were sampled at 25 Hz and recorded for 15 seconds. The data were processed offline using a method based on a mathematical dynamic neural network. Iris gray-level detection was used to obtain the iris print of each patient. To examine spontaneous nystagmus (in degrees), the slow phase velocity of horizontal, vertical and torsional components was measured when the participants gazed at a target. Eye movement processing has been described in detail by 1,24 = 12.28; p<0.01) and Session  were found for the pointing error in the left-right direction. The pointing error was significantly higher for patients than for healthy participants. A significant interaction between Group and Session  showed that the pointing error differed over time more in patients than in healthy participants. Planned comparisons revealed that the effect of Session was obvious only for the patients , indicating a drastic impairment in target location after unilateral vestibular loss. An overall effect of Space was also found , with a higher pointing error observed in the near space than in the far space. A major effect of Condition  and significant effects of the Group x Condition interaction  and the Group x Space x Condition interaction  were observed, indicating that the pointing error was differentially affected in patients and healthy participants as a function of space and condition  and far  spaces. The difference between the groups was obvious during the early stages following unilateral vestibular loss at D+7 for the near (p<0.001) and far (p<0.001) spaces; mean lateral deviations toward the operated side of 3.4±0.8° (Mean±CI) and 2.6±0.5° relative to the target location in the near and far spaces, respectively, were observed for patients, and deviations of 0.1±0.1° and 0.1±0.1° were observed in the healthy group. For patients, the pointing error regained preoperative values at D+30. Under light conditions, the mean pointing error was less than that observed in darkness. At D+7, only a slight difference was reported between patients (0.4±0.5°) and healthy participants (0.1±0.1°) for the near space (p<0.01).Significant effects of Group ; the pointing error was higher in the near space than in the far space. Significant effects of the Group x Session interaction  and the Group x Condition x Session interaction  were observed, suggesting changes in perceived target location after unilateral vestibular loss. These effects indicated that the pointing error was affected differentially over time for patients and healthy participants and only in darkness. In the fore-aft direction, the only factor affecting the mean pointing error was Space  was observed in the up-down direction. An additional ANOVA comparing the pointing error recorded in both planes revealed no significant differences between the up-down pointing error in the vertical plane and the fore-aft pointing error in the horizontal plane for both groups of participants. Moreover, an ANOVA was performed to assess whether the left-right pointing error in the vertical plane differed from that observed in the horizontal plane. The lack of significant differences revealed that the left-right pointing error was similar in both planes.The pointing error was also measured in the vertical plane to clarify whether the error in the fore-aft direction in the horizontal plane (i.e. an error on the target distance estimate) is associated with an error in the up-down direction (i.e. an error on the target height estimate). For the fore-aft direction in the horizontal plane, an interaction between Group and Session  as a function of the experimental session . A significant interaction between Group and Session  revealed that SSA was differentially affected over time in the two groups of participants. The patients showed a rightward deviation , while the healthy participants exhibited no such deviation (session 2: −1.3±1.1° and session 3: −0.4±1.4°). In addition, the patient SSA significantly decreased between D+7 and D+30 (p<0.001). During the first session, the directions indicated for the healthy participants and patients were close to the sagittal fore-aft orientation and did not significantly differ between the two groups.The variations in SSA for each session are illustrated in A spontaneous nystagmus was observed in the dark and in the light during the early stages following unilateral vestibular loss (D+7), particularly for the horizontal component. Because participants were fixing a target, the spontaneous nystagmus velocity was weak . In darkness, the pointing accuracy and spontaneous nystagmus responses were associated: both nystagmoid eye movements and spatial errors peaked at D+7, while no spontaneous nystagmus was observed at D−1 or at D+30, where there were no significant spatial errors. In addition, the data indicated that, for each visual condition, the velocity of the spontaneous nystagmus did not significantly differ according to target location. In darkness, the correlation between the velocity of spontaneous nystagmus and spatial error at D+7 for both spaces and each target location was not significant . No significant correlations were found when patients were looking either left (r = 0.41) or right (r = 0.45), in near space (r = 0.44) or in far space (r = 0.13). Finally, all patients displayed a static ocular torsion after UVN resulting from a rotation of the upper pole of both eyes toward the operated side. The mean ocular torsion was 7.5±1.6° at D+7 and 3.9±1.1° at D+30 toward the operated side.This study assessed the representation of external space in patients with a complete unilateral vestibular loss. In the dark, the configuration of targets was shifted toward the lesioned side and compressed toward the contralesional hemifield. Patients underestimated the location of targets in both the fore-aft and up-down directions. Representation was impaired for both near and far spaces, while error was higher in the near space. In addition, this impairment was associated with an impairment of representation of body pointing direction.For the first time, data show that patients examined during the early stages following unilateral vestibular loss experience impairment in the spatial location of visual memorized targets in the dark, i.e., when participants rely on egocentric strategies Target location deviates toward the affected side for both near and far spaces, with a deviation that is higher for near space. This dissociation, together with the previously reported asymmetric distortion of the represented space, suggests that the disorders of the extrapersonal space representation are similar in patients with unilateral vestibular loss and in patients with spatial neglect Pointing error is also reported in the fore-aft and in the up-down directions. In these cases, it might result from an underestimation of either the distance between the body and the target location or the target height. However, because errors in these two dimensions did not differ, our data do not distinguish distance and height errors in the target representation. The role of vestibular information has been previously reported for travelled distance representation in healthy subjects Targets are accurately located in a structured visual background, supplying references and leading participants to rely on allocentric strategies. Under these conditions, error is either much weaker than in the dark or completely absent. Similarly, the perceptual bias in the judgment of the visual target location resulting from ocular partial paralysis Since the representation of body pointing direction is involved in egocentric pointing, the effect of unilateral vestibular loss on SSA has also been investigated. In the present study, patients with unilateral vestibular loss show an ipsilesional rotation of the SSA in the horizontal plane. This is consistent with previous data reported acutely Considering the potential role of eye movements, we especially address the question about whether changes in external space representation are secondary  to temporary nystagmoid eye movements and torsion or are simultaneous effects appearing in parallel. Data indicate that both nystagmoid eye movements and spatial errors peak at D+7, whereas no spontaneous nystagmus is observed at D−1 or at D+30, where there are no significant spatial errors. However, at D+7, the nystagmus velocity is not correlated with the extent of the spatial errors, either in the left versus right hemifields or in near versus far space. It should be noted that the nystagmus velocity is weak, which is probably due to the fact that patients were requested to fix a visual target. Dissociation between perceptive responses and eye movements has also been reported in similar patient populations, as the SSA deviation remains during the chronic stage following unilateral vestibular loss, when no patients have residual spontaneous nystagmus Finally, the fact that the deficits observed in the representation of external space and of body pointing direction may be partially associated with decreases in the attentional resources allocated to the task cannot be ignored see . SeveralThese results provide strong evidence that vestibular signals are involved in external space representation for spatial location of visual memorized targets in the dark, i.e., when participants are relying on an egocentric strategy. This evidence further supports the hypothesis that deviations in the representation of body pointing direction are partially responsible for the inaccurate location of targets and suggests that vestibular signals are necessary for the precise integration of sensory information to maintain accurate spatial representation. Consequently, the present study stresses the importance of the vestibular system in spatial cognition. Therefore, vestibular syndrome may also be considered a disorder of spatial representation and is thereby comparable to spatial neglect.


Veterans of Operation Enduring Freedom and Operation Iraqi Freedom (OEF/OIF) are at increased risk for alcohol misuse, and innovative methods are needed to improve their access to alcohol screening and brief interventions (SBI). This study adapted an electronic SBI (e-SBI) website shown to be efficacious in college students for OEF/OIF veterans and reported findings from interviews with OEF/OIF veterans about their impressions of the e-SBI.a priori as well as emergent domains.Outpatient veterans of OEF/OIF who drank ≥3 days in the past week were recruited from a US Department of Veterans Affairs (VA) Deployment Health Clinic waiting room. Veterans privately pretested the anonymous e-SBI then completed individual semistructured audio-recorded interviews. Their responses were analyzed using template analysis to explore domains identified a priori domains). They also experienced challenges with portions of the e-SBI assessment and viewed feedback regarding alcohol risk and normative drinking as problematic, but described potential benefits derived from the e-SBI (emergent domains). The most appealing e-SBIs would ensure anonymity and provide personalized transparent feedback about alcohol-related risk, consideration of the context for drinking, strategies to reduce drinking, and additional resources for veterans with more severe alcohol misuse.During interviews, all nine OEF/OIF veterans (1 woman and 8 men) indicated they had received feedback for risky alcohol consumption. Participants generally liked the standard-drinks image, alcohol-related caloric and monetary feedback, and the website’s brevity and anonymity (Results of this qualitative exploratory study suggest e-SBI may be an acceptable strategy for increasing OEF/OIF veteran access to evidenced-based alcohol SBI. Individuals deployed for combat in Iraq and Afghanistan for Operation Enduring Freedom and Operation Iraqi Freedom (OEF/OIF), a group now numbering more than 2.2 million , are at Evidence-based alcohol screening and brief alcohol intervention (SBI) can reduce drinking . The VA Web-based alcohol screening and BI (e-SBI) can increase access to evidenced-based care for alcohol misuse by providing anonymous personalized interventions to large numbers of people at low cost -17 and cElectronically delivered SBIs vary widely in length, therapeutic intensity, design, and populations targeted, and systematic reviews have arrived at different conclusions about their overall efficacy -26. A rehttp://www.DrinkCheck.org) used a briefer alcohol screen, the 3-item AUDIT-C (consumption questions only) questionnaire  doesn’t tell me maybe I should do something different. Doesn’t motivate me to.""Veteran F explained how greater personalization could encourage a decrease in drinking:If you saw that and were thinking about what could make my relationship better, and it says ‘less drinking,’ you’d be like, “Oh, well, there’s an eye-opener.” …[I]f it said something personal that could really only apply to you, by taking this, then I think people would think twice about going and drinking a lot.""[Referring to risk assignment]: Further, for some veterans, the risk feedback implied a judgment about them and their drinking:[People] use this and it just turns back and says, “Yeah, you’re drinking too much.” And, all of a sudden, what has that done? Just one more person accusing them.""[Veteran B]: Although the risk categories were intended to highlight the continuum of alcohol-related risk, some veterans viewed alcohol risk as “all or nothing”:If society says that you drink too much, then you’re deemed an alcoholic or a drunk. And, you know, it’s like there’s no real safe gray area in there that you could play with all that much.""[Veteran I]: Participants did not typically consider their alcohol consumption as risky or inappropriate for the occasion and thought the context for drinking and whether problems were present, not just the level of consumption, were relevant to the assessment of risk:Maybe it‘s just me, but maybe the wording ‘risky drinking.’ You know, you’re doing risky drinking. Well, it doesn’t seem risky when I’m doing it. (Laughter) I’m not going to hurt anybody. I’m not going to do anything wrong.""[Veteran A]: The context for risky drinking was perceived as particularly relevant if drinking was associated with a festive occasion:[It’s been] my birthday, then Christmas, then New Year’s, all in two weeks. And, I’m always responsible with my drinking. I always make sure that if I, I’ve had more than three drinks total in the night, I always take it easy at least an hour before I drive. Make sure I’m calmed down.""[Veteran E]: Context was also considered particularly relevant for a difficult occasion:For me, once a month going out and doing 13 drinks over 9 hours. Like, that’s not much compared to the two or three drinks I’ll have once or twice a week… Usually when I do have those binge nights, there’s a very specific reason I go out and drink like that, whether I’m celebrating… but this last time it was—I didn’t get a job, and I was very upset, and I thought I was gonna get it and it’s been two months, and I’m just like, uh. So, there was a reason; it was, “I’m just gonna go out and do this, and get all my frustrations out.”""[Veteran G]: Individual differences were also important for explaining heavy episodic drinking:That didn’t scare me into drinking any less ‘cause, honestly, I think people’s recommended limits aren’t everybody’s. ‘Cause people are, like, “Oh, don’t drink any more than a six pack.” And, that’s pretty much what I like. And, I don’t really seem to be having a problem with it."": All participants voluntarily reported receiving normative feedback for heavy episodic drinking, which attempted to highlight the incongruence between participants’ perception of “normal” drinking and actual norms. However, they had difficulty accepting veteran outpatients as an appropriate comparison group given differences in experiences among veterans:If you’re just taking into account all VA patients 30 to 39 over the entire country, doesn’t necessarily mean that they spent as much time on deployment, doesn’t mean that they went to the same places that we did, or that I have.""[Veteran E]: Moreover, a few were skeptical of the comparison group data, believing it underrepresented OEF/OIF veterans’ drinking, and that other OEF/OIF veterans would think so as well. Participants felt they knew what their peers were consuming, and it was as much or more than their own consumption:This one here is one of the ones that I thought, “Oh, bull.” “I drink more drinks in a single day than 99 percent of male VA patients my age.” I don’t believe that for a second, okay?… I mean, I know a lot of guys, a lot of Vets… you know, and, they drink just as much as I do, if not more.""[Veteran H]: Although the interview was designed to elicit participant opinions about DrinkCheck and not private information about themselves, every participant volunteered personal details about their experience with alcohol. Personal stories were interwoven throughout the interviews and were often the byproduct of a participant’s review of a specific e-SBI feature. In reference to “managing pain” listed among the health concerns, Veteran F shared this:I’ll have to say, I do drink sometimes cause of that. Like, I separated my shoulder last week, and I can’t just go to the doctors anytime and pick up medication. And, so, a six pack helps. And, a six pack and a hot tub does a pretty good job on that one. So, I have to say I do drink for that sometimes.""Veterans also spoke openly about their experience with alcohol without responding to a specific e-SBI feature. In explaining his tolerance for alcohol, Veteran E offered:My family has a history of alcoholism, and I remember being seven years old and having to drive my dad home drunk from the bar. At seven. It’s kind of screwy. Since then, my father has severely cut back on his drinking. He’s 74 now. He’ll go out, you know, maybe once, twice, a week.... He’ll go out with his buddy, and they’ll have six, seven beers… another night during the week, they’ll do the same thing.""Despite elicited criticism regarding specific e-SBI features, all nine OEF/OIF veterans found the program somewhat helpful in encouraging consideration of their drinking. Specifically, some veterans appreciated being provided the recommended drinking limits  and feltI’ve seen a whole bunch of, like, drinking videos and stuff like that, and they really don’t say, you know, what the actual limit is for, you know, not really harming yourself.""[Veteran C]: Further, despite some doubts and dissatisfaction with the feedback, a few OEF/OIF veterans expressed interest in changing their drinking:You know, it definitely makes me reflect. I think, “Well, hold on a second, you know, it would be better not to drink quite so much at those times.”""[Veteran A]: Some were surprised by their consumption and suggested other veterans using the program might be as well:I liked this page, because I was like “Oh my god” (laughs). Cause I just didn’t realize… putting it on the most that you drank in one night, I was like, “Wow, remind me not to do that very often.” I’ll think about that a lot more.""[Veteran G referring to normative feedback]: Lastly, several participants indicated interest in assistance with their drinking beyond what DrinkCheck had to offer:I don’t want to stop drinking alcohol. And I think I can safely have a drink or two, but maybe I should look at some strategies. Not for quitting, ‘cause I saw [the “Strategies for Cutting Down” link on the Resource page] and I thought, “That’s it.”""[Veteran D]: In particular, Veteran B, who described a family history of problem drinking and experience with Alcoholics Anonymous, suggested offering additional resources:Maybe resource links if the VA has resources… “These are healthy drinking habits. These are resources that are available to you to help you get there.”…[I]f it takes more of a, a corrective approach, giving them steps, giving them resources, giving them options… those types of things are going to cause people to want to come back, to want to re-evaluate themselves, to continue to use the system.""[Veteran B]: This qualitative observational study explored the acceptability of an efficacious web-based Australian e-SBI for college students  adapted Some findings from this study are specific to development of e-SBIs for OEF/OIF veterans. Assessment of alcohol consumption should allow for possible recent changes in drinking. Some participants reported averaging their drinking over the entire year, including time when they were not drinking due to deployment. To avoid underestimation of recent alcohol use, future e-SBI adaptations for returned veterans may want to account for deployment periods by assessing recent drinking or using the AUDIT-C questions without a timeframe . AdditioSome findings from this study of nine OEF/OIF veterans may be applicable to the development of e-SBIs and possibly clinically delivered BIs for other populations. Results suggest that feedback needs to be transparent, as participants were nearly universal in their wish to know how they were assigned to their particular risk category. Tailored personal feedback could include a summary of a participant’s drinking and whether, and the extent to which, weekly or daily limits were exceeded. Electronically delivered SBIs could also be used to educate patients about the risks of heavy episodic drinking. Several participants felt their heavy drinking episodes were not problematic if they were not taking physical risks  or experiencing adverse alcohol-related consequences. The e-SBI could inform such patients of the association of heavy episodic drinking with cognitive deficits  and deveFinally, this study suggests that following e-SBI with an opportunity for participants to debrief could serve as a method to engage veterans around their drinking. Although all nine OEF/OIF veterans easily engaged in a detailed candid discussion of the website as anticipated, it was not anticipated that participants would spontaneously volunteer personal information about their own drinking. This finding suggests that providing returning veterans anonymous opportunities to complete and then discuss their alcohol-use assessment and feedback may be a useful method for engaging them in conversations about their drinking.Results of this study also suggest areas for further research. Normative feedback is a common component of e-SBI and, although no systematic review of the effectiveness of normative feedback has been conducted, it has reduced drinking in college students  and was This study has important limitations, the greatest being that enrollment was limited to a convenience sample of nine OEF/OIF veterans, all of whom were outpatients at a single medical center. Therefore, analyses were limited to descriptive exploration of responses, and it is unlikely that saturation of themes was reached. Further, the small sample was likely inadequate for observing the variation that exists among the general population of OEF/OIF veterans. It is possible that different findings may have emerged had purposive sampling been used to identify patients from different veteran subgroups, additional sites and regions, and veterans who were not receiving care from the VA. Results from this study may best be interpreted as a useful starting point for informing future development of e-SBI for OEF/OIF and other veterans.To summarize, this qualitative exploratory study of nine OEF/OIF veterans found that e-SBI was useful in promoting consideration of their own drinking. Such interventions may be most appealing to OEF/OIF veterans if they ensured anonymity, provided personalized transparent feedback about alcohol-related risk, consider the context for drinking, and provide strategies to reduce drinking and additional resources for veterans with more severe alcohol misuse. Results also highlight the importance of educating patients about the risks associated with heavy episodic drinking. Offering an e-SBI program that is relevant and attractive to OEF/OIF veterans could be an effective strategy for increasing their access to evidenced-based care for alcohol misuse.Topic 1: General Reactions to Web-based Screening and Intervention 1.1 First to begin, can you tell me your general response and impression of the website? [Probe: How did you feel about receiving computerized feedback?] [Probe: How did you feel about the feedback you received?]  [Probe: What did you think of your drinking compared to others? Surprised?] [Probe: Was the summary information about your drinking clear and credible enough?] [Probe: What information on the website was of interest or useful or new to you?] [Probe: What information on the website was uninteresting or bothersome?] [Probe: Did it cause you to think about your drinking?]Topic 2: Barriers/Facilitators to Web-based Screening and Intervention 1.1 What are your thoughts about a web-based intervention generally? For tobacco use? Alcohol use? [Probe: What would make you most likely want to use such a website?] [Probe: Do you have any concerns about answering questions about your alcohol use?] [Probe: Do you feel it would be easier to respond more openly to a computer than a VA provider?] [Probe: Is there anything about your drinking that you would feel comfortable sharing on the web that you would not be willing to share with your provider?] [Probe: Would you want to talk to your provider about the feedback you received from this website?] [Probe: Would you want your responses to this website made available to your provider?] 1.2 Would you be willing to complete this web-based program in a kiosk in the waiting room of the clinic? Through MyHealtheVet?[Probe: Would you be willing to complete the web-based program if you were not compensated for your time?]Topic 3: Impressions of Web Page Layout 2.1 What were your impressions of this page? [Probe: What did you like?] [Probe: What did you dislike?]  [Probe: Which feedback if any was unclear?] [Probe: Which questions or feedback bothered you] [Probe: What do you like or dislike about the tables/figures?] [Probe: Does the placement of tables/figures complement or distract from the text?]  [Probe: How do you think others would respond to this page?] [Probe: How could we improve this page?]  [Probe: Is there anything about the pictures that you like or dislike?]Topic 4: Reactions to Web Page Content 3.1 What sort of reactions did you have when you read this page?  [Probe: Is it relevant to you or other OEF/OIF veterans?] [Probe: Was the feedback based on peer drinking believable?] [Probe: Can you think of any other information it should include?]Topic 5: General Questions 4.1 In general, how was the website helpful to you? [Probe: How could you see it being helpful to others?] [Probe: Was there anything you’d like to see in the website that wasn’t there?] [Probe: What would help make it a better website?] [Probe: What did you think the program was going to be about?] [Probe: Did you want to stop the website? Where in the website did you want to stop? [Probe: What other health topics should the website include?] 4.2 What thoughts do you have on how the VA should help veterans who drink an unhealthy amount of alcohol?1) more web-based interventions2) provide list of self-help or community resources3) provide medications to reduce drinking4) telephone counseling5) review of recovery goals and commitment to change6) individual counselingWe certify that none of the authors has an affiliation with, or financial involvement in, any organization or entity with a direct financial interest in the subject matter or materials discussed in the manuscript . We declare that no financial or nonfinancial competing interests exist concerning the authors and the contents of this article.All authors made substantial contributions to the execution of the study, have been involved from the beginning in drafting the manuscript, and have given approval of the final draft. Specifically, GTL carried out project management, completed patient interviews, participated in iterative review of patient interview coding, and drafted the manuscript. EJH participated in study design, coded the patient interviews, participated in iterative review of patient interview coding, and helped draft the manuscript. LJC coded patient interviews, participated in iterative review of patient interview coding, and helped draft the manuscript. CEL participated in study design, iterative review of patient interview coding, and helped draft the manuscript. ECW participated in study design, iterative review of patient interview coding, and helped draft the manuscript. RMT reviewed transcripts and coding for accuracy, participated in iterative review of patient interview coding, and helped draft the manuscript. EJL provided expertise on qualitative study design and analyses, participated in iterative review of patient interview coding, and helped draft the manuscript. KK provided the web-interface and programming of the Australian e-SBI for adaptation in this study as well as expertise on study design and e-SBI development, participated in iterative review of patient interview coding, and helped draft the manuscript. SCH provided access to the Deployment Health Clinic for patient recruitment as well as expertise on OEF/OIF veterans, participated in study design and review of patient interview coding, and helped draft the manuscript. KAB conceived of the study, obtained funding, and oversaw and participated in all aspects of the study including drafting of the manuscript. All authors read and approved the final manuscript.


Gene regulation underlies fungal physiology and therefore is a major factor in fungal biodiversity. Analysis of genome sequences has revealed a large number of putative transcription factors in most fungal genomes. The presence of fungal orthologs for individual regulators has been analysed and appears to be highly variable with some regulators widely conserved and others showing narrow distribution. Although genome-scale transcription factor surveys have been performed before, no global study into the prevalence of specific regulators across the fungal kingdom has been presented.In this study we have analysed the number of members for 37 regulator classes in 77 ascomycete and 31 basidiomycete fungal genomes and revealed significant differences between ascomycetes and basidiomycetes. In addition, we determined the presence of 64 regulators characterised in ascomycetes across these 108 genomes. This demonstrated that overall the highest presence of orthologs is in the filamentous ascomycetes. A significant number of regulators lacked orthologs in the ascomycete yeasts and the basidiomycetes. Conversely, of seven basidiomycete regulators included in the study, only one had orthologs in ascomycetes.This study demonstrates a significant difference in the regulatory repertoire of ascomycete and basidiomycete fungi, at the level of both regulator class and individual regulator. This suggests that the current regulatory systems of these fungi have been mainly developed after the two phyla diverged. Most regulators detected in both phyla are involved in central functions of fungal physiology and therefore were likely already present in the ancestor of the two phyla. Gene regulation is of major importance for physiology of all organisms, and has been intensively studied in fungi. It ensures that the required genes are switched on and act under the circumstances they are needed, and allows fungi to respond to changing conditions. Thirty-seven classes of regulator proteins have been identified in fungi, such asSaccharomyces cerevisiae,,].].RegulatClick here for filePresence of homologs of the 64 selected regulators in 108 fungal genomes. Presence or absence of homologs of each regulator are tabulated for the 108 fungal genomes analysed.Click here for fileGene IDs of the homologs of the 64 regulators. Gene IDs are displayed as indicated in the genome downloads. In the case of mutiple homologs, the gene ID’s are in the same cell separated by a comma.Click here for file


Background: Although histologic chorioamnionitis (HCA) is known to be associated with poor outcomes in preterm infants, its clinical significance among term infants is not clearly known.Objectives: To investigate the utility of HCA in determining early onset clinical sepsis (EOCS) among term newborns.Methods: The incidence of HCA and EOCS in term infants born during 2008–2009 was evaluated in a single center retrospective study (n = 3417). The predictive value of HCA for determining EOCS in term infants admitted to the neonatal intensive care unit (NICU) for suspected sepsis (n = 388) was quantified. Outcome of otherwise healthy term infants in the nursery with HCA was also investigated.Results: Overall, 11% of term infants with HCA also had EOCS. HCA was associated with increased risk for EOCS  among term infants admitted to the NICU for suspected sepsis. No cases of EOCS were found among otherwise well-appearing infants in the nursery with HCA. Multiple logistic regression analysis indicated that addition of HCA does not increase the power of a model combining C-reactive protein (CRP) and immature to total neutrophil ratio in determining EOCS.Conclusion: Although HCA in term infants is associated with EOCS, it did not improve the ability of CRP and immature to total neutrophil ratio to predict EOCS. Routine placental examination may not contribute to the diagnosis of EOCS in term infants. The potin utero –17.Histologic chorioamnionitis is also identified on placental examination after up to 23.6% of full-term deliveries , but itsThis single center, retrospective cohort study was conducted at Winthrop University Hospital, a tertiary center in Mineola, NY, USA with approximately 5000 deliveries annually. Institutional review board approval was obtained prior to the start of this study.n = 3417). Data regarding presence or absence of HCA were obtained from placental pathology reports, and HCA positive cases were reviewed by blinded pathologists for staging of HCA based on the Redline classification system  colonization with inadequate intrapartum antibiotic prophylaxis, prolonged rupture of membranes) or signs and symptoms suggestive of EOCS . Data were obtained regarding the clinical course of infants, including complete blood counts (CBC), immature to total neutrophils (IT) ratio, serial C-reactive protein (CRP) levels, blood culture, and duration of antibiotic therapy. CBC and blood culture were obtained immediately upon admission to the NICU for suspected EOCS. Repeat CBCs were done at 24 and 48 h after admission. Serial CRPs were obtained at 12, 24, and 48 h of life. Abnormal CRP was defined as at least two consecutive CRP values ≥10 g/L and a high IT ratio was defined as ≥0.20.n = 11) or (2) data obtained from chart review were incomplete (n = 13).Our primary outcome of interest is diagnosis of EOCS, which is defined as a composite of proven sepsis (positive blood culture) or probable sepsis  . InfantsP < 0.05.Descriptive statistics were presented as proportions. Fisher’s exact test was used to evaluate association between sepsis and HCA. Stepwise multiple logistic regression models were used and areas under receiver operator characteristic (ROC) curves were computed using the method of Hanley and McNeil , 22 and P < 0.001, Fisher’s exact test)  had HCA and none had EOCS. Of the infants admitted to the NICU for suspected sepsis, 105 (27.1%) had HCA and 100 (25.8%) had EOCS. The incidence of EOCS was observed to be higher among infants with HCA compared to infants without HCA  and IT ratio  compared to infants without HCA. Blood culture was positive in only three patients (one with HCA and two without HCA) . The PPV of HCA for EOCS was 40% (95% CI = 31–50%) and the NPV was 80% (95% CI = 74–84%).Using Fisher’s exact test, HCA was found to be associated with EOCS . A model adding HCA to CRP and IT ratio did not significantly improved predicting power  of 0.774. A model using both CRP and IT ratio combined significantly improved predictive power with AUC of 0.796 (er Table .P = 0.507).Subgroup analysis was also done to evaluate whether classifying HCA based on severity using the Redline classification would improve its ability to predict EOCS. Of the 105 with HCA, 6 were classified as stage 1, 96 as stage 2, and 3 as stage 3. There was no association between increasing staging of HCA and incidence of EOCS  (P = 0.04)  . Another= 0.001) . Likewis = 0.04) . Most ofIn our study, 389 of the 3417 term placentas (11.4%) examined during the study period had HCA, suggesting that HCA is also a common finding among term infants. This is consistent with other studies that have reported the incidence of HCA in term placentas anywhere from 11.5%  to as hiThe majority of HCA cases  were found among well-appearing term infants, representing almost 10% of infants admitted to the nursery. With current recommendations for discharge as early as 48 h following an uncomplicated vaginal delivery, a number of infants with HCA are likely already discharged home from the nursery by the time placental pathology reports are available. Currently, there is unclear evidence on how to approach this clinical dilemma. Our study is reassuring, in that, none of the well-appearing term infants in the nursery with HCA had EOCS. This suggests that in otherwise healthy-appearing infants, an isolated finding of HCA does not warrant further diagnostic testing or treatment with antibiotics. It also further highlights the importance of relying on good clinical assessment in identifying newborn infants at risk for early onset sepsis.The remaining HCA cases  were found among term infants with signs and/or risk factors suspicious for sepsis. Our study demonstrated that although HCA was associated with EOCS in this population, it did not provide any additional benefit in predicting risk for EOCS beyond that derived from the current recommended diagnostic testing . Though results of placental examination are not available at the time when the decision to start antibiotics is made, they are often reported at around the time when clinicians determine whether to continue antibiotic therapy for presumed EOCS. Our study suggests that it is safe and reasonable to not include placental finding of HCA in decision-making regarding continued treatment for suspected EOCS. It is important to emphasize that our study does not suggest that placental examinations are not needed in term infants. Indeed, a number of important findings can be gleaned from the placenta that could prove useful for other diagnoses, including perinatal asphyxia, intrauterine growth restriction, and twin to twin transfusion.Our study is consistent with that of Roberts et al., who correlated histologic and microbiological evaluation of term placentas. He observed that although 34% (67/195) of placentas had evidence of HCA, only 4% (8/195) had evidence of any infection based on culture and PCR analysis, demonstrating that HCA in term infants is often a result of non-infectious inflammatory process . In anotThere are a number of limitations to our study. The lack of follow-up, especially of well-appearing term infants in the nursery with isolated HCA, prevented us from determining if any infant subsequently developed sepsis after discharge. The inclusion of only term infants with available placental examination, although necessary to determine HCA, likely resulted in a selection bias toward a higher risk population, as term placentas are generally not submitted for examination unless indicated . The rate of HCA among well-infants in our study may thus be an overestimation and likely does not reflect the general term population. The inclusion of both proven and probable sepsis in our definition of EOCS is also an important limitation. Although the most objective evidence for diagnosis of sepsis is microbiological isolation of bacteria from culture, false-positives from contamination by skin flora and false-negatives from small sampling volumes or pre-treatment with antibiotics are not uncommon. There are thus no simple criteria that can be used for the diagnosis of sepsis in newborns, and the combination of clinical and laboratory findings suggestive of sepsis despite negative cultures have been considered an acceptable surrogate . It is aIn conclusion, our study demonstrates that although HCA is associated with EOCS, its presence in placental examination adds little to the current practice of clinical observation and limited laboratory evaluation in detecting and managing early onset sepsis in term infants. Routine placental examination may not contribute to the management of early onset sepsis in term infants.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Structural equation modeling demonstrated the excellent fit of a model linking hopelessness to excessive drinking indirectly via depressive symptoms and drinking to cope. Bootstrapping indicated that this indirect effect was significant. Both depressive symptoms and drinking to cope should be intervention targets to prevent/decrease excessive drinking among Aboriginal youth high in hopelessness.Canadian Aboriginal youth show high rates of excessive drinking, hopelessness, and depressive symptoms. We propose that Aboriginal adolescents with higher levels of hopelessness are more susceptible to depressive symptoms, which in turn predispose them to drinking to cope—which ultimately puts them at risk for excessive drinking. Adolescent drinkers ( AmeIn nonAboriginals, hopelessness has been linked to alcohol use and abuse in both adults and adolescents e.g., , 19), fo, fo19]),self-medication [A number of theoretical models have been proposed to explain the relation between depression and alcohol misuse . Ondication , which pdication ), alcohodication ), and aldication ), for exdication . Few studies, however, have tested the underlying mechanism posited in self-medication theory—namely, drinking-to-cope with negative emotions. In two separate groups of nonAboriginal Canadian adolescents, it was shown that consistent with self-medication theory predictions, drinking to cope mediated the relations of both hopelessness and depressive symptoms with alcohol-related problems . It was The purpose of the present study was to examine the relations among four variables  assessed cross-sectionally in a large group of Canadian Aboriginal adolescent drinkers. We propose that Aboriginal adolescents with higher levels of hopelessness are more susceptible to depressive symptoms, which in turn predispose them to drinking to cope—a drinking motivation that puts them at greater risk for excessive drinking. Thus, we specified a structural equation model in which we hypothesized (a) hopelessness would be directly linked to depressive symptoms, (b) depressive symptoms would be directly linked to drinking to cope, and (c) drinking to cope would be directly linked to excessive drinking. We also specified three meditational hypotheses. We expected (d) hopelessness to be indirectly related to drinking to cope through depressive symptoms, (e) depressive symptoms to be indirectly related to excessive drinking through drinking to cope, and (f) hopelessness to be indirectly related to excessive drinking through depressive symptoms and drinking to cope. The hypothesized structural model is depicted in n = 2) and rural (n = 8) schools were represented. Of the total 837 students, 286 (34%) indicated that they had not consumed any alcohol in the last four months, leaving 551 participants (66%) to be classified as “drinkers” . Of the 551 drinkers, 178 (32%) self-identified as Cree, 91 (16%) as Ojibway, 57 (10%) as Metis, 32 (6%) as Oji-Cree, and 25 (5%) as Dakota. A further 29 (5%) were classified as “Other Aboriginal”.2 Also included among the drinkers, due to their attendance at the participating schools, were 63 (12%) students who self-identified as Caucasian or Black.3 A further 76 (14%) drinkers skipped the race/ethnicity question altogether.4 Of the 551 drinkers, 326 (59%) were from schools in Saskatchewan, 168 (31%) from schools in Manitoba, and 57 (10%) from schools in Quebec. Only the data provided by the 551 drinkers were employed in all subsequent analyses. Abstainers were excluded from completing the measure of drinking to cope because the measure used requires that respondents have at least some relatively recent drinking experience to answer items enquiring about why they drink. The inclusion of even infrequent, light drinkers  ensured that the results would be applicable to a wide range of adolescent drinkers from participating schools . Both urban (ols (see ).α = .78.The SURPS  is a 23-α = .86.The BSI-DEP  is a 7-iα = .81.The DMQ-R  is a 20-quantity. Participants were informed with visual and verbal cues that one drink equals one bottle/can of beer, one small glass of wine, one shot of hard liquor, or one cooler. Next, participants reported how often they normally consumed alcohol; this index was referred to as drinking frequency. Finally, participants indicated how often they normally consumed 5+ drinks in a single sitting (4+ drinks for girls); this index was referred to as binge drinking. Each item was rated on a scale of 1–5, such that high scores on each of the three measures index higher levels of excessive  drinking. Response options for the quantity item were 1 = 1 or 2, 2 = 3 or 4, 3 = 5 or 6, 4 = 7 to 9, and 5 = 10 or more. Response options for the frequency item were 1 = less than monthly, 2 = once per month, 3 = 2 to 3 times per month, 4 = weekly, and 5 = daily or almost daily. Response options for the binge item were 1 = never, 2 = less than monthly, 3 = monthly, 4 = weekly, and 5 = daily or almost daily. Previous research has found adequate reliability of self-reported alcohol consumption measures across a broad range of response formats [α = .79).Excessive drinking was indexed with three items assessing degree of alcohol use in the last four months . First, formats . Nonethe formats . Specifi formats , studentThis study was part of a larger project on alcohol abuse prevention in the 10 participating schools. The project received approval from the Dalhousie Health Sciences Human Research Ethics Board (protocol no.: 2007-1628) and Health Canada's Research Ethics Board (protocol no.: 2007-0026). Recruitment occurred through the active process of relationship- and partnership-building with the communities involved. Policing partners and other community members  approached the investigators upon learning of our previous alcohol abuse prevention work in Aboriginal  communities in Nova Scotia . EssentiSchool administration partners advised as to the method of distributing information about the study to parents/guardians of students in grades 7–12 in participating schools, prior to administration of the survey. An information sheet describing the study was sent in a mail-out directly to parents/guardians. Parents/guardians were encouraged to contact the researchers or school principal for any further information they desired about the study. Parents/guardians were asked to let the researchers/school principal know if they did not consent to having their child participate . Parents/guardians were provided with a toll-free number to contact the researchers for additional information, if they so desired. School administration partners at each site also advised as to whether announcements describing the study should be delivered school-wide through the loud-speaker system along with regular morning announcements and/or delivered by individual classroom teachers. Prior to survey administration, students were informed about the nature of the study, and willing students provided written informed consent at the time of the survey. Consent forms were maintained separately from the completed questionnaires to ensure confidentiality and anonymity. Students were informed that the purpose of the survey was to investigate individual differences in reasons for alcohol use. The student consent form provided potential participants with information about the purpose of the survey, as well as the voluntary and confidential nature of the questions. Students were told that they were free to decline to participate and free to withdraw at any time. Those who declined participation were invited to the school library (under the librarian's supervision) or were asked to remain seated and read while their classmates were completing the survey. All students in grades 7–12 in participating schools were invited to take part in the survey. Approximately 20 students in total declined study participation resulting in a very high response rate (approximately 98% from among eligible students who were attending school on the day of survey administration). Data collection was conducted on a grade-by-grade basis during class time, with the permission and input of the school principal. Data collection was led by one of the coauthors (MNC). Following survey administration, the researcher leading the data collection offered a brief presentation on psychology research. No feedback was given to parents, teachers, or students regarding individual students' scores. Teachers had the option of remaining in the classroom at the time of the survey. Measures were administered in a standard order as follows: demographics, excessive drinking indices, BSI-DEP, DMQ-R, and SURPS. During questionnaire completion, students were permitted to ask questions of the researchers. The small minority of students who had difficulties with reading were offered assistance in reading the survey questions by trusted teachers or classroom aides. Translation into French was provided for the students in the Cree school in Quebec.In order to protect any student from being singled out and labeled, the survey was anonymous. To maintain anonymity and confidentiality, students were asked not to write their names on the forms. The toll-free number mentioned above that was established for parent communication was also offered to students in case they had had any questions about the survey forms in particular, or about the research project more generally. The privacy of each call was ensured because one of the coauthors (M. N. Comeau) was the only person who took the calls. Although participants were not compensated financially, survey administrations were concluded with a snack or meal. By integrating an educational component into data collection and engaging the students, they had the opportunity to participate in a project where the ultimate goal was to develop future culturally relevant alcohol abuse prevention efforts that are more meaningful to the lives of youth in their communities.Mean (and SD) scores on each of the study measures for the total group of 551 drinkers are displayed in the right hand column of r = .42, P < .01) but did not approach the strength at which there would be concern about multicollinearity or redundancy. This result lends support to our conceptualization of hopelessness and depressive symptoms as distinct constructs.5Bivariate correlations between the various study measures are also shown in χ2/df) around 2, a comparative fit index (CFI) and an incremental fit index (IFI) around .95, and a root-mean-square error of approximation (RMSEA) around .06 [SEM was conducted using AMOS 7.0 . Full inound .06 . We repoχ2 = 12.93, P < .05; χ2/df = 2.16; CFI = .99; IFI = .99; RMSEA = .05 . The final model is depicted in P < .05; see Three manifest variables were selected to represent the excessive drinking latent variable: drinking quantity, drinking frequency, and binge drinking. Each observed variable showed substantial and significant loadings (ranging from .63 to .89) on the excessive drinking latent variable. Fit indices also suggested the structural model fit the data well: n = 551) bootstrap samples. These samples were then utilized to estimate bias-corrected standard errors for each hypothesized indirect effect in question. In the case of the indirect path from hopelessness to excessive drinking, the indirect effect was based on all paths and was computed by multiplying (a) path coefficients from the predictor to mediators and (b) path coefficients from mediators to the criterion. In addition, CIs were computed. An indirect effect may be described as significant (P < .05) when the 95% CI for this indirect effect does not include zero.A significant indirect effect indicates that mediation has taken place . We usedβ = .143, B = .167, , and SE = .025. That is, the indirect effect of hopelessness on drinking to cope through depressive symptoms was significant , and SE = .025. Put differently, the indirect effect of depressive symptoms on excessive drinking through drinking to cope was significant , and SE = .029. That is, the indirect effect of hopelessness on excessive drinking through depressive symptoms and drinking to cope was significant .Consistent with hypotheses, hopelessness was directly linked to depressive symptoms. This finding replicates, in an Aboriginal adolescent group, much previous work linking hopelessness with depression in nonAboriginal groups –19. ThisThe present finding linking depressive symptoms directly to drinking to cope replicates, in Canadian Aboriginal adolescents, previous findings from nonAboriginal adolescents showing that depressive symptoms were related to drinking to cope with negative emotions . The strThe third hypothesized direct effect in the current study was a path from drinking to cope to excessive drinking. Cooper's model of adolescent drinking motives contends that drinking to cope is a particularly risky motivation for drinking that sets teens up for greater rates of excessive drinking and drinking-related problems . The preAs hypothesized, the relation of depressive symptoms to excessive drinking was indirect—an effect mediated through drinking to cope. This finding contributes to the understanding of the significant overlap between depression and alcohol use disorders in adults and adolescents alike  by suggePrevious cross-sectional and longitudinal research with nonAboriginal youth suggests a relationship between hopelessness and excessive drinking , 27, 28.Two comments should be made on the composition of our study group. Although data were collected across three Canadian provinces, from both urban and rural communities, and included distinct Aboriginal groups , national representation and ability to generalize across all Canadian Aboriginal groups were nonetheless limited. While initially this may appear a limitation of the present study, this criticism would be misguided. For example, in Canada, there are 11 major Aboriginal language families and 65 distinct dialects . To expeSecond, participants in the present study did not entirely consist of Aboriginal youth, and included at least 12% of student drinkers who were nonAboriginal (see Endnotes 3 and 4). However, the 10 participating schools were schools with high proportions of Aboriginal students, and the large majority of the study participants were Aboriginal. The decision to include all student drinkers regardless of ethnicity/race was consistent with the wishes of our community partners (see Endnote 3) and enhanced our ability to generalize the findings to a wide variety of students attending such schools in Canada.Several potential limitations to the present study should be acknowledged, each of which suggests important avenues for future research. The study was cross-sectional in nature and lies in contrast with the time frame of the theoretical model, which implies unfolding of relations between the study variables over time . While this cross-sectional analysis is a first step in testing the utility of the proposed structural model, the model still requires further investigation within a multiwave longitudinal design . Second, depression from drinking to cope with anxiety [A third possible limitation is that study measures were developed for use with nonAboriginal adolescents and not all have been investigated in terms of their psychometric properties when used with Aboriginal youth. Nonetheless, all showed good internal consistency in the present study and some  have be anxiety ; this reFifth, the present study did not consider potential moderators. For example, given known gender differences in depressive symptoms , drinkiThe present findings of an indirect relation between hopelessness and excessive drinking suggest that targeted interventions for Aboriginal youth who are high in hopelessness are needed to prevent or decrease excessive drinking. The meditational results can be helpful for informing the content of such preventative or early interventions . MediatiA cognitive-behavioral secondary prevention program focusing on hopelessness, depressive symptoms, and drinking to cope has been tested in the form of a school-based intervention among nonAboriginal youth via two randomized controlled trials (RCTs). The intervention targeting adolescents with high levels of hopelessness was shown to increase alcohol abstinence, decrease problem drinking , and redIn sum, we used structural equation modeling to demonstrate the excellent fit of a model which links hopelessness to excessive drinking indirectly via depressive symptoms and drinking to cope in Canadian Aboriginal youth. Bootstrapping indicated that this indirect effect of hopelessness on excessive drinking was significant. Both depressive symptoms and drinking to cope should be intervention targets in school-based programs designed to prevent or decrease excessive drinking among Aboriginal youth with high levels of hopelessness.


In the United States, video game playing is an immensely popular form of entertainment; the majority of adults will have had some experience with video games during their lives , as well as how often they fail (negative feedback), and the associated competitive outcomes.Ericsson et al.  found thWhen Ericsson et al.  studied Deliberate practice requires that individuals engage in training at the limits of their ability, where they often fail. It is not as enjoyable as tasks, like play, where performance can be generated easily and effortlessly. One method to make the activity attractive is to mix a short duration of deliberate practice (10–15 min for beginners) with the majority of time being spent on play. Many video games are structured in a similar manner by having the player spend most time on already mastered activities until they reach challenges, but after a few they are sent back to familiar territory.Video games offer researchers an opportunity to study skill development in a relatively well-controlled environment, where they can manipulate specific parameters of the game. It is relatively easy to maintain a minimum level of experimental control and log data in such a way that specific behaviors can be isolated and related to subsequent performance gains. However, it is difficult to draw generalizable conclusions about the nature of skill development when examining games in which positive feedback outweigh the instances of negative .We believe that applying the expert-performance approach to skill in video game environments is essential for understanding the mechanisms of superior performance as well as the shared components that may eventually account for discovered correlations between video games and general ability measures. Furthermore, we believe that it is unlikely that classic game environments are optimally designed to foster continued improvement in an ecologically valid way.Many of the video games that have been proposed in the literature as vehicles for academic and real-world improvement are those such as Space Fortress, Medal of Honor, Rise of Nations, and Tetris that seem to be highly self-motivating, primarily because they give more positive feedback than informative negative feedback. We argue that meaningful improvement is only achieved through principles of deliberate practice, which include negative feedback , and that many traditionally studied games do not adequately incorporate these components. Superior skill is the product of many years and decades of intense training under conditions that are very different from those in a typical video game. If meaningful real-world tasks were designed like a casual or recreational video game environment, it is unlikely that individuals would improve beyond a relatively low and automated state, which is the observed outcome of extended experience in most professional environments (Ericsson,


Cip1 (p21) plays a crucial role in restraining cellular proliferation and maintaining cellular quiescence. Using in vivo and in vitro approaches the present study shows that p21 is expressed in the subgranular zone of the dentate gyrus of the hippocampus in early neuronal progenitors and in immature neurons, but not in mature neurons or astroglia. In vitro, proliferation is higher in neuronal progenitor cells derived from p21-/- mice compared to cells derived from wild-type mice. Proliferation is increased in neuronal progenitor cells after suppression of p21 using lentivirus expressing short hairpin RNA against p21. In vivo, chronic treatment with the non-selective antidepressant imipramine as well as the norepinephrine-selective reuptake inhibitor desipramine or the serotonin-selective reuptake inhibitor fluoxetine all decrease p21 expression, and this was associated with increased neurogenesis. Chronic antidepressant treatment did not affect the expression of other Cdk inhibitors. Untreated p21-/- mice exhibit a higher degree of baseline neurogenesis and decreased immobility in the forced swim test. Although chronic imipramine treatment increased neurogenesis and reduced immobility in the forced swim test in wild-type mice, it reduced neurogenesis and increased immobility in p21-/- mice. These results demonstrate the unique role of p21 in the control of neurogenesis, and support the hypothesis that different classes of reuptake inhibitor-type antidepressant drugs all stimulate hippocampal neurogenesis by inhibiting p21 expression.The relationships among hippocampal neurogenesis, depression and the mechanism of action of antidepressant drugs have generated a considerable amount of controversy. The cyclin-dependent kinase (Cdk) inhibitor p21 The subgranular zone (SGZ) of the dentate gyrus of the hippocampus and the subventricular zone, which lines the border between the striatum and the lateral ventricle, are known to produce new neurons throughout life. Neuronal stem cells express glial fibrillary acidic protein (GFAP) The relationships among hippocampal neurogenesis, depression and the mechanism of action of antidepressants have generated a considerable amount of interest and controversy. Most antidepressant drugs produce a rapid increase in synaptic levels of norepinephrine and/or serotonin; however, the onset of clinical improvement usually takes 3–4 weeks Cip1 (p21) plays a crucial role in restraining proliferation and maintaining cellular quiescence The end-point molecular mechanisms regulating hippocampal neurogenesis are not clear. In mammalian cells, the control of proliferation primarily occurs in the G1 phase of the cell cycle Previously, we showed that p21 is expressed in the SGZ of the hippocampus tm1Tyj) mice were originally purchased from the Jackson Laboratory, but currently are bred in our laboratory. p21-/- mice were backcrossed to the C57Bl/6 genetic background 6 times prior to testing. p21+/- females and males were used for breeding, and both WT and p21-/- animals were obtained from the same litters. Two month old male mice were used for the experiments. For Western blot analyses and for obtaining NPC, the mice were sacrificed by cervical dislocation, the brains removed and rapidly cooled in ice-cold saline and the hippocampi were dissected out This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Institutional Animal Care and Use Committee at Cedars-Sinai Medical Center (project# 2263). C57Bl/6 and p21-/- , increase neurogenesis and/or produce neurochemical changes reflective of antidepressant action BrdU incorporation was assessed as described previously Analyses were conducted as described previously 4 cells/ml were plated on ECL cell Attachment Matrix coated coverslips  in 24 well plates and cultured for 3 days (to detect DCX-positive cells) or for 8 days. Cells were then fixed in 4% paraformaldehyde and immunocytochemistry performed to detect neuronal markers.Cultures were prepared and conducted according to published protocols NPC cells were isolated from 2 month old WT and p21-/- mice. After the culture was established, at passage 2 the spheres were collected, dispersed and 0.5 cells/well were seeded in 96 well plates in quadruplicate. Ten days later the number of spheres/well was counted as a percentage of cells capable of forming spheres. All spheres were then collected, dispersed and the total number of cells derived from the spheres was counted in each culture.7 transducing units. At passage 2 or 3 the spheres were collected, dispersed and plated at 5000 cell/well in 24-well plates in duplicate. Transducing lentiviral particles expressing p21 shRNA or control non-targeting shRNA were added in the amount calculated to reach optimal multiplicity of infection (MOI) of 20 units. After 48 hr the cells were re-infected with the same amount of lentiviral particles. Cells were fixed in 4% paraformaldehyde 48 hr after the second infection.Lentiviral-based particles permit efficient infection and integration of specific shRNA constructs into non-dividing cells such as neuronal precursors. Mission®Lentiviral Transduction Particles (Sigma-Aldrich) consist of sequence-verified p21 shRNA cloned into pLKO.1-puro lentiviral vectors. The viral titer was greater than 1.5×10Brain paraffin sections (5 µM) or coverslips were double-labeled with primary antibodies conjugated with Alexa 488 and Alexa 568 fluorescent dyes (Molecular Probe)The FST is used primarily as a screen for detecting antidepressant activity, but it also has been used to study animal models of depression Independent-samples t-tests were used to analyze paired and planned comparisons. Groups with more than 2 comparisons were analyzed using one-way ANOVA (to determine the effects of treatments) followed by the Dunnett's test for pairwise comparisons to controls, or two-way ANOVA (to determine the effects of genotype and treatments) followed by the Newman-Keuls posthoc test for all pairwise comparisons. In one instance where homogeneity of variance was not met, the non-parametric Kruskal-Wallis test was used followed by a multiple comparisons of means to detect differences between groups. The level of significance for all analyses was set at p<0.05.+/SOX2+ or GFAP+/nestin+ type 1 cells), transit amplifying progenitors , and neuroblasts (DCX+/βIII-tubulin+), but not mature neurons  +/SOX2+ or in GFAP+/nestin+ cells (data not shown), indicating that p21 is not expressed in radial neuronal stem cells. In the SGZ, p21 is co-localized with SOX2 and with nestin in amplifying neuronal progenitors  of nestin+ cells expressed p21. p21 is expressed in DCX+ neuroblasts   . Thus, ptroglia) .+ and SOX2+ cells co-expressing Ki67 was higher in NPC derived from p21-/- mice compared to WT mice , and at the same time the percentage of p21+ cells was decreased  . The cells were collected, fixed and double-stained for p21 and Ki67. Although the same number of cells was initially plated , at the time of collection there were significantly more cells in NPC cultures treated with p21 shRNA than in the control cultures  , confirmPreviously we showed that chronic treatment with the nonselective tricyclic antidepressant imipramine markedly decreased hippocampal p21 mRNA and protein levels + cells in the SGZ was significantly higher in antidepressant-treated compared to the saline-treated mice, showing that antidepressant treatment increased cellular proliferation  were determined. The number of BrdU+/DCX+ cells was increased in all three antidepressant-treated groups compared to saline-treated controls , the mice were injected with BrdU and then sacrificed 24 hr later. The number of BrdUferation . To detecontrols . Therefo+ cells were counted in the SGZ. Compared to WT mice that received saline, the number of BrdU+ and BrdU+/DCX+ co-labeled cells was significantly higher in the saline-treated p21-/- mice, indicating increased neurogenesis. Treatment with imipramine increased number of BrdU+ and BrdU+/DCX+ co-labeled cells in WT mice, but did not increase the number BrdU+ cells in p21-/- mice. There is a tendency for imipramine to decrease BrdU labeling in p21-/- mice, although this effect did not reach statistical significance. However, impramine significantly reduced the number of BrdU+/DCX+ cells in p21-/- mice  in imipramine-treated p21-/- mice.Caspase 3 is a member of the caspase family, and in apoptotic cell caspase is cleaved to shorter isoforms to trigger the apoptotic process For the FST, WT and p21-/- mice were treated chronically with saline or imipramine as described above. Twenty four hr after the end of antidepressant  treatment the mice underwent the FST. Compared to WT mice that received saline, there was less immobility in the saline-treated p21-/- mice . Althoug+ astrocytes do not express p21, whereas p21 expression is evident in early stage both DCX+ and immature βIII-tubulin+ (Tuj-1+) neurons. Co-labeling for p21 and Ki67 show that these two proteins rarely co-localize. Therefore, when p21 is present, proliferation is restrained in these cells.The data indicate that p21 is involved in the fundamental molecular mechanisms underlying the control of neurogenesis in the hippocampus. The present study confirmed previous findings that p21 is expressed exclusively in the SGZ of hippocampus, but not in granular cell layer where the mature neurons reside in vitro studies support this hypothesis. Compared to neuropheres derived from WT mice, spheres derived from p21-/- mice were larger in size, the spheres were comprised of more cells and they exhibited higher spherogenic ability and increased proliferation. In addition, the percentage of both SOX2+ and nestin+ cells expressing Ki67 (proliferating cells) was higher in NPC derived from p21-/- mice. The experiments using conditional knock down of p21 in NPC from WT mice showed that when p21 expression is decreased, proliferation of NPC is increased. Taken together, these results strongly support the hypothesis that p21 plays a fundamental role in regulating neuronal lineage cell proliferation in the hippocampus and in keeping these cells in a quiescent state. In vivo, saline-treated p21-/- mice showed greater cellular proliferation in the SGZ (as detected by increased BrdU labeling) and increased neurogenesis (as indicated by more BrdU+/DCX+ co-labeled cells) compared to WT mice. These results are in agreement with the finding of increased proliferation in the lateral ventricle wall and in the forebrain stem cells in young p21-/- mice The Other published studies Tissue specific factors can start or stop the proliferation of mitotic cells. Out of three tasted major Cdk inhibitors , it is only p21 that changed in response to antidepressant treatment. We can speculate that in the hippocampus, some tissue-specific factors selectively affect p21. Similar tissue specificity has been demonstrated for other brain regions. For example, TGFβ exerts its anti-proliferative properties in the cerebellum, but not in the hippocampus + and DCX+/BrdU+ cells in response to chronic imipramine treatment. These observations were supported by the results showing that imipramine decreased protein levels of both DCX and NeuN in the p21-/- mice. The mechanism underlying the decrease is not clear. Cleaved caspase 3 is a protein implicated in cell death signaling pathways An important finding is that neurogenesis was not increased in p21-/- mice chronically treated with imipramine, suggesting that p21 suppression is not simply a correlate of increased neuronal cell proliferation, but that down-regulation of p21 is required for the stimulation of neurogenesis produced by this class of antidepressants. In fact, in p21-/- mice there was a decrease, rather than an increase in the number of both BrdUAlthough there is evidence pointing to a mechanistic link between neurogenesis and the mechanism of action of antidepressant drugs, the role of neurogenesis in the pathophysiology of depression remains unclear. Suppression of neurogenesis by hippocampus-directed irradiation in vitro were not found (data not shown), it is likely that antidepressants suppress p21 secondary to enhancing neurotransmission. Similar effects was shown for the long term lithium treatment which suppress p53 expression in cerebellum granular cells In summary, p21 regulates the proliferation of neuronal progenitors in the hippocampus, and the stimulation of neurogenesis produced by reuptake inhibitor-type antidepressants might be a consequence of decreased p21 expression and the subsequent release of neuronal progenitor cells from the blockade of proliferation. Because many antidepressants stimulate neurogenesis, it is possible that their shared common mechanism of action is suppression of p21. At this point the mechanisms underlying this effect are not known. As direct effects of the antidepressants on p21 expression in NPC


Video S1 and Video S3 have been switched in the Supporting Information. Please view the correct videos and their description below.Video S1First day culture of normal newborn ovary. A square indicates somatic cell invasion of oocyte cyst.(MP4)Click here for additional data file.Video S3First day culture of Figla null newborn ovary.(MP4)Click here for additional data file.