This work was supported by National Science Foundation Award #1257162 to AB, and NIH/NIMH BRAINS Innovation award #MH087495 to DK. “
“It is well established that prolonged or chronic exposure to stress can lead to a variety of adverse physiological and psychological consequences, including obesity, drug abuse, and mood disorders (McEwen, 2005, McEwen, 2007 and de Kloet Akt inhibitor et al., 1998). Furthermore, a growing body of evidence indicates that periods marked by significant brain maturation and plasticity, such as perinatal and adolescent development, may be especially vulnerable to these disruptive effects of stress (Romeo et al., 2009 and Eiland

and Romeo, 2013). Less appreciated, however, is the fact that not all individuals exposed to extended or repeated stressors necessarily go on to develop neurobehavioral dysfunctions. The factors that mediate this resilience to stress-induced vulnerabilities are unclear, but likely involve an interaction between genetic and environmental variables (Rutter, 2013 and Southwick and Charney, 2012). The purpose of this review is to discuss possible mechanisms that may contribute to stress resilience, particularly during the adolescent stage of development. Given

the scarcity of data that directly addresses stress resilience during adolescence, this review will also suggest potential future lines of research to help fill this gap in our understanding. An emergent body of research has begun to show the Venetoclax short- and long-term effects of exposure to stress during adolescence on a

diverse set of negative physiological and neurobehavioral outcomes (Eiland and Romeo, 2013, McCormick and Green, 2013, McCormick, 2010, Hollis et al., 2013, McCormick and Mathews, 2010 and McCormick et al., 2010). It has been proposed that all adolescents may show a heightened sensitivity to stressors based on at least three converging factors (Romeo, 2013). First, animal studies have indicated that peripubertal individuals display greater hormonal stress responses compared to adults following a variety of physical and psychological stressors (Romeo, 2010a, Romeo, 2010b and McCormick and Mathews, 2007). Second, neuroanatomical studies have reported that the brain areas known to be highly sensitive to stressors in adulthood, namely the amygdala, hippocampus, and prefrontal cortex, all continue to mature during adolescence (Giedd and Rapoport, 2010). Third, the adolescent brain may be more responsive to the stress-related hormones than the more mature brain, as a previous study in rats showed that exposure to similar levels of corticosterone increased gene expression for glutamate receptor subunits to a greater degree in the adolescent compared to adult hippocampus (Lee et al., 2003).

rhizome powder (1000 mg/kg body weight) Full-size table Table options View in workspace Download as CSV Body weight, food and water intake were monitored daily for 21 days. To detect blood glucose level on 0, 7th and 14th day, blood was collected in heparinized eppendroff tubes, from retro-orbital venous plexus under light ether anaesthesia, using capillary tubes. After the last dose (21st day) of Aqueous slurry of C. orchioides Gaertn. rhizome powder (ASCO) or Glibenclamide had been administered, 16 h Selleck BAY 73-4506 fasted rats were sacrificed by cervical dislocation and cardiac blood was collected. The serum was separated by

centrifugation (5 min, 5000 rpm) and stored in the refrigerator until it was analysed. Blood glucose was estimated using Crest Biosystems kit (Enzymatic glucose oxidase peroxidase (GOD-POD) method by Trinder, 1969). 15 On 21st day, the animals were sacrificed by cervical dislocation and pancreas, kidney and liver were taken and

fixed in 10% buffered formalin, embedded in paraffin wax, serial sections of 5 μm were cut, stained with ZD1839 purchase hematoxylin-eosin, mounted on glass slides, photomicrographed and the observations made were recorded. Values are expressed as the mean ± SE for six animals in each group. Statistical analysis was done using One-way ANOVA followed by Dunnett’s multiple comparison tests at 5% level of significance. Preliminary phytochemical analysis of various solvent extracts of C. orchioides Gaertn. rhizome showed the presence of saponins, glycosides, tannins, phenols, flavonoid and mucilage. Phytochemical analysis by TLC of C. orchioides Gaertn. rhizome extracts showed presence of anthracene, arbutin, cardiac glycosides, bitter drugs, coumarins, essential oils, lignans, pungent–tasting principles, saponins, triterpenes and valepotriates. No toxic effect of ASCO was observed on treatment up to 2000 mg/kg

body weight as the physical health and behaviour of the treated rats appeared normal and no death occurred. The first ASCO was found to be safe till the dose of 2000 mg/kg body weight in rats. The serum glucose level was estimated in normal control, diabetic control, Glibenclamide treated (modern drug control) and ASCO treated rats. The serum glucose level of diabetic control group showed increase from zero day to twenty first day, whereas the groups treated with Glibenclamide showed decrease in the serum glucose level after 21 days of treatment. Animals treated with ASCO showed significant decrease in serum glucose level, which was at par with Glibenclamide treated group (P < 0.05) ( Fig. 1). Each value is mean ± SEM. for 6 rats in each group; *:- Shows significant decrease at P ≤ 0.05 compared to diabetic control Pancreatic islet cells from control rat exhibited normal histoarchitecture.

La seconde étape est la mesure de la vitesse de conduction motrice et de la latence distale : elles sont normales au début de la maladie. Ensuite, la perte importante en axones moteurs peut retentir sur la vitesse de conduction qui ne devient cependant pas inférieure à 80 % de la limite inférieure des valeurs normales. Au-delà, la coexistence d’une neuropathie périphérique doit être évoquée. Lors de l’étude des ondes F, les anomalies sont variables, incluant une augmentation de la latence, en général inférieure à 125 % de la limite supérieure de la normale.

L’amplitude des ondes F varie suivant la prédominance LY294002 de l’atteinte centrale (augmentée) et périphérique (diminuée).

Des blocs de conduction moteurs sont recherchés au cours de l’évaluation des vitesses de conduction motrice par des stimulations étagées comparant les amplitudes des aires proximales et distales. Il est raisonnable d’affirmer qu’il n’existe pas de vrai bloc de conduction au cours d’une SLA certaine. La learn more constatation de blocs de conduction motrice multiples est capitale. Elle doit amener à évoquer le diagnostic de neuropathie motrice multifocale. Il s’agit d’un diagnostic différentiel majeur en raison des possibilités thérapeutiques et d’un meilleur pronostic. La stimulation répétitive est un test diagnostique d’anomalie de la jonction neuromusculaire, il peut être altéré au cours de la SLA. Le décrément observé témoigne d’une instabilité de la conduction et de la transmission neuromusculaires dans les axones dénervés. Il serait un élément de mauvais pronostic. Cette technique est très utile au diagnostic différentiel avec la myasthénie dans les formes bulbaires : l’examen est alors en faveur d’une myasthénie si le décrément s’accompagne de potentiels d’unités motrices de forme normale. Étude de la conduction sensitive périphérique : les vitesses de conduction sensitive et surtout les amplitudes des potentiels sensitifs

sont normales au cours de la SLA, y compris dans les territoires très déficitaires sur le plan moteur. Des anomalies sensitives incitent à rechercher une plexopathie, une polyneuropathie ou une maladie de Kennedy. Si certaines études électrophysiologiques Non-specific serine/threonine protein kinase font état d’altérations sensitives discrètes, celles-ci restent stables alors que la dénervation motrice progresse. Ainsi, les anomalies discrètes ne doivent pas remettre en cause la règle générale d’une absence d’anomalies de la conduction des fibres sensitives périphériques au cours de la SLA. L’ENMG conventionnel joue un rôle essentiel dans le diagnostic de la SLA, cependant de nouvelles techniques ont été proposées dans un but d’évaluation ou de meilleure compréhension de la physiopathologie de cette affection.

The gene products were ligated to the pGEMT-easy vector (Promega), and the sequences were confirmed by DNA sequencing. The pGEMTeasy-pspA constructs were digested with the appropriate restriction endonucleases

and the resulting fragments were ligated to the linearized pAE-6xHis vector [24]. Competent E. coli BL21(DE3) (Invitrogen) were transformed with the pAE-6xHis vectors containing the pspA gene fragments. Protein expression was induced in the mid-log-phase cultures by 1 mM IPTG (Sigma). The recombinant proteins, bearing an N-terminal histidine tag, were purified PLX-4720 order from the soluble fraction through affinity chromatography with Ni2+ charged chelating Sepharose resin (HisTrap Chelating HP; GE HealthCare)

in an Akta Prime apparatus (GE HealthCare). Elution was carried out with 500 mM imidazole. The purified BMS 354825 fractions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), dialyzed against 10 mM Tris–HCl (pH 8) – 20 mM NaCl, and stored at −20 °C. All strains used in this study are described in Table 1. Pneumococci were maintained as frozen stocks (−80 °C) in Todd-Hewitt broth supplemented with 0.5% yeast extract (THY) with 10% glycerol. In each experiment, the isolates were plated on blood agar prior to growth in THY. Female BALB/c mice from Instituto Butantan (São Paulo, Brazil) were immunized intraperitoneally with 5 μg of recombinant PspA derivatives in saline solution 0.9% with 50 μg of Al(OH)3 as adjuvant (500 μl per mouse). The adjuvant alone was used as a control. The animals were given three doses of protein at 7-day intervals. Sera were collected from mice at 14 and 21 days by retro-orbital bleeding. The antibody titers were examined by ELISA [21]. Cross-reactivity of anti-PspA antibodies was analyzed by immunoblot. of S. pneumoniae

strains were grown in 50 ml of THY to mid- to late-log phase. Bacteria were harvested by centrifugation and the pellets were washed 3× in phosphate-buffered saline (PBS), suspended in 1 ml of 2% choline chloride (Sigma) in PBS (pH 7.0), incubated for 10 min at room temperature and centrifuged to recover the eluates [25]. Choline extracts (2 μg) from pneumococcal strains bearing PspAs of clades 1 and 2 were separated by SDS-PAGE and transferred to nitrocellulose membranes (GE Healthcare). Pooled anti-PspA sera (six mice per group) generated against the recombinant PspA fragments of clades 1 and 2 were added at a dilution of 1:1000 (sera collected after the second immunization), followed by incubation with horseradish peroxidase-conjugated goat anti-mouse IgG (diluted 1:1000; Sigma). Detection was performed with an ECL kit (GE Healthcare). S. pneumoniae strains ( Table 1) were grown in THY to a concentration of 108 CFU/ml (optical density of 0.4–0.5) and harvested by centrifugation at 2000 × g for 3 min.

It demonstrated that the likelihood of emergence and spread of equine influenza viruses was dependent on the immunity landscape characterizing the horse population, HDAC cancer and for the first time the relationship between immune escape and epidemic potential was quantified. The impact of pre-existing cellular immunity on influenza virus epidemiological and evolutionary

dynamics is less clear yet likely non-negligible. This calls for further quantitative studies on pre-existing herd immunity—both antibody- and cell-mediated—as a major component of human-to-human transmission barriers. Although acquisition of transmissibility is necessary for the crossing of the human-to-human transmission barriers, it is not sufficient to guarantee sustained spread and maintenance of influenza viruses in a susceptible

human population. The ability of influenza viruses to spread in a host population can be measured by the basic reproduction number R0, which corresponds to the number of secondary cases that arise from one infected individual in a well-mixed susceptible population [181]. R0 is defined mathematically by the product of the transmission rate and the length of the infectious period (Eq. (1)). equation(1) R0=βα+γ Here β is the transmission rate, α is the virus induced-mortality/morbidity rate BVD-523 manufacturer and γ is the recovery rate. The length of the infectious period is defined by 1/(α + γ). Only viruses with R0 above 1 will successfully spread in a well-mixed susceptible population and result in an epidemic. As the epidemic unfolds, the proportion of susceptible individuals (s) decreases as they become infected, recovered and immune, and the effective

reproductive number (Re) of the virus declines (Eq. (2)) equation(2) Re=sR0.Re=sR0. At the peak of the epidemic, Re = 1. Thereafter, Re < 1, and local stochastic extinction of the virus may occur during the epidemic trough [182]. As seen previously, the presence of pre-existing immunity in heptaminol the human population can impact influenza virus probability of emergence and epidemic dynamics. In addition, variability in susceptibility to infection and in infectiousness, e.g., associated with host age or predisposing factors, as well as variability in host behaviour that can affect transmission or infectious period can have dramatic consequences on the epidemic dynamics and maintenance of influenza virus in the human population [183]. For example, schoolchildren are considered to play a primary role in influenza virus transmission [184] and [185], and school terms and holidays in association with heterogenous mixing patterns of individuals of different age classes can be considered important drivers of influenza epidemic dynamics [186] and [187].

During evolution, HPVs have adapted to specific epithelial niches, with different

types having different disease associations and disease prevalence [13], [14] and [23]. Amongst cutaneous HPVs, the diversity within the Alpha (species 2, 3, 4 and 14; see Fig. 1), Beta and Gamma genera contrasts sharply to what is seen in the apparently less successful Mu and Nu genera. The most well studied HPV types are, however, the mucosal Alpha types that cause cervical cancer (see Fig. 2A) [24], and for these the biology of disease is relatively well understood [3]. This is certainly the case for HPV16 (Fig. 2B) infections of the ectocervix and the cervical transformation zone where the majority of HPV16-associated NVP-AUY922 chemical structure cervical cancers develop (Fig. 3). The life-cycle organisation of HPV16 (and Alpha types in general) at other important epithelial sites, such as the anus, the endocervix, the penis [25] and [26] and the oropharynx [27] is, however, still poorly understood [28]. The Alpha PVs are divided into cutaneous and mucosal types, and the mucosal types are further subdivided into high-risk

and low-risk groups [1]. The cutaneous Olaparib cell line Alpha types are also ‘low-risk’, and include HPV2 and 57, which cause common warts, and HPV3 and 10, which cause flat warts [1] and [20]. The low-risk mucosal types (Fig. 2A), which despite their name can also cause cutaneous genital lesions, share a low-risk HPV life-cycle organisation and do not typically cause neoplasia [29] (Figs. 4B and 5). Cutaneous lesions caused by Alpha, Beta, Gamma and Mu types can become difficult to manage in patients with SCID (severe combined immunodeficiency) [30] and EV (epidermodysplasia verruciformis) and in organ transplant recipients and others who are pharmacologically immunosuppressed [31], with certain Beta

types being associated with the appearance of neoplastic precursors (Bowen’s disease, actinic keratosis) [32] and the development of non-melanoma skin cancer at sun-exposed sites in these Ketanserin individuals [6], [31], [33] and [34]. A predisposition to HPV-associated disease and cancer progression is also seen in WHIM syndrome (warts, hypogammaglobulinemia, infections, and myelokathexis) patients, which is associated with defective CXCR4 signalling [35]. The molecular defects that underlie these conditions are known [36], but it is not yet clear (in most cases) exactly how they predispose to disease and whether it is the infected keratinocyte [37] and [38] or the immune system that is primarily compromised [39] and [40]. Thus, the low-risk viruses are occasionally found to be associated with human cancers and can in some instances be associated with papillomatosis, especially in individuals with immune defects. Carcinomas associated with the high-risk HPV types are, however, a far more significant burden [4] and [24].

BALB/c mice (6–8 Ruxolitinib order weeks), free of specific pathogens, were maintained in individually ventilated cages, housed in autoclaved cages and fed on OVA-free diets, in an

air-conditioned room on a 12 h light/dark cycle. Sterile special processing forage and water were provided adequately. Cages, bedding, food, and water were sterilized before use. Pregnant mice went into labor on 20 day of pregnancy and newborn mice were raised and maintained in the same conditions. Mice were divided into the following groups: (1) sensitizations and challenges with ovalbumin (OVA group); (2) treatment with PCV7 immunization in infant, sensitizations and challenges with OVA in adult (PCV7 + OVA group); (3) the control group. On day 21, mice in the PCV7 + OVA group were administered 7-valent pneumococcal conjugate vaccine (PCV7, Wyeth, USA) 33 μl intranasally every 12 h for

three doses [8]. The mice in the OVA and PCV7 + OVA groups were sensitized intraperitoneally with 100 μg ovalbumin (OVA, Sigma) diluted in 50% aluminum hydroxide (Pierce) to a total volume of 200 μl on day 28 and day 42. From day 49 to 52, the mice were challenged with OVA aerosolized for 30 min every day lasting for 4 days. The control group mice were sensitized and challenged with Dasatinib mw sterile PBS at the same time. AAD was assessed 24 h after the final challenge. In our experiment, each experiment was repeated three times. Two to three mice were used in every experimental test described hereafter. This study was approved by the Institutional Animal Care and Research Advisory Committee at the Chongqing Medical University. All experimental animals were used in accordance not with the guidelines issued by the Chinese Council on Animal Care. AHR was assessed in vivo by measuring changes in transpulmonary

resistance using a mouse plethysmograph and methods similar to those previously described [12]. Briefly, 24 h after the final challenge, AHR was assessed in conscious, unrestrained mice by means of whole-body plethysmography (Emca instrument; Allmedicus, France). Each mouse was placed into a plastic chamber and exposed to aerosolized PBS followed by increasing concentrations of an aerosolized methacholine (Sigma-Aldrich, St. Louis, Mo. USA) solution (3.125, 6.25, 12.5, 25, and 50 mg/ml; Sigma) in PBS for 3 min exposures and then the mice had a rest for 2 min, after which a computer program was used to calculate Penh from the continuously recorded pressure and flow data for 5 min. Then take the average of the 5 min records as the value of Penh of this concentration. Penh is a dimensionless value and correlates with pulmonary airflow resistance. It represents a function of the ratio of peak expiratory flow to peak inspiratory flow and a function of the timing of expiration. After formalin fixation, the left lung was dissected and embedded in paraffin. Sections of 4 μm thickness were cut and stained with hematoxylin and eosin (H&E; Sigma).

One shoulder should always point in the direction of movement. Always take off and land on the balls of the feet. Don’t let knees buckle inwards. Complete course twice. 10. Bounding Bound forward, bringing the knee of the trailing leg up as high as possible and bend the opposite arm in front of the body when bounding. Land softly on the ball of the foot with a slightly bent knee. Don’t let knee buckle inwards during take-off or landing. Cover 30 metres twice. Full-size table Table options View in workspace Download as CSV The control group continued their regular warm-up exercises, which usually consists of running exercises,

dynamic and static stretching, and sprinting. The control group was not informed about the injury prevention program implemented in the intervention group and received no further instructions. The control teams were also randomly visited to observe and record this website possible selfinitiated Selleck Dabrafenib preventive measures in their warm-up, specifically those included in the intervention program. All injuries occurring during the competition season were

recorded weekly in a web-based injury registration system by the paramedical staff of the team. An injury was defined as a physical complaint sustained by a participant that resulted from a soccer training session or soccer match, irrespective of the need for medical attention or time lost from soccer activities (Fuller et al 2006, van Beijsterveldt et al 2012). Information about the date of injury, diagnosis, origin, recurrence, and possible contributing factors was collected. After full recovery, defined as participation for the entire duration of a soccer training session or match (van

Beijsterveldt et al 2012), an online recovery form was completed. This recovery form recorded healthcare use, work or school absenteeism, and the purchase of secondary preventive devices (eg, tape and insoles) for the entire injury episode. Economic analysis was performed from the societal perspective, which means that all significant costs associated with the injury were considered, regardless of who pays them (Hakkaart-van Roijen et al 2011). Mean costs first per participant and mean costs per injured participant were calculated. The economic evaluation was designed as a cost-effectiveness analysis to determine the costs of preventing an injury by means of the intervention program, compared to the control group. The incremental cost-effectiveness ratio presents the incremental costs of using the intervention program to prevent one injury, in comparison with regular warm-up. Incremental cost-effectiveness ratios were calculated by dividing the difference in mean total costs per participant between the intervention group and control group by the difference in numbers of injuries between the two groups, corrected for the difference in the number of participants between the groups.

4). Compound no. 1 (10–50 μg/ml) & compound no. 2 (10–50 μg/ml) was able to inhibit the gastric Hydrogen Potassium ATPase activity in comparison to omeprazole with an IC50 value of 101.22 μg/ml & 55.4 μg/ml respectively. Positive control used during experiment was omeprazole (10–50 μg/ml) and it was able to reduce the enzyme activity with an IC50 value of 30.24 μg/ml (Table 4). A. squamosa is known for its different types of medicinal properties, but still a lot of work is required to establish its antiulcer activity. In our present work, we have tested antiulcer activity of ethanolic extract of A. squamosa

whole plant and have established a better antiulcer activity. The results obtained are comparable to selleck kinase inhibitor standard drug omeprazole. Isolated compounds (compound no.1&2) were tested for Hydrogen Potassium ATPase activity & they are showing a very good antiulcer activity. All authors have none to declare. The author gratefully acknowledges the expert guidance of Dr. Y. Kumar and Dr. S. Sadish Kumar for their valuable suggestions. Author also acknowledges the necessary platform & financial assistance for research provided by I.T.S Paramedical (Pharmacy) College, Muradnagar, Ghaziabad. “
“Isoxazoles are one of the five membered categorised heterocycles having two different hetero atoms in their cyclic Bak apoptosis skeleton. In recent years there has been renewed interest in them due to their uses as pharmaceutical1 and pesticide.2 and 3

Analeptic activity associated with toxicity has been found

in numerous N-substituted amides of some isoxazole carboxylic acids. A number of 5-isoxazolone and 4-isoxazolone dyes have been reported in the literature as photographic sensitizers and super sensitizers.4, 5, 6, 7, 8 and 9 According to Barnes et al,10 the highly enolised diketones which posses alternative H-bonded (tautomeric structures),a rigorous study on the direction of enolisation was a governing factor in the ratio of the products obtained as well as the site selectivity,11 and there is a possibility of the formation of the two regioisomers of isoxazoles by the nucleophillic attack of hydroxylamine either on α or γ-carbonyl of diketoester. We report herein a convenient, rapid and general method for the synthesis of 5-(substituted phenyl)-4-methyl-3yl-(imidazolidine-1yl Electron transport chain methyl, 2-ylidene nitro imine)-isoxazole 6a–k (Table 1) using 5 synthetic stages in the Scheme 1. 5-(substituted phenyl)-4-methyl-3yl-(imidazolidine-1yl methyl, 2-ylidene nitro imine)-Isoxazole were obtained in good to excellent yields, and screened for fungal activity (Table 2). 1-Phenyl-propan-1-one (13 g, 0.1 mol) was added drop wise over a period of 10 min to a solution of sodium methoxide (5.5 g, 0.1 mol) was added drop wise without external cooling. Freshly prepared hydroxylamine hydrogen-sulphate (HAS) ins sulfuric acid was added to the above solution and the reaction mixture was heated to reflux for 2 h and the reaction was monitored by TLC (hexane: EtOAc, 90:10).

, 2005) and results in a stronger immune response in younger versus older adolescents (Dobson et al., 2013). There is evidence, as well, that HPV vaccine induces robust immune memory (Olsson et al., 2007) Selleck FG 4592 and that sufficient antibody levels may last for at least 12 years and perhaps much longer in most vaccinated individuals (Fraser et al., 2007). Evidence has also suggested that, if needed, an additional dose of vaccine administered years after the initial series may boost the sustained effectiveness of vaccination (Olsson et al., 2007). A communication challenge posed by HPV vaccination is that while both

vaccines are very efficacious, they do not protect against all types of HPV responsible for cervical and other anogenital cancers. This kind of complexity (high efficacy against vaccine types, but more modest efficacy when the whole range of oncogenic HPV is considered)

may be difficult to communicate in a health care setting and difficult for parents to understand. Visual aids, such as the use of charts and graphs, may help to most effectively deliver this kind of information (Chua et al., 2006). In the context of such communication, the need for sexually active females who have been vaccinated to nonetheless have periodic cervical cancer screening must remain learn more an emphasis. Although the strong evidence for efficacy and safety of HPV vaccine dispels many concerns that have been associated with a new vaccine, it is also important to note that HPV vaccine has been licensed in the U.S. and Canada since 2006 and in Australia since 2007 (Centers for Disease Control and Prevention, 2007, Garland and Smith, 2010 and National Advisory Committee on Immunization, 2012). Clinicians who are influential in vaccine uptake, therefore, should no longer consider this vaccine new. Content analysis studies about the media’s representation of the HPV vaccine demonstrate that the

tone associated with the vaccine is inconsistent, ranging from negative to neutral to positive (Briones et al., 2012, Habel et al., 2009 and Keelan et al., 2010). Unfortunately, it is often the unrealistic, negative vaccine fears that become salient to Histone demethylase the public, which then tends to sensationalize potential side effects of vaccination. These rumors then filter down to adolescents and become further exaggerated (Brabin et al., 2009). In order to overcome this type of misinformation, clinicians and public health officials need to advocate for more accurate vaccine information and evidence-based media coverage (Cooper et al., 2008). Further, using social media tools (e.g. Facebook, Twitter) is another key strategy to disseminate accurate information and dispel some of misinformation that is spread by the anti-vaccine movement (Betsch et al., 2012 and Keelan et al., 2010).