Comparing the effectiveness of robot-based to human-based intervention in improving joint attention in autistic children.

Background: Children with autism have impairments in initiation of joint attention (IJA) and response to joint attention (RJA). Aims: The present study compared the learning effectiveness of robot-based intervention (RBI) with that of content-matched human-based intervention (HBI) in improving joint attention (JA). We examined whether RBI would enhance RJA, in comparison to HBI. We also examined whether RBI would increase IJA, in comparison to HBI. Methods and procedures: Thirty-eight Chinese-speaking children with autism aged 6 to 9 years were randomly assigned to RBI and HBI groups. Before intervention, their autism severity, cognitive abilities, and language skills were assessed. Each child received six 30-min training sessions over 3 weeks. During training, he/she watched one or two robot/human dramas twice where two robot/human actors demonstrated eye contact and RJA. Outcomes and results: Children in the RBI (but not HBI) group produced more RJA and IJA behaviors in the delayed post-test than in the pre-test. Parents of the RBI children rated the program more positively than those of the HBI children. Conclusions and implications: RBI may be more effective than HBI in promoting JA in autistic children with high support needs. Our findings shed light on the application of robot dramas in enhancing social communication skills.

Robot dramas may improve joint attention of Chinese-speaking low-functioning children with autism: stepped wedge trials.

INTRODUCTION: Children with autism spectrum disorder (ASD), especially those with low cognitive functioning, have deficits in joint attention. Previous research has found that these children are interested in engaging with social robots. PURPOSE: In the present study, we designed a robot drama intervention for promoting responses to joint attention abilities (RJA) of children with low-functioning autism (IQs < 70). MATERIALS AND METHODS: Using a stepped wedge design, Chinese-speaking children aged six to eight were randomly assigned to three tiers (N = 18). Children of all three tiers had comparable autism severity, language and cognitive function, and joint attention abilities. Tier 1 first received intervention, followed by Tiers 2 and 3. They watched six dramas in which social robots demonstrated RJA behaviours. RESULTS: The RJA of children of all tiers improved after intervention and such improvement was maintained over time. Despite initiation of joint attention (IJA) not being explicitly taught, IJA of all children was found to improve after intervention. CONCLUSIONS: It was, therefore, concluded that a robot drama could enhance the joint attention of children with low-functioning ASD.Implications for rehabilitationIn comparison to typically developing children, children with autism spectrum disorder (ASD), especially those with low cognitive functioning, have deficits in joint attention.Robot-based drama intervention program was developed to promote responses to joint attention (RJA) abilities of children with low-functioning autism (IQs < 70).Modelling RJA in robot dramas can promote RJA abilities in these children.An improvement in RJA also yielded an increase in the initiation of joint attention (IJA) abilities.Children with low-functioning autism might be able to extract the initiation of joint attention skills from the drama, even though these behaviours were not explicitly taught.

What affects gestural learning in children with and without Autism? The role of prior knowledge and imitation.

The present study examined whether prior knowledge to the learning target and imitation during learning affected learning outcomes in preschool children with autism spectrum disorder (ASD, N = 22) compared to their typically developing (TD, N = 15) peers. Children's gestural skills in recognizing and producing the target gestures before and after the training, as well as their imitative behavior during the training were coded. Results showed that consistent prior knowledge benefited gestural learning in both groups. Besides, only children with ASD were hindered by inconsistent prior knowledge. Notably, the effect of imitation was not significant in the ASD group. In conclusion, the learning process in children with ASD may differ from those with typical development, suggesting special-designed interventions are required.

Gesture Development in Chinese-Speaking Preschool Children With Autism and the Roles of Parental Input and Child-Based Factors.

PURPOSE: Children with autism are found to have delayed and heterogeneous gesture abilities. It is important to understand the growth of gesture abilities and the underlying factors affecting its growth. Addressing these issues can help to design effective intervention programs. METHOD: Thirty-five Chinese-speaking preschoolers with autism spectrum disorder (M age = 4.89 years, SD = 0.91; four girls) participated in four play sessions with their parents over 9 months. Their child-based factors including autism severity, intellectual functioning, and expressive language abilities were assessed. The gestures (deictic, iconic, and conventional) of the children and their parents were coded. Growth curve analyses were conducted to examine individual growth trajectories and the roles of child-based factors and parental input in shaping the children's gesture development. RESULTS: Child-based factors and parental input predicted gesture development differently. Parents' gestures positively predicted their children's gestures of the same type. Autism severity negatively predicted iconic and conventional gestures. Overall growth was found in deictic rather than iconic and conventional gestures. Subgroup variation was also found. Specifically, children with better expressive language ability showed a decrease in deictic gestures. An increase in iconic and conventional gestures was found in children with more severe autism and those with poorer expressive language ability and intellectual functioning, respectively. CONCLUSIONS: Different types of gestures may have different growth trajectories and be predicted by different child-based factors. Particular attention should be given to children who never produced iconic gestures, which is more challenging and may not develop over a short period, and hence require direct instruction.

Conversation Skills in Chinese-Speaking Preschoolers with Autism: The Contributing Role of Parents' Verbal Responsiveness.

Children with autism spectrum disorder (ASD) have conversation deficits, yet the growth of conversation abilities is understudied, especially in Chinese-speaking populations. Little is known about whether their parents' verbal responsiveness and redirectives are related to their conversation skills. Children with ASD (N = 37; M = 5;5) and their parents contributed their language samples. These children interacted with their parents at four time points over nine months. The number of conversational turns and the proportion of child-initiated conversation (but not the proportion of children's appropriate responses) grew over nine months. After controlling for time, autism severity, and language skills, parents' verbal responsiveness positively predicted children's appropriate responses. Parents' redirectives negatively predicted the proportion of children's appropriate responses and the number of conversational turns.

Structural Insight into Integrin Recognition and Anticancer Activity of Echistatin.

Echistatin (Ech) is a short disintegrin with a long 42NPHKGPAT C-terminal tail. We determined the 3-D structure of Ech by X-ray crystallography. Superimposition of the structures of chains A and B showed conformational differences in their RGD loops and C-termini. The chain A structure is consistent with our NMR analysis that the GPAT residues of the C-terminus cannot be observed due to high flexibility. The hydrogen bond patterns of the RGD loop and between the RGD loop and C-terminus in Ech were the same as those of the corresponding residues in medium disintegrins. The mutant with C-terminal HKGPAT truncation caused 6.4-, 7.0-, 11.7-, and 18.6-fold decreases in inhibiting integrins αvß3, αIIbß3, αvß5, and α5ß1. Mutagenesis of the C-terminus showed that the H44A mutant caused 2.5- and 4.4-fold increases in inhibiting αIIbß3 and α5ß1, and the K45A mutant caused a 2.6-fold decrease in inhibiting αIIbß3. We found that Ech inhibited VEGF-induced HUVEC proliferation with an IC50 value of 103.2 nM and inhibited the migration of A375, U373MG, and Panc-1 tumor cells with IC50 values of 1.5, 5.7, and 154.5 nM. These findings suggest that Ech is a potential anticancer agent, and its C-terminal region can be optimized to improve its anticancer activity.

Gestures in Storytelling by Preschool Chinese-Speaking Children With and Without Autism.

Previous findings on gestural impairment in autism are inconsistent, while scant evidence came from Chinese-speaking individuals. In the present study, preschool Chinese-speaking children with typical development and with autism were asked to generate stories from a set of wordless Cartoon pictures. Two groups were matched in chronological age and language developmental age. Their speech and gestures were coded. Compared to children with typical development, children with autism produced fewer gestures and showed lower gesture rate. Besides, children with autism produced fewer emblems and fewer supplementary gestures compared to their TD peers. Unlike children with typical development, children with autism tend to produce emblems for reinforcing, rather than supplementing information not conveyed in speech. Results showed the impairments in integrating the cross-modal semantic information in children with autism.

A Robot-Based Play-Drama Intervention May Improve the Joint Attention and Functional Play Behaviors of Chinese-Speaking Preschoolers with Autism Spectrum Disorder: A Pilot Study.

Children with autism spectrum disorder (ASD) have deficits in joint attention and play behaviors. We examined whether a robot-based play-drama intervention would promote these skills. Chinese-speaking preschool children were randomly assigned to an intervention group (N = 12) and a waitlist control group (N = 11). Children in the intervention group watched three robot dramas and engaged in role-plays with both robots and human experimenters over the course of 9 weeks. There were significant improvements in joint attention initiations and functional play behaviors in the intervention group. Parents of this group of children also reported less severe social impairments. It was therefore concluded that a robot-based play-drama intervention can enhance the joint attention and play behaviors of children with ASD.

Robot-based play-drama intervention may improve the narrative abilities of Chinese-speaking preschoolers with autism spectrum disorder.

BACKGROUND: Children with autism spectrum disorder (ASD) have deficits in their narrative skills and gestural communication. Very few intervention studies have been conducted with the aim of improving these skills. AIMS: We examined whether children with ASD who received the robot-based drama intervention had better narrative abilities and gestured more often than their peers who did not receive the intervention. METHODS AND PROCEDURES: Preschool children were randomly assigned to the intervention group (N = 13) and waitlist control group (N = 13). Children in the intervention group watched three robot dramas and engaged in roleplays with both robots and human experimenters. Children in both groups took the pre-tests, immediate post-tests, and, two week later, delayed post-tests, in which they narrated three stories. OUTCOMES AND RESULTS: There were significant improvements in various narrative measures, including narrative length, syntactic complexity, narrative structure, and cognitive inferences, in the intervention group. There was also an improvement in the average number of overall gestures per clause in this condition. These learning outcomes were maintained in the delayed post-test. These patterns were not found in the waitlist control group. CONCLUSIONS AND IMPLICATIONS: A robot-based play-drama intervention can enhance the narrative abilities and gestural communication of children with ASD.

Who is a better teacher for children with autism? Comparison of learning outcomes between robot-based and human-based interventions in gestural production and recognition.

BACKGROUND: Individuals with autism spectrum disorder (ASD) tend to show deficits in engaging with humans. Previous findings have shown that robot-based training improves the gestural recognition and production of children with ASD. It is not known whether social robots perform better than human therapists in teaching children with ASD. AIMS: The present study aims to compare the learning outcomes in children with ASD and intellectual disabilities from robot-based intervention on gestural use to those from human-based intervention. METHODS AND PROCEDURES: Children aged six to 12 with low-functioning autism were randomly assigned to the robot group (N = 12) and human group (N = 11). In both groups, human experimenters or social robots engaged in daily life conversations and demonstrated to children 14 intransitive gestures in a highly-structured and standardized intervention protocol. OUTCOMES AND RESULTS: Children with ASD in the human group were as likely to recognize gestures and produce them accurately as those in the robot group in both training and new conversations. Their learning outcomes maintained for at least two weeks. CONCLUSIONS AND IMPLICATIONS: The social cues found in the human-based intervention might not influence gestural learning. It does not matter who serves as teaching agents when the lessons are highly structured.

Robot-based intervention may reduce delay in the production of intransitive gestures in Chinese-speaking preschoolers with autism spectrum disorder.

Background: Past studies have shown that robot-based intervention was effective in improving gestural use in children with autism spectrum disorders (ASD). The present study examined whether children with ASD could catch up to the level of gestural production found in age-matched children with typical development and whether they showed an increase in verbal imitation after the completion of robot-based training. We also explored the cognitive and motor skills associated with gestural learning. Methods: Children with ASD were randomly assigned to two groups. Four- to 6-year-old children with ASD in the intervention group (N = 15) received four 30-min robot-based gestural training sessions. In each session, a social robot, NAO, narrated five stories and gestured (e.g., both hands clapping for an awesome expression). Children with ASD were told to imitate the gestures during training. Age-matched children with ASD in the wait-list control group (N = 15) and age-matched children with typical development (N = 15) received the gestural training after the completion of research. Standardized pretests and posttests (both immediate and delayed) were administered to assess the accuracy and appropriateness of gestural production in both training and novel stories. Children's language and communication abilities, gestural recognition skills, fine motor proficiencies, and attention skills were also examined. Results: Children with ASD in the intervention condition were more likely to produce accurate or appropriate intransitive gestures in training and novel stories than those in the wait-list control. The positive learning outcomes were maintained in the delayed posttests. The level of gestural production accuracy in children with ASD in the delayed posttest of novel stories was comparable to that in children with typical development, suggesting that children with ASD could catch up to the level of gestural production found in children with typical development. Children with ASD in the intervention condition were also more likely to produce verbal markers while gesturing than those in the wait-list control. Gestural recognition skills were found to significantly predict the learning of gestural production accuracy in the children with ASD, with such relation partially mediated via spontaneous imitation. Conclusions: Robot-based intervention may reduce the gestural delay in children with ASD in their early childhood.

Dynamics and functional differences between dendroaspin and rhodostomin: insights into protein scaffolds in integrin recognition.

Dendroaspin (Den) and rhodostomin (Rho) are snake venom proteins containing a PRGDMP motif. Although Den and Rho have different 3D structures, they are highly potent integrin inhibitors. To study their structure, function, and dynamics relationships, we expressed Den and Rho in Pichia pastoris. The recombinant Den and Rho inhibited platelet aggregation with the K(I) values of 149.8 and 83.2 nM. Cell adhesion analysis showed that Den was 3.7 times less active than Rho when inhibiting the integrin αIIbß3 and 2.5 times less active when inhibiting the integrin αvß3. In contrast, Den and Rho were similarly active when inhibiting the integrin α5ß1 with the IC50 values of 239.8 and 256.8 nM. NMR analysis showed that recombinant Den and Rho have different 3D conformations for their arginyl-glycyl-aspartic acid (RGD) motif. However, the comparison with Rho showed that the docking of Den into integrin αvß3 resulted in a similar number of contacts. Analysis of the dynamic properties of the RGD loop in Den and Rho showed that they also had different dynamic properties. These results demonstrate that protein scaffolds affect the function, structure, and dynamics of their RGD motif.

Expression in Pichia pastoris and characterization of echistatin, an RGD-containing short disintegrin.

Echistatin (Ech) is a potent inhibitor of integrins and was isolated from snake Echis carinatus. To facilitate the study on the molecular determinants of integrin-ligand interactions, we expressed recombinant Ech and its mutants in the Pichia pastoris (P. pastoris) expression system and purified them to homogeneity with the yields of 2-7 mg/L. Ech produced in P. pastoris inhibited platelet aggregation with the IC(50) value of 210.5 nM. The sequential assignment and structure analysis of recombinant Ech were obtained by 2D and 3D (15)N-edited NMR spectra. These data suggests that Ech produced in P. pastoris retained its function and native fold. The results presented here provide the evidences that four disulfide-bonded peptide inhibitor of integrin, Ech, can be expressed in P. pastoris with correct fold and high yield. Platelet aggregation analysis of Ech mutants showed that the length of C-terminus and the K45 residue of Ech are important for interacting with integrin αIIbß3. We also found that recombinant Ech can inhibit the migration of human A375 melanoma cell. These findings may serve as the basis for understanding the activities of Ech.

Expression in Pichia pastoris and characterization of APETx2, a specific inhibitor of acid sensing ion channel 3.

Acid sensing ion channels (ASICs) are family of proteins predominantly present in the central and peripheral nervous system. They are known to play important roles in the pathophysiology of pain and ischemic stroke. APETx2 is a potent and selective inhibitor of ASIC3-containing channels and was isolated from sea anemone Anthopleura elegantissima. To facilitate the study on the molecular determinants of ASIC3-ligand interactions, we expressed recombinant APETx2 in the Pichia pastoris (P. pastoris) expression system and purified it to homogeneity. Recombinant APETx2 produced in P. pastoris inhibited the acid-evoked ASIC3 current with the IC(50) value of 37.3 nM. The potency of recombinant toxin is similar to that of native APETx2. The sequential assignment and structure analysis of APETx2 were obtained by 2D and 3D (15)N-edited NMR spectra. Our NMR data suggests that APETx2 produced in P. pastoris retained its native fold. The results presented here provide the first direct evidence that highly disulfide bonded peptide inhibitor of ASIC3, APETx2, can be expressed in P. pastoris with correct fold and high yield. We also showed that the R17A mutant exhibited a decrease in activity, suggesting the feasibility of the use of this expression system to study the interactions between APETx2 and ASIC3. These evidences may serve as the basis for understanding the selectivity and activity of APETx2.

Association between plasma alkaline phosphatase and C-reactive protein in Hong Kong Chinese.

BACKGROUND: Alkaline phosphatase (ALP) is a biomarker for hepatobiliary and skeletal diseases. It is also raised in sepsis. In atherosclerotic plaques, ALP is expressed. Similar to C-reactive protein (CRP), it may be another marker of systemic inflammation. Therefore, we investigated their association in a Hong Kong Chinese population. METHODS: Plasma ALP and CRP were measured in 205 subjects (110 men, 95 women; age 55.2+/-11.6 years) in the Hong Kong Cardiovascular Risk Factor Prevalence Study-2 cohort. RESULTS: The blood levels of ALP and CRP were significantly correlated (r=0.30, p<0.001), which was due to a significant correlation in women (r=0.43, p<0.001). In a multivariate model, CRP level was related to ALP (beta=0.18, p=0.008). After adjusting for confounding factors and other liver enzymes, the relationship between ALP and CRP remained significant in women (beta=0.28, p=0.019), but in men, ALP was not an independent determinant of CRP levels. CONCLUSIONS: ALP may be another marker of systemic inflammation, especially in women. Whether it provides clinical information additional to CRP requires further study.

Components of the metabolic syndrome predictive of its development: a 6-year longitudinal study in Hong Kong Chinese.

OBJECTIVE: To investigate which of the components of the metabolic syndrome best predict its development. DESIGN: Long-term cohort of randomly selected adults. PATIENTS: One thousand five hundred and forty-eight subjects from the Hong Kong Cardiovascular Risk Factor Prevalence Study who did not have the metabolic syndrome by the US National Cholesterol Education Program (NCEP) or International Diabetes Federation (IDF) criteria at baseline. MEASUREMENTS: Waist circumference, blood pressure, glucose, triglycerides and high-density lipoprotein-cholesterol (HDL). RESULTS: After a median interval of 6.4 years, there were 219 and 143 new cases (21.9 and 14.3 per 1000 person-years) of the metabolic syndrome by the NCEP and IDF criteria, respectively. The odds ratio for the NCEP metabolic syndrome was highest for low HDL, 4.08 [95% confidence interval (CI): 2.90-5.73] and that for the IDF metabolic syndrome was highest for central obesity, 5.94 [95% CI: 3.98-8.87]. Low HDL, found in 27.8% men and 34.3% women, had the highest sensitivity for the NCEP metabolic syndrome (48% in men and 57% in women) and the IDF metabolic syndrome (41% in men and 54% in women). Central obesity had the highest positive predictive values except that triglycerides had the highest positive predictive value for the NCEP metabolic syndrome in women. The areas under the receiver operator characteristic curve for waist circumference, triglycerides and HDL were similar. A model that included waist circumference and HDL predicted the metabolic syndrome as well as a model that included all five metabolic syndrome components. CONCLUSION: Obese Chinese adults should be periodically screened for the metabolic syndrome and have waist and HDL measurement.

Development of diabetes in Chinese with the metabolic syndrome: a 6-year prospective study.

OBJECTIVE: We investigated the association of the metabolic syndrome with new-onset diabetes in the Hong Kong Cardiovascular Risk Factor Prevalence Study cohort. RESEARCH DESIGN AND METHODS: We followed up on 1,679 subjects without diabetes at baseline. Those with a previous diagnosis of diabetes or those who were receiving drug treatment were considered to be diabetic. The remaining subjects underwent a 75-g oral glucose tolerance test (OGTT). Diabetes was defined by plasma glucose > or =7.0 mmol/l with fasting and/or > or =11.1 mmol/l at 2 h. RESULTS: The prevalences of the metabolic syndrome at baseline were 14.5 and 11.4%, respectively, according to U.S. National Cholesterol Education Program (NCEP) and International Diabetes Federation (IDF) criteria. After a median of 6.4 years, there were 66 and 54 new cases of diabetes in men and women, respectively. The metabolic syndrome at baseline predicted incident diabetes. Hazard ratios (HRs) for the NCEP and IDF definitions of the syndrome were 4.1 [95% CI 2.8-6.0] and 3.5 [2.3-5.2], respectively. HRs for fasting plasma glucose (FPG) > or =6.1 or 5.6 mmol/l were 6.9 [4.1-11.5] and 4.1 [2.8-6.0], respectively. The NCEP and IDF criteria had 41.9 and 31.7% sensitivity and 87.5 and 90.2% specificity, respectively. Their positive predictive values were low, approximately 20%, but their negative predictive values were approximately 95%. CONCLUSIONS: The metabolic syndrome, particularly its component, elevated FPG, predicts diabetes in Chinese. An individual without the metabolic syndrome is unlikely to develop diabetes, but one who has it should practice therapeutic lifestyle changes and have periodic FPG measurements to detect new-onset diabetes.

Preparation of amino-acid-type selective isotope labeling of protein expressed in Pichia pastoris.

We report the culture conditions for successful amino-acid-type selective (AATS) isotope labeling of protein expressed in Pichia pastoris (P. pastoris). Rhodostomin (Rho), a six disulfide-bonded protein expressed in P. pastoris with the correct fold, was used to optimize the culture conditions. The concentrations of [alpha-15N] selective amino acid, nonlabeled amino acids, and ammonium chloride, as well as induction time, were optimized to avoid scrambling and to increase the incorporation rate and protein yield. The optimized protocol was successfully applied to produce AATS isotope-labeled Rho. The labeling of [alpha-15N]Cys has a 50% incorporation rate, and all 12 cysteine resonances were observed in HSQC spectrum. The labeling of [alpha-15N]Leu, -Lys, and -Met amino acids has an incorporation rate greater than 65%, and the expected number of resonances in the HSQC spectra were observed. In contrast, the labeling of [alpha-15N]Asp and -Gly amino acids has a low incorporation rate and the scrambling problem. In addition, the culture condition was successfully applied to label dendroaspin (Den), a four disulfide-bonded protein expressed in P. pastoris. Therefore, the described condition should be generally applicable to other proteins produced in the P. pastoris expression system. This is the first report to present a protocol for AATS isotope labeling of protein expressed in P. pastoris for NMR study.