Transcript-specific enrichment enables profiling rare cell states via scRNA-seq.

Single-cell genomics technologies have accelerated our understanding of cell-state heterogeneity in diverse contexts. Although single-cell RNA sequencing (scRNA-seq) identifies many rare populations of interest that express specific marker transcript combinations, traditional flow sorting limits our ability to enrich these populations for further profiling, including requiring cell surface markers with high-fidelity antibodies. Additionally, many single-cell studies require the isolation of nuclei from tissue, eliminating the ability to enrich learned rare cell states based on extranuclear protein markers. To address these limitations, we describe Programmable Enrichment via RNA Flow-FISH by sequencing (PERFF-seq), a scalable assay that enables scRNA-seq profiling of subpopulations from complex cellular mixtures defined by the presence or absence of specific RNA transcripts. Across immune populations (n = 141,227 cells) and fresh-frozen and formalin-fixed paraffin-embedded brain tissue (n = 29,522 nuclei), we demonstrate the sorting logic that can be used to enrich for cell populations via RNA-based cytometry followed by high-throughput scRNA-seq. Our approach provides a rational, programmable method for studying rare populations identified by one or more marker transcripts.

Environment-based approaches to improve participation of young people with physical disabilities during COVID-19.

AIM: To examine the effects of the Pathways and Resources for Engagement and Participation (PREP) intervention during the COVID-19 pandemic on (1) activity performance and satisfaction, and (2) motor, cognitive, and affective body functions. METHOD: An interrupted time-series design with multiple baselines across 21 young people (13 females, eight males) aged 16 to 25 years (median = 21 years 5 months) with physical disabilities was employed. The young people engaged in an 8-week self-chosen leisure activity (e.g. football, piano, photography) at their home or community. The Canadian Occupational Performance Measure (COPM) assessed activity performance and satisfaction weekly. Mental health problems, including affective and cognitive outcomes, were assessed weekly using the Behavior Assessment System for Children, Third Edition. Motor functions (e.g. trunk control, reaching, strength) were assessed biweekly. Linear mixed-effects models were used. RESULTS: The intervention had large effects on activity performance (0.78) and satisfaction (0.88) with clinically significant change in COPM scores (2.6 [95% confidence interval {CI}: 2.0-3.2] and 3.2 points [95% CI: 2.4-3.9] respectively). Young people without mental health problems at baseline benefited more from the intervention (p = 0.028). Improvements in at least one domain of body function occurred in 10 young people especially for motor outcomes. INTERPRETATION: Results demonstrate the effectiveness of PREP during adverse times and suggest benefits going beyond participation, involving outcomes at the body-function level. WHAT THIS PAPER ADDS: Environmental-based interventions can improve participation even during adverse times such as the COVID-19 pandemic. Significant improvement with large effect sizes occurred in both activity performance and satisfaction. Intervention was effective for all; those without mental health problems benefited more. Improvements in body-function outcomes were partially observed, especially in motor-related outcomes. Body functions may improve through participation even if not targeted by the intervention.

Feasibility of an eye-gaze technology intervention for students with severe motor and communication difficulties in Taiwan.

Eye-gaze technology provides access to a computer through the control of eye movements, thus allowing students with severe motor and communication difficulties to communicate and participate in curriculum activities and leisure; however, few studies have investigated whether any challenges exist to its implementation. This study examines the feasibility for teachers, parents, and therapists of applying an eye-gaze technology intervention for students with severe motor and communication difficulties in everyday settings. A mixed-method design was applied, focusing on the acceptability, demands, implementation, and practicality of the technology applications. Data was collected from 16 participants who assisted five students using eye-gaze technology in a previous 6-month intervention. The intervention comprised (a) use of eye-gaze devices with individualized content; and (b) services including training in use, team meetings, and bi-monthly support on implementation problems. The results showed that the participants perceived the technology as appropriate to enhance interaction and understanding of the students' learning and communication messages. Portable and easy-to-adjust systems were crucial to apply eye-gaze technology in different contexts. Improving eye-gaze services was required to afford in-service education, follow-up services, and loaning programs for sustainable implementation. The facilitators and barriers could guide researchers and practitioners to enhance the implementation of eye-gaze technology.

Single-cell multi-omics of mitochondrial DNA disorders reveals dynamics of purifying selection across human immune cells.

Pathogenic mutations in mitochondrial DNA (mtDNA) compromise cellular metabolism, contributing to cellular heterogeneity and disease. Diverse mutations are associated with diverse clinical phenotypes, suggesting distinct organ- and cell-type-specific metabolic vulnerabilities. Here we establish a multi-omics approach to quantify deletions in mtDNA alongside cell state features in single cells derived from six patients across the phenotypic spectrum of single large-scale mtDNA deletions (SLSMDs). By profiling 206,663 cells, we reveal the dynamics of pathogenic mtDNA deletion heteroplasmy consistent with purifying selection and distinct metabolic vulnerabilities across T-cell states in vivo and validate these observations in vitro. By extending analyses to hematopoietic and erythroid progenitors, we reveal mtDNA dynamics and cell-type-specific gene regulatory adaptations, demonstrating the context-dependence of perturbing mitochondrial genomic integrity. Collectively, we report pathogenic mtDNA heteroplasmy dynamics of individual blood and immune cells across lineages, demonstrating the power of single-cell multi-omics for revealing fundamental properties of mitochondrial genetics.

Ride-on car training using sitting and standing postures for mobility and socialization in young children with motor delays: a randomized controlled trial.

PURPOSE: To examine the effects of ride-on car (ROC) training using different postures on mobility and social function in children with motor delays in comparison with conventional therapy. MATERIALS AND METHODS: Thirty-eight children (22 males, 16 females) with motor delays were recruited and randomly assigned to three groups: ROC training while sitting (ROC-Sit, n = 15; mean age, 20.25 months; standard deviation [SD], 5.29), ROC training while standing (ROC-Stand, n = 12; mean age, 24.80 months; SD, 8.42), and conventional therapy (control, n = 11; mean age: 20.25 months, SD: 5.37). All groups underwent 2-h training sessions twice weekly over a 12-week intervention phase. The Pediatric Evaluation of Disability Inventory and Goal Attainment Scaling were performed before and after the intervention and at follow-up sessions after 12 weeks. RESULTS: Mobility and goal achievement improved significantly in all groups after the intervention (p < 0.001; p < 0.0001). However, social function improved significantly only in the ROC-Stand group (p = 0.001), which had the highest number of participants showing clinically meaningful changes in mobility and social function. CONCLUSION: Increased practice and caregivers' involvement can improve children's mobility and goal achievement. Adopting a standing posture in an ROC can enhance social function.IMPLICATIONS FOR REHABILITATIONProviding active exploratory experience through ride-on cars or practicing specific skills can improve children's mobility function.Ride-on car training in a standing posture allows children to access their distal environment visually, resulting in improved social function.Setting goals with caregivers maximizes the effects of treatment on goal achievement.

Exact Enumeration Approach to Estimate the Theta Temperature of Interacting Self-Avoiding Walks on the Simple Cubic Lattice.

We compute the exact root-mean-square end-to-end distance of the interacting self-avoiding walk (ISAW) up to 27 steps on the simple cubic lattice. These data are used to construct a fixed point equation to estimate the theta temperature of the collapse transition of the ISAW. With the Bulirsch-Stoer extrapolation method, we obtain accurate results that can be compared with large-scale long-chain simulations. The free parameter ω in extrapolation is precisely determined using a parity property of the ISAW. The systematic improvement of this approach is feasible by adopting the combination of exact enumeration and multicanonical simulations.

Increasing participation in computer activities using eye-gaze assistive technology for children with complex needs.

PURPOSE: Eye-gaze assistive technology offers children with severe motor and communication difficulties the opportunity to access and control a computer through eye movements. The aim of this study was to examine the impact of eye-gaze assistive technology intervention on participation in computer activities and technology usability among children with complex needs in Taiwan. MATERIALS AND METHODS: This study involved a multiple baseline design across individuals. The participants were four children aged three to six years with severe motor and communication difficulties and low eye-control skills. The six-month intervention consisted of two collaborative team meetings and 12 individual supports to facilitate the use of eye-gaze assistive technology at home or in educational environments. Participation in computer activities (diversity, frequency, and duration) was repeatedly measured through a computer use diary. Other outcomes included assessments of goal achievements and parents/teachers' ratings on children's performance in computer activities. RESULTS: The young children increased the diversity of their computer activities and their frequency and duration of computer use from baseline to the intervention phase. The children attained six of eight predefined goals related to play, communication, and school learning. Parents and teachers perceived the children's changes in performance as meaningful. CONCLUSION: This study strengthens the evidence that eye-gaze assistive technology is useful in everyday contexts for children with complex needs in Taiwan. The findings add knowledge that children with weak eye-control skills increased participation in computer activities as a result of the eye-gaze assistive technology. Implications for RehabilitationEye-gaze assistive technology (EGAT) as an access method to control a computer can provide opportunities for children with severe motor and communication difficulties to participate in computer activities.Children with severe motor and communication difficulties and low eye-control skills with sufficient practice can learn to use EGAT for communication and learning, with support from stakeholders and collaborative service.EGAT could be introduced for children with complex needs at early ages as a means of using computers for play, communication, and school learning, which could be helpful for later education and learning.Stakeholders in educational environments could include EGAT in educational computer systems so that pupils with severe motor and communication difficulties could interact with a computer, thereby enhancing their engagement and learning.

Communicative Interaction with and without Eye-Gaze Technology between Children and Youths with Complex Needs and Their Communication Partners.

Use of eye-gaze assistive technology (EGAT) provides children/youths with severe motor and speech impairments communication opportunities by using eyes to control a communication interface on a computer. However, knowledge about how using EGAT contributes to communication and influences dyadic interaction remains limited. AIM: By video-coding dyadic interaction sequences, this study investigates the impacts of employing EGAT, compared to the Non-EGAT condition on the dyadic communicative interaction. METHOD: Participants were six dyads with children/youths aged 4-19 years having severe physical disabilities and complex communication needs. A total of 12 film clips of dyadic communication activities with and without EGAT in natural contexts were included. Based on a systematic coding scheme, dyadic communication behaviors were coded to determine the interactional structure and communicative functions. Data were analyzed using a three-tiered method combining group and individual analysis. RESULTS: When using EGAT, children/youths increased initiations in communicative interactions and tended to provide more information, while communication partners made fewer communicative turns, initiations, and requests compared to the Non-EGAT condition. Communication activities, eye-control skills, and communication abilities could influence dyadic interaction. CONCLUSION: Use of EGAT shows potential to support communicative interaction by increasing children's initiations and intelligibility, and facilitating symmetrical communication between dyads.

The Regulatory Effects of a Formulation of Cinnamomum osmophloeum Kaneh and Taiwanofungus camphoratus on Metabolic Syndrome and the Gut Microbiome.

The number of people with metabolic syndrome (MetS) is increasing year by year, and MetS is associated with gut microbiota dysbiosis. The demand for health supplements to treat or prevent MetS is also growing. Cinnamomum osmophloeum Kaneh (CO) and Taiwanofungus camphoratus (TC) are endemic to Taiwan. Both have been shown to improve the symptoms of MetS, such as dyslipidemia and hyperglycemia. Herein, we investigated the effect of CO, TC and their formulations on diet-induced obese mice. Male C57BL/6J mice were fed with a high-fat diet (HFD) for 10 weeks to induce MetS. After that, the mice were fed with HFD supplemented with CO, TC, and various CO/TC formulations, respectively, for 14 weeks. The changes in physiological parameters and the composition of the gut microbiome were investigated. The results indicated that CO, TC, and their formulations effectively reduced hyperglycemia, and tended to alleviate MetS in obese mice. Moreover, we also observed that CO, TC, and their formulations improved gut microbiota dysbiosis by decreasing the Firmicutes-to-Bacteroidetes ratio and increasing the abundance of Akkermansia spp. Our results revealed that CO and TC might have potential for use as a prebiotic dietary supplement to ameliorate obesity-related metabolic disorders and gut dysbiosis.

Collaborative Home-Visit Program for Young Children With Motor Delays in Rural Taiwan: A Pilot Randomized Controlled Trial.

BACKGROUND: Caregiver engagement and collaborative team early childhood intervention (ECI) services are international trends; however, relevant evidence of collaborative home-visiting ECI in rural areas is as yet undetermined. OBJECTIVE: The study aimed to investigate the effectiveness of a collaborative ECI program in a rural area of Taiwan. DESIGN: The study was a pilot randomized control led trial. METHODS: Children aged 6 to 33 months experiencing motor delays and their caregivers were enrolled in Taitung, Taiwan. Using stratified randomization, 24 participants were allocated to either experimental or control groups, and both received 5 home visits within 3 months. The experimental group received ECI services based on the International Classification of Functioning, Disability and Health framework and family-centered approaches. The control group received regular home visits by local social workers. Child outcomes included Pediatric Evaluation of Disability Inventory Chinese Version and Peabody Developmental Motor Scale, 2nd edition. Family outcomes included the Disability-Adapted Infant-Toddler version of Home Observation for Measurement, and Chinese versions of the Knowledge of Infant Development Inventory and Parental Stress Index-Short Form. A tester blinded to the study conducted assessments at baseline, postintervention, and 3-month follow-up. Two-way mixed analysis of variance was used with α = .05 (2-tailed). RESULTS: The experimental group improved scores on the Disability-Adapted Infant-Toddler version of Home Observation for Measurement significantly more than the control group with an effect size of 0.64 at follow-up. In other outcomes, both groups showed no significant differences. The follow-up rate was 69%, and adherence to the ECI program was acceptable. LIMITATIONS: A limitation of the study was the heterogeneity of the sample. CONCLUSIONS: This pilot study revealed possible effectiveness in implementing collaborative ECI programs based on family-centered approaches and the International Classification of Functioning, Disability and Health in rural areas. Larger field studies are needed to confirm our findings.

Validating the ICF core set for cerebral palsy using a national disability sample in Taiwan.

Purpose: To validate the activities and participation (d) codes of two age-specific brief International Classification of Functioning, Disability, and Health (ICF) core sets for school-aged children with cerebral palsy (CP), using national dataset of the child version of the Functioning Scale of the Disability Evaluation System (FUNDES) in Taiwan.Methods: Students with CP aged 6-17.9 years (n = 546) in the national dataset were analyzed. Items of the child version of the FUNDES were linked to the ICF d-codes and matched to two brief ICF core sets for CP. The restriction rate of the linked d-codes were calculated. Random Forest regression was applied to select the important linked d-codes for predicting school participation frequency.Results: The vast majority of the content of the Taiwanese dataset was covered by two core sets. The matched d-codes represent high restriction rates (80%) and most were important for predicting school participation. One important code, d740 (formal relationships, such as relationship with teachers), identified in this study were not included in two ICF core sets.Conclusions: Two brief ICF core sets for CP capture the majority of relevant functional information collected by the child version of the FUNDES. Some additional codes not covered in the international ICF core sets should be considered for inclusion in the revised Taiwanese version.Implications for rehabilitationCerebral palsy (CP) is the most common cause of severe physical disability in childhood. ICF core sets for CP promote a comprehensive assessment and service provision.To ensure applicability, ICF core sets for CP were validated in Taiwan using the child and youth national dataset of the child version of the Functioning Scale of the Disability Evaluation System. This study shows content validity and proposes new ICF codes additions for the Taiwanese version.Among top five ICF-based predictors for school participation frequency, four of them were consistent in both children and youth groups as d310-d350 (basic communication), d750 (informal social relationships), d820 (school education), and d710-d720, d880 and d920 (social play), which could be taken into consideration in clinical application.

Folic acid ameliorates homocysteine-induced angiogenesis and portosystemic collaterals in cirrhotic rats.

INTRODUCTION AND OBJECTIVES: Liver cirrhosis is characterized by increased intrahepatic resistance, splanchnic vasodilation/angiogenesis, and formation of portosystemic collateral vessels. Collaterals can cause lethal complications such as gastroesophageal variceal hemorrhage. Homocysteine is linked to vascular dysfunction and angiogenesis and higher levels have been reported in cirrhotic patients. It is also known that folic acid supplementation reverses the effects of homocysteine. However, the treatment effect in cirrhosis has yet to be investigated. MATERIAL AND METHODS: Liver cirrhosis was induced in Sprague-Dawley rats with common bile duct ligation (CBDL). The CBDL rats randomly received (1) vehicle; (2) dl-homocysteine thiolactone (1g/kg/day); (3) dl-homocysteine thiolactone plus folic acid (100mg/kg/day); or (4) folic acid. On the 29th day, hemodynamic parameters, liver and renal biochemistry, protein expressions of proangiogenic factors, mesenteric vascular density and portosystemic shunting were evaluated. RESULTS: In the cirrhotic rats, homocysteine increased mesenteric vascular density and the severity of shunting. It also up-regulated the protein expressions of mesenteric vascular endothelial growth factor (VEGF) and phosphorylated-endothelial nitric oxide synthase (p-eNOS). These effects were reversed by folic acid treatment (P<0.05). CONCLUSION: Folic acid ameliorated the adverse effects of homocysteine in the cirrhotic rats, which may be related to down-regulation of the VEGF-NO signaling pathway.

Nucleos(t)ide Analogs Do Not Independently Influence Hepatic Fibrosis and Portal Hypertension beyond Viral Suppression in CBDL-Induced Cirrhotic Rat.

Chronic hepatitis is the major cause of liver cirrhosis and portal hypertension. Several factors affect portal pressure, including liver fibrosis, splanchnic vasodilatation, and pathologic angiogenesis. Nucleos(t)ide analogs (NUCs), the oral antiviral agents, effectively attenuate chronic hepatitis B-related liver cirrhosis and portal hypertension via viral suppression and alleviation of hepatitis. On the other hand, NUCs affect tumor necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF), and nitric oxide, which participate in fibrogenesis, vasodilatation, and angiogenesis. However, whether NUCs independently influence liver fibrosis and portal hypertension beyond viral suppression is unknown. This study thus aimed to evaluate the influences of three frequently used NUCs in rats with nonviral cirrhosis. Male Sprague-Dawley rats received common bile duct ligation (CBDL) to induce cholestatic cirrhosis and portal hypertension. The rats were randomly allocated into four groups, treated by mouth with lamivudine (30 mg/kg per day), entecavir (0.09 mg/kg per day), tenofovir (50 mg/kg per day), or distilled water (vehicle control) from the 15th day after CBDL. On the 29th day, liver cirrhosis- and portal hypertension-related parameters were evaluated. The results showed that chronic NUCs treatment did not affect hemodynamic parameters, plasma TNF-α concentration, and hepatic fibrogenesis protein expressions in rats with nonviral cirrhosis. Though the mesenteric VEGF receptor 2 phosphorylation was downregulated in NUCs-treated groups, the splanchnic angiogenesis was not influenced. In conclusion, lamivudine, entecavir, and tenofovir had no additional effects on liver cirrhosis and portal hypertension in rats with nonviral cirrhosis.

Modified ride-on cars and mastery motivation in young children with disabilities: Effects of environmental modifications.

BACKGROUND: Independent mobility is important for children's psychological development. Modified ride-on cars are innovative, alternative options to enhance independent mobility, socialization, and motivation in young children with disabilities. AIM: We compared the effects of combining ride-on car use and a social interaction training program on mastery motivation and home affordances with a home education program in young children with disabilities. METHODS AND PROCEDURES: Twenty-nine children with disabilities aged 1-3 years were recruited. The treatment group (n = 15) received two 2-h sessions/week for 9 weeks of ride-on car training in a hospital environment in Taiwan. The control group (n = 14) underwent similar home education programs. No treatment except regular therapy was administered during the 9-week follow-up period. Assessments included the Revised Dimensions of Mastery Questionnaire-Chinese version and the Affordance in the Home Environment for Motor Development-Toddler version-Chinese version. OUTCOMES AND RESULTS: The treatment group (compared to controls) had significantly greater improvements in object persistence during the intervention. Both groups showed significant improvements in mastery pleasure and home affordances during the intervention. CONCLUSIONS AND IMPLICATIONS: This novel study showed the potential use of modified ride-on cars to enhance mastery motivation in a hospital environment.

Augmented reality (AR) and virtual reality (VR) applied in dentistry.

The OSCE is a reliable evaluation method to estimate the preclinical examination of dental students. The most ideal assessment for OSCE is used the augmented reality simulator to evaluate. This literature review investigated a recently developed in virtual reality (VR) and augmented reality (AR) starting of the dental history to the progress of the dental skill. As result of the lacking of technology, it needs to depend on other device increasing the success rate and decreasing the risk of the surgery. The development of tracking unit changed the surgical and educational way. Clinical surgery is based on mature education. VR and AR simultaneously affected the skill of the training lesson and navigation system. Widely, the VR and AR not only applied in the dental training lesson and surgery, but also improved all field in our life.

Extrahepatic angiogenesis hinders recovery of portal hypertension and collaterals in rats with cirrhosis resolution.

Liver cirrhosis is characterized by portal hypertension. However, the alteration of portal hypertension-related derangements during cirrhosis resolution is not well known. The present study aimed to establish animal models with cirrhosis resolution and to investigate the relevant changes during this process. Male Sprague-Dawley rats were applied. In reverse thioacetamide (rTAA) model, rats were randomly allocated into four groups with control, thioacetamide (TAA) cirrhosis and rTAA groups that discontinued TAA for 4 or 8 weeks after cirrhosis induction. In reverse bile duct ligation (rBDL) model, rats received choledochoduodenal shunt surgery upon the establishment of cirrhosis and 4, 8, or 16 weeks were allowed after the surgery. At the end, portal hypertension-related parameters were evaluated. Cirrhosis resolution was observed in rTAA groups. Portal pressure (PP) decreased after cirrhosis resolution but remained higher than control group (control, TAA, rTAA4, rTAA8 (mmHg): 5.4 ± 0.3, 12.9 ± 0.3, 8.6 ± 0.4, 7.6 ± 0.6). Further survey found the increased splanchnic blood flow did not reduce during cirrhosis resolution. The extrahepatic pathological angiogenesis was not ameliorated (% of mesenteric window area: 1.2 ± 0.3, 7.3 ± 1.1, 8.3 ± 1.0, 11.3 ± 2.7). In collateral system, the shunting degree reduced while the vessels structure remained. The vascular contractility of all systems and nitric oxide (NO) production were normalized. In rBDL series, PP decreased in rBDL16 groups but the extrahepatic angiogenesis persisted. In conclusion, cirrhosis resolution attenuates but not completely normalizes portal hypertension because of persistently high splanchnic inflow and angiogenesis. In clinical setting, vascular complications such as varices could persist after cirrhosis resolution and further investigation to define the follow-up and treatment strategies is anticipated.

The beneficial effects of curcumin in cirrhotic rats with portal hypertension.

In liver cirrhosis with portal hypertension, the uneven distribution of vasoactive substances leads to increased intrahepatic vascular resistance and splanchnic vasodilatation. Angiogenesis also induces increased portal inflow and portosystemic collaterals. The collaterals may induce lethal complications such as gastroesophageal variceal hemorrhage, but the therapeutic effect of vasoconstrictors is still suboptimal due to poor collateral vasoresponsivenss. Curcumin has aroused much attention for its antifibrosis, vasoactive, and anti-angiogenesis actions. However, whether it affects the aforementioned aspects is unknown. Liver cirrhosis was induced by common bile duct ligation (CBDL) in Sprague-Dawley rats. Sham-operated rats were controls. CBDL and sham rats were randomly allocated to receive curcumin (600 mg/kg per day) or vehicle since the 15th day after BDL. On the 29th day, portal hypertension related parameters were surveyed. Portosystemic collateral in situ perfusion was performed to evaluate vascular activity. Chronic curcumin treatment decreased portal pressure (PP), cardiac index (CI) and increased systemic vascular resistance (SVR) in cirrhotic rats. In splanchnic system, curcumin decreased superior mesenteric artery (SMA) flow and increased SMA resistance. Mesenteric angiogenesis was attenuated by curcumin. Acute administration of curcumin significantly induced splanchnic vasoconstriction. The mesenteric protein expressions of p-endothelial nitric oxide synthase (eNOS), cyclooxygenase (COX) 2 (COX2), vascular endothelial growth factor (VEGF), p-VEGF receptor 2 (VEGFR2), and p-Erk were down-regulated. In collateral system, curcumin decreased portosystemic shunting and induced vasoconstriction. In conclusion, chronic curcumin administration in cirrhotic rats ameliorated portal hypertension related hemodynamic derangements and portosystemic collaterals. Curcumin also attenuated splanchnic hyperdynamic circulation by inducing vasoconstriction through inhibition of eNOS activation and by decreasing mesenteric angiogenesis via VEGF pathway blockade.

Microbiota, metabolome, and immune alterations in obese mice fed a high-fat diet containing type 2 resistant starch.

SCOPE: We examined the intestinal and systemic responses to incorporating a type 2 resistant starch (RS) into a high fat diet fed to obese mice. METHODS AND RESULTS: Diet-induced obese, C57BL/6J male mice were fed an HF diet without or with 20% (by weight) high-amylose maize resistant starch (HF-RS) for 6 weeks. Serum adiponectin levels were higher with RS consumption, but there were no differences in weight gain and adiposity. With HF-RS, the expression levels of ileal TLR2 and Reg3g and cecal occludin, TLR2, TLR4, NOD1 and NOD2 were induced; whereas colonic concentrations of the inflammatory cytokine IL-17A declined. The intestinal, serum, liver, and urinary metabolomes were also altered. HF-RS resulted in lower amino acid concentrations, including lower serum branched chain amino acids, and increased quantities of urinary di/trimethylamine, 3-indoxylsulfate, and phenylacetylglycine. Corresponding to these changes were enrichments in Bacteroidetes (S24-7 family) and certain Firmicutes taxa (Lactobacillales and Erysipelotrichaceae) with the HF-RS diet. Parabacteroides and S24-7 positively associated with cecal maltose concentrations. These taxa and Erysipelotrichaceae, Allobaculum, and Bifidobacterium were directly correlated with uremic metabolites. CONCLUSION: Consumption of RS modified the intestinal microbiota, stimulated intestinal immunity and endocrine-responses, and modified systemic metabolomes in obese mice consuming an otherwise obesogenic diet.

Modified Ride-On Cars and Young Children with Disabilities: Effects of Combining Mobility and Social Training.

BACKGROUND: Research has shown that the use of power mobility devices is safe and beneficial for motor and cognitive development in children with motor disabilities; nevertheless, strong evidence of the benefits for social skill development is limited. This study aimed to examine the effects of combining ride-on car training with an adult-directed, social interaction program in a hospital-based environment on mobility and social functions in young children with motor disabilities. METHODS: This study used a prospective, nonequivalent pretest-posttest control group design. Twenty-nine young children with motor disabilities, aged between 1 and 3 years, were recruited from local hospitals in Taiwan. The treatment group (n = 15) underwent 2-h ride-on car training sessions twice per week for a total of 9 weeks in the hospital environment. The control group (n = 14) underwent a 9-week home education program (mean: 200 min/week) focusing on mobility and social skills training. The Chinese version of the Pediatric Evaluation of Disability Inventory, Parenting Stress Index, and Goal Attainment Scaling were administered to all participants before and after the intervention, and at the end of the 9-week follow-up phase. RESULTS: Mobility and social functions significantly improved in both groups after the 9-week intervention, but this improvement was not maintained at the follow-up phase. The treatment group showed significantly better improvement in social function, parenting stress levels, and goal achievement than the control group at posttest. CONCLUSION: This two-group design study showed the benefits of combining a ride-on car use with a family-centered, structured, social interaction program for positive impacts on mobility, social function, and parenting stress levels. The combination of a modified ride-on car and a social training program has the potential to enhance socialization in young children with motor disabilities. CLINICAL TRIAL REGISTRATION: www.ClinicalTrials.gov, identifier NCT02527499.

Impact of Different Fecal Processing Methods on Assessments of Bacterial Diversity in the Human Intestine.

The intestinal microbiota are integral to understanding the relationships between nutrition and health. Therefore, fecal sampling and processing protocols for metagenomic surveys should be sufficiently robust, accurate, and reliable to identify the microorganisms present. We investigated the use of different fecal preparation methods on the bacterial community structures identified in human stools. Complete stools were collected from six healthy individuals and processed according to the following methods: (i) randomly sampled fresh stool, (ii) fresh stool homogenized in a blender for 2 min, (iii) randomly sampled frozen stool, and (iv) frozen stool homogenized in a blender for 2 min, or (v) homogenized in a pneumatic mixer for either 10, 20, or 30 min. High-throughput DNA sequencing of the 16S rRNA V4 regions of bacterial community DNA extracted from the stools showed that the fecal microbiota remained distinct between individuals, independent of processing method. Moreover, the different stool preparation approaches did not alter intra-individual bacterial diversity. Distinctions were found at the level of individual taxa, however. Stools that were frozen and then homogenized tended to have higher proportions of Faecalibacterium, Streptococcus, and Bifidobacterium and decreased quantities of Oscillospira, Bacteroides, and Parabacteroides compared to stools that were collected in small quantities and not mixed prior to DNA extraction. These findings indicate that certain taxa are at particular risk for under or over sampling due to protocol differences. Importantly, homogenization by any method significantly reduced the intra-individual variation in bacteria detected per stool. Our results confirm the robustness of fecal homogenization for microbial analyses and underscore the value of collecting and mixing large stool sample quantities in human nutrition intervention studies.