Evaluation on the Nutrition Quotient Scores of Elderly People Living Alone in Korea.

As the Korean society is aging rapidly, the issues on physical, social, economic, and mental disabilities of single-person households aged 65 years or older has also increased. This study aimed to investigate the nutrition-related dietary conditions of elderly people living alone and determine their dietary behavior by calculating the nutrition quotient for elderly (NQ-E). One hundred and three elderly people living alone who were basic living recipients were recruited from six senior welfare centers in Seoul, and the data were collected using a questionnaire from 19 July 2016 to 17 August 2016, with a 1:1 in-depth interview using the modified version of the NQ-E questionnaire. The data were analyzed using SPSS 27.0 for Mac (IBM Corp., Armonk, NY, USA); a p value of <0.05 was considered significant. The nutrition-related dietary conditions of the elderly living alone were limited, and many of them received support from the government, which helped improve their diet. The nutrition quotient score of the elderly living alone was 50.14, which was lower than the NQ-E mean score (57.6) of the Korean elderly and the NQ-E (62 points), which is the top 25% of the national survey subjects according to the criteria value presented by the Korean Nutrition Society. Elderly people living alone often have poor dietary habits and nutritional status. The NQ-E presented in this study can be used to evaluate the dietary conditions of the elderly and is expected to be used as an indicator for developing community programs for health promotion and evaluating their effectiveness.

You Are Not Alone: A Serial Mediation of Social Attraction, Privacy Concerns, and Satisfaction in Voice AI Use.

The popularity of voice-activated artificial intelligence (voice AI) has grown rapidly as people continue to use smart speakers such as Amazon Alexa and Google Home to support everyday tasks. However, little is known about how loneliness relates to voice AI use, or the potential mediators in this association. This study investigates the mediating roles of users' perceptions (i.e., social attraction, privacy concerns, and satisfaction) in the relationship between users' social loneliness and intentions to continue using voice AI. A serial mediation model based on survey data from current voice AI users showed that users' perceptions were positively associated with behavioral intentions. Several full serial mediations were observed: people who felt lonely perceived (1) voice AI as a more socially attractive agent and (2) had fewer privacy concerns. These aspects were each tied to satisfaction and subsequent usage intention. Theoretical and practical implications are discussed.

Efficacy of a Hip Brace for Hip Displacement in Children With Cerebral Palsy: A Randomized Clinical Trial.

Importance: There is no consensus on interventions to slow the progress of hip displacement in patients with cerebral palsy. Objective: To investigate the efficacy of a novel hip brace in preventing progressive hip displacement in patients with cerebral palsy. Design, Setting, and Participants: This 2-group randomized clinical trial was conducted at 4 tertiary hospitals in South Korea from July 2019 to November 2021. Participants included children aged 1 to 10 years with nonambulatory cerebral palsy (Gross Motor Function Classification System level IV or V). Block randomization was used to assign an equal number of patients to the study and control groups via computerized random allocation sequences. Data were analyzed from November to December 2021. Interventions: The intervention group wore the hip brace for at least 12 hours a day for the study duration (ie, 12 months). Follow-up evaluations were performed after 6 and 12 months of wearing the brace. Both groups proceeded with conventional rehabilitation therapy during the trial. Main Outcomes and Measures: The primary outcome was the Reimers migration index (MI) on radiography, as assessed by 3 blinded investigators. Primary outcome variables were analyzed using linear mixed models. Secondary outcomes include change in the Caregiver Priorities & Child Health Index of Life with Disabilities, on which lower scores indicate better quality of life. Results: A total of 66 patients were included, with 33 patients (mean [SD] age, 68.7 [31.6] months; 25 [75.8%] boys) randomized to the intervention group and 33 patients (mean [SD] age, 60.7 [24.9] months; 20 [60.6%] boys) randomized to the control group. The baseline mean (SD) MI was 37.4% (19.3%) in the intervention group and 30.6% (16.3%) in the control group. The mean difference of the MI between the intervention group and control group was -8.7 (95% CI, -10.2 to -7.1) percentage points at 6 months and -12.7 (95% CI, -14.7 to -10.7) percentage points at 12 months. The changes in the Caregiver Priorities & Child Health Index of Life with Disabilities were favorable in the study group and reached statistical significance at the 6-month follow-up compared with the control group (difference, -14.2; 95% CI, -25.2 to -3.3). Conclusions and Relevance: In this randomized clinical trial, the novel hip brace was significantly effective in preventing the progression of hip displacement, compared with the control group. It effectively improved quality of life in patients with nonambulatory cerebral palsy. Therefore, hip brace use could be a promising treatment method to delay hip surgery and improve the quality of life of patients with nonambulatory cerebral palsy. Trial Registration: ClinicalTrials.gov Identifier: NCT04033289.

Evaluation of the nutrient quotient for the elderly (NQ-E) using congregate meal services according to their oral health conditions.

BACKGROUND/OBJECTIVES: The purposes of this study were to evaluate the nutritional status and dietary habits of the elderly using the nutrition quotient for the elderly (NQ-E) and to analyze the differences in the NQ-E according to their levels of oral health. SUBJECTS/METHODS: The survey was administered to 123 elderly people receiving congregate meal services in Seoul. The questionnaire comprised 3 domains: oral health status, general characteristics, and the NQ-E for the elderly. RESULTS: The respondents were divided into 2 groups based on the average score of their levels of oral health (the group with high oral health scores: 4.42 points and the group with low oral health scores: 2.89 points). As a result of evaluating nutritional status using the NQ-E, it was found that the average NQ-E score was 58.7 points, with 46.0 points in the balance domain, 47.0 points in the diversity domain, 72.9 points in the moderation domain, and 61.8 points in the dietary behavior domain. The NQ-E score (62.3 points) of the group with high oral health scores is significantly higher than the NQ-E score (54.7 points) of the group with low oral health scores (P < 0.001). Concerning the NQ domain scores, the elderly with good oral health status had "favorable" results in terms of balance and dietary behavior, and the elderly with poor oral health status had "favorable" results only in terms of balance. CONCLUSIONS: Overall, several dietary areas needed improvement in general. Those with poor oral health conditions urgently needed to improve related factors to minimize the risk of increasing imbalanced nutrition and comorbidities due to insufficient nutrition and undesirable eating habits.

Specialized neurons in the right habenula mediate response to aversive olfactory cues.

Hemispheric specializations are well studied at the functional level but less is known about the underlying neural mechanisms. We identified a small cluster of cholinergic neurons in the dorsal habenula (dHb) of zebrafish, defined by their expression of the lecithin retinol acyltransferase domain containing 2 a (lratd2a) gene and their efferent connections with a subregion of the ventral interpeduncular nucleus (vIPN). The lratd2a-expressing neurons in the right dHb are innervated by a subset of mitral cells from both the left and right olfactory bulb and are activated upon exposure to the odorant cadaverine that is repellent to adult zebrafish. Using an intersectional strategy to drive expression of the botulinum neurotoxin specifically in these neurons, we find that adults no longer show aversion to cadaverine. Mutants with left-isomerized dHb that lack these neurons are also less repelled by cadaverine and their behavioral response to alarm substance, a potent aversive cue, is diminished. However, mutants in which both dHb have right identity appear more reactive to alarm substance. The results implicate an asymmetric dHb-vIPN neural circuit in the processing of repulsive olfactory cues and in modulating the resultant behavioral response.

Generation of foxn1/Casper Mutant Zebrafish for Allograft and Xenograft of Normal and Malignant Cells.

Cell transplantation into immunodeficient recipients is a widely used approach to study stem cell and cancer biology; however, studying cell states post transplantation in vivo is inconvenient in mammals. Here, we generated a foxn1/Casper mutant zebrafish that is transparent and exhibits T cell deficiency. By employing the line for hematopoietic stem cell (HSC) transplantation (HSCT), we could achieve nonconditioned transplantation. Meanwhile, we found that fetal HSCs from 3 days post fertilization zebrafish embryos produce a better transplant outcome in foxn1/Casper mutants, compared with adult HSCs. In addition to HSCT, the foxn1/Casper mutant is feasible for allografts of myelodysplastic syndrome-like and muscle cells, as well as xenografts of medaka muscle cells. In summary, foxn1/Casper mutants permit the nonconditioned engraftment of multiple cell types and visualized characterization of transplanted cells in vivo.

Loss of abcd4 in zebrafish leads to vitamin B12-deficiency anemia.

ABCD4, a member of the ATP-binding cassette transporter superfamily, is associated with the transport of vitamin B12 which is crucial for the development of red blood cells (RBCs) and may also be involved in its metabolism. However, the molecular function of ABCD4 during RBC development in zebrafish is mostly unknown. Using a morpholino-based knockdown approach, we found that abcd4-knockdown resulted in abnormal RBCs of irregular shapes and various sizes. o-Dianisidine staining, as an indicator of hemoglobin in RBCs, further confirmed that abcd4 morphants possessed fewer hemoglobinized cells and impaired blood circulation. Multiple protein sequence alignment revealed that the amino acid sequence for residues 13-292, which is the domain of vitamin B12 transport, of the zebrafish Abcd4 was highly conserved compared to that of other species. Accordingly, the abcd4 morphants can be rescued with human ABCD4, demonstrating a conserved role of ABCD4 in vertebrates. Notably, the vitamin B12-deficient phenotype in abcd4 morphants, which causes anemia, was recapitulated in the newly-established abcd4 mutant, indicating the possibility that the abcd4 mutant could be used as a disease model of vitamin B12-deficiency anemia. Our study provides an insight that the analysis of the newly-established abcd4 mutant may contribute to understanding its roles in ABCD4-related vitamin B12-deficiency anemia and the associated pathogeneses in humans.

A new species of bomolochid Orbitacolax brevispinus n. sp. and redescription of taeniacanthid Cirracanthus inimici (Yamaguti et Yamasu, 1959) (Crustacea: Copepoda: Cyclopoida) parasitic on marine fishes of Korea.

Two species of cyclopoid copepods are recorded in this study. (1) A new species of bomolochid, Orbitacolax brevispinus n. sp. (Crustacea) is described based on adult females collected from the gill filaments and inner surface of the opercula of red barracuda, Sphyraena pinguis Günther (Perciformes: Sphyraenidae), captured in Korean waters. The new species differs from its congeners by the possession of two pairs of spines on the dorsal surface of the cephalothorax located just posterior to the rostrum and a different setal formula on the distal exopodal segments of legs 2-4. (2) A taeniacanthid Cirracanthus inimici (Yamaguti et Yamasu, 1959) (Crustacea) is redescribed based on the specimens collected from the gill filaments and inner surface of the opercula of devil stinger, Inimicus japonicus (Cuvier) (Scorpaeniformes: Synancellidae). This finding is the first record in Korean waters and the first description of male. A checklist of parasitic copepods of the families Bomolochidae Sumpf, 1871 and Taeniacanthidae Wilson, 1911of Korea is also provided.

Targeted knockout of a chemokine-like gene increases anxiety and fear responses.

Emotional responses, such as fear and anxiety, are fundamentally important behavioral phenomena with strong fitness components in most animal species. Anxiety-related disorders continue to represent a major unmet medical need in our society, mostly because we still do not fully understand the mechanisms of these diseases. Animal models may speed up discovery of these mechanisms. The zebrafish is a highly promising model organism in this field. Here, we report the identification of a chemokine-like gene family, samdori (sam), and present functional characterization of one of its members, sam2 We show exclusive mRNA expression of sam2 in the CNS, predominantly in the dorsal habenula, telencephalon, and hypothalamus. We found knockout (KO) zebrafish to exhibit altered anxiety-related responses in the tank, scototaxis and shoaling assays, and increased crh mRNA expression in their hypothalamus compared with wild-type fish. To investigate generalizability of our findings to mammals, we developed a Sam2 KO mouse and compared it to wild-type littermates. Consistent with zebrafish findings, homozygous KO mice exhibited signs of elevated anxiety. We also found bath application of purified SAM2 protein to increase inhibitory postsynaptic transmission onto CRH neurons of the paraventricular nucleus. Finally, we identified a human homolog of SAM2, and were able to refine a candidate gene region encompassing SAM2, among 21 annotated genes, which is associated with intellectual disability and autism spectrum disorder in the 12q14.1 deletion syndrome. Taken together, these results suggest a crucial and evolutionarily conserved role of sam2 in regulating mechanisms associated with anxiety.

Chd7 Is Critical for Early T-Cell Development and Thymus Organogenesis in Zebrafish.

Coloboma, heart defect, atresia choanae, retarded growth and development, genital hypoplasia, ear anomalies/deafness (CHARGE) syndrome is a congenital disorder affecting multiple organs and mainly caused by mutations in CHD7, a gene encoding a chromatin-remodeling protein. Immunodeficiency and reduced T cells have been noted in CHARGE syndrome. However, the mechanisms underlying T lymphopenia are largely unexplored. Herein, we observed dramatic decrease of T cells in both chd7knockdown and knockout zebrafish embryos. Unexpectedly, hematopoietic stem and progenitor cells and, particularly, lymphoid progenitor cells were increased peripherally in nonthymic areas in chd7-deficient embryos, unlikely to contribute to the T-cell decrease. Further analysis demonstrated that both the organogenesis and homing function of the thymus were seriously impaired. Chd7 might regulate thymus organogenesis through modulating the development of both neural crest cell-derived mesenchyme and pharyngeal endoderm-derived thymic epithelial cells. The expression of foxn1, a central regulator of thymic epithelium, was remarkably down-regulated in the pharyngeal region in chd7-deficient embryos. Moreover, the T-cell reduction in chd7-deficient embryos was partially rescued by overexpressing foxn1, suggesting that restoring thymic epithelium may be a potential therapeutic strategy for treating immunodeficiency in CHARGE syndrome. Collectively, the results indicated that chd7 was critical for thymic development and T-lymphopenia in CHARGE syndrome may be mainly attributed to the defects of thymic organogenesis. The current finding may benefit the diagnosis and therapy of T lymphopenia and immunodeficiency in CHARGE syndrome.

Deficiency of a brain-specific chemokine-like molecule, SAM3, induces cardinal phenotypes of autism spectrum disorders in mice.

Chemokines are small secreted signaling proteins produced by a broad range of cells, including immune cells. Several studies have recently suggested potential roles of chemokines and their receptors in the pathophysiology of autism spectrum disorders (ASDs). SAM3 is a novel brain-specific chemokine-like molecule with an unknown physiological function. We explored the relevance of chemokines in the development of ASD in mice, with a focus on SAM3. We generated Sam3 gene knockout (KO) mice and characterized their behavioral phenotypes, with a focus on those relevant to ASD. Sam3-deficient mice displayed all three core phenotypes of ASD: impaired responses to social novelty, defects in social communication, and increased repetitive behavior. In addition, they showed increased anxiety. Interestingly, gender differences were identified for several behaviors: only male Sam3 KO mice exhibited increased anxiety and increased repetitive behaviors. Sam3 KO mice did not exhibit changes in other behaviors, including locomotor activities, fear learning and memory, and object recognition memory. These findings indicate that a deficiency of SAM3, a novel brain-specific chemokine-like molecule, may lead to the pathogenesis of ASDs and suggest the possibility that SAM3, a soluble factor, could be a novel therapeutic target for ASD treatment.

Ottogi Inhibits Wnt/ß-catenin Signaling by Regulating Cell Membrane Trafficking of Frizzled8.

Wnt signaling controls critical developmental processes including tissue/body patterning. Here we report the identification of a novel regulator of Wnt signaling, OTTOGI (OTG), isolated from a large-scale expression screening of human cDNAs in zebrafish embryos. Overexpression of OTG in zebrafish embryos caused dorso-anteriorized phenotype, inhibited the expression of Wnt target genes, and prevented nuclear accumulation of ß-catenin. Conversely, knockdown of zebrafish otg using specific antisense morpholino promoted nuclear accumulation of ß-catenin and caused ventralization. However, OTG failed to rescue headless-like phenotype induced by inhibition of GSK-3ß activity, suggesting that OTG acts upstream of GSK-3ß. OTG bound specifically to Frizzled8 (Fz8) receptor and caused retention of Fz8 in the endoplasmic reticulum possibly by preventing N-linked glycosylation of Fz8. Taken together, our data indicate that OTG functions as a novel negative regulator of Wnt signaling during development by the modulation of cell surface expression of Fz receptor.

Zebrafish knockout of Down syndrome gene, DYRK1A, shows social impairments relevant to autism.

BACKGROUND: DYRK1A maps to the Down syndrome critical region at 21q22. Mutations in this kinase-encoding gene have been reported to cause microcephaly associated with either intellectual disability or autism in humans. Intellectual disability accompanied by microcephaly was recapitulated in a murine model by overexpressing Dyrk1a which mimicked Down syndrome phenotypes. However, given embryonic lethality in homozygous knockout (KO) mice, no murine model studies could present sufficient evidence to link Dyrk1a dysfunction with autism. To understand the molecular mechanisms underlying microcephaly and autism spectrum disorders (ASD), we established an in vivo dyrk1aa KO model using zebrafish. METHODS: We identified a patient with a mutation in the DYRK1A gene using microarray analysis. Circumventing the barrier of murine model studies, we generated a dyrk1aa KO zebrafish using transcription activator-like effector nuclease (TALEN)-mediated genome editing. For social behavioral tests, we have established a social interaction test, shoaling assay, and group behavior assay. For molecular analysis, we examined the neuronal activity in specific brain regions of dyrk1aa KO zebrafish through in situ hybridization with various probes including c-fos and crh which are the molecular markers for stress response. RESULTS: Microarray detected an intragenic microdeletion of DYRK1A in an individual with microcephaly and autism. From behavioral tests of social interaction and group behavior, dyrk1aa KO zebrafish exhibited social impairments that reproduce human phenotypes of autism in a vertebrate animal model. Social impairment in dyrk1aa KO zebrafish was further confirmed by molecular analysis of c-fos and crh expression. Transcriptional expression of c-fos and crh was lower than that of wild type fish in specific hypothalamic regions, suggesting that KO fish brains are less activated by social context. CONCLUSIONS: In this study, we established a zebrafish model to validate a candidate gene for autism in a vertebrate animal. These results illustrate the functional deficiency of DYRK1A as an underlying disease mechanism for autism. We also propose simple social behavioral assays as a tool for the broader study of autism candidate genes.

Development of zebrafish medulloblastoma-like PNET model by TALEN-mediated somatic gene inactivation.

Genetically engineered animal tumor models have traditionally been generated by the gain of single or multiple oncogenes or the loss of tumor suppressor genes; however, the development of live animal models has been difficult given that cancer phenotypes are generally induced by somatic mutation rather than by germline genetic inactivation. In this study, we developed somatically mutated tumor models using TALEN-mediated somatic gene inactivation of cdkn2a/b or rb1 tumor suppressor genes in zebrafish. One-cell stage injection of cdkn2a/b-TALEN mRNA resulted in malignant peripheral nerve sheath tumors with high frequency (about 39%) and early onset (about 35 weeks of age) in F0 tp53e7/e7 mutant zebrafish. Injection of rb1-TALEN mRNA also led to the formation of brain tumors at high frequency (58%, 31 weeks of age) in F0 tp53e7/e7 mutant zebrafish. Analysis of each tumor induced by somatic inactivation showed that the targeted genes had bi-allelic mutations. Tumors induced by rb1 somatic inactivation were characterized as medulloblastoma-like primitive neuroectodermal tumors based on incidence location, histopathological features, and immunohistochemical tests. In addition, 3' mRNA Quanti-Seq analysis showed differential activation of genes involved in cell cycle, DNA replication, and protein synthesis; especially, genes involved in neuronal development were up-regulated.

The effect of weight bearing on bone mineral density and bone growth in children with cerebral palsy: A randomized controlled preliminary trial.

BACKGROUND: The present study aims to explore the effect of weight bearing exercise on bone mineral density (BMD) and bone growth in children with cerebral palsy (CP). METHODS: Twelve children with CP of functional level of gross motor functional classification scale (GMFCS) V and 6 healthy children (control group) were included in the study. Participants underwent a dual-energy X-ray absorptiometry scan to measure the BMD of the femur and full-length anteroposterior radiography to measure the bone length of the femur and tibia at baseline and after 6 months. Patients were randomly divided into 2 groups: group A with programmed standing exercises and assisted standing for more than 2 hours a day, more than 5 days a week; and group B with conventional physiotherapy with a standing program for 20 minutes a day, 2 to 3 days a week. RESULTS: A 6-month follow-up showed significantly increased BMD on the femur neck in the control group. Although the changes in BMD were not significant in both groups, group A demonstrated an increased trend of BMD, whereas group B showed a decreased trend. Bone length was significantly increased in all 3 groups at the 6-month follow-up. Although this increase was not significant, the change in bone length was greatest in the control group. The smallest changes were observed in group B. CONCLUSIONS: Weight bearing exercise may play an important role in increasing or maintaining BMD in children with CP and is also expected to promote bone growth. Programmed standing may be used as an effective treatment method to increase BMD in children with CP. However, further studies with a larger cohort and longer follow-up period are required to reveal further information on the benefit of weight bearing exercise and to develop a detailed program.

Thoracic Radiculopathy Resulting From Retrograde Neuronal Degeneration.

We report a case of thoracic radiculopathy caused by retrograde degeneration from an intercostal nerve mass. A 74-year-old woman presented with thoracic radicular pain in the T4 dermatome. Needle electromyography revealed abnormal spontaneous activity in the left paraspinal muscle. Magnetic resonance imaging of the thoracic spine showed no signs of a herniated thoracic disk or root compression but revealed a mass along the intercostal space. The pathologic findings included perineural infiltration. A mass located along the intercostal space approximately 1.8 cm from the dorsal root ganglion may cause thoracic radiculopathy via retrograde degeneration.

Complete mitochondrial genome of Kishi velvet shrimp, Metapenaeopsis dalei (Rathbun, 1902) (Crustacea: Decapoda: Penaeidae) in the East China Sea.

We determined the complete mitochondrial genome of Metapenaeopsis dalei (Rathbun, 1902), which is collected from East China Sea (124°E and 33.5°N). Total mitochondrial genome length of M. dalei was 15 939 bp, in which 13 proteins, two ribosomal RNAs, 22 transfer RNAs and a putative control region were encoded. The gene arrangement of M. dalei was well conserved with nine known Penaeidae mitochondrial genomes from COX1 to tRNATyr . The protein-coding genes started with ATN except for COX1 in which ACG is used. Four genes (COX2, COX3, ND3 and ND5) exhibited an incomplete stop codon. Nucleotide sequence identity of M. dalei mitochondrial genome to those of nine Penaeidae species ranged from 78% to 80%. Based on the COI region, M. dalei is most closely related to Penaeus notialis.

Improved Dysphagia After Decannulation of Tracheostomy in Patients With Brain Injuries.

OBJECTIVE: To investigate improved dysphagia after the decannulation of a tracheostomy in patients with brain injuries. METHODS: The subjects of this study are patients with brain injuries who were admitted to the Department of Rehabilitation Medicine in Myongji Hospital and who underwent a decannulation between 2012 and 2014. A video fluoroscopic swallowing study (VFSS) was performed in order to investigate whether the patients' dysphagia had improved. We measured the following 5 parameters: laryngeal elevation, pharyngeal transit time, post-swallow pharyngeal remnant, upper esophageal width, and semisolid aspiration. We analyzed the patients' results from VFSS performed one month before and one month after decannulation. All VFSS images were recorded using a camcorder running at 30 frames per second. An AutoCAD 2D screen was used to measure laryngeal elevation, post-swallow pharyngeal remnant, and upper esophageal width. RESULTS: In this study, a number of dysphagia symptoms improved after decannulation. Laryngeal elevation, pharyngeal transit time, and semisolid aspiration showed no statistically significant differences (p>0.05), however after decannulation, the post-swallow pharyngeal remnant (pre 37.41%±24.80%, post 21.02%±11.75%; p<0.001) and upper esophageal width (pre 3.57±1.93 mm, post 4.53±2.05 mm; p<0.001) showed statistically significant differences. CONCLUSION: When decannulation is performed on patients with brain injuries who do not require a ventilator and who are able to independently excrete sputum, improved esophageal dysphagia can be expected.

Effectiveness of Rehabilitative Balloon Swallowing Treatment on Upper Esophageal Sphincter Relaxation and Pharyngeal Motility for Neurogenic Dysphagia.

OBJECTIVE: To investigate the relationship between dysphagia severity and opening of the upper esophageal sphincter (UES), and to assess the effect of balloon size on functional improvement after rehabilitative balloon swallowing treatment in patients with severe dysphagia with cricopharyngeus muscle dysfunction (CPD). METHODS: We reviewed videofluoroscopic swallowing studies (VFSS) conducted in the Department of Physical Medicine and Rehabilitation, Myongji Hospital from January through December in 2012. All subjects diagnosed with CPD by VFSS further swallowed a 16-Fr Foley catheter filled with barium sulfate suspension for three to five minutes. We measured the maximum diameter of the balloon that a patient could swallow into the esophagus and subsequently conducted a second VFSS. Then, we applied a statistical technique to correlate the balloon diameter with functional improvement after the balloon treatment. RESULTS: Among 283 inpatients who received VFSS, 21 subjects were diagnosed with CPD. It was observed that the degree of UES opening evaluated by swallowing a catheter balloon had inverse linear correlations with pharyngeal transit time and post-swallow pharyngeal remnant. Videofluoroscopy guided iterative balloon swallowing treatment for three to five minutes, significantly improved the swallowing ability in terms of pharyngeal transit time and pharyngeal remnant (p<0.005 and p<0.001, respectively). Correlation was seen between balloon size and reduction in pharyngeal remnants after balloon treatment (Pearson correlation coefficient R=-0.729, p<0.001), whereas there was no definite relationship between balloon size and improvement in pharyngeal transit time (R=-0.078, p=0.738). CONCLUSION: The maximum size of the balloon that a patient with CPD can swallow possibly indicates the maximum UES opening. The iterative balloon swallowing treatment is safe without the risk of aspiration, and it can be an effective technique to improve both pharyngeal motility and UES relaxation.

ZC4H2, an XLID gene, is required for the generation of a specific subset of CNS interneurons.

Miles-Carpenter syndrome (MCS) was described in 1991 as an XLID syndrome with fingertip arches and contractures and mapped to proximal Xq. Patients had microcephaly, short stature, mild spasticity, thoracic scoliosis, hyperextendable MCP joints, rocker-bottom feet, hyperextended elbows and knees. A mutation, p.L66H, in ZC4H2, was identified in a XLID re-sequencing project. Additional screening of linked families and next generation sequencing of XLID families identified three ZC4H2 mutations: p.R18K, p.R213W and p.V75in15aa. The families shared some relevant clinical features. In silico modeling of the mutant proteins indicated all alterations would destabilize the protein. Knockout mutations in zc4h2 were created in zebrafish and homozygous mutant larvae exhibited abnormal swimming, increased twitching, defective eye movement and pectoral fin contractures. Because several of the behavioral defects were consistent with hyperactivity, we examined the underlying neuronal defects and found that sensory neurons and motoneurons appeared normal. However, we observed a striking reduction in GABAergic interneurons. Analysis of cell-type-specific markers showed a specific loss of V2 interneurons in the brain and spinal cord, likely arising from mis-specification of neural progenitors. Injected human wt ZC4H2 rescued the mutant phenotype. Mutant zebrafish injected with human p.L66H or p.R213W mRNA failed to be rescued, while the p.R18K mRNA was able to rescue the interneuron defect. Our findings clearly support ZC4H2 as a novel XLID gene with a required function in interneuron development. Loss of function of ZC4H2 thus likely results in altered connectivity of many brain and spinal circuits.