Whole-Exome Sequencing Identified Rare Genetic Variants Associated with Undervirilized Genitalia in Taiwanese Pediatric Patients.

Disorders/differences of sex development (DSDs) are a group of rare and phenotypically variable diseases. The underlying genetic causes of most cases of 46XY DSDs remains unknown. Despite the advent of genetic testing, current investigations of the causes of DSDs allow genetic-mechanism identification in about 20-35% of cases. This study aimed primarily to establish a rapid and high-throughput genetic test for undervirilized males with and without additional dysmorphic features. Routine chromosomal and endocrinological investigations were performed as part of DSD evaluation. We applied whole-exome sequencing (WES) complemented with multiplex ligation-dependent probe amplification to seek explainable genetic causes. Integrated computing programs were used to call and predict the functions of genetic variants. We recruited 20 patients and identified the genetic etiologies for 14 (70%) patients. A total of seven of the patients who presented isolated DSD phenotypes were found to have causative variants in the AR, MAP3K1, and FLNA genes. Moreover, the other seven patients presented additional phenotypes beyond undervirilized genitalia. Among them, two patients were compatible with CHARGE syndrome, one with Robinow syndrome, and another three with hypogonadotropic hypogonadism. One patient, who carried a heterozygous FLNA mutation, also harbored a heterozygous PTPN11 mutation and thus presented some phenotypes of Noonan syndrome. We identified several genetic variants (12 nonsense mutations and one microdeletion) that account for syndromic and nonsyndromic DSDs in the Taiwanese population. The identification of these causative genes extended our current understanding of sex development and related congenital disorders.

[The Reliability and Validity of the Chinese Version of the Experienced Level of Existential Emptiness].

BACKGROUND: Psychiatric patients are affected by diseases and mental symptoms that may worsen their ability to adjust emotionally. Being unable to respond to the emptiness, increases the risk of suicidal behaviors. PURPOSE: This study was designed to translate the Experienced Level of Existential Emptiness (ELEE) scale, developed by Hazell in 1984, from the original English into Chinese and then to test its reliability and validity. METHODS: This research adopted a cross-sectional design and collected data using convenience sampling and a structured questionnaire. The subjects of this study were psychiatric outpatients in the acute and chronic wards of a psychiatric hospital in Taiwan. The instruments used in this research included a demographic datasheet; the ELEE; the University of California, Los Angeles Loneliness Scale, version 3; the Beck Depression Inventory-II; the State-Trait Anxiety Inventory Y form; and the Oxford Happiness Inventory. After the data were obtained, the reliability and validity of the Chinese-version scale was tested. RESULTS: Three hundred subjects were surveyed. The content validity index value of the ELEE was between .88 and 1, and the retest reliability and intrinsic consistency were good. From the analysis of criterion-related validity, a higher degree of emptiness was shown to correlate with more-obvious feelings of loneliness, depression, and anxiety. Conversely, a higher sense of happiness was shown to correlate with less-obvious feelings of these three variables. After the construct validity test, which used confirmatory factor analysis and regarded the co-variation coefficient of association and model fit index as the basis of consideration, the scale was reduced to two sub-scales of seven questions each. After the deletion of items, the scale retained good retest reliability and intrinsic consistency, supporting the retention of the 14 questions in the scale. CONCLUSIONS / IMPLICATIONS FOR PRACTICE: The findings of this study support using the Chinese version of the ELEE to measure emptiness in patients with mental illness and then providing appropriate medical assistance based on the measured results.

New Structural and Single Nucleotide Mutations in Type I and Type II Collagens in Taiwanese Children With Type I and Type II Collagenopathies.

Collagenopathy is a rare genetic condition characterized by abnormality in either collagen structure or metabolism. Variations in its clinical presentations highlight diversity in the genetic causes and potential existence of concurrent mutations. Through whole exome sequencing (WES) complemented with multiplex ligation-dependent probe amplification, we identified the genetic etiologies for six cases with osteogenesis imperfecta (OI) in COL1A1 (p.T1298N, p.Q1280Pfs∗51, and p.G557Vfs∗23) and COL1A2 (c.1-1677_133-441del) as well as three cases with spondyloepiphyseal dysplasia congenita in COL2A1 (p.G1041S, p.G654S, and p.G441A). Co-occurrence of COL1A1 and WNT1 mutations was found in a patient with a mild OI phenotype but severe osteoporosis. These findings extended the pathogenic variant spectrum of COL1A1, COL1A2, and COL2A1 for type I and type II collagenopathies. Although WES provides a fast and accurate method to identify the genetic causes in most of the patients with type I and type II collagenopathies, its limitation of detecting CNVs because of variable capturing uniformity should be kept in mind when interpreting the results. Taken together, we demonstrate that multiple genetic characterizing technologies can provide an accurate and efficient molecular diagnostic of new genetic variants in disease-causing genes that are compatible with clinical phenotypes.

Identification of a delayed leaf greening gene from a mutation of pummelo.

Delayed greening of young leaves is an unusual phenomenon of plants in nature. Citrus are mostly evergreen tree species. Here, a natural mutant of "Guanxi" pummelo (Citrus maxima), which shows yellow leaves at the young stage, was characterized to identify the genes underlying the trait of delayed leaf greening in plants. A segregating population with this mutant as the seed parent and a normal genotype as the pollen parent was generated. Two DNA pools respectively from the leaves of segregating seedlings with extreme phenotypes of normal leaf greening and delayed leaf greening were collected for sequencing. Bulked segregant analysis (BSA) and InDel marker analysis demonstrated that the delayed leaf greening trait is governed by a 0.3 Mb candidate region on chromosome 6. Gene expression analysis further identified a key candidate gene (Citrus Delayed Greening gene 1, CDG1) in the 0.3 Mb region, which showed significantly differential expression between the genotypes with delayed and normal leaf greening phenotypes. There was a 67 bp InDel region difference in the CDG1 promoter and the InDel region contains a TATA-box element. Confocal laser-scanning microscopy revealed that the CDG1-GFP fusion protein signals were co-localized with the chloroplast signals in the protoplasts. Overexpression of CDG1 in tobacco and Arabidopsis led to the phenotype of delayed leaf greening. These results suggest that the CDG1 gene is involved in controlling the delayed leaf greening phenotype with important functions in chloroplast development.

Topographic Analysis of the Isthmus in Mesiobuccal and Mesial Roots of First Molars in a South Korean Population.

The purpose of this study was to evaluate the incidence and microscopic anatomy of the isthmus to provide more precise anatomical information about the mesiobuccal (MB) roots of the maxillary first molars and the mesial (M) roots of the mandibular first molars. Twenty-eight maxillary and 31 mandibular first molars were embedded, sectioned, stained, and observed at 30× magnification to evaluate the incidence and microscopic anatomy of the isthmus. The incidence of an isthmus 3 mm from the apex was 89.3% and 100% in the MB roots of the maxillary first molars and in the M roots of the mandibular first molars, respectively. The mean dentin thickness between the isthmus and the distal root surface was <1 mm at a distance of 3 mm from the apex in both types of roots. In this study, whenever two main canals were located in the MB roots of the maxillary first molars and in the M roots of the mandibular first molars, the likelihood of the presence of an isthmus increased. Therefore, clinicians should be aware of the thinnest dimensions in the distal surface of the MB roots of the maxillary first molars and the M roots of the mandibular first molars during nonsurgical and surgical root canal treatment.

A novel pathogenic mutation on Interleukin-7 receptor leading to severe combined immunodeficiency identified with newborn screening and whole exome sequencing.

BACKGROUND: Patients with severe combined immunodeficiency (SCID), which is caused by genetic defects in immune-related genes involved in the development or activation of the adaptive immune system, often died in infancy due to severe infections before definite molecular diagnosis could be made. Although recent improvement in early diagnosis has been achieved by newborn screening, the genetic basis of many of the patients is still unknown. METHODS: Here we performed whole exome sequencing (WES) to investigate the underlying genetic causes of SCID in a proband identified with newborn screening. Inheritance of the mutation was confirmed with targeted sequencing of the parents. Homozygosity mapping from the WES was used to investigate the consanguinity of the parents. Immunoblotting was used to confirm the loss of expression of the mutant protein. RESULTS: A novel homozygous frameshift mutation of IL7R was identified through WES. Both parents are carriers for this 1-bp deletion. HLA typing and exome-wide homozygous stretch mapping suggested that the parents are consanguineous. Immunoblotting showed no expression of IL7Rα isoform in the whole blood sample of the proband. The proband received peripheral blood stem cell transplantation and her general condition became stable. Our results suggest that IL7R is essential for T cell development but dispensable for the development of certain human NK cells B cells and suggest that WES can be a useful tool for precise genetic diagnosis of SCID following newborn screening in the index patient without the need to screen other members of the whole family.

[Nursing Experience With Reconstructing Self-Control Using Rational-Emotive Behavior Therapy on a Patient With Schizophrenia and Obsessive-Compulsive Symptoms].

This case report describes a nursing experience caring for a patient with schizophrenia and obsessive-compulsive symptoms. This patient suffered from symptoms of being controlled, obsessive thoughts, and compulsive behaviors. In addition, the patient showed no interest in implementing strategies for dealing with anxiety, no motivation for changing this suffering, and an inability to receive a higher level of rehabilitative job training in daycare. These problems impeded this patient's reintegration into the community. Therefore, the authors employed a five-dimension assessment (physical, emotional, cognitive, social, and spiritual) in order to address the two major nursing problems. The period of nursing care was from October 21, 2016 to January 10, 2017. The two nursing problems addressed included: 1) altered thought processes and 2) ineffective coping. The author provided potentially helpful nursing processes based on the theory of Rational-Emotive Behavior Therapy in order to help the patient cope with symptoms, including being controlled and obsessive-compulsive behaviors. Meanwhile, a relaxation technique was applied to reduce the patient's feelings of discomfort during the nursing processes. As a result, the patient's coping skills to deal with symptoms of being controlled, obsession, and compulsion were improved through refutation of irrational beliefs. In addition to showing rational emotions and appropriate behavior to handle pressures, the patient was also able to apply the relaxation technique to relieve the discomfort from anxiety and pain as needed. This case report suggests that nurses may implement the irrational beliefs refutation training regimen under Rational-Emotive Behavior Therapy for similar cases at the beginning of nursing-patients relationships. Furthermore, providing relaxation techniques in the nursing process may assist patients to deal with stressful life events. The results of this nursing experience are expected to help nursing colleagues apply the above theory and skills with schizophrenia patients with obsessive-compulsive symptoms.

Rare Compound Heterozygous Frameshift Mutations in ALMS1 Gene Identified Through Exome Sequencing in a Taiwanese Patient With Alström Syndrome.

Alström syndrome (AS) is a rare autosomal recessive disorder that shares clinical features with other ciliopathy-related diseases. Genetic mutation analysis is often required in making differential diagnosis but usually costly in time and effort using conventional Sanger sequencing. Herein we describe a Taiwanese patient presenting cone-rod dystrophy and early-onset obesity that progressed to diabetes mellitus with marked insulin resistance during adolescence. Whole exome sequencing of the patient's genomic DNA identified a novel frameshift mutation in exons 15 (c.10290_10291delTA, p.Lys3431Serfs*10) and a rare mutation in 16 (c.10823_10824delAG, p.Arg3609Alafs*6) of ALMS1 gene. The compound heterozygous mutations were predicted to render truncated proteins. This report highlighted the clinical utility of exome sequencing and extended the knowledge of mutation spectrum in AS patients.

Using a Visible Light-Polymerized Resin to Fabricate an Interim Partial Removable Dental Prosthesis.

An interim partial removable dental prosthesis (RDP) is any dental prosthesis that replaces some teeth in a partially dentate arch designed to enhance esthetics, stabilization, and/or function for a limited period of time, after which it is to be replaced by a definitive dental prosthesis. This article describes a technique that uses a visible light-polymerized (VLP) resin as the base material for an interim partial RDP. This technique can be easily accomplished in a dental office or laboratory and results in a predictable dental prosthesis. This technique eliminates the need for laboratory processing.

Next-generation sequencing identifies rare variants associated with Noonan syndrome.

Noonan syndrome (NS) is a relatively common genetic disorder, characterized by typical facies, short stature, developmental delay, and cardiac abnormalities. Known causative genes account for 70-80% of clinically diagnosed NS patients, but the genetic basis for the remaining 20-30% of cases is unknown. We performed next-generation sequencing on germ-line DNA from 27 NS patients lacking a mutation in the known NS genes. We identified gain-of-function alleles in Ras-like without CAAX 1 (RIT1) and mitogen-activated protein kinase kinase 1 (MAP2K1) and previously unseen loss-of-function variants in RAS p21 protein activator 2 (RASA2) that are likely to cause NS in these patients. Expression of the mutant RASA2, MAP2K1, or RIT1 alleles in heterologous cells increased RAS-ERK pathway activation, supporting a causative role in NS pathogenesis. Two patients had more than one disease-associated variant. Moreover, the diagnosis of an individual initially thought to have NS was revised to neurofibromatosis type 1 based on an NF1 nonsense mutation detected in this patient. Another patient harbored a missense mutation in NF1 that resulted in decreased protein stability and impaired ability to suppress RAS-ERK activation; however, this patient continues to exhibit a NS-like phenotype. In addition, a nonsense mutation in RPS6KA3 was found in one patient initially diagnosed with NS whose diagnosis was later revised to Coffin-Lowry syndrome. Finally, we identified other potential candidates for new NS genes, as well as potential carrier alleles for unrelated syndromes. Taken together, our data suggest that next-generation sequencing can provide a useful adjunct to RASopathy diagnosis and emphasize that the standard clinical categories for RASopathies might not be adequate to describe all patients.