Clinical Application of Sequential Epigenetic Analysis for Diagnosis of Silver–Russell Syndrome

Background@#Silver-Russell syndrome (SRS) is a pre- or post-natal growth retardation disorder caused by (epi)genetic alterations. We evaluated the molecular basis and clinical value of sequential epigenetic analysis in pediatric patients with SRS. @*Methods@#Twenty-eight patients who met ≥ 3 Netchine-Harbison clinical scoring system (NH-CSS) criteria for SRS were enrolled;26 (92.9%) were born small for gestational age, and 25 (89.3%) showed postnatal growth failure. Relative macrocephaly, body asymmetry, and feeding difficulty were noted in 18 (64.3%), 13 (46.4%), and 9 (32.1%) patients, respectively. Methylation-specific multiplex ligation-dependent probe amplification (MSMLPA) on chromosome 11p15 was performed as the first diagnostic step. Subsequently, bisulfite pyrosequencing (BP) for imprinting center 1 and 2 (IC1 and IC2) at chromosome 11p15, MEST on chromosome 7q32.2, and MEG3 on chromosome 14q32.2 was performed. @*Results@#. Seventeen (60.7%) patients exhibited methylation defects, including loss of IC1 methylation (N = 14; 11 detected by MS-MLPA and three detected by BP) and maternal uniparental disomy 7 (N = 3). The diagnostic yield was comparable between patients who met three or four of the NH-CSS criteria (53.8% vs 50.0%). Patients with methylation defects responded better to growth hormone treatment. @*Conclusions@#NH-CSS is a powerful tool for SRS screening. However, in practice, genetic analysis should be considered even in patients with a low NH-CSS score. BP analysis detected additional methylation defects that were missed by MS-MLPA and might be considered as a first-line diagnostic tool for SRS.

Management of Osteogenesis Imperfecta: A Multidisciplinary Comprehensive Approach

Osteogenesis imperfecta (OI) is characterized by recurring fractures and limb and spine deformities. With the advent of medical therapeutics and the discovery of causative genes, as well as the introduction of a newly devised intramedullary rod, the general condition and ambulatory function of patients diagnosed with OI have been improved over the past decades. This review covers recent developments in research and management of OI.

Functional Outcomes of Hip Arthroscopy for Pediatric and Adolescent Hip Disorders

BACKGROUND: There is a paucity of literature on the use of hip arthroscopy for pathologic conditions in skeletally immature patients. Thus, the indications and safety of the procedure are still unclear. The purpose of this study was to investigate the safety and functional outcomes of hip arthroscopy for pediatric and adolescent hip disorders. We further attempted to characterize arthroscopic findings in each disease.METHODS: We retrospectively reviewed 32 children and adolescents with hip disorders who underwent 34 hip arthroscopic procedures at a tertiary care children's hospital from January 2010 to December 2016. We evaluated functional limitations and improvement after operation by using the modified Harris hip score (HHS), the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), subjective pain assessment with a visual analog scale (VAS), and range of hip motion as well as the complications of hip arthroscopy. Arthroscopic findings in each disease were recorded.RESULTS: Hip arthroscopy was performed for Legg-Calvé-Perthes disease (n = 6), developmental dysplasia of the hip (n = 6), slipped capital femoral epiphysis (n = 5), idiopathic femoroacetabular impingement (n = 6), sequelae of septic arthritis of the hip (n = 3), hereditary multiple exostosis (n = 2), synovial giant cell tumor (n = 3), idiopathic chondrolysis (n = 2), and posttraumatic osteonecrosis of the femoral head (n = 1). Overall, there was a significant improvement in the modified HHS, WOMAC, VAS, and range of hip motion. Symptom improvement was not observed for more than 18 months in four patients who had dysplastic acetabulum with a labral tear (n = 2) or a recurrent femoral head bump (n = 2). There were no complications except transient perineal numbness in five patients.CONCLUSIONS: Our short-term follow-up evaluation shows that hip arthroscopy for pediatric and adolescent hip disorder is a less invasive and safe procedure. It appears to be effective in improving functional impairment caused by femoroacetabular impingement between the deformed femoral head and acetabulum or intra-articular focal problems in pediatric and adolescent hip disorders.

Mycobacterium bovis Osteitis Following Immunization with Bacille Calmette-Guérin (BCG) in Korea

BACKGROUND: Mycobacterium bovis Bacille Calmette-Guérin (BCG) osteitis, a rare complication of BCG vaccination, has not been well investigated in Korea. This study aimed to evaluate the clinical characteristics of BCG osteitis during the recent 10 years in Korea. METHODS: Children diagnosed with BCG osteitis at the Seoul National University Children's Hospital from January 2007 to March 2018 were included. M. bovis BCG was confirmed by multiplex polymerase chain reaction (PCR) in the affected bone. BCG immunization status and clinical information were reviewed retrospectively. RESULTS: Twenty-one patients were diagnosed with BCG osteitis and their median symptom onset from BCG vaccination was 13.8 months (range, 6.0–32.5). Sixteen children (76.2%) received Tokyo-172 vaccine by percutaneous multiple puncture method, while four (19.0%) and one (4.8%) received intradermal Tokyo-172 and Danish strain, respectively. Common presenting symptoms were swelling (76.2%), limited movement of the affected site (63.2%), and pain (61.9%) while fever was only accompanied in 19.0%. Femur (33.3%) and the tarsal bones (23.8%) were the most frequently involved sites; and demarcated osteolytic lesions (63.1%) and cortical breakages (42.1%) were observed on plain radiographs. Surgical drainage was performed in 90.5%, and 33.3% of them required repeated surgical interventions due to persistent symptoms. Antituberculosis medications were administered for a median duration of 12 months (range, 12–31). Most patients recovered without evident sequelae. CONCLUSION: Highly suspecting BCG osteitis based on clinical manifestations is important for prompt management. A comprehensive national surveillance system is needed to understand the exact incidence of serious adverse reactions following BCG vaccination and establish safe vaccination policy in Korea.

Diagnosis and Management of Hip Dislocation in Patients with Kabuki Syndrome

BACKGROUND: Kabuki syndrome is a rare genetic disorder characterized by distinct dysmorphic facial features, growth deficiency, intellectual disabilities, unusual dermatoglyphic patterns, and skeletal abnormalities. The incidence of hip dislocation in Kabuki syndrome ranges from 18% to 62%. We reviewed the outcomes of management of hip dislocations in patients with Kabuki syndrome with special attention to the diagnostic processes for hip dislocation and Kabuki syndrome. METHODS: Among 30 patients with mutation-confirmed Kabuki syndrome, we selected six patients who had hip dislocations and reviewed their medical records and plain radiographs. The modes of presentation and diagnostic processes for both hip dislocations and Kabuki syndrome were investigated. The management and treatment outcomes of hip dislocations in patients with Kabuki syndrome were evaluated. RESULTS: The average age of patients at the time of diagnosis of hip dislocation was 7.7 months (range, 1 week to 22 months). None of the patients were diagnosed as having Kabuki syndrome at that time. Two patients were treated with a Pavlik harness; one, with closed reduction; two, with open reduction and later pelvic and/or femoral osteotomies; and one, with open reduction combined with pelvic osteotomy. The patients were followed up for 5.8 years on average (range, 2.0 to 10.5 years). The radiologic outcome was graded as Severin IA or IB for three patients who were older than 6 years at the latest follow-up (mean age, 9.9 years; range, 7.8 to 12.4 years). In the remaining three patients younger than 6 years (mean age, 3.8 years; range, 2.7 to 5.3 years), the lateral center edge angle was more than 15°. The clinical diagnosis of Kabuki syndrome was made during follow-up after hip dislocation treatment and confirmed by mutational analysis at a mean age of 4.7 years. The mean interval between the diagnosis of hip dislocation and Kabuki syndrome was 4.0 years. CONCLUSIONS: The management of hip dislocation by conservative or surgical method showed successful results. Awareness of Kabuki syndrome could lead to an early diagnosis of this rare disease in patients with hip dislocation and allow for early detection of other underlying conditions and multidisciplinary management.

Regarding Camurati-Engelmann Disease: In Reply

No abstract available.

Orthopedic Manifestations of Type I Camurati-Engelmann Disease

BACKGROUND: Camurati-Engelmann disease (CED) is a rare genetic skeletal disorder characterized by limb pain, muscle emaciation and weakness, and cortical thickening of the diaphysis of long bones. It is caused by mutations in the transforming growth factor beta 1 (TGFB1) (type I) or other unknown gene(s) (type II). We present 8 consecutive patients with type I CED. METHODS: We retrospectively reviewed medical records and radiographs of type I CED patients with special reference to the mode of presentation, process of diagnostic work-up, and disease course. They were 4 sporadic patients, and two pairs of mother and son. RESULTS: We categorized the mode of presentation into three groups. Group I had 4 patients who mainly presented with motor disturbances in young age. They drew medical attention for waddling gait, awkward ambulation or running, difficulty in going upstairs, or a positive Gower's sign at age 4 to 6 years. Subsequent development of limb pain and radiographic abnormality led to the diagnosis of CED at age 6 to 29 years. Group II had 3 patients who mainly presented with limb pain at age 15, 20, and 54 years, respectively. Radiographic evaluation and molecular genetic test led to the diagnosis of CED. The remaining 1 patient (group III) was asymptomatic until age 9 years when bony lesions at the tibiae were found incidentally. For the last 10 years, he intermittently complained of leg pain in the morning or after sports activities, which did not interfere with daily life. All the patients in group I showed a body mass index in the underweight range (< 18.4 kg/m²). At the latest follow-up, 4 patients in groups I and II required medication for the limb pain. CONCLUSIONS: CED presents with a wide range of severity. Awareness of this rare disease entity may be the key to timely correct diagnosis. This disease entity should be considered in the differential diagnosis of limb pain or motor disturbance in children to avoid unnecessary diagnostic work-up.

New Records of Xylaria Species in Korea: X. ripicola sp. nov. and X. tentaculata

During a Korean mushroom diversity survey from 2011 to 2014, we found one new Xylaria species (X. ripicola sp. nov.) and one Xylaria species that had not been previously observed in Korea (X. tentaculata). To confirm the phylogenetic placement of the new species, we conducted a phylogenetic investigation based on internal transcribed spacer regions of ribosomal DNA sequences. Additionally, the new species, X. ripicola, was subsequently analyzed for RNA polymerase II subunit sequences. We also evaluated the macroscopic and microscopic features of this species. Herein, X. ripicola is described as a new species that was collected from a natural beach habitat and X. tentaculata is formally reported as newly found in Korea.

Mushroom Flora of Ulleung-gun and a Newly Recorded Bovista Species in the Republic of Korea

We conducted five times surveys, in June, September and October in 2012; June and September 2013, to catalog the mushroom flora in Ulleung-gun, Republic of Korea. More than 400 specimens were collected, and 317 of the specimens were successfully sequenced using the ribosomal DNA internal transcribed spacer barcode marker. We also surveyed the morphological characteristics of the sequenced specimens. The specimens were classified into 2 phyla, 7 classes, 21 orders, 59 families, 122 genera, and 221 species, and were deposited in the herbarium of Korea National Arboretum. Among the collected species, 72% were saprophytic, 25% were symbiotic, and 3% were parasitic. The most common order was Agaricales (189 specimens, 132 species), followed by Polyporales (47 specimens, 27 species), Russulales (31 specimens, 22 species), Boletales (10 specimens, 7 species), and so on. Herein, we also reported the first Bovista species in Korea, which was collected from Dokdo, the far-eastern island of Korea.

CO/HO-1 Induces NQO-1 Expression via Nrf2 Activation

BACKGROUND: Carbon monoxide (CO) is a cytoprotective and homeostatic molecule with important signaling capabilities in physiological and pathophysiological situations. CO protects cells/tissues from damage by free radicals or oxidative stress. NAD(P)H:quinone oxidoreductase (NQO1) is a highly inducible enzyme that is regulated by the Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway, which is central to efficient detoxification of reactive metabolites and reactive oxygen species (ROS). METHODS: We generated NQO1 promoter construct. HepG2 cells were treated with CO Releasing Molecules-2 (CORM-2) or CO gas and the gene expressions were measured by RT-PCR, immunoblot, and luciferase assays. RESULTS: CO induced expression of NQO1 in human hepatocarcinoma cell lines by activation of Nrf2. Exposure of HepG2 cells to CO resulted in significant induction of NQO1 in dose- and time-dependent manners. Analysis of the NQO1 promoter indicated that an antioxidant responsible element (ARE)-containing region was critical for the CO-induced Nrf2-dependent increase of NQO1 gene expression in HepG2 cells. CONCLUSION: Our results suggest that CO-induced Nrf2 increases the expression of NQO1 which is well known to detoxify reactive metabolites and ROS.

A Case of a Extrasynovial Ossifying Chondrolipoangioma in the Infrapatellar Fat Pad: Advanced Stage of Hoffa's Disease

Hoffa's disease refers to inflammation, subsequent hypertrophy and impingement of the infrapatellar fat pad, and this is all triggered by acute trauma or repetitive microtrauma. Patients with Hoffa's disease manifest with anterior knee pain and functional impairment. The chronic impingement of the infrapatellar fat pad, which contains many progenitor cells, may promote fibrocartilaginous formation and osteochondral metaplasia of adipose tissue, so osteochondroma is sometimes found in patients with advanced Hoffa's disease. We report here on an ossifying chondrolipoangioma as an advanced form of Hoffa disease that occurred in the infrapatellar fat pad.

The Effect of Taekwondo Training on Aerobic Capacity, MVO2, ST Slope and QRS Duration / 체질인류학회지

The purpose of this study was to investigate the effects of taekwondo training on VO2max, MVO2 and electrocardiogram ST slope, QRS duration in college woman. Subject were divided into two groups, one is taekwondo group (n=11) and the other is control group (n=10). They participated in graded exercise test (Balke protocol) and measured aerobic capacity and electrocardiogram. The aerobic capacity (VO2max and AT) was measured by autogas analyser. The resting and exercise rate-pressure product (RPP) could be measured with HR and systolic BP for MVO2 measurement. The electrocardiogram ST slope, QRS duration was calculated using automated software program (QMC). The measured data were analyzed by using paired t -test. The results of this study were as follows : There was a significant difference of VO2max and AT between taekwondo group and control group. VO2max of taekwondo group was 4~5 mL/kg/min higher than control group (p<0.01) and AT also taekwondo group was 3.4% higher than control group (p<0.05). There was no significant difference of maximal exercise MVO2 between taekwondo and control group. There was no significant difference of electrocardiogram ST slope between group but rest QRS duration in taekwondo group was longer than the control group (p<0.01). Conclusively, these data suggest that regular taekwondo training will strengthen on aerobic capacity and resting slow heart beats.

Predisposition of genetic disease by modestly decreased expression of GCH1 mutant allele

Recently it was shown that single nucleotide polymorphisms (SNPs) can explain individual variation because of the small changes of the gene expression level and that the 50% decreased expression of an allele might even lead to predisposition to cancer. In this study, we found that a decreased expression of an allele might cause predisposition to genetic disease. Dopa responsive dystonia (DRD) is a dominant disease caused by mutations in GCH1 gene. The sequence analysis of the GCH1 in a patient with typical DRD symptoms revealed two novel missense mutations instead of a single dominant mutation. Family members with either of the mutations did not have any symptoms of DRD. The expression level of a R198W mutant allele decreased to about 50%, suggesting that modestly decreased expression caused by an SNP should lead to predisposition of a genetic disease in susceptible individuals.

Isokinetic Strength of Quadriceps and Hamstring, Hamstrings/Quadriceps Ratios in Elite Sports Athletes / 체질인류학회지

The purpose of this study was to investigate of isokinetic strength of quadriceps and hamstring, hamstrings/quadriceps ration in elite sports athletes to provide the data for injury prevention, fitness level evaluation and training methodology. The subjects selected for this study were 104 elite sports athletes. They didn't have any medical problem. Isokinetic peak torque, flexor/extensor ratio at 60 deg/sec of the knee extension and flexion were measured using an cybex 770. The data were analyzed utilizing One way ANOVA. The level of statistical significance was set at p<.05. On the basis of the results analyzed in this study, conclusion were drawn as follow: 1. Dominant knee extension peak torque (Nm) were no significant differences in man but man elite athletes were higher than woman elite players in 60 deg/sec (p<.05). 2. Dominant knee extension peak torque (% body weight) of man elite athletes were higher than woman elite players in 60 deg/sec except only man volleyball athletes (p<.05). 3. Dominant knee flexion peak torque (Nm) were no significant differences in man but man elite athletes were higher than woman elite players in 60 deg/sec (p<.05). 4. Dominant knee flexion peak torque (% body weight) of man elite athletes were higher than woman elite players in 60 deg/sec (p<.05). 5. Knee flexion/extension peak torque ratio (H : Q) were no significant differences according to sex and proper H : Q ratio was 54~64%.

Increased lysine N-methylation of a 23-kDa protein during hepatic regeneration

The methylation of a 23-kDa nuclear protein increased after partial hepatectomy and methylation returned to basal levels after the initial stage of regeneration. The methylating enzyme was partially purified from rat liver by ammonium sulfate precipitation, DEAE-anion exchange chromatography and Butyl-Sepharose chromatography. The 23-kDa protein was purified from a nuclear fraction of liver tissue with SP-Sepharose. When the 23-kDa protein was methylated with the partially purified methyltransferase and analyzed on C18 high performance liquid chromatography (HPLC), the methylated acceptor amino acid was monomethyl lysine (MML). Previously, only arginine N-methylation of specific substrate proteins has been reported during liver regeneration. However, in this report, we found that lysine N-methylation increased during early hepatic regeneration, suggesting that lysine N-methylation of the 23-kDa nuclear protein may play a functional role in hepatic regeneration. The methyltransferase did not methylate other proteins such as histones, hnRNPA1, or cytochrome C, suggesting the enzyme is a 23-kDa nuclear protein- specific lysine N-methyltransferase.

Detection and Typing of HSV-1, HSV-2, CMV and EBV by Quadruplex PCR

The development of a multiplex polymerase chain reaction (PCR) method for rapid and accurate detection and typing of herpes simplex virus type 1 (HSV-1), and type-2 (HSV-2), cytomegalovirus (CMV) and Epstein-Barr virus (EBV) is very important for clinical diagnosis to allow the deliver of therapy as early as possible. Large scale amplifications by multiplex PCR of viral DNA can lower the cost and time for viral diagnosis. In this study, therefore sensitive quadruplex PCR was achieved by optimizing parameters such as primers, and 1.5 mM magnesium and 200 uM dNTPs concentrations. The concentrations of HSV-1, HSV-2, CMV and EBV primers were 0.5, 0.3, 0.25 and 0.25 pmoles, respectively. Optimal annealing temperature was 54 degrees C. Employing these conditions, we could detect 10 copies of reconstructed template plasmid DNA, which were cloned to vectors containing target sequences of viral DNA. PCR products of 271 bp for HSV-1, 231 bp for HSV-2, 368 bp for CMV, and 326 bp for EBV were separated on 5.0% polyacrylamide gel electrophoresis and confirmed by direct sequencing. The present study showed that the quadruplex PCR assay described herein has potential application in clinical diagnosis, when rapid, accurate detection and typing of viruses HSV-1, HSV-2, CMV or EBV are necessary.

A Family with A Missense Mutation in the SCN5A Gene

Brugada syndrome, an autosomal dominantly inherited form of ventricular fibrillation, is characterized by ST-segment elevation in leads V1-3 and right bundle-branch block on surface electrocardiogram. It is caused by mutations in the cardiac sodium channel gene, SCN5A, and to the best of our knowledge, there has been no report of this mutation in Korea. Three members of a family were heterozygous for a G to T substitution at the nucleotide position 5851 in exon 28 of the SCN5A gene. This nucleotide alteration makes a missense mutation, leading to a valine to leucine substitution (V1951L), in the carboxy terminal region of the sodium channel a subunit. We report here a missense mutation in a Korean family with Brugada-type electrocardiogram.

Quadruplex PCR for 8 Autosomal STR Loci in Korean / 대한법의학회지

Two quadruplex PCR reactions were designed for 7 tetrameric (D3S2406, D4S2368, D5S818, D7S821, D9S925, D13S317, D19S253) and one trimeric (D6S1043) short tandem repeats loci to study the allele frequency and the applicability of genetic variation in these loci in forensic case works. For 310 unrelated Koreans DNA was isolated from peripheral blood using phenol/chloroform method. Quadruplex I was consisted of D4S2368, D6S1043, D7S821, D9S925 and quadruplex II D3S2406, D5S818, D13S317, D19S253. The amplified products were analyzed by 5%polyacrylamide gel electrophoresis followed by silver staining. The heterozygosity in each loci ranged 92.91-66.13%, and PD(Power of Discrimination) was above 0.85 in each loci. Every loci except D6S1043 followed hardy-Weinberg expectation. The cumulative PI was low as 1.65x10-10. Two mutations were noted, one in D19S253 and the other in D9S925 among 234 gametes. With these results above eight STR loci studied here proved to be highly polymorphic enough to be used in forensic field. This study provides valuable population data in these loci for Korean.

Quadruplex PCR for 8 Autosomal STR Loci in Korean / 대한법의학회지

Two quadruplex PCR reactions were designed for 7 tetrameric (D3S2406, D4S2368, D5S818, D7S821, D9S925, D13S317, D19S253) and one trimeric (D6S1043) short tandem repeats loci to study the allele frequency and the applicability of genetic variation in these loci in forensic case works. For 310 unrelated Koreans DNA was isolated from peripheral blood using phenol/chloroform method. Quadruplex I was consisted of D4S2368, D6S1043, D7S821, D9S925 and quadruplex II D3S2406, D5S818, D13S317, D19S253. The amplified products were analyzed by 5%polyacrylamide gel electrophoresis followed by silver staining. The heterozygosity in each loci ranged 92.91-66.13%, and PD(Power of Discrimination) was above 0.85 in each loci. Every loci except D6S1043 followed hardy-Weinberg expectation. The cumulative PI was low as 1.65x10-10. Two mutations were noted, one in D19S253 and the other in D9S925 among 234 gametes. With these results above eight STR loci studied here proved to be highly polymorphic enough to be used in forensic field. This study provides valuable population data in these loci for Korean.

Study on DXS7132 and GATA31D10 Loci in Korean Population / 대한법의학회지

The validation study for two STR loci on X-chromosome, DXS7132 and GATA31D10, was done including allelic distribution and frequency of each allele to use these results for individual identification and paternity testing. For 496 unrelated Koreans, above two STR loci were amplified simultaneously using duplex PCR amplification method. The amplified products were analyzed by polyacrylamide gel electrophoresis followed by silver staining. In male DXS7132 locus revealed 7 different alleles ranging from 276bp to 300bp. The largest allele was consisted of 14 repetition of [TCTA] unit and took 0.3417. The allele 15 followed next as 0.3165 and allele 13 as 0.1726. In female general distribution was same except one allele, allele 18 was found additionally. The heterozygosity was 0.7706 and 23 different genotypes were found. Polymorphism information content(PIC) was 0.727. Two cases of mutation were noted in DXS7132 locusIn both male and female 7 different alleles were noted in GATA31D10 locus and the alleles ranged from 195bp to 231bp. The allele 15(199bp) took the majority of all as 0.825. The other alleles showed rather relatively low frequency. The heterozygosity was 0.2385 and 11 different genotypes were found. PIC was 0.2521, and no mutation was noted in GATA31D10 locus. Considering these two loci together, 22 different halpotype were noted.