Therapeutic Drug Monitoring of 6 New-Generation Antiseizure Medications Using a Mass Spectrometry Method: Analysis of 2-Year Experience in a Large Cohort of Korean Epilepsy Patients.

CONTEXT.­: New-generation antiseizure medications (ASMs) are increasingly prescribed, and therapeutic drug monitoring (TDM) has been proposed to improve clinical outcome. However, clinical TDM data on new-generation ASMs are scarce. OBJECTIVE.­: To develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for therapeutic drug monitoring (TDM) of 6 new-generation ASMs in serum and analyze the clinical TDM data from a large cohort of Korean patients with epilepsy. DESIGN.­: Stable isotope-labeled internal standards were added to protein precipitations of serum. One microliter of sample was separated on Agilent Poroshell EC-C18 column, and lacosamide, perampanel, gabapentin, pregabalin, vigabatrin, and rufinamide were simultaneously quantified by Agilent 6460 triple-quad mass spectrometer in multiple-reaction monitoring mode. Linearity, sensitivity, precision, accuracy, specificity, carryover, extraction recovery, and matrix effect were evaluated. TDM data of 458 samples from 363 Korean epilepsy patients were analyzed. RESULTS.­: The method was linear with limit of detection less than 0.05 µg/mL in all analytes. Intraassay and interassay imprecisions were less than 5% coefficient of variation. Accuracy was within ±15% bias. Extraction recovery ranged from 85.9% to 98.8%. A total of 88% (403 of 458) were on polypharmacy, with 29% (118 of 403) using concomitant enzyme inducers. Only 38% (175 of 458) of the concentrations were therapeutic, with 53% (244 of 458) being subtherapeutic. Drug concentration and concentration-to-dose ratio were highly variable among individuals in all 6 ASMs. CONCLUSIONS.­: A simple and rapid LC-MS/MS method for TDM of 6 ASMs was developed and successfully applied to clinical practice. This large-scale TDM data could help establish an effective monitoring strategy for these drugs.

Identification of two novel COL3A1 variants in patients with vascular Ehlers-Danlos syndrome.

BACKGROUND: Vascular Ehlers-Danlos syndrome (vEDS) is an autosomal dominant disease caused by aberrations in COL3A1, which encodes type III collagen. Sanger sequencing has limitations for diagnosis since exon deletion/duplication and splicing alterations are not uncommon in COL3A1. We report 2 patients with vEDS who were not diagnosed by conventional Sanger sequencing. METHODS: We performed either targeted panel or whole-genome sequencing. Complementary DNA (cDNA) sequencing was performed using cultured skin fibroblasts. Sanger sequencing of DNA was performed for the confirmation of breakpoints in the case of exon deletion. We also evaluated the sensitivity of the splicing prediction tool, SpliceAI. RESULTS: An exon 27 deletion was suspected on targeted panel sequencing of 1 patient. The deletion was confirmed using cDNA sequencing (r.1870_1923del) and breakpoints were confirmed (c.1870-109_1923+10del). On targeted panel sequencing in the other patient, we found a novel intronic variant of c.1149+6T>C that leads to skipping of exon 16 (r.1051_1149del) by cDNA sequencing. SpliceAI showed 98.8% sensitivity for known splicing variants in COL3A1. CONCLUSION: Our study highlights the necessity of a comprehensive approach to the genetic diagnosis of vEDS. In addition, cDNA sequencing was useful as an auxiliary method, especially considering the limited sensitivity of the splicing prediction tool.

Whole-genome sequencing in clinically diagnosed Charcot-Marie-Tooth disease undiagnosed by whole-exome sequencing.

Whole-genome sequencing is the most comprehensive form of next-generation sequencing method. We aimed to assess the additional diagnostic yield of whole-genome sequencing in patients with clinically diagnosed Charcot-Marie-Tooth disease when compared with whole-exome sequencing, which has not been reported in the literature. Whole-genome sequencing was performed on 72 families whose genetic cause of clinically diagnosed Charcot-Marie-Tooth disease was not revealed after the whole-exome sequencing and 17p12 duplication screening. Among the included families, 14 (19.4%) acquired genetic diagnoses that were compatible with their phenotypes. The most common factor that led to the additional diagnosis in the whole-genome sequencing was genotype-driven analysis (four families, 4/14), in which a wider range of genes, not limited to peripheral neuropathy-related genes, were analysed. Another four families acquired diagnosis due to the inherent advantage of whole-genome sequencing such as better coverage than the whole-exome sequencing (two families, 2/14), structural variants (one family, 1/14) and non-coding variants (one family, 1/14). In conclusion, an evident gain in diagnostic yield was obtained from whole-genome sequencing of the whole-exome sequencing-negative cases. A wide range of genes, not limited to inherited peripheral neuropathy-related genes, should be targeted during whole-genome sequencing.

Comprehensive Evaluation of the NeoBase 2 Non-derivatized MSMS Assay and Exploration of Analytes With Significantly Different Concentrations Between Term and Preterm Neonates.

Background: Despite the popularity of the NeoBase 2 Non-derivatized MSMS assay (PerkinElmer, Turku, Finland), there are no reports of its comprehensive evaluation, including the ability to distinguish transient tyrosinemia of the newborn (TTN) from tyrosinemia type 1 (TYR 1) using succinylacetone (SUAC). No newborn screening (NBS) cutoffs for preterm neonates in the Korean population have been suggested. We evaluated the NeoBase 2 assay and identified analytes requiring different cutoffs in preterm neonates. Methods: Residual NBS dried blood spot samples and proficiency testing (PT) materials of the Newborn Screening Quality Assurance Program and the Korean Association of External Quality Assessment Service were used. Precision, accuracy, limit of detection (LOD), lower limit of quantification (LLOQ), linearity, recovery, carryover, and performance of SUAC were evaluated. Cutoffs were determined, and analytes requiring different cutoffs in preterm neonates were investigated. Results: Mean CVs for within-run and between-day precision were within 15%. Accuracy analysis indicated high agreement with in-house derivatized assay results and results of other PT participants. All analytes demonstrated acceptable LOD, LLOQ, and linearity. Recoveries were acceptable, except for SUAC. Carryover was negligible. Cutoffs were established for all analytes; Tyr, adenosine, and C20:0-lysophosphatidylcholine required different cutoffs in preterm neonates. Differential diagnosis of TYR 1 and TTN was successful with simultaneous Tyr and SUAC measurement. Conclusions: The NeoBase 2 assay demonstrated satisfactory performance. The additional analytes provide a wider diagnostic coverage, and the simultaneous measurement of Tyr and SUAC is efficient in excluding TYR 1. The new cutoffs for preterm neonates may decrease false-positive rates, without compromising diagnostic sensitivity.

Analysis of ABO grouping discrepancies among patients from a tertiary hospital in Korea.

BACKGROUND: Accurate ABO typing is essential for preventing ABO incompatibility reactions. However, the causes of ABO grouping discrepancy has not been sufficiently studied, and it may vary among different ethnic populations. Thus, the aim of this retrospective study was to investigate the causes of ABO discrepancy in the East Asian population. MATERIALS AND METHODS: A retrospective observational study on ABO typing discrepancy among patients in a tertiary hospital was carried out using the electronic medical record database of Samsung Medical Center (Seoul, Korea) between July 2016 and May 2019. RESULTS: ABO grouping was performed on 551,959 blood samples during the study period; 1468 events of serologic ABO discrepancy were determined from 1334 (0.24 %) samples. A total of 134 samples (0.02 %) presented multiple causes of ABO discrepancy. Weak/missing serum reactivity (594, 40.5 %) was the most frequent reason for ABO discrepancy, followed by extra serum reactivity (370, 25.2 %), weak/missing red cell reactivity (267, 18.2 %), mixed-field red cell reactivity (176, 12.0 %), and extra red cell reactivity (61, 4.2 %). In the category of weak/missing red cell reactivity, ABO subgroup was the most common reason, and using ABO genotyping, 26.2 % of the cases genotyped were found to be related to the cis-AB allele. CONCLUSIONS: Our results suggest that the incidence and cause of ABO typing discrepancies vary among institutes and ethnic groups. Our data helps to better understand and facilitate the resolution of ABO typing discrepancies in patients.

Anthropometric comparison of the idealized youth and hideous old man of Leonardo's profile drawings.

The aim of this study is to compare the old and ugly with the young and beautiful in Leonardo's profile drawings using soft tissue anthropometry.Thirty-one of Leonardo's profile drawings and portraits of the lateral view were collected and scanned. Among the 29, 9 young and 20 hideous old man scans had a measurable nasion-subnasale (n-sn) distance, and 49 anthropometric items (43 distances and 6 angles) were used for measurement as relative distances to the n-sn distance.The measured distances or angles did not differ significantly between the young and old in the 39 anthropometric items. However, the remaining 10 items were statistically significant. The young group had a significantly greater (P < 0.05) upper face height (n-stomion [sto]), nasolabial angle, and upper lip height (sn-sto) compared with the old group. However, the supraorbital depth (glabella-tragion), mandible height (sto-gnathion), nasal bridge height (n-pronasale), cutaneous lower lip height (labiale inferius-sublabiale), ear length (superaurale-subaurale), Rickett line-upper lip distance, and facial inclination were significantly greater (P < 0.05) in the old group compared with the young group.The difference of soft tissue cephalometric results between the young and old subjects can be the index in rejuvenating surgeries.

Correction of an alar web with a feather-edge rolled-in flap.

The aim of this study was to see the histological nature of the alar web and to introduce a featheredged rolled-in flap to reduce the alar web.On a cadaver, the perpendicular section of the alar web revealed a thickened dermis portion on both the skin side and the nasal side distal to the alar cartilage. According to histological results, we thought the thinning and rolling in of the distal margin of the end of the open rhinoplasty incision could reduce the alar web. An open rhinoplasty incision was made just distal to the hair-bearing vestibular skin and a V-Y shape incision created at the alar base. After the cartilage work, the skin of the distal end of the flap was featheredged to a 0.5-mm thickness. The distal margin was rolled in and sutured to the nasal lining. A bolster dressing was applied using a 4-0 nylon suture.Thirteen patients (8 males, 5 females) were operated on, and 8 patients were followed up for more than 12 months. Their preoperative and postoperative worm's eye views were compared. Four anthropometric distances were measured preoperatively and postoperatively. The columellar length increased significantly after the operation (P = 0.001 [independent 2-sample t test]) on the cleft side. Preoperatively, the alar width was significantly greater (P = 0.02 [paired-samples t test]) on the cleft side (0.17 ± 0.03 of an intercanthal distance) than the noncleft side (0.14 ± 0.03). After the operations, they became similar (0.16 ± 0.03 on cleft side, 0.16 ± 0.04 on the noncleft side; P = 1.00 [paired-samples t test]).We think this featheredged rolling-in flap might be a good method for the correction of an alar web since this technique increased the columellar length and decreased the alar width on the cleft side.

Would medical students enter an exclusion zone in an infected district with a high mortality rate? An analysis of book reports on 28 (secondary publication).

This study aimed to ascertain whether medical students would enter a closed area where there was a raging epidemic of an infectious disease with a high fatality rate, and includes reasons for the students entering or refusing to enter. Participants included 50 second-year medical students. They were assigned to read a novel entitled 28, written by Youjeong Jeong, and discuss it in groups. Using their book reports, their decisions of whether or not to enter Hwayang, the city from the novel, and the reasons for their decisions were analyzed; we furthermore investigated the factors affecting their decisions. Among the 50 respondents, 18 students (36%) answered that they would enter, and the remaining 32 students (64%) answered that they would not enter the zone. The reasons given for entering were responsibility (44%), sense of ethics (33%), social duty (17%), and sense of guilt (6%). The reasons the students provided for not entering were inefficiency (44%), worry regarding family (28%), needlessness of sacrifice (19%), and safety not ensured (9%). Students who had four or fewer family members were more likely to enter Hwayang than were students who had five or more family members (odds ratio, 1.85). Students who had completed over 100 hours of volunteer work were more likely to enter Hwayang than were students who had volunteered less than 100 hours (odds ratio, 2.04). Owing to their "responsibility" as a doctor, 36% of medical students answered that they would enter an exclusion zone in an infected district with a high fatality rate. However, 64% answered they would not enter because of "inefficiency." For the medical students it is still a question 'To enter or not to enter?'