Search

Objective: The objective of this study was to assess the longitudinal relationship of weight change on incidence and remission of insulin resistance (IR)., Methods: We performed a cohort study in apparently healthy Korean men, 30 to 59 years of age, who underwent a health checkup and were followed annually or biennially between 2002 and 2009. The computer model of homeostasis model assessment, HOMA2-IR, was obtained at each visit, and IR was defined as HOMA2-IR ≥75th percentile., Results: For IR development, 1,755 of the 6,612 IR-free participants at baseline developed IR (rate 5.1 per 100 person-years) during 34,294.8 person-years of follow-up. The hazard ratios (95% confidence intervals) for incident IR with weight changes of <-0.9 kg, 0.6-2.1 kg and ≥2.2 kg from visit 1 to visit 2 (average 1.8 years) compared to weight change of -0.9-0.5 kg (reference) were 0.78 (0.68-0.90), 1.19 (1.04-1.35) and 1.26 (1.11-1.44), respectively. This association persisted in normal-weight individuals or those without any metabolic syndrome traits and remained significant after introducing weight categories and confounders as time-dependent exposures (P-trend <0.001). For IR remission, 903 of 1,696 IR participants had no IR (remission rate 10.3 per 100 person-years) during 8,777.4 person-years of follow-up. IR remission decreased with increasing quartiles of weight change (P-trend <0.001) and this association persisted in normal-weight individuals., Conclusions: Weight gain was associated with increased IR development and decreased IR remission regardless of baseline BMI status. Preventing weight gain, even in healthy and normal-weight individuals, is an important strategy for reducing IR and its associated consequences.

Ataxia-telangiectasia (A-T) is a rare autosomal recessive neurodegenerative disorder. It is characterized by early-onset, progressive cerebellar ataxia, oculomotor apraxia, choreoathetosis, conjunctival telangiectasias, immunodeficiency, and an increased risk of malignancy. Although A-T is known to be the most common cause of progressive cerebellar ataxia in childhood, there have been no confirmed cases in Korea. We report the clinical and genetic findings of Korean siblings who presented with limb and truncal ataxia, oculomotor apraxia, choreoathetosis, and telangiectasias of the eyes. Sequence analysis of the ataxia-telangiectasia mutated (ATM) gene revealed a known missense mutation (c.8546G>C; p.Arg2849Pro) and a novel intronic variant of intron 17 (c.2639-19&#95;2639-7del13). Reverse-transcription PCR and sequencing analysis revealed that the c.2639-19&#95;2639-7del13 variant causes a splicing aberration that potentiates skipping exon 18. Because A-T is quite rare in Korea, the diagnosis of A-T in Korean patients can be delayed. We recommend that a diagnosis of A-T should be suspected in Korean patients exhibiting the clinical features of A-T.

The expansion of a noncoding hexanucleotide repeat (GGGGCC) in the chromosome 9 open reading frame (C9orf72) gene has been identified as the most common cause of familial and sporadic amyotrophic lateral sclerosis (ALS) in Caucasian populations. The role of the C9orf72 repeat expansion in Korean ALS patients, however, has not been reported. We therefore investigated the frequency of the C9orf72 repeat expansion in 254 Korean patients with familial (n = 8) and sporadic (n = 246) ALS and found that none of the patients had the expansion. The number of hexanucleotide repeats ranged from 2 to 11 in the 254 ALS patients without the expansion. Our results suggest that the C9orf72 repeat expansion is not the main cause of ALS in the Korean population., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Hyperekplexia is a rare inherited neurologic disorder that is characterized by hypertonia and an exaggerated startle response to sudden external stimuli. Until now, 5 genes are known to be associated with hyperekplexia: GLRA1, SLC6A5, GLRB, GPHN, and ARHGEF9. In this report, we performed a clinical and genetic analysis of 4 Korean children with hyperekplexia. Two patients had typical clinical manifestations of hyperekplexia that initially were misdiagnosed as epilepsy. Direct sequencing of the GLRB and GLRA1 genes revealed 2 novel mutations, GLRB c.298-1G>A and c.1028C>T (p.S343F), in patient 1 and 1 novel mutation, GLRA1 c.895C>T (p.R299X), in patient 2. The other 2 familial cases, patients 3 and 4, exhibited startle responses, which appeared at the age of 1 year, and had global developmental delay. Those patients showed negative results for the 5 genes.

Pyridoxine-dependent epilepsy (PDE) is a rare autosomal recessive disorder that causes intractable seizures, especially in neonates and infants. Patients are typically resistant to typical antiepileptic drugs (AEDs) but respond dramatically to pyridoxine. Mutations in the ALDH7A1 gene are associated with the pathogenesis of PDE. Herein, we report the clinical phenotypes and disease-causative mutations in the ALDH7A1 gene in three Korean patients with PDE. We reviewed the medical records, electroencephalography (EEG), brain magnetic resonance imaging (MRI) findings, and the results of molecular genetic tests for the patients who were diagnosed with PDE in our institution between Jan. 1996 and Dec. 2010. In all patients, the first seizures began during the first week of life. The seizures were not fully controlled with multiple AEDs, but disappeared immediately after administration of pyridoxine and returned after it was transiently discontinued. Before the use of pyridoxine, interictal EEGs showed multifocal epileptiform discharges, which became normalized with pyridoxine. Direct sequencing analyses revealed two mutant alleles in all three patients. Patient 1 was compound heterozygous with two different missense mutations, c.1061A>G (p.Y354C) and c.1232C>T (p.P411L). Patient 2 was homozygous for a missense mutation, c.1279G>C (p.E427Q). Patient 3 was compound heterozygous for two different missense mutations, c.1061A>G (p.Y354C) and c.1279G>C (p.E427Q), and her parents and younger brother were heterozygous carriers of each one of the mutations. All three mutations had not previously been reported. Herein, we report three Korean patients with three novel mutations who presented with PDE.

Tricho-rhino-phalangeal syndrome type I (TRPSI) is a rare autosomal dominant hereditary disorder characterized by sparse hair, bulbous nose, long philtrum, thin upper lip, and skeletal abnormalities including cone-shaped epiphyses, shortening of the phalanges, and short stature. TRPSI is caused by mutations in the TRPS1 gene. Herein, we report two Korean cases of TRPSI. Although both patients (a 17-year-old-female and a 14-year-old male) had typical clinical findings, Patient 1 had an additional growth hormone (GH) deficiency. Treatment with recombinant human growth hormone (rhGH) 0.7 IU/kg/week led to an increase in growth velocity. Over 10 years of GH therapy, the mean growth velocity was 5.7 ± 0.9 cm/year. However, the patient 2 did not show apparent GH deficiency by GH stimulation test, had a poor response with rhGH therapy and GH therapy was discontinued after 6 months. Upon genetic analysis of the TRPS1 gene, two mutations were found. Patient 1 had a heterozygous mutation c.2520dupT (p.Arg841LysfsX3) which had not been previously reported. Patient 2 had a known nonsense mutation c.1630C>T (p.Arg544X). In summary, we were the first to report Korean patients with mutation of TRPS1.

Rabson-Mendenhall syndrome (RMS) is a rare syndrome manifested by extreme insulin resistance with hyperinsulinemia, acanthosis nigricans, tooth dysplasia and growth retardation. Our patient was first noted at the age of 8 months due to pigmentations on skin-folded areas. Initial laboratory tests showed normal fasting glucose (69 mg/dL). Fasting insulin level was severely elevated, up to 554.6 µIU/mL, and c-peptide level was increased, up to 13.81 ng/mL. However, hemoglobin A1c was within normal range (4.8%). He is now 11 yr old. His growth development followed the 5-10th percentile and oral hypoglycemic agents are being administered. The last laboratory results showed insulin 364.1 µIU/mL, C-peptide 4.30 ng/mL, and hemoglobin A1c 7.6%. The boy was a compound heterozygote for the c.90C > A and c.712G > A mutations of the insulin receptor gene, INSR, which are nonsense and missense mutations. In summary, we report the first Korean case of RMS, which was confirmed by two novel mutations of the INSR.

About 5% of amyotrophic lateral sclerosis (ALS) cases are known to be familial (fALS) and mutations in SOD1 and other genes are found in more than 20% of fALS patients and in 2%-4% of apparently sporadic ALS (sALS) cases. However, there are few reports on the proportion of fALS and the frequency of mutations in Korean patients with ALS. We screened mutations in the SOD1, FUS, TARDBP, ANG, and OPTN genes in 258 consecutively enrolled Korean patients with ALS from October 2006 to November 2010. The frequency of fALS was estimated to be 3.5% (9/258), and mutations were identified in 88.9% (8/9) of fALS patients but only in 2.8% (7/249) of sALS patients. Seven fALS and 3 sALS patients had mutations in SOD1 gene while all the others had FUS gene. The proportion of fALS was lower than that reported in Caucasian populations but the frequency of SOD1 gene mutations in Korean fALS patients (77.8%, 7/9) was much higher than that reported in other ethnic groups. These findings might suggest that there is an ethnic difference in the proportion of fALS and the genetic background of ALS., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background/aims: Endoscopic submucosal dissection (ESD) is accepted as a standard treatment of early gastric cancer (EGC) and gastric adenoma. Occasionally, tumorous lesion is not found and pathologic discrepancies can occur after ESD. The aim of this study was to analyze the factors affecting the negative pathologic results after ESD., Methods: We retrospectively reviewed the data from all patients with gastric neoplasm (276 EGC and 516 gastric adenomas) who were treated with ESD during past 3 years and enrolled the patients who had negative pathologic results., Results: Out of 792 patients treated with ESD, 27 patients (3.4%) were eligible for inclusion. Among the 27 patients, factors affecting the negative pathologic results were, most commonly, the focal lesion (n=13, 48.2%) which was small enough to be removed completely during pre-ESD biopsy, followed by pathologic discrepancies (n=11, 40.7%) between pathologists and lastly the operator factor (n=3, 11.1%) dissecting incorrect lesions. Of the focal lesions, the initial pathologic diagnoses were adenocarcinoma in 11 cases (84.6%). In cases with pathologic discrepancies, all the pretreatment diagnoses were adenoma with low grade dysplasia. In cases caused by operator factors, intestinal metaplasia was accompanied by elevated adenoma in all cases., Conclusions: To decrease negative pathologic results after ESD, an endoscopist should perform ESD after sufficient communication with pathologists, especially for adenoma with low grade dysplasia, and choose correct lesion, especially located at the antrum and associated with intestinal metaplasia. The possibility of total removal of small lesions even by forcep biopsy should be considered.

The frequencies and spectra of germline mutations in the BRCA1 and BRCA2 genes vary among populations. In the present study, the mutation spectra of the BRCA1/BRCA2 genes in Korean breast cancer patients were investigated using whole-gene sequencing method. A total of 134 unrelated Korean breast cancer patients who were identified as being at high risk of carrying BRCA1/BRCA2 mutations were included. PCR amplification and direct sequencing were performed covering all exons and flanking intronic sequences of the BRCA1/BRCA2 genes. A total of 26 mutations were detected in 31 of 134 patients (23.1%). The mutation detection rate in the present study is higher than those of previous studies using screening methods (2.5-11.3%) and similar to that of a recent study, which used whole-gene sequencing (21.2%). The BRCA2: c.7480C>T mutation, which has been suggested to be a founder mutation in Koreans, was detected in only one patient. Five mutations were recurrent but observed in no more than two patients. Given that the mutation detection rates using whole-gene sequencing were much higher than for screening methods and that there were no consistent observations of founder mutations, whole-gene sequencing of both BRCA1 and BRCA2 genes should be the method of choice to identify mutations in high-risk Korean patients.

The formation of glial scars impedes growth of regenerating axons after CNS injuries such as spinal cord injury (SCI). Hepatocyte growth factor (HGF), originally identified as a mitogen for hepatocytes, exerts pleiotropic functions in the nervous system. HGF has been implicated in peripheral wound healing via regulation of the transforming growth factor beta (TGFβ), which is also a potent inducer of glial scar formation in CNS. In the present study, we found that HGF completely blocked secretion of TGFβ1 and β2 from activated astrocytes in culture. HGF also prevented expression of specific chondroitin sulfate proteoglycan (CSPG) species. To determine whether HGF inhibits glial scar formation in an in vivo SCI model, HGF overexpressing mesenchymal stem cells (HGF-MSCs) were transplanted into hemisection spinal cord lesions at C4. Transplantation of HGF-MSCs markedly diminished TGFβ isoform levels and reduced the extent of astrocytic activation. In addition, HGF-MSCs also significantly decreased neurocan expression and glycosaminoglycan chain deposition around hemisection lesions. Furthermore, animals treated with HGF-MSCs showed increased axonal growth beyond glial scars and improvement in recovery of forepaw function. Our results indicate that anti-glial scar effects of HGF, together with its known neurotrophic functions, could be utilized to ameliorate functional deficits following SCI., (Copyright © 2011 Elsevier Inc. All rights reserved.)

We investigated the relationships between demographics, lifestyle characteristics, and serum total prostate specific antigen (PSA) concentration and examined the population-based distribution of total PSA by age among 2,246 Korean men with a median age of 45 years. We obtained data about demographic and lifestyle characteristics based on self-reporting using a questionnaire. We also performed physical examinations, anthropometric measurements, and biochemical measurements. The PSA concentration increased with age and there was a significant difference in total PSA concentration between the age groups of 21-60 years and >60 years. Age>60 years, height≥1.8 m, a low frequency of alcohol consumption, and taking nutritional supplements showed a significantly increased odds ratio for increased PSA when 3.0 ng/ mL was chosen as the PSA cut-off level. Smoking status, BMI, percent body fat, diabetes mellitus, fatty liver, herbal medicine use, vitamin use, and diet were not significantly associated with total PSA regardless of the cut-off level. When interpreting a single PSA test, height, alcohol consumption, and nutritional supplement use should be considered, in addition to age.

Background: Cytochrome P450 2C9 (CYP2C9) is the major isoform of the CYP2C subfamily, and is involved in the metabolism of many clinically important therapeutic agents. Here we describe a patient who was intolerant of warfarin treatment because of impaired drug metabolism related to the CYP2C9*3/*13 genotype and high warfarin sensitivity associated with the VKORC1 1173TT genotype., Case: A 64-year-old Korean man had taken warfarin for treatment of an occluded left subclavian artery and atrial fibrillation. Although the warfarin doses prescribed were not high (14 mg/week) during the induction of anticoagulation, the prothrombin time (PT) was elevated to over 120 s after two weeks of warfarin therapy. For this patient, a very low dose of warfarin (4 mg/week) was required in order to achieve target INR values., Conclusions: To the best of our knowledge, this is the first report of a patient with CYP2C9*3/*13 and VKORC1 1173TT genotypes who had slower than normal warfarin metabolism, resulting in the need to administer an extremely low dose of warfarin in order to achieve the target INR value. Our report reinforces the relevance of pharmacogenetic testing to explain warfarin dose variability and to enable individualized dosage adjustments for improved warfarin treatment outcomes., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Objective: To investigate thyroid-stimulating hormone receptor (TSHR) and thyroid peroxidase (TPO) mutations in Korean patients with primary congenital hypothyroidism (CH)., Context: Congenital hypothyroidism is a common genetic disorder in which the majority of mutations occur in the TSHR and TPO genes., Design: We examined the frequencies of TSHR and TPO mutations among Korean patients with primary CH. Furthermore, we explored the relationships between imaging findings and mutation status., Patients: A total of 193 paediatric patients with nonsyndromic CH were enrolled in the present study., Measurements: Patients with decreased (99m) Tc uptake were screened for TSHR mutations using Sanger sequencing, and those with increased uptake were screened for TPO mutations. The relationships between scintigraphic and ultrasonographic findings and mutation status were analysed., Results: Thirteen (16·5%) of 79 patients with decreased (99m) Tc uptake were found to harbour TSHR mutations including G132R, G245S, R450H, R519C and F525S. The R450H mutation was present in 13 (72·2%) of 18 disease alleles. Seven (10·3%) of 68 patients with increased (99m) Tc uptake harboured TPO mutations including R189Q, K439E, G493S, C808LfsX72, A863T, R875Hfs and P883S. The TSHR and TPO mutations were observed only in patients with normal to slightly enlarged thyroid glands., Conclusions: This study identified underlying TSHR and TPO mutations in Korean patients with CH and revealed a possible relationship between imaging findings and mutation status. In addition, the low rate of mutation positivity suggests significant genetic heterogeneity of CH in the Korean population., (© 2011 Blackwell Publishing Ltd.)

Congenital central hypoventilation syndrome (CCHS), also known as Ondine's curse, is characterized by idiopathic failure of autonomic breathing and is often associated with neurocristopathies such as Hirschsprung disease (HSCR). CCHS is caused by mutations in the paired-like homeobox 2B (PHOX2B) gene, often manifest as polyalanine repeat expansions. Herein, we report the cases of two unrelated Korean patients with Ondine-Hirschsprung disease. The patient's clinical manifestations were apnea and cyanosis requiring immediate endotracheal intubation, recurrent hypoventilation with hypercapnia, hypoxia after ventilator removal, and abdominal distension since birth. Intestinal biopsies were performed and the absence of ganglion cells in the colon was consistent with HSCR. We performed direct sequencing analysis in the PHOX2B and RET genes and fluorescence polymerase chain reaction in order to determine the polyalanine tract expansion in exon 3 of the PHOX2B gene. Expansion mutations were detected in both patients; one had 20/24 repeats and the other had 20/27 repeats. The 20/24 genotype has not been previously described in severe CCHS phenotypes and associated HSCR. We believe that the information in this report will improve our understanding of the phenotypic and genotypic heterogeneities of CCHS and HSCR.

Chronic progressive external ophthalmoplegia (CPEO) is a mitochondrial myopathy commonly caused by deleterious changes in the mitochondrial DNA (mtDNA). We describe a 45-year-old man who was referred to us for investigation of progressive ptosis. We performed a multiplex ligation-dependent probe amplification (MLPA) assay of mtDNA from muscle tissue and peripheral blood leukocytes, and followed up with gap-polymerase chain reaction (PCR) and direct sequence analysis. Results showed a deletion of a 4,407 bp segment in the mtDNA region, ranging from nucleotide position 8,577 in the MT-ATP6 gene to nucleotide position 12,983 in the MT-ND5 gene. To the best of our knowledge, this is the first report of a CPEO patient with a large novel deletion of mtDNA genetically confirmed by MLPA assay. MLPA can be a feasible platform for clinical laboratories to detect large deletion mutations in the mtDNA for suspected cases.

We identified a novel missense mutation in the Cu/Zn superoxide dismutase (SOD1) gene in a 47-year-old woman with familial amyotrophic lateral sclerosis (ALS). The heterozygous mutation, in exon 1 of the SOD1 gene, is a GC to TT transversion in nucleotide positions 13 and 14 leading to an alanine 4 to phenylalanine (A4F) amino acid substitution. It was found in six family members. The effect of the A4F mutation was of similar severity to that of the A4V mutation. We discuss structural instability as a possible pathogenic mechanism in the case of this SOD1 mutation. The proband displayed upper motor neuron signs not observed in individuals with other codon 4 mutations. This could be because longer survival allows UMN dysfunction to become evident. We also provide a literature review., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Carrier woman of Duchenne muscular dystrophy (DMD) can mimic the inflammatory myositis in presenting symptoms. Two diseases should be differentiated by the clinical history, muscle biopsy and genetic study. There are few reports in which both histochemical and genetic study showed the possible link of overlapping inflammatory pathophysiology with dystrophinopathy. We report a 40-yr-old woman who presented with subacute proximal muscle weakness and high serum level of creatine kinase. She had a history of Graves' disease and fluctuation of serum liver aminotransferase without definite cause. MRI, EMG and NCV were compatible with proximal muscle myopathy. Muscle biopsy on vastus lateralis showed suspicious perifascicular atrophy and infiltration of mono-macrophage lineage cells complicating the diagnosis. Dystrophin staining showed heterogeneous diverse findings from normal to interrupted mosaic pattern. Multiple ligation probe amplification and X chromosome inactivation test confirmed DMD gene deletion mutation in exon 44 and highly skewed X inactivation.

Coagulation factor VII (FVII) deficiency is a rare hereditary coagulopathy caused by mutations in the F7 gene. The aims of this study were to characterize the molecular defect of F7 in Korean patients with FVII deficiency and to find genotype-phenotype correlations. Study individuals consisted of 14 unrelated Korean patients with FVII deficiency with residual FVII activities ranging from 1 to 34%. To identify causative mutations, we performed PCR amplification and direct sequencing of all exons and flanking sequences of F7 gene. In all 14 patients, one (N = 4) or two (N = 10) mutant alleles were identified. A total of 11 unique mutations were detected, of which four were novel (c.-1C>T, p.V54RfsX53, p.R59&#95;R60dupRR, and p.L314 V). Four recurrent mutations were observed in 86% of patients (12 of 14) (C389G, C115X, G343S, and c.572-1G>A) and accounted for 71% of all mutant alleles (17 of 24). The residual FVII activity was more than 5% in all six asymptomatic patients (21%, range 6-34%), whereas it was 5% or less in all eight symptomatic patients (2%, range 1-5%). In addition, the mean residual FVII activity in four patients with a single mutant allele was 23.6% (range 3-34%), which was significantly higher than that in 10 patients with two mutant alleles that was 4.6% (range 1-19%) (P = 0.023). In summary, the data from this study on the largest number of FVII deficiency in Korea showed recurrent mutations in this population and suggested genotype-phenotype correlations.

Hereditary spastic paraplegia (HSP) is a neurodegenerative disease characterized by progressive spasticity in the lower extremities. Mutations in the atlastin GTPase 1 (ATL1) gene cause approximately 10% of autosomal dominantly inherited HSP. For many subjects with an ATL1 mutation, spastic gait begins in early childhood and does not significantly worsen, even over many years; such cases resemble spastic diplegic cerebral palsy. Herein we report a heterozygous R239C mutation in the ATL1 gene in a Korean family. The family members exhibited early onset pure spastic paraplegia and had been previously diagnosed with the diplegic form of cerebral palsy. We suggest that spastic paraplegia type 3 (SPG3A) be included in the differential diagnosis of early onset spastic paraplegia. To the best of our knowledge, this is the first report of a genetically confirmed family affected with SPG3A in Korea.

Purpose: The aim of this study was to evaluate the periodontal and prosthodontic complications of multiple freestanding implants in the posterior jaws for up to 1 year of function., Materials and Methods: Eight patients received 20 implants posterior to canines. Two or more implants were consecutively inserted to each patient. Single crowns were delivered onto the implants. Marginal bone loss, implant mobility, probing depth, and screw loosening were examined to evaluate the clinical success of such restorations for maximum 1 year of functional loading., Results: All the implants performed well during the observation period. Neither periodontal nor prosthodontic complications were found except a slight porcelain chipping. While the marginal bone level was on average 0.09 mm lower around the implant after 6 months of loading, it was 0.15 mm higher after 1 year., Conclusion: Within the limits of this investigation, separate single-tooth implant restorations to replace consecutive missing teeth may clinically function well in the posterior jaw.

Coagulation factor XII (FXII) participates in the initiation of blood coagulation, fibrinolysis, complement systems, and bradykinin generation. Hereditary FXII deficiency is caused by mutations in the F12 gene. In this report, we describe three Korean patients with FXII deficiency. In all three patients, prolonged activated partial thromboplastin time (aPTT) was detected on preoperative coagulation screening, without a history of bleeding tendency. Patient 1 was a 4-year-old girl with congenital heart anomaly. Her aPTT was prolonged at 67.5 s (reference range, 29-42 s) and the FXII activity was 5% (48-177%). Patient 2 was a 50-year-old man with anal fistula. His aPTT was 95.2 s and FXII activity was less than 0.5% (below the detection limit). Patient 3 was a 53-year-old woman with pituitary macroadenoma. Her aPTT was 238.8 s and FXII activity was less than 0.5%. Direct sequencing analyses of F12 revealed two mutations in all three patients. Patient 1 was compound heterozygous for two missense mutations, c.1583C>T (p.S528F) and c.1744G>A (p.G582S). Patient 2 was homozygous for a nonsense mutation, c.405C>A (p.C135X). Patient 3 was compound heterozygous for a frameshift mutation and a missense mutation, c.1093&#95;1094insC (p.K365QfsX69) and c.1744G>A (p.G582S). All four mutations were novel. To the best of our knowledge, these are the first genetically confirmed cases of FXII deficiency in Korea. FXII deficiency should be included in differential diagnoses of prolonged aPTT, particularly in those without bleeding tendency.

Myotonic dystrophy type 1 (DM1) is caused by unstable expansion of the CTG repeat in the DMPK gene. According to the hypothesis that expanded CTG repeat alleles originated from larger normal alleles, a correlation exists between prevalence of DM1 and frequency of large normal alleles. We examined the number of CTG repeats in a group of 481 non-DM1 individuals and 116 DM1 patients. Among these subjects, we analyzed the haplotype in 76 unrelated non-DM1 individuals and 14 unrelated DM1 patients using 8 bialleleic markers on the DM1 locus. Different CTG repeats from 5 to 36 and variable allele frequencies were observed. The most common allele was 12 CTG repeats (27.3%), and the frequency of larger normal alleles (>19 CTG repeats) was 3.7%. Haplotype analysis revealed that 100% of alleles with 5 and >19 CTG repeats were haplotype A. In this study, we provide the first haplotypic molecular evidence for a founder effect of DM1 mutations in Korea, and reinforce the hypothesis that out-of-Africa DM1 alleles were derived by expansion from a pool of non-DM1 alleles with haplotype A.

Background: Warfarin is a widely used oral anticoagulant with broad within- and between-individual dose requirements. Warfarin concentrations can be monitored by assessing its pharmacologic effects on International Normalized Ratio (INR). However, this approach has not been applied in the routine clinical management of patients receiving warfarin therapy. We performed a plasma warfarin assay using high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) to determine if such an assay can be utilized in routine clinical practice., Methods: We included a total of 105 patients with atrial fibrillation, and who were receiving warfarin for more than 1 yr. The plasma concentrations of total warfarin and 7-hydroxywarfarin were determined by HPLC-MS/MS (Waters, UK). We assessed the association between warfarin dose, concentration, and INR as well as the effects of these factors on warfarin concentrations., Results: The mean maintenance dose of warfarin in 105 patients was 4.1 +/-1.3 mg/day (range, 1.7-8.0 mg/day) and their mean plasma warfarin concentration was 1.3+/-0.5 mg/L. We defined a concentration range of 0.6-2.6 mg/L (corresponding to the 2.5th to 97.5th percentile range of the Plasma warfarin levels in the 74 patients showing INR within target range) as the therapeutic range for warfarin. The correlation of warfarin dose with warfarin concentration (r(2)=0.259, P<0.001) was higher than that with INR (r(2)=0.029, P=0.072)., Conclusions: There was a significant correlation between warfarin dose and plasma warfarin concentrations in Korean patients with atrial fibrillation. Hence, plasma warfarin monitoring can help determine dose adjustments and improve our understanding of individual patient response to warfarin treatment.

Branchio-oto-renal (BOR) and branchio-oto (BO) syndromes are autosomal dominant hereditary disorders characterized by the presence of hearing loss and branchial fistulae and cysts, with (BOR syndrome) or without (BO syndrome) renal malformations of varying degrees of severity. Mutations in the human homologous of the Drosophila eyes absent (EYA1) gene are frequently the cause of BOR/BO syndrome. Here we describe a Korean family with BO syndrome; the proband had preauricular pit, cup-shaped auricles, branchial fistula, and hearing loss, without renal involvement. Molecular genetic study revealed a novel mutation occurring in the consensus acceptor splice site of intron 8 (c.868-2A > G) in the EYA1 gene. To the best of our knowledge, this is the first report of a splice site mutation in a family with BO syndrome without renal involvement, further extending the phenotypic-genotypic heterogeneity of BOR/BO syndrome.

Hereditary factor XI (FXI) deficiency is a rare bleeding disorder inherited in an autosomal recessive manner. The genetic background of FXI deficiency is the mutations in the F11 gene on the chromosome band 4q35. The prevalence is known to be particularly high in Ashkenazi Jews with well documented recurrent mutations; however, founder mutations in F11 have also been reported in non-Jewish patients. In this report, we describe a Korean patient with severe FXI deficiency whose causative mutations were identified by molecular genetic tests. The patient was a 33-year-old pregnant woman. Routine gynecologic workup revealed prolonged activated partial thromboplastin time. Her FXI level was severely decreased at 1% (reference range, 60-140%). Direct sequencing analysis of the F11 gene was performed to identify the causative mutations. The patient was shown to have two different mutations, c.1546 G>A (Val498Met) and c.1560dupG (Tyr503ValfsX32) in the F11 gene. Val498Met is a novel missense mutation, and the Tyr503ValfsX32 mutation was recently reported in a Japanese patient. Both mutations occurred in the exon 13 of F11 and were believed to disrupt the catalytic domain of the FXI protein, leading to severe FXI deficiency. To the best of our knowledge, this is the first genetically confirmed case of severe FXI deficiency in Korea, and more cases are needed to find any signature of founder effect in the Korean population and its potential relationship with other Asian populations.

Peptide oral delivery is still a significant challenge because of two major impediments, low absorption efficiency and instability in gastrointestinal tract. The aim of this study was to design, prepare, and evaluate an intestine enzyme-resistant, superior intestine absorptive, and biologically preserved glucagon-like peptide-1 (GLP-1) derivative using the site-specific modification of biotin, which can take advantage of the carrier-mediated active intestine transport. Two series of site-specific Lys34- and Lys26,34-biotin-GLP-1 derivatives were prepared, and their intestine membrane permeabilities, proteolytic stabilities against the intestine enzymes, and bioactivities were then evaluated. Especially, Lys26,34-biotin-GLP-1 was found to have the most promising scores: (i) it displayed a 5.6-fold higher Caco-2 cell monolayer permeability than GLP-1; (ii) it showed an 8.5- and 3.5-fold longer half-life than GLP-1 in rat intestine fluid and homogenate, respectively; and interestingly (iii) it had a well-preserved insulinotropic activity (94.5% vs. GLP-1) in the rat islets. Finally, Lys26,34-biotin-GLP-1 showed a 9-fold higher oral hypoglycemic efficacy (25.3%) than native GLP-1 (2.7%) (P<0.005) after direct peroral administration into type 2 diabetic db/db mice. This study highlights the oral hypoglycemic potential of site-specific Lys26,34-biotinylated GLP-1, and this orally available analogue would find a role in the treatment of type 2 diabetes.

Background: Point-of-care testing (POCT) glucometers are increasingly being used for making important therapeutic decisions and managing diabetes. We examined the analytical performance of GLUCOCARD X-METER (ARKRAY Global Business Inc., Japan) against three other glucometers and a reference laboratory method., Methods: We evaluated the analytical performance of GLUCOCARD X-METER in comparison with three other glucometers. Studies on precision, linearity, the analysis time, and effects of hematocrit and temperature were carried out and the results were compared with those of the laboratory reference method (hexokinase method by Hitachi 760, Hitachi Co., Japan)., Results: GLUCOCARD X-METER showed a good linearity and within-run and total-run precision. Comparison between each glucometer and the Hitachi 7600 showed a good correlation. Although differences with the reference method were within an allowable range, all glucometers showed variable bias. Application of an insufficient amount of blood could produce some changes in test results. Changes in hematocrit were found to cause overestimation or underestimation of glucose values. For some test strips, the results were affected by prolonged exposure to room temperature or 4 degrees C refrigerator., Conclusions: GLUCOCARD X-METER showed a good analytical performance in linearity, precision, and comparison. The effect of hematocrit, sample volume, and storage condition for test strips were noted and glucometers had variable deviations to both directions from laboratory reference values (<20%). The GLUCOCARD X-METER provided rapid and reliable measurements of blood glucose. It could be appropriate for monitoring blood glucose values in diabetic patients.

The purpose of this study was to demonstrate the biological potentials of PEGylated salmon calcitonin (PEG-sCT) derivatives administered intratracheally and their dependences on PEG Mw (1, 2, 5 kDa). Initially, three different PEG-sCT derivatives were site-specifically synthesized by attaching PEG to the Lys(18)-amine. In an attempt to examine the pulmonary feasibilities of these derivatives, the following evaluations were undertaken to determine their; (i) proteolytic resistances to pulmonary enzymes, (ii) bioactivities, and (iii) pulmonary pharmacokinetic and pharmacologic profiles. The results obtained showed that the pulmonary stabilities and pharmacokinetic properties of these derivatives were greatly improved by increasing PEG Mw. PEG-sCTs had 10.5-, 40.1-, and 1066.0-fold greater stabilities than that of sCT in rat lung homogenates. Moreover, all pharmacokinetic parameters (AUC(inf), C(max), t(1/2), and others) of these derivatives in endotracheally cannulated rats were significantly improved by PEGylation. Specifically, C(max) values increased on increasing PEG Mw, i.e., 78.1+/-21.1, 102.9+/-9.1, and 115.2+/-5.7 for 1, 2, 5 kDa, respectively, vs. 54.8+/-3.9 ng/mL for sCT. Their circulating t(1/2) values also increased to 53.9+/-6.0, 100.7+/-21.7, and 119.4+/-13.7 min, respectively, vs. 34.6+/-7.6 min for sCT. Despite having the best properties, Lys(18)-PEG(5k)-sCT was found to have significantly lower hypocalcemic efficacy than other PEG-sCTs, probably due to its reduced intrinsic bioactivity ( approximately 30% vs. sCT). Rather, Lys(18)-PEG(2k)-sCT showed the most promising pulmonary potential because of its well-preserved bioactivity (>80% of sCT). Taken together, our findings suggest that the site-specific substitution to peptides like sCT with a PEG of an appropriate size offers optimized therapeutic potential by dual advantages, i.e., (i) increased proteolytic stability and (ii) extended circulating half-life in terms of intrapulmonary delivery.

A 47-year-old man presented with headache, nausea, vomiting and fever. Laboratory findings including analysis of cerebrospinal fluid suggested bacterial meningitis. Erysipelothrix rhusiopathiae was identified in cultures of cerebrospinal fluid. The patient recovered without any neurological sequelae after antimicrobial treatment. It is interesting that intracranial infection by E. rhusiopathiae reappeared after scores of years and that it presented with absence of an underlying cause or bacteraemia.

Rapid translation of genome sequences into meaningful biological information hinges on the integration of multiple experimental and informatics methods into a cohesive platform. Despite the explosion in the number of genome sequences available, such a platform does not exist for filamentous fungi. Here we present the development and application of a functional genomics and informatics platform for a model plant pathogenic fungus, Magnaporthe oryzae. In total, we produced 21,070 mutants through large-scale insertional mutagenesis using Agrobacterium tumefaciens-mediated transformation. We used a high-throughput phenotype screening pipeline to detect disruption of seven phenotypes encompassing the fungal life cycle and identified the mutated gene and the nature of mutation for each mutant. Comparative analysis of phenotypes and genotypes of the mutants uncovered 202 new pathogenicity loci. Our findings demonstrate the effectiveness of our platform and provide new insights on the molecular basis of fungal pathogenesis. Our approach promises comprehensive functional genomics in filamentous fungi and beyond.

This study investigated the porous-microparticle (PM) with low mass density and large size for pulmonary drug delivery. PM was prepared by the water-in-oil-in-water (W(1)/O/W(2)) multi-emulsion method with cyclodextrin derivative as a porogen and a stabilizer of peptide drugs. Herein, sucrose ethyl acetate (SAIB) was incorporated in PM for long acting protein release. In vitro release studies, the rapid release rate of proteins from PM was reduced due to the high viscosity of the added SAIB. As a result, BSA release from PM continued up to 7 days. This result suggests that PM having sustained release characteristics may be successfully applied for long-term pulmonary administration of protein or peptide drug. In addition, it is expected that these particles arrive at a deep lung epithelium due to low density (high porosity) and limit macrophage recognition because of big particle size (more than 5 microm).

Porous microparticles (PMs) with a low density (<0.4 g/cm3) for pulmonary protein delivery were prepared by the water-in-oil-in-water (W1/O/W2) multi-emulsion method using a cyclodextrin derivative as a porogen. The complexation of positively charged lysozyme (Lys) and negative-charged hyaluronate (HA) was investigated for long-term protein release from PMs. The interaction of Lys and HA not only increased protein encapsulation efficiency but also stabilized Lys against a denaturing organic solvent (dichloromethane). Furthermore, PMs with Lys/HA complexes increased the Lys release period up to 7 days, as opposed to a 4h Lys release time from PMs without Lys/HA complexes. In particular, PMs containing 10mg of HA and 50mg of Lys showed almost zero-order Lys release kinetic for 7 days and preserved the bioactivity of Lys more than 98% during its entire release period. This result suggests that PMs with Lys/HA complexes may be applied in long-term pulmonary administration of protein or peptide drugs, including those that require particles to arrive at a deep lung epithelium with the help of low density (high porosity) of PMs.

In stabilizing proteins during microsphere fabrication, the viscous solid-in-water-in-oil-in-water (S/W/O/W) method was compared to the conventional multi-emulsion W/O/W and S/O/W method. Solid proteins lyophilized with cyclodextrin derivatives and polyethylene glycol (PEG) pass through an organic solvent phase and are then embedded in aqueous microdroplets of first emulsion. Proteins were stabilized at the water/organic solvent interface by an internal aqueous phase containing viscous polysaccharides, and then can be safely encapsulated without degradation. In addition, these microspheres showed a long-term protein release followed by nearly zero-order kinetics with minimal initial burst. This means that the viscous S/W/O/W method provides a safe strategy for microsphere fabrication and has promising properties, involving the preservation of protein bioactivity, the inhibition of protein denaturation or agglomeration, and long-term protein release.

This study investigated the suitability of microsphere formulations for extended protein delivery and complete protein release. These microspheres were prepared by a multi-emulsion method and prepared using a mixture of poly(lactide-co-glycolide) (PLGA), RG 502H (lactide:glycolide=50:50, M(W) 9300) and sucrose acetate isobutyrate (SAIB). SAIB embedded into the microspheres and mixed with PLGA, improved the efficiency of enzyme encapsulation. The in vitro release rate of lysozyme (Lys) from the microspheres was reduced due to the high viscosity of the added SAIB and less degradation of PLGA by SAIB. These properties enabled prolonged release of Lys for up to 2 months, characterized by a minimal initial burst of Lys and nearly zero-order protein release kinetics result from co-administration of sorbitan monooleate 80. When it is considered that degradation products of SAIB are inactive for labile proteins, SAIB may be regarded as a promising candidate for long-acting protein delivery.