Otoconial Degeneration After Transient Ischemia Induced by Four-Vessel Occlusion in Rats

Background@#and Purpose Ischemia of the inner ear may damage the otoconia. However, no study has explored any changes in the configuration of otoconia after transient ischemia of the labyrinth. @*Methods@#Nineteen 7-week-old Sprague-Dawley rats were randomly assigned to either the sham (n=5) or the experimental group (n=14). The rats in the experimental group were subjected to global ischemia for 20 minutes using a four-vessel occlusion model, and were sacrificed seven days after the procedure. The rats in the sham group were sacrificed without any procedure. The otolithic organs (utricle and saccule) were dissected out for scanning electron microscope. @*Results@#The otolithic organs in the sham group showed their normal gross configuration with a dense clumping of otoconia with a normal hexagonal morphology and a smooth surface. The otolithic organs in the experimental group also maintained a grossly normal configuration, but each otoconia showed irregular surfaces with numerous cracks or furrows, especially in the periphery of the otoconial bed. @*Conclusions@#The current study showed that otoconial degeneration may occur even after transient ischemia of the labyrinth. This finding supports an association between cerebral ischemia and benign paroxysmal positional vertigo.

Whole Exome Sequencing in Patients with Phenotypically Associated Familial Intracranial Aneurysm

Objective@#Familial intracranial aneurysms (FIAs) are found in approximately 6%–20% of patients with intracranial aneurysms (IAs), suggesting that genetic predisposition likely plays a role in its pathogenesis. The aim of this study was to identify possible IA-associated variants using whole exome sequencing (WES) in selected Korean families with FIA. @*Materials and Methods@#Among the 26 families in our institutional database with two or more IA-affected first-degree relatives, three families that were genetically enriched (multiple, early onset, or common site involvement within the families) for IA were selected for WES. Filtering strategies, including a family-based approach and knowledge-based prioritization, were applied to derive possible IA-associated variants from the families. A chromosomal microarray was performed to detect relatively large chromosomal abnormalities. @*Results@#Thirteen individuals from the three families were sequenced, of whom seven had IAs. We noted three rare, potentially deleterious variants (PLOD3 c.1315G>A, NTM c.968C>T, and CHST14 c.58C>T), which are the most promising candidates among the 11 potential IA-associated variants considering gene-phenotype relationships, gene function, co-segregation, and variant pathogenicity. Microarray analysis did not reveal any significant copy number variants in the families. @*Conclusion@#Using WES, we found that rare, potentially deleterious variants in PLOD3, NTM, and CHST14 genes are likely responsible for the subsets of FIAs in a cohort of Korean families.

A de novo Microdeletion of ANKRD11 Gene in a Korean Patient with KBG Syndrome

KBG syndrome is a very rare genetic disorder characterized by macrodontia of upper central incisors, global developmental delay, distinctive craniofacial features, short stature, and skeletal anomalies. Ankyrin repeat domain 11 gene (ANKRD11) has recently been identified as a causal factor of this syndrome. We describe a 6-yr-old Korean boy with features of KBG syndrome. The patient had a short stature, macrodontia, dysmorphic facial features, speech and motor delay with intellectual disability, and partial seizures as indicated by the electroencephalogram, but he was neither autistic nor had autism spectrum disorders. Using high-resolution oligonucleotide array comparative genomic hybridization, we identified a heterozygous 240-kb deletion at 16q24.3 corresponding to ANKRD11. This patient provided additional evidence on the influence of ANKRD11 in KBG syndrome and suggested that deletion limited to ANKRD11 is unlikely to cause autism.

Clinical Significance of Previously Cryptic Copy Number Alterations and Loss of Heterozygosity in Pediatric Acute Myeloid Leukemia and Myelodysplastic Syndrome Determined Using Combined Array Comparative Genomic Hybridization plus Single-Nucleotide Polymo

The combined array comparative genomic hybridization plus single-nucleotide polymorphism microarray (CGH+SNP microarray) platform can simultaneously detect copy number alterations (CNA) and copy-neutral loss of heterozygosity (LOH). Eighteen children with acute myeloid leukemia (AML) (n=15) or myelodysplastic syndrome (MDS) (n=3) were studied using CGH+SNP microarray to evaluate the clinical significance of submicroscopic chromosomal aberrations. CGH+SNP microarray revealed CNAs at 14 regions in 9 patients, while metaphase cytogenetic (MC) analysis detected CNAs in 11 regions in 8 patients. Using CGH+SNP microarray, LOHs>10 Mb involving terminal regions or the whole chromosome were detected in 3 of 18 patients (17%). CGH+SNP microarray revealed cryptic LOHs with or without CNAs in 3 of 5 patients with normal karyotypes. CGH+SNP microarray detected additional cryptic CNAs (n=2) and LOHs (n=5) in 6 of 13 patients with abnormal MC. In total, 9 patients demonstrated additional aberrations, including CNAs (n=3) and/or LOHs (n=8). Three of 15 patients with AML and terminal LOH>10 Mb demonstrated a significantly inferior relapse-free survival rate (P=0.041). This study demonstrates that CGH+SNP microarray can simultaneously detect previously cryptic CNAs and LOH, which may demonstrate prognostic implications.

Prenatal Diagnosis of der(X)t(X;Y)(p22.31;q11.22) in a Male Fetus by Using Array Comparative Genomic Hybridization

Xp/Yq translocations are rare chromosomal rearrangements, and the phe-notype of male carriers varies according to the segment of the Xp region that is deleted. In this case report, we describe a der(X)t(X;Y)(p22.31;q11.22) translocation, detected by conventional cytogenetic analysis, in a male fetus at a gestational age of 16 weeks. Chromosomal analysis of parental blood confirmed that this chromosomal aberration had been maternally inherited. Array comparative genomic hybridization (CGH) analysis of fetal blood further indicated a nullisomy of Xp22.31-pter and a breakpoint between the STS and KAL1 genes. The STS, NLGN4, ARSE, CSF2RA, and SHOX genes are present in the region that was deleted, and are known to be related to conditions such as X-linked ichthyosis, chondrodysplasia punctata, mental retardation, and facial dysmorphism in humans. Prenatal ultrasonographic findings and autopsy results were consistent with Xp22.31-pter deletion phenotypes. Genetic counseling was provided for the mother. The observations from this case study indicate that advanced molecular techniques can provide a more precise prenatal diagnosis of chromosomal anomalies than conventional cytogenetics can.

Molecular Cytogenetic Characterization of Supernumerary Marker Chromosomes by Chromosomal Microarray

PURPOSE: Supernumerary marker chromosome (SMC) could be associated with various phenotypic abnormalities based on the chromosomal origin of SMCs. The present study aimed to determine the genomic contents of SMCs using chromosomal microarray and to analyze molecular cytogenetic characterizations and clinical phenotypes in patients with SMCs. MATERIALS AND METHODS: Among patients with SMCs detected in routine chromosomal analysis, SMCs originating from chromosome 15 were excluded from the present study. CGH-based oligonucleotide chromosomal microarray was performed in 4 patients. RESULTS: The chromosomal origins of SMCs were identified in 3 patients. Case 1 had a SMC of 16.1 Mb in 1q21.1-q23.3. Case 2 showed 21 Mb gain in 19p13.11-q13.12. Case 3 had a 4.5 Mb-sized SMC rearranged from 2 regions of 2.5 Mb in 22q11.1-q11.21 and 2.0 Mb in 22q11.22-q11.23. CONCLUSION: Case 1 presented a wide range of phenotypic abnormalities including the phenotype of 1q21.1 duplication syndrome. In case 2, Asperger-like symptoms are apparently related to 19p12-q13.11, hearing problems and strabismus to 19p13.11 and other features to 19q13.12. Compared with cat-eye syndrome type I and 22q11.2 microduplication syndrome, anal atresia in case 3 is likely related to 22q11.1-q11.21 while other features are related to 22q11.22-q11.23. Analyzing SMCs using high-resolution chromosomal microarray can help identify specific gene contents and to offer proper genetic counseling by determining genotype-phenotype correlations.

Identification of a Novel Deletion Region in 3q29 Microdeletion Syndrome by Oligonucleotide Array Comparative Genomic Hybridization / 대한진단검사의학회지

BACKGROUND: The 3q29 microdeletion syndrome is a genomic disorder characterized by mental retardation, developmental delay, microcephaly, and slight facial dysmorphism. In most cases, the microdeletion spans a 1.6-Mb region between low-copy repeats (LCRs). We identified a novel 4.0- Mb deletion using oligonucleotide array comparative genomic hybridization (array CGH) in monozygotic twin sisters. METHODS: G-banded chromosome analysis was performed in the twins and their parents. Highresolution oligonucleotide array CGH was performed using the human whole genome 244K CGH microarray (Agilent Technologies, USA) followed by validation using FISH, and the obtained results were analyzed using the genome database resources. RESULTS: G-banding revealed that the twins had de novo 46,XX,del(3)(q29) karyotype. Array CGH showed a 4.0-Mb interstitial deletion on 3q29, which contained 39 genes and no breakpoints flanked by LCRs. In addition to the typical characteristics of the 3q29 microdeletion syndrome, the twins had attention deficit-hyperactivity disorder, strabismus, congenital heart defect, and gray hair. Besides the p21-activated protein kinase (PAK2) and discs large homolog 1 (DLG1) genes, which are known to play a critical role in mental retardation, the hairy and enhancer of split 1 (HES1) and antigen p97 (melanoma associated; MFI2) genes might be possible candidate genes associated with strabismus, congenital heart defect, and gray hair. CONCLUSIONS: The novel 4.0-Mb 3q29 microdeletion found in the twins suggested the occurrence of genomic rearrangement mediated by mechanisms other than nonallelic homologous recombination. Molecular genetic and functional studies are required to elucidate the contribution of each gene to a specific phenotype.

Development of Trans Attack Anticancer Gene Vaccine Interact in HPV Oncoprotein Expression Inhibition / 대한산부인과학회잡지

OBJECTIVE: Adeno-associated virus Rep 78 protein is known to inhibit the promoter site of several onco-genes and viral gene, including the human papillomavirus type 16 E6 transforming genes. In this study, we investigated AAV Rep 78 mediated inhibition of HPV 16 E6 promotor activity. METHODS: pcDNA3.1/V5/His-Topo vector, cloned by AAV mediated Rep 78, is transfected into cervical cancer cell line (Caski). After that, we confirmed HPV16 derived E6 expression and cell growth inhibition. RESULTS: Transfection rate of Rep78 GFP-vector, approximately from 30 to 60 per-cent, is highly expressed at first day. But E6 expression is lower at this day. The growth of CaSki and HeLa cervical cancer cell lines was inhibited by Rep78 (p<0.05). But, the other cervical cancer cells were unaffected by Rep78 transfection. CONCLUSION: In spite of the high Rep78 transfection efficiency and expression rate, we could not show the cervical cancer cell growth inhibition. In our data, long term expression of Rep78 strategy is needed for cervical carcinoma gene therapy using adeno-associated virus vector.

Retrospective Analysis of Treatment Outcome and Prognostic Factors in Multiple Myeloma / 대한혈액학회지

BACKGROUND: The prognostic outlook for multiple myeloma has markedly improved in recent decades, which is probably related to the introduction of chemotherapy and the development of supportive care for various complications. In the present work we analysed retrospectively the therapeutic outcomes of newly diagnosed patients with multiple myeloma treated at Korea Cancer Center Hospital (KCCH). And we studied to identify prognostic factors influencing the therapeutic outcome of the disease. METHODS: Between January 1987 and December 1998, eighty three patients were diagnosed as multiple myeloma by the criteria of Southwestern Oncology Group at KCCH. Of these patients, clinical analysis was performed retrospectively for sixty-one patients who were treated with combination chemotherapy. RESULTS: The median age at diagnosis was 55 years of age, which was lower than that of western countries and 48% of patients were in their 4th decade. Male to female ratio was 1: 1.1. The main chief complaint at diagnosis was bone pain. Ninety-one percent of the patients were clinical stage III. Serum immuno-electrophoresis revealed M-protein as IgG in 41%. The ratio of micro to lambda light chain was 1.5:1. The response rate to initial chemotherapy was 75% (95% C. I.=63.4~86.6%) and median progression free interval was 20.9 months. The median overall survival of total 61 patients was 28.5 months. The patients' age at diagnosis and the response to initial chemotherapy were statistically significant prognostic factors influencing the overall survival of patients. CONCLUSION: This study showed that we could get relatively high response rate with conventional chemotherapy for the patients with multiple myeloma, but, that most patients eventually progressed and the cure was nearly impossible. To obtain cure or to have much longer survival time, we need more delicate strategies including more intensive chemotherapy with stem cell transplantation for the treatment of multiple myeloma patients.

Implications of Serum Levels of Basic Fibroblast Growth Factor and Vascular Endothelial Growth Factor in Chronic Liver Diseases and Hepatocellular Carcinoma / 대한간학회지

BACKGROUND/AIMS: Angiogenesis occurs in response to tissue damage, and is of vital importance for tumor growth and metastasis. Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) are potent angiogenic factors, and have been suggested to be useful diagnostic markers in certain hypervascular tumors. However, little is known of serum bFGF and VEGF in patients with hepatocellular carcinoma (HCC). We attempted to measure serum bFGF and VEGF in patients with chronic liver diseases (CLD) and HCC to assess their pathogenetic role and usability as tumor markers. METHODS: Serum bFGF and VEGF were measured in 8 patients with chronic hepatitis (CH), 15 patients with liver cirrhosis (LC), and 49 patients with HCC. bFGF was measured in 33, and VEGF was measured in 50, healthy blood donors. RESULTS: Serum bFGF was 3.8+/-1.9, 2.0+/-1.4, 4.2+/-6.0, 17.4+/-30.0 pg/mL in normal control, CH, LC, HCC, respectively. The serum bFGF level was significantly increased in patients with HCC when compared with normal control or patients with CLD. No difference, however, was observed in serum VEGF levels among the four groups. The serum levels of bFGF and VEGF were not significantly different in patients with HCC according to tumor type, size and stage. Serum bFGF showed good sensitivity (90%), specificity (87%), and positive predictive value (94%) in differentiating patients with HCC from those with CLD at the cut-off value of 4.6 pg/mL. CONCLUSIONS: bFGF might play a role in the growth of HCC and its serum level might be used as a tumor marker. On the other hand, serum VEGF does not seem to be an adequate tumor marker.

Clinical Characteristics of Female Hepatocellular Carcinoma / 대한간학회지

BACKGROUND/AIMS: The incidence of hepatocellular carcinoma has been universally lower in female than in male. The aims of our study are to define whether there are any difference between female and male patients with hepatocellular carcinoma in terms of clinical characteristics and results of treatment. METHODS: Retrospective analyses of medical history, physical findings, laboratory results, etiological factors, characteristics of tumor, and therapeutic results were performed in 80 female patients with hepatocellular carcinoma compared to 160 male patients. RESULTS: Asymptomatic presentation and family history of liver disease were found more frequently in female patients than in male patients. A history of smoking and alcohol drinking were found less frequently in female patients than in male patients. The detection rate of spider angioma was significantly lower in female patients than in male patients. There was no difference in laboratory results, characteristics of tumor, and therapeutic results between female and male patients. CONCLUSIONS: Environmental factors such as smoking and alcohol drinking could contribute the sexual difference of hepatocarcinogenesis. However, clinical characteristics at the time of diagnosis and therapeutic results were not significantly different between female and male patients with hepatocellular carcinoma.

Necessity and Safety of Fine-needle Aspiration Cytology for Diagnosis of Hepatocellular Carcinoma / 대한간학회지

BACKGROUNDS/AIMS: The fine-needle aspiration (FNA) is a useful method for diagnosis of hepatocellular carcinoma (HCC). The aims of our study are to assess diagnostic accuracy of FNA, to define proper indications of FNA for diagnosis of HCC, and to evaluate the complications of FNA. SUBJECTS AND METHODS: To assess diagnostic accuracy we compared the results of preoperative FNA with postoperative pathology in 38 resected cases with primary liver cancer. To define proper indications and complications of FNA, we prospectively followed 138 patients received FNA for their liver tumors which were suspicious of primary liver tumor. RESULTS: The sensitivity, specificity, positive and negative predictive values of FNA were 100%, 97%, 100% and 66% respectively. All patients with serum alpha-fetoprotein (AFP) level over 1000 ng/ml were having HCC on FNA result. Among 36 patients with AFP level ranged 15-1000 ng/ml and hypervascular mass on angiography, 96% were having HCC. Among 50 patients with normal AFP level and hypervascular mass on angiography, 92% were having HCC. The major complications after FNA such as hemoperitoneum, pneumothorax, and iatrogenic arterioportal shunt developed in 2%, 2%, and 7% of subjects, respectively. We did not find any case of needle-tract seeding of cancer during a mean 4.7 months of follow-up. CONCLUSIONS: Although the FNA is an accurate method for diagnosis of HCC, FNA was usually not indicated for patients with serum AFP level over 1000 ng/ml or patients with hypervascular mass on angiography when they were suspected of having primary liver cancer. Major complications were hemoperitoneum, pneumothorax and iatrogenic arterioportal shunt. Iatrogenic arterioportal shunt may influence the efficacy of subsequent transcatheter arterial embolization.