Clinical, Biochemical, and Genetic Characterization of Glycogen Storage Type IX in a Child with Asymptomatic Hepatomegaly / 대한소아소화기영양학회지

Glycogen storage disease type IX (GSD IX) is caused by a defect in phosphorylase b kinase (PhK) that results from mutations in the PHKA2, PHKB, and PHKG2 genes. Patients usually manifest recurrent ketotic hypoglycemia with growth delay, but some may present simple hepatomegaly. Although GSD IX is one of the most common causes of GSDs, its biochemical and genetic diagnosis has been problematic due to its rarity, phenotypic overlap with other types of GSDs, and genetic heterogeneities. In our report, a 22-month-old boy with GSD IX is described. No other manifestations were evident except for hepatomegaly. His growth and development also have been proceeding normally. Diagnosed was made by histologic examination, an enzyme assay, and genetic testing with known c.3210_3212del (p.Arg1070del) mutation in PHKA2 gene.

Rapid genotyping of Plasmodium vivax Pvs25 and Pv38 genes by using mismatch specific endonuclease

Mismatch specific endonuclease (MSE) method was used to detect natural polymorphisms in Pvs25 and Pv38 genes of Plasmodium vivax. Eighty seven patients with P. vivax were recruited in the Republic of Korea (ROK). Pvs25 and Pv38 genes were amplified by polymerase chain reaction (PCR), and the PCR amplicons were mixed with reference DNA sequences. Following the denaturation and gradual annealing, the product mixtures were cleaved by the MSE. Heteroduplex types were readily detected by gel electrophoresis, where extra bands with shorter sizes would appear from the cleavage. After MSE cleavage of 657- bp product from Pvs25 mixtures, three genotypes were detected, while Pv38 mixtures with 1220-bp products presented two genotypes in ROK isolates. After the MSE cleavage, the mismatched samples of Pvs25 and Pv38 were completely sequenced, and the results were in complete agreement with the MSE analyses. In conclusion, genotyping of Pvs25 and Pv38 with MSE cleavage could be a potential method for the high-throughput screening of the large field samples.

Laparoscopic surgery for pelvic floor disorder / 한양의대학술지

Pelvic organ prolapse(POP) is a major health care problem. Up to 50% of parous women have some degree of pelvic organ prolapse although only 10?20% are symptomatic. The first line of treatment is surgical repair. Many surgical procedures have been described to correct pelvic organ prolapse. The majority of these procedures are performed either vaginally or abdominally, or with a combined abdominovaginal approach. With recent advancements of laparoscopic instruments and surgical techniques, interest in laparoscopic treatment of pelvic organ prolepse has surged. Beyond the well-known advantages of laparoscopy - less postoperative discomfort, shorter hospital stay, the laparoscopic approach offers the superior visualization of the pelvis, thereby allowing better exposure of the pelvic floor anatomy and more exact identification of the defect. Moreover, laparoscopic surgery provides a magnified view of the operative field and enables to perform an easier and more precise dissection. This enhances the identification of pelvic floor defects and allows more precise suture placement and improved correction of specific site defects. Laparoscopic pelvic floor repair is an effective procedure and enables to combine the advantages of laparotomy with the low morbidity of the vaginal route. This article reviews pelvic support anatomy and various laparoscopic surgical techniques currently available for reconstructive pelvic surgery.

Gene expression profiling of oxidative stress on atrial fibrillation in humans

Atrial Fibrillation (AF) is thought be caused by oxidative stress. Oxidative stress at the cellular level results from many factors, including exposure to alcohol, medications, cold, toxins or radiation. In this study we investigated gene transcriptional profiles on the human myocardial tissues from AF and oxidative stress conditions. Right atrial appendages were obtained from AF patients (n = 26) undergoing the Maze procedure, and from control patients (n = 26) who were in normal sinus rhythm and undergoing coronary artery bypass graft operation. To examine the effects of oxidative stress on AF, we used radioactive complementary DNA (cDNA) microarrays to evaluate changes in the expression of 1,152 known genes. This technology, which monitors thousands of genes simultaneously, gives us a better picture of the interactions between AF and oxidative stress. Total RNAs prepared from the retrieved tissues were used to synthesize(33)P-labeled cDNAs by reverse transcription and hybridized to cDNA microarrays. Gene expression profiles showed that 30 genes were upregulated and 25 were downregulated in AF patients compared with control patients. Moreover, comparison rank analysis revealed that the expression of five genes related to reactive oxygen species (ROS)-including flavin containing monooxygenase 1, monoamine oxidase B, ubiquitin specific protease 8, tyrosinase-related protein 1, and tyrosine 3-monooxygenase-increased by more than 2.0 of the Z-ratio, and two genes related to anti-oxidants including glutathione peroxidase 1, and heme oxygenase 2-decreased to the Z-ratio levels of <= -2.0. Apparently, a balanced regulation of pro- and anti-oxidation can be shifted toward pro-oxidation and can result in serious damage similar to that of human AF. Western blotting analysis confirmed the upregulation of tyrosinase-related protein 1 and tyrosine 3-monooxygenase and the downregulation of heme oxygenase 2. These results suggested that the gene expression pattern of myocardial tissues in AF patients can be associated with oxidative stress, resulting in a significant increase in ROS. Thus, the cDNA microarray technique was useful for investigating transcription profiles in AF. It showed that the intracellular mechanism of oxidative stress plays a pivotal role in the pathologic progression of AF and offers novel insight into potential treatment with antioxidants.

A retrospective comparison of four different procedures for extracting dermoid cyst by laparoscopy / 대한산부인과학회잡지

OBJECTIVES: To compare results of 4 different extraction methods in laparoscopic management of dermoid cyst. STUDY DESIGN: This article is a retrospective, multicenter study for 247 patients with benign dermoid cyst in period of 1995-1998. Dermoid cyst was extracted by Endopouch (99 cases), puncture-irrigation-extraction (69 cases), colpotomy (35 cases), and dermoid cyst as a "pouch bag" (44 cases). RESULTS: We analyzed irrigation amount, operative time, postoperative hospital stay and complications by four different extraction methods. Endopouch extraction method needed less amount of irrigation fluid for cleaning the abdominal cavity and had a shorter postoperative hospital stay (ANOVA, p=0.0001). There were no significant differences in operative times among groups. There were four cases of morbidity in puncture-irrigation-extraction method (6%), three had fever (> 38degrees C) and one intraabdominal abscess. One incisional hernia was noted in "pouch bag" method (2%). CONCLUSIONS: We recommend minimal spillage method for extraction of dermoid and careful irrigation of abdominal cavity to prevent potential risk of chemical peritonitis such as Endopouch, "pouch bag", and colpotomy with the exception of puncture-irrigation-extraction method.

Expression and biological activity of two recombinant polypeptides related to subunit 1 of the interferon-alpha receptor

Abnormal production of interferon alpha (IFN-a) has been found in certain autoimmune diseases and can be also observed after prolonged therapy with IFN-a. IFN-a can contribute to the pathogenesis of allograft rejection in bone marrow transplants. Therefore, the development of IFN-a inhibitors as a soluble receptor protein may be valuable for the therapeutic control of these diseases. We have expressed two polypeptides encoding amino acids 93-260 (P1) and 261-410 (P2) of the extracellular domain of subunit 1 of the interferon-a receptor (IFNAR 1-EC) in E. coli. The activities of the recombinant polypeptides and of their respective antibodies were evaluated using antiproliferative and antiviral assays. Expression of P1 and P2 polypeptides was achieved by transformation of cloned plasmid pRSET A into E. coli BL21(DE3)pLysS and by IPTG induction. P1 and P2 were purified by serial sonication steps and by gel filtration chromatography with 8 M urea and refolded by dialysis. Under reducing SDS-PAGE conditions, the molecular weight of P1 and P2 was 22 and 17 kDa, respectively. Polyclonal anti-P1 and anti-P2 antibodies were produced in mice. P1 and P2 and their respective polyclonal antibodies were able to block the antiproliferative activity of 6.25 nM IFN-aB on Daudi cells, but did not block IFN-aB activity at higher concentrations (>6.25 nM). On the other hand, the polypeptides and their respective antibodies did not inhibit the antiviral activity of IFN-aB on Hep 2/c cells challenged with encephalomyocarditis virus.