Recently, next generation sequencing (NGS) has received attention as the ultimate genotyping method to overcome the limitations of capillary electrophoresis (CE)-based short tandem repeat (STR) analysis, such as the limited number of STR loci that can be measured simultaneously using fluorescent-labeled primers and the maximum size of STR amplicons. In this study, we analyzed 15 autosomal STR markers via the NGS method and evaluated their effectiveness in STR analysis. Using male and female standard DNA as single-sources and their 1:1 mixture, we sequentially generated sample amplicons by the multiplex polymerase chain reaction (PCR) method, constructed DNA libraries by ligation of adapters with a multiplex identifier (MID), and sequenced DNA using the Roche GS Junior Platform. Sequencing data for each sample were analyzed via alignment with pre-built reference sequences. Most STR alleles could be determined by applying a coverage threshold of 20% for the two single-sources and 10% for the 1:1 mixture. The structure of the STR in each allele was accurately determined by examining the sequences of the target STR region. The mixture ratio of the mixed sample was estimated by analyzing the coverage ratios between assigned alleles at each locus and the reference/variant ratios from the observed sequence variations. In conclusion, the experimental method used in this study allowed the successful generation of NGS data. In addition, the NGS data analysis protocol enables accurate STR allele call and repeat structure determination at each locus. Therefore, this approach using the NGS system will be helpful to interpret and analysis the STR profiles from singe-source and even mixed samples in forensic investigation.
Alleles ; DNA ; Electrophoresis, Capillary ; Female ; Gene Library ; Humans ; Ligation ; Male ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction ; Statistics as Topic

BACKGROUND: Silver has been used for disinfection and sterilization. We aimed to confirm the in-vitro antibacterial effects of nanocrystalline silver-coated gauze. METHODS: Fourteen clinical isolates each of Escherichia coli and Acinetobacter baumannii were used. Bacterial suspensions made in tryptic soy broth were exposed to Ordinary and silver-coated gauze. Bacteria were then harvested from the gauze immediately and after 24 h incubation, cultured on blood agar plates and eunmerated for viable counts. The number of colonies was converted into common logarithms for comparison. RESULTS: The number of colonies recovered from silver-coated gauze was significantly lower than those recovered from ordinary gauze when harvested immediately after exposure (E. coli, 3.06 vs 1.73; A. baumannii, 3.13 vs 1.98; P<0.001). After 24 h incubation of exposed gauze, silver-coated gauze produced less than 1 CFU/mL, whereas ordinary gauze produced a number of colonies significantly higher than it did immediately after exposure (E. coli, 4.13; A. baumannii, 4.46; P<0.001). Conclusion: Compared with ordinary gauze, silver-coated gauze was shown to have 99.99% antibacterial effect.
Acinetobacter baumannii ; Agar ; Bacteria ; Disinfection ; Escherichia coli ; Silver ; Sterilization ; Suspensions

No abstract available.
Animals ; Cricetinae* ; Mice*

PURPOSE: To investigate the effect of estrogen on the expression of calcium-activated potassium (KCa) channels in an overactive bladder rat model. To this end, mRNA and protein levels of KCa channel subtypes in the bladder of ovariectomized rats were measured by reverse transcription polymerase chain reaction and western blotting, respectively. METHODS: Ten-week-old female Sprague-Dawley rats were divided randomly into 3 groups: sham-operated control group (n=11), ovariectomy group (n=11), and the group treated with estrogen after ovariectomy (n=12). Rats in the last group were subcutaneously injected with 17β-estradiol (50 µg/kg) every other day for 2 weeks, whereas rats in the other 2 groups received vehicle (soybean oil) alone. Two weeks after treatment, the whole bladder was excised for mRNA and protein measurements. RESULTS: Protein levels of the large-conductance KCa (BK) channels in the ovariectomy group were 1.5 folds higher than those in the sham-operated control group. However, the protein levels of the other KCa channel subtypes did not change significantly upon bilateral ovariectomy. Treatment with 17β-estradiol after ovariectomy restored BK channel protein levels to the control value. In contrast, BK channel mRNA levels were not significantly affected by either ovariectomy alone or 17β-estradiol treatment. The small-conductance KCa type 3 channel (SK3) mRNA and protein levels decreased to 75% of control levels upon 17β-estradiol treatment. CONCLUSIONS: These results suggest that 17β-estradiol may influence urinary bladder function by modulating BK and SK3 channel expression.
Animals ; Blotting, Western ; Estrogens ; Female ; Humans ; Models, Animal ; Ovariectomy ; Polymerase Chain Reaction ; Potassium ; Potassium Channels, Calcium-Activated* ; Rats* ; Rats, Sprague-Dawley ; Reverse Transcription ; RNA, Messenger ; Urinary Bladder* ; Urinary Bladder, Overactive

OBJECTIVES: E-cadherin is a transmembrane protein that is one of the key players involved in cell to cell adhesion. Loss of E-cadherin expression is suggested to promote tumor invasion and distant metastasis in tumor development. Recently, it has been proposed E-cadherin function requires its linkage to the cytoskeleton through catenins. So defects in catenins may cause defective E-cadherin function and promote tumor invasion. We intend to evaluate the expression of E-cadherin and alpha-, beta-, gamma- catenin in tissues of human endometrial carcinoma to analyze the patterns of cell adhesion molecules' expression in endometrial carcinoma and to investigate the relationship between status of cell adhesion molecules and various clinicopathological factors. MATERIALS AND METHODS: The present study investigated the immunohistochemical expression of E-cadherin and alpha-, beta-, gamma- catenin in 33 paraffin embedded formalin fixed tissues of endometrial carcinomas. RESULTS: Aberrant E-cadherin, alpha-, beta-, gamma- catenin expression was observed in 33.3(11 of 33), 27.3(9 of 33), 18.2 (6 of 33), and 51.5(17 of 33) % of the specimens, respectively. Statistically significant correlation was found between aberrant expression of E-cadherin and lymph node metastasis and cell types other than endometrioid adenocarcinoma. Aberrant pattern of gamma- catenin expression also correlated with deep myometrial invasion. But alpha-, beta- catenin expression were not correlated with any clinicopathological parameters. Using Kaplan-Meier curves, abnormal expression of E-cadherin correlated closely with poor survival (p<0.05). CONCLUSION: We revealed aberrant expression of these cell adhesion molecules in part of patients with endometrial carcinoma. Aberrant expression of E-cadherin was correlated with lymph node metastasis and cell types other than endometrioid adenocarcinoma and aberrant expression of gamma-catenin was related with deep myometrial invasion.
Cadherins* ; Carcinoma, Endometrioid ; Catenins* ; Cell Adhesion ; Cell Adhesion Molecules ; Cytoskeleton ; Endometrial Neoplasms* ; Female ; Formaldehyde ; gamma Catenin ; Humans ; Lymph Nodes ; Neoplasm Metastasis ; Paraffin

No abstract available.
Animals ; Brain* ; Mice* ; RNA, Messenger*

No abstract available.
Pulmonary Atelectasis*

Leukemia is a rare disease in the newborn infant. We have presented an autopsy case of congenital acute myelocytic leukemia in a female neonate and discussecd with review of literature. At birth, she was relatively in good health with 4.2 kg in body weight except a large cephalhematoma on left parietal scalp and multiple subcutaneous nodules with ecchymosis on entire body surface. Hemoglobin concentration was 12.0 gm/, Hct 34.6% and erythrocyte count was 2.24 millions. Of 212,400 leukocytes/mm2, 47% were myeloblast. Biopsy of skin nodules reveal leukemia cutis, which disappear dramatically with anticancer drug. The infant was expired 12 days after admission due to intracerebral hemorrhage and acute renal failure.
Infant ; Male ; Female ; Infant, Newborn ; Humans ; Biopsy

Leukemia is a rare disease in the newborn infant. We have presented an autopsy case of congenital acute myelocytic leukemia in a female neonate and discussecd with review of literature. At birth, she was relatively in good health with 4.2 kg in body weight except a large cephalhematoma on left parietal scalp and multiple subcutaneous nodules with ecchymosis on entire body surface. Hemoglobin concentration was 12.0 gm/, Hct 34.6% and erythrocyte count was 2.24 millions. Of 212,400 leukocytes/mm2, 47% were myeloblast. Biopsy of skin nodules reveal leukemia cutis, which disappear dramatically with anticancer drug. The infant was expired 12 days after admission due to intracerebral hemorrhage and acute renal failure.
Infant ; Male ; Female ; Infant, Newborn ; Humans ; Biopsy

Teratocarcinoma is a form of mixed germ cell tumor composed of a mixture of teratoma and embryonal carcinoma. It is rarely found in the brain, but when present it is most commonly found in the pineal region. We investigated a case of primary teratocarcinoma in the pineal region. The patient was a 10 year old boy who had suffered from a bitemporal headache for one month. Brain CT and MRI revealed a rather well defined ovoid heterogenous mass in the pineal gland region, measuring 4.3 x 3.8 x 3.0 cm in size. Microscopically the tumor contained areas of immature teratoma and embryonal carcinoma. Immunohistochemical staining revealed positive reactions for cytokeratin, epithelial membrane antigen, carcinoembryonic antigen, and alpha-fetoprotein in the embryonal carcinoma component.