Assays of clinical diagnosis and species identification using molecular markers are performed according to a quantitative method in consideration of sensitivity, cost, speed, convenience, and specificity. However, typical polymerase chain reaction (PCR) assay is difficult to quantify and have various limitations. In addition, to perform quantitative analysis with the quantitative real-time PCR (qRT-PCR) equipment, a standard curve or normalization using reference genes is essential. Within the last a decade, previous studies have reported that the digital PCR (dPCR) assay, a third-generation PCR, can be applied in various fields by overcoming the shortcomings of typical PCR and qRT-PCR assays. We selected Stilla Naica System (Stilla Technologies), Droplet Digital PCR Technology (Bio-Rad), and Lab on an Array Digital Real-Time PCR analyzer system (OPTOLANE) for comparative analysis among the various droplet digital PCR platforms currently in use commercially. Our previous study discovered a molecular marker that can distinguish Hanwoo species (Korean native cattle) using Hanwoo-specific genomic structural variation. Here, we report the pros and cons of the operation of each dPCR platform from various perspectives using this species identification marker. In conclusion, we hope that this study will help researchers to select suitable dPCR platforms according to their purpose and resources.

The gastrointestinal tract is the first organ directly affected by fasting. However, little is known about how fasting influences the intestinal immune system. Intestinal dendritic cells (DCs) capture antigens, migrate to secondary lymphoid organs, and provoke adaptive immune responses. We evaluated the changes of intestinal DCs in mice with short-term fasting and their effects on protective immunity against Listeria monocytogenes(LM). Fasting induced an increased number of CD103 + CD11b − DCs in both small intestinal lamina propria (SILP) and mesenteric lymph nodes (mLN). The SILP CD103 + CD11b − DCs showed proliferation and migration, coincident with increased levels of GM-CSF and C-C chemokine receptor type 7, respectively. At 24 h post-infection with LM, there was a significant reduction in the bacterial burden in the spleen, liver, and mLN of the short-term-fasted mice compared to those fed ad libitum. Also, short-term-fasted mice showed increased survival after LM infection compared with ad libitum-fed mice. It could be that significantly high TGF-β2 and Aldh1a2 expression in CD103 + CD11b - DCs in mice infected with LM might affect to increase of Foxp3 + regulatory T cells. Changes of major subset of DCs from CD103 + to CD103 - may induce the increase of IFN-γ–producing cells with forming Th1-biased environment.Therefore, the short-term fasting affects protection against LM infection by changing major subset of intestinal DCs from tolerogenic to Th1 immunogenic.

BACKGROUND: Study on epigenetics of urothelial carcinomas has expanded and allowed better understanding of their correlation with clinicopathologic features. The aim of this study was to determine reliable predictive epigenetic markers for patients with urothelial carcinoma of urinary bladder. METHODS: In 64 urothelial carcinomas of the urinary bladder, methylationspecific polymerase chain reaction with RAS association domain family 1A (RASSF1A), adenomatous polyposis coli (APC), death-associated protein-kinase (DAPK), runt-related transcription factor 3 (RUNX3), p14, p16 and MGMT was performed and correlated the results with p53 mutations, DNA ploidy, clinicopathologic parameters and recurrences. RESULTS: Hypermethyation of RASSF1A, APC, DAPK, RUNX3, p14, p16 and MGMT promoters was observed in 35 (54.7%), 29 (45.3%), 18 (28.1%), 18 (28.1%), 9 (14.1%), 2 (3.1%), and 6 (9.4%) cases, respectively. Hypermethylation of RUNX3 and APC was significantly associated with high histologic grades and aneuploidy. Methylation of DAPK was significantly associated with muscle invasion. Methylation of DAPK and RUNX3 genes was significantly associated with recurrence. In survival analyses, methylation of RUNX3 gene and methylation-high (methylation at two or more loci) phenotype was significantly associated with poor recurrence-free survival. CONCLUSIONS: Methylation of RUNX3 gene and methylation-high phenotype are significant indicator of recurrence.
Adenomatous Polyposis Coli ; Aneuploidy ; DNA ; Epigenomics ; Genes, Tumor Suppressor ; Humans ; Methylation ; Muscles ; Phenotype ; Ploidies ; Polymerase Chain Reaction ; Prognosis ; Recurrence ; Transcription Factor 3 ; Urinary Bladder

OBJECTIVES: In addition to the main symptoms of posttraumatic stress disorder (PTSD), which include reexperience, avoidance and hyperarousal, many patients complain of cognitive deficits and especially in attention and memory. This study was conducted to evaluate the cognitive deficits of survivors of a tragic shooting incident at a frontline guard post. All of the survivors experienced the same accident, and they were homogeneous in terms of age and education level, which are closely associated with cognitive performance. METHODS: We recruited 12 survivors who suffered from PTSD following the same traumatic incident and we also recruited 12 normal volunteers, and we assessed their neurocognitive functions with using a vigilance test, a continuous attention test, a reaction unit test and the Corsi block tapping test in the computerized Vienna Test System, as well as an auditory verbal learning test and complex figure test in the Rey-Kim Memory Test. Standardized clinical scales, including the Clinician-Administered PTSD Scale, the Hamilton Depression Rating Scale, the Hamilton Anxiety Scale and the State-Trait Anxiety Scale I and II, were used to assess the involved mental areas and the severity of the PTSD symptoms. RESULTS: The patient group showed significant impairments in continuous attention, and the visual and auditory information processing time on the Vienna test when compared to the control group. The patient group also showed significant impairments in the verbal memory, visual memory and visuospatial function on the Rey-Kim Memory Test. CONCLUSION: In this study, the author reports on the cognitive impairments in patients with PTSD as measured by computerized neurocognitive tests and memory tests. Future studies are needed to determine the changes in cognitive functioning that are related to symptom improvement, as well as the influence of the therapeutic effects on the cognitive improvement.
Anxiety ; Automatic Data Processing ; Cognition ; Depression ; Humans ; Memory ; Neuropsychological Tests ; Psychomotor Performance ; Stress Disorders, Post-Traumatic ; Survivors ; Verbal Learning ; Weights and Measures

PURPOSE: This study was designed to compare the CT findings and clinical manifestations in children and adolescents with acute sialadenitis according to the involved salivary glands. MATERIALS AND METHODS: The study included fifty children and adolescents (34 boys, 16 girls) with acute sialadenitis that was diagnosed during the past five years. All of the subjects were divided into three groups: group I (parotid gland involvement, n = 16), group II (submandibular gland involvement, n = 20) and group III (involvement of both glands, n = 14). We analyzed the presence of an abscess, sialolith, bilaterality, cellulitis and lymphadenopathy on CT scans. The analyzed clinical data were age, sex, lymphadenopathy, pain, swelling, presence of a mass, tonsillitis, treatment period and surgical treatment if it was performed. RESULTS: The presence of an abscess, sialolith, cellulitis, swelling, age, presence of a palpable mass and treatment period were statistically significant factors for the patients in the three groups. An abscess was combined only in group I patients. There was a high rate of sialolith in group II patients and cellulitis in group III patients as seen on CT scans. Swelling in group II patients and group III patients and the presence of a palpable mass in group I patients were identified as clinical manifestations. Age was younger in group I patients (mean age, 5.3 years) than in group II patients (mean age, 12.9 years) and group III patients (mean age, 15.2 years). The treatment period was longer for group I patients. CONCLUSION: For acute sialadenitis in children and adolescents, age, presence of an abscess, sialolith, cellulitis, swelling, presence of a palpable mass and treatment period were different according to the involved salivary glands.
Abscess ; Adolescent ; Cellulitis ; Child ; Humans ; Lymphatic Diseases ; Palatine Tonsil ; Salivary Gland Calculi ; Salivary Glands ; Sialadenitis ; Tonsillitis

Subperiosteal hematoma of the orbit is a rare disease and most of the cases occur in young adult males as a result of direct facial or orbital trauma. In the absence of direct facial or orbital trauma, nontranmatic subperiosteal orbital hematoma has rarely been reported in association with a sudden elevation of cranial venous pressure or venous congestion, systemic diseases associated with a bleeding diathesis and paranasal sinusitis. We report here on a rare case of subperiosteal orbital hematoma associated with ethmoid sinusitis, as was seen on CT imaging.
Disease Susceptibility ; Ethmoid Sinus* ; Ethmoid Sinusitis* ; Hematoma* ; Hemorrhage ; Humans ; Hyperemia ; Male ; Orbit* ; Rare Diseases ; Sinusitis ; Venous Pressure ; Young Adult

OBJECTIVE: Cervical cancer has long been linked to the sexually transmitted human papillomavirus (HPV), and the oncoproteins E6 and E7 disrupt the functions of tumour suppressor genes, resulting in genetic alteration. It was shown that loss of heterozygosity at 6p is a common genetic alteration in cervical cancer. However, the molecular genetics of cancer have only recently been understood, and for the development of cervical cancer additional genetic alterations in host cell genes are required. The present study has identified the differential changes of the cervical cancer-associated genetic alterations by a genome-wide array based comparative genomic hybridization (array-CGH). METHODS: We analyzed 15 cases of cervical cancer from St. Mary's hospital of The paraffin-fixed tissue samples were microdissected under microscope and DNA was extracted by the procedures of proteinase K digestion and chloroform extraction. Array-based CGH and genomic PCR were carried out with statistical analyses such as hierarchical clustering and Gene Ontology. The BAC array used in this study consisted of 1,440 human BACs, the space among the clones were approximately 2.08 megabase (Macrogen, Seoul, Korea). RESULTS: All of 15 cases of cervical cancer showed specific gains and losses. The analysis limit of average gains and losses was 53%. A significant positive correlation was found between 1p36.32, 3p14.2, 3q27.1, 7p21.1, 8q24.3 and 11q13.1 changes through the cervical carcinogenesis. The high-level of gain regions, BAC clones encoded GSDMDC1, RECQL4, TP73, ABCF3, ALG3, HDAC9, ESRRA and RPS6KA4 genes. Frequently gained BAC clones encoded genes were PRSS8, FUS, COL18A1, PCOLN3, MAFG and ASPSCR1. The genes encoded by frequently lost BAC clones were PTPRG, GRM7, ZDHHC3, EXOSC7, LRP1B and NR3C2. Also, hierarchical clustering of the expression data readily distinguished genomic alterations in cervical cancer. A subset of cellular processes from each gene was clustered by Gene Ontology database. CONCLUSION: Using Array-CGH, genomic alterations related to cervical cancer were identified to determine whether induction of chromosomal imbalances occurs prior to carcinogenesis. The high resolution of array-CGH combined with human genome database would give a chance to find out possible target genes present in the gained or lost clones.
Carcinogenesis ; Chloroform ; Clone Cells ; Comparative Genomic Hybridization ; Digestion ; DNA ; Endopeptidase K ; Gene Ontology ; Genes, Suppressor ; Genome, Human ; Humans ; Loss of Heterozygosity ; Molecular Biology ; Oncogene Proteins ; Polymerase Chain Reaction ; Seoul ; Uterine Cervical Neoplasms*

OBJECTIVES: The Korean College of Neuropsychopharmacology and the Korean Academy of Schizophrenia developed the Korean algorithm project for schizophrenia to aid clinical decisions. The purpose of this study was to assess the feasibility of Korean Medication Algorithm for Schizophrenia patients in clinical settings in Korea. METHODS: A total of 108 schizophrenia and schizophreniform disorder patients were enrolled at 19 centers and treated according to the algorithm. PANSS (Positive and Negative Symptom Scale) and CGI (Clinical Global Impression) were used to evaluate symptom severity. Also UKU (UKU side effect rating scale) and LUNSERS (Liverpool University Neuroleptic Side Effect Rating Scale), DAI-10 (Drug Attitude Inventory-10), PPS (Patient Preference Scale), SWN (Subjective Well-Being under Neuroleptic treatment) and WHOQOL (World Health Organization Quality of Life) were used to evaluate tolerability and satisfaction of patient respectively. RESULTS: Overall ratings including symptom severity, compliance of medication, side effect of medication, quality of life were favorable. The treatment response (PANSS improvement > or = 20%) rate was 63%, 75% at the first Clinical decision point (CDP) and 4 month respectively. CONCLUSION: Symptom improvement, tolerability and quality of life were all favorable. These results suggest that this algorithm can be useful in clinical practices.
Compliance ; Humans ; Korea ; Psychotic Disorders ; Quality of Life ; Schizophrenia ; World Health Organization

PURPOSE: Screening in cervical cancer is now progressing to discover candidate genes and proteins that may serve as biological markers and that play a role in tumor progression. We examined the protein expression patterns of the squamous cell carcinoma (SCC) tissues from Korean women with using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer. MATERIALS AND METHODS: Normal cervix and SCC tissues were solubilized and 2-DE was performed using pH 3~10 linear IPG strips of 17 cm length. The protein expression was evaluated using PDQuest 2-D software(TM). The differentially expressed protein spots were identified with a MALDI-TOF mass spectrometer, and the peptide mass spectra identifications were performed using the Mascot program and by searching the Swiss-prot or NCBInr databases. RESULTS: A total of 35 proteins were detected in SCC. 17 proteins were up-regulated and 18 proteins weredown-regulated. Among the proteins that were identified, 12 proteins (pigment epithelium derived factor, annexin A2 and A5, keratin 19 and 20, heat shock protein 27, smooth muscle protein 22 alpha, alpha-enolase, squamous cell carcinoma antigen 1 and 2, glutathione S-transferase and apolipoprotein a1) were protein previously known to be involved in tumor, and 21 proteins were newly identified in this study. CONCLUSION: 2-DE offers the total protein expression profiles of SCC tissues; further characterization of these differentially expressed proteins will give a chance to identify the badly needed tumor-specific diagnostic markers for SCC.
Annexin A2 ; Apolipoproteins ; Biomarkers ; Carcinoma, Squamous Cell ; Cervix Uteri ; Databases, Protein ; Electrophoresis, Polyacrylamide Gel ; Epithelium ; Female ; Glutathione Transferase ; HSP27 Heat-Shock Proteins ; Humans ; Hydrogen-Ion Concentration ; Keratin-19 ; Mass Screening ; Muscle, Smooth ; Phosphopyruvate Hydratase ; Uterine Cervical Neoplasms*

PURPOSE: The tumor suppressor gene, p53, has been established as an essential component for the suppression of tumor cell growth. In this study, we investigated the time-course anticancer effects of adenoviral p53 (Adp53) infection on human ovarian cancer cells to provide insight into the molecular-level understanding of the growth suppression mechanisms involved in Adp53-mediated apoptosis and cell cycle arrest. MATERIALS AND METHODS: Three human cervical cancer cell lines (SiHa, CaSki, HeLa and HT3) were used. The effect of Adp53 infection was studied via cell count assay, cell cycle analysis, FACS, Western blot and macroarray assay. RESULTS: Adp53 exerts a significant role in suppressing cervical cancer cell growth. Adp53 also showed growth inhibitory effects in each cell line, and it induced apoptosis and cell cycle arrest. Adp53 differentially regulated the expression of genes and proteins, and the gene expression profiles in the SiHa cells revealed that the p21, p53 and mdm2 expressions were significantly up-regulated at 24 and 48 hr. Western blot shows that the p21 and p53 expressionlevels were significantly increased after Adp53 infection. In addition, in all cell lines, both the CDK4 and PCNA protein expression levels were decreased 48 h after Adp53 infection. Cell cycle arrest at the G1 phase was induced only in the SiHa and HeLa cells, suggesting that exogenous infection of Adp53 in cancer cells was significantly different from the other HPV-associated cervical cancer cells. CONCLUSION: Adp53 can inhibit cervical cancer cell growth through induction of apoptosis and cell cycle arrest, as well as through the regulation of the cell cycle-related proteins. The Adp53-mediated apoptosis can be employed as an advanced strategy for developing preferential tumor cell-specific delivery.
Adenoviridae* ; Apoptosis ; Blotting, Western ; Cell Count ; Cell Cycle Checkpoints ; Cell Cycle* ; Cell Line ; G1 Phase ; Genes, Tumor Suppressor ; Genetic Therapy ; HeLa Cells ; Humans* ; Ovarian Neoplasms ; Papilloma* ; Proliferating Cell Nuclear Antigen ; Transcriptome ; Uterine Cervical Neoplasms*