Evaluation of Multiplex Polymerase Chain Reaction Assay for the Simultaneous Detection of Sexually Transmitted Infections Using Swab Specimen

Sexually transmitted infections (STIs) are caused by the spread of pathogens via sexual activity and can cause serious complications if left untreated, regardless of their symptoms. Therefore, early diagnosis of STI is important, and molecular diagnostic methods for rapid detection and monitoring are needed. In this study, we evaluated a multiplex polymerase chain reaction (PCR) kit for simultaneously detecting 13 different bacterial, fungal, and viral microorganisms that cause STIs. The kit performance was evaluated for its sensitivity, lot-to-lot variation, and interference in detecting different pathogens. Additionally, its clinical usefulness was evaluated by estimating its sensitivity and specificity for clinical samples. The limit of detection (LOD) was 0.021–50.104 copies for each pathogen. In the tests of lot-to-lot, 100% of positive samples were detected at low concentrations and negative samples all showed negative results. This result confirms that there is no the variation of lot-to-lot. In the test for interference between pathogens, the efficiency of amplification for each pathogen was not significantly reduced and no nonspecific amplification product was formed. We tested 322 vaginal swab samples using the multiplex PCR kit and confirmed that its clinical sensitivity and specificity were 100% for all pathogens. This multiplex PCR kit can be used widely for rapid diagnosis and monitoring of STIs.

Diagnosis of Viral Infection Using Real-time Polymerase Chain Reaction

The laboratory-based diagnosis of viral infection has been evolving over the years, to increase objectivity, accuracy, and sensitivity via the continuous development of various technologies. Cell culture, which is one of the methods used for the diagnosis of viral infection, is a “gold-standard” approach; however, it is time consuming and is associated with a high risk of contamination. To overcome these shortcomings, molecular biology methods, such as conventional polymerase chain reaction (cPCR), real-time PCR, and sequencing, have been used recently for virus diagnosis. Realtime PCR has higher accuracy and sensitivity compared with cPCR. Moreover, realtime PCR can quantify viral nucleic acids by confirming the amplification using the threshold cycle, which is the initial amplification point. Real-time PCR applications for the detection of various types of viruses in clinical settings should be based on the use of appropriate samples, nucleic acid extraction according to virus characteristics, and selection of diagnostic methods using sensitivity and specificity targets. In addition, the implementation of real-time PCR requires to evaluate the performance of the test protocol by measuring sensitivity, specificity, accuracy, and reproducibility. The verified real-time PCR method is an easy, fast, and accurate method for monitoring the diagnosis and treatment outcomes in a clinical setting. In this review, we summarize the characteristics of the typical diagnostic methods for viral infection, especially of the advanced real-time PCR method, to detect human pathogenic viruses.

Evaluation of EZplex MTBC/NTM Real-Time PCR kit: diagnostic accuracy and efficacy in vaccination

PURPOSE: Tuberculosis (TB) is mainly caused by Mycobacterium tuberculosis, which is a pathogenic mycobacterial species grouped under Mycobacterium tuberculosis complex (MTBC) with four other pathogenic mycobacterial species. The mycobacteria not included in MTBC are known as nontuberculous mycobacteria (NTM), and cause several pulmonary diseases including pneumonia. Currently, NTM occurrences in TB-suspected respiratory specimens have increased, due to which, precise detection of MTBC and NTM is considered critical for the diagnosis and vaccination of TB. Among the various methods available, real-time PCR is frequently adopted for MTBC/NTM detection due to its rapidness, accuracy, and ease of handling. In this study, we evaluated a new real-time PCR kit for analytical and clinical performance on sputum, bronchial washing, and culture specimens. MATERIALS AND METHODS: For assessing its analytical performance, limit of detection (LOD), reactivity, and repeatability test were performed using DNA samples. To evaluate clinical performance, 612 samples were collected and clinically tested at a tertiary hospital. RESULTS: LOD was confirmed as 0.584 copies/µL for MTBC and 47.836 copies/µL for NTM by probit analysis (95% positive). For the reactivity test, all intended strains were detected and, in the repeatability test, stable and steady results were confirmed with coefficient of variation ranging from 0.36 to 1.59. For the clinical test, sensitivity and specificity were 98.6%–100% and 98.8%–100% for MTBC and NTM, respectively. CONCLUSION: The results proved the usefulness of the kit in TB diagnosis. Furthermore, it could be adopted for the assessment of vaccine efficacy.

Impact of Glycemic Control and Metformin Use on the Recurrence and Progression of Non-Muscle Invasive Bladder Cancer in Patients with Diabetes Mellitus

The purpose of the present study was to determine the potential relationships of glycemic control and use of metformin with non-muscle invasive bladder cancer characteristics. We reviewed data from 645 patients with non-muscle invasive bladder cancer between January 2004 and May 2015. We analyzed the association of pre and post-operative glycemic control and use of metformin with clinical characteristics of bladder tumors. We also analyzed the association of glycemic control and use of metformin with recurrence-free and progression-free survivals. Diabetes mellitus patients showed decreased recurrence-free survival (hazard ratio 1.42; 95% confidence interval 1.1-1.9; P = 0.021) and progression-free survival (hazard ratio 1.79; 95% confidence interval 1.1-2.8; P = 0.013). Diabetes mellitus patients with a HbA1c ≥ 7.0% demonstrated a higher rate of progression (P = 0.026). Kaplan-Meier analysis showed that progression-free survival rate was associated with poor baseline glycemic control (P = 0.026) and post-operative glycemic control (P = 0.025). However, use of metformin had no impact on the recurrence (P = 1.00) and progression (P = 0.282). In conclusion, poor baseline and post-operative glycemic control was related with shorter progression-free survival of patients with non-muscle invasive bladder cancer. Use of metformin had no impact on the recurrence and progression. Therefore, tight glycemic control and close follow-up for bladder tumor may be beneficial in patients with poor glycemic control.

Stigma of Mental Illnesses as Perceived by North Korean Defectors Living in South Korea

OBJECTIVE: This study aims to provide the information of the stigmas of mental illness such as psychosis, alcoholism, attempt suicide, and depression among North Korean defectors. METHODS: We examined stigma for the mental illnesses of 639 North Korean defectors aged 19 to 65 years who live in the Settlement Support Center for North Korean Refugees. The stigmas of mental illnesses were assessed using the Perceived Devaluation-Discrimination Scale We directly compared the stigma level between North Korean defectors and the general population of South Korea. RESULTS: North Korean defectors had higher perceived stigmas of psychosis and alcoholism and lower perceived stigmas of depression than South Koreans. Perceived stigma associated with attempted suicide was similar for North Korean defectors and South Koreans. Only marital status in sociodemographic variables had associations with higher perceived stigma of psychosis, alcoholism, and depression in the North Korean defectors. North Korean defectors, who spent more than one year in transit country, had associations with lower perceived stigma of psychosis and alcoholism. North Korean defectors, who had the experience of compulsory repatriation to North Korea or North Korean family in South Korea, had an association with higher perceived stigma of depression. CONCLUSION: North Korean defectors had higher perceived stigmas of psychosis and alcoholism and lower perceived stigmas of depression than South Koreans. Further studies are needed to document serial changes in stigmas for mental illnesses associated with the receipt of education at the Settlement Support Center for North Korean defectors.

Effect of Multi-Walled Carbon Nanotubes on MUC5AC and MUC5B Expression in Airway Epithelial Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Multi-walled carbon nanotubes (MWCNT) are one of the most commonly used nanomaterials to date. Recent studies have demonstrated that MWCNT increase immune response and allergic inflammation in airway epithelial cells. However, the effects of MWCNT on mucin in human airway epithelial cells have not been reported. Therefore, in the present study, the effect of MWCNT on MUC16, MUC5AC, and MUC5B expressions were investigated in human airway epithelial cells. SUBJECTS AND METHOD: In mucin-producing human NCI-H292 airway epithelial cells and primary cultures of normal nasal epithelial cells, the effects of MWCNT on MUC16, MUC5AC, and MUC5B expression were analyzed by reverse transcription polymerase chain reaction, real-time polymerase chain reaction, and enzyme-linked immunosorbent assay. RESULTS: In human NCI-H292 airway epithelial cells, MWCNT significantly induced the expression MUC5AC and MUC5B mRNA and the production of MUC5AC and MUC5B protein. However, MWCNT did not induce the expression of MUC16 mRNA. In the primary cultures of normal nasal epithelial cells, MWCNT also induced the expression of MUC5AC and MUC5B mRNA and the production of MUC5AC and MUC5B proteins. CONCLUSION: The results of this study demonstrate that MWCNT induces MUC5AC and MUC5B expression in human airway epithelial cells. These findings provide important information about the biological role of MWCNT on mucus-secretion in human airway epithelial cells.

Acute esophageal necrosis occurring in a patient undergoing percutaneous coronary intervention

Acute esophageal necrosis is uncommon in the literature. Its etiology is unknown, although cardiovascular disease, hemodynamic compromise, gastric outlet obstruction, alcohol ingestion, hypoxemia, hypercoagulable state, infection, and trauma have all been suggested as possible causes. A 67-year-old female underwent a coronary angiography (CAG) for evaluation of chest pain. CAG findings showed coronary three-vessel disease. We planned percutaneous coronary intervention (PCI). Coronary arterial dissection during the PCI led to sudden hypotension. Six hours after the index procedure, the patient experienced a large amount of hematemesis. Emergency gastrofibroscopy was performed and showed mucosal necrosis with a huge adherent blood clot in the esophagus. After conservative treatment for 3 months, the esophageal lesion was completely improved. She was diagnosed with acute esophageal necrosis. We report herein a case of acute esophageal necrosis occurring in a patient undergoing percutaneous coronary intervention.

Effect of Anthocyanidin on MUC5AC and MUC5B Expression in Airway Epithelial Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Naringenin and delphinidin are types of anthocyanidin, which are flavonoids and thus have anti-inflammatory property. Moderate consumption of natural dietary naringenin and delphinidin is believed to do anti-inflammatory action, but the action mechanism is unclear. Therefore, this study aimed to investigate the effects of naringenin and delphinidin on interleukin-1beta (IL-1beta)- and lipopolysaccharide (LPS)-induced MUC5AC and MUC5B expressions in airway epithelial cells. MATERIALS AND METHOD: In NCI-H292 cells and cultured nasal polyp epithelial cells, the effects of naringenin and delphinidin on IL-1beta- and LPS-induced MUC5AC and MUC5B expressions were analyzed by real-time polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: Delphinidin attenuated IL-1beta- and LPS-induced MUC5AC and MUC5B mRNA and glycoprotein expression in a dose-dependent pattern in NCI-H292 cells and in cultured nasal polyp epithelial cells. Naringenin partially attenuated IL-1beta- and LPS-induced MUC5AC and MUC5B mRNA and glycoprotein expression at a high dose. CONCLUSION: These results suggest that delphinidin attenuates MUC5AC and MUC5B expressions in the airway epithelial cells. Between anthocyanidin and delphinidin, delphinidin exhibits greater potential as an ideal therapeutic agent for the control of mucus-hypersecretion in the treatment of airway inflammatory diseases.

Effect of Udenafil on MUC5B Expression in Human Airway Epithelial Cells / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Mucus hypersecretion in the airway may lead to increased frequency and duration of infection, declined lung function, and increased morbidity and mortality in inflammatory respiratory diseases. Udenafil, a phosphodiesterase (PDE) 5 inhibitor, is an oral medication for erectile dysfunction. Recent studies show that PDE5 inhibitor has various anti-inflammatory properties. However, the effect of udenafil on mucus secretion in human airway epithelial cells is unclear. Therefore, the effect and brief signaling pathway of udenafil on MUC5B expression were investigated in human airway epithelial cells. MATERIALS AND METHOD: We analyzed the effects and brief signaling pathway of udenafil on the lipopolysaccharide (LPS) induced MUC5B expression in mucin-producing NCI-H292 epithelial cells using reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay, and immunoblot analysis. RESULTS: Udenafil attenuated the LPS-induced MUC5B mRNA expressions and glycoprotein production in NCI-H292 epithelial cells. It also attenuated LPS-induced toll like receptor 4 (TLR4) mRNA expression and phosphorylation of extracellular regulated kinase1/2 (ERK1/2) and p38 in NCI-H292 epithelial cells. CONCLUSION: These results suggest that udenafil attenuates the LPS induced MUC5B expression via TLR4, ERK1/2 and p38 mitogen activated protein kinase (MAPK) pathway in human airway epithelial cells, and that it could be a novel therapeutic agent for controlling chronic airway disease.

Comparison of the ambulatory blood pressure with the clinical blood pressure and electrocardiographic left ventricular hypertrophy / 대한내과학회지

BACKGROUND: This study compared the results of 24 hour ambulatory blood pressure monitoring with the clinical blood pressure measurements, and we investigated the relationship of the blood pressure measurement and left ventricular hypertrophy, as determined by routine 12 lead electrocardiography. METHODS: We studied 204 healthy adults with no prior history of heart disease or antihypertensive medication. The clinic blood pressure was measured 3 times and the average was taken. We compared the clinic blood pressure with the daytime blood pressure of the 24 hour ambulatory blood pressure monitoring, and we compared the blood pressure with the sum of the voltage of the S wave on V1 and the R wave on V5. RESULTS: The average of the daytime ambulatory blood pressure of all the patients was 135.33+/-13.73 mmHg for the systolic pressure and 86.55+/-10.14 mmHg for the diastolic pressure. The average of the clinic blood pressure measurement was 140.10+/-17.41 mmHg for the systolic pressure and 88.84+/-10.14 mmHg for the diastolic pressure. The clinic blood pressure averaged higher than the daytime ambulatory blood pressure by 5 mmHg on the systolic pressure and 2 mmHg on the diastolic pressure (p<0.001). The normal ambulatory blood pressure limits were estimated as those that best correlated with 140/90 mmHg at the clinic. The estimated value was 135/87 mmHg for the daytime ambulatory blood pressure (p+/-0.001). The incidence of white coat hypertension was 10.8%. The sum of the voltage on electrocardiography showed a positive linear relationship with all the blood pressure measurements. The daytime systolic blood pressure showed the strongest correlation with the 24 hour ambulatory blood pressure monitoring (r=0.283, p+/-0.001). CONCLUSIONS: We found a linear relation and we analyzed the differences between the clinical and 24 hour ambulatory blood pressure. A daytime ambulatory blood pressure value of 135/87 mmHg was a suitable upper normal limit for the corresponding cutoff value of the clinic blood pressure. Left ventricular hypertrophy showed the strongest relationship with the daytime systolic blood pressure among the results of the 24 hour ambulatory blood pressure monitoring.

The change of serum level of total antioxidant status and cytokine, and in-stent restenosis after supplementation of antioxidant / 대한내과학회지

BACKGROUND: Oxidative stress might be a role in atherosclerosis and increased intake of antioxidant appear to be protective and modify neointimal formation. An antioxidant and probucol prevents endothelial dysfunction and low density lipoprotein oxidation and also inhibits the secretion of cytokine by macrophages. We aimed 1) to study the effects of antioxidant (Vitamin C, E and probucol) supplementation on serum level of antioxidant status (TAS), P-selectin, MCP-1, IL-6 and IL-10 and 2) to investigate the effects of antioxidant intake on in-stent restenosis. METHODS: Total 90 patients were assigned to control or antioxidant group (probucol; 500 mg, vitamin C; 1,000 mg, vitamin E; 400 mg). We performed follow up coronary angiography in 35 patients of antioxidant group and 36 patients of control group after 6 months of coronary bare metal stent implantation. We counted the stenotic lesions more than 50% of implanted stent lumen as a restenosis by quantitative coronary angiography. The serum levels of total antioxidant status, P-selectin, MCP-1, IL-6 and IL-10 were measured. RESULTS: The serum levels of total antioxidant status was not elevated in antioxidant group. Antioxidant supplementation did not change the serum levels of P-selectin, MCP-1, IL-6 and IL-10. The 6-month angiographic in-stent restenosis rate was 27% versus 30% (p=NS) with an associated late loss of 0.76+/-1.01 mm versus 0.91+/-1.00 mm (p=NS) for antioxidant group and control group. The serum levels of total antioxidant status did not correlate with the restenosis or late loss after stent implantation. CONCLUSIONS: Vitamin C, E and probucol did not elevate the serum level of antioxidant status and could not prevent in-stent restenosis after bare metal stent implantation.

Three Cases of Stress Induced Transient LV Dysfunction: Stress Induced Cardiomyopathy

A recently reported cardiac syndrome of transient left ventricular dysfunction, clinically resembles acute myocardial infarction and presents with chest pain, ECG changes and minimal elevation of cardiac enzymes in absence of myocardial ischemia or injury. The clinical presentation includes a wide range of symptoms and left ventricular function is normalized completely within days to weeks. This syndrome is likely a non-ischemic, metabolic-dependent syndrome caused by stress-induced activation of the cardiac adrenoreceptors. We report three cases of stress-induced transient LV dysfuction.