Monitoring Glutathione Dynamics and Heterogeneity in Living Stem Cells

Glutathione (GSH) is a major antioxidant in cells, and plays vital roles in the cellular defense against oxidants and in the regulation of redox signals. In a previous report, we demonstrated that stem cell function is critically affected by heterogeneity and dynamic changes in cellular GSH concentration. Here, we present a detailed protocol for the monitoring of GSH concentration in living stem cells using FreSHtracer, a real-time GSH probe. We describe the steps involved in monitoring GSH concentration in single living stem cells using confocal microscopy and flow cytometry. These methods are simple, rapid, and quantitative, and able to demonstrate intracellular GSH concentration changes in real time. We also describe the application of FreSHtracer to the sorting of stem cells according to their GSH content using flow cytometry. Typically, microscopic or flow cytometric analyses of FreSHtracer and MitoFreSHtracer signals in living stem cells take ~2~3 h, and the fractionation of stem cells into subpopulations on the basis of cellular GSH levels takes 3~4.5 h. This method could be applied to almost every kind of mammalian cell with minor modifications to the protocol described here.

Simultaneous Determination and Recognition Analysis of Coumarins in Angelica decursiva and Peucedanum praeruptorum by HPLC-DAD

Peucedani Radix is the root of Angelica decursiva Franchet et Savatier (=Peucedanum decursivum Maximowicz) or Peucedanum praeruptorum Dunn in several Asian countries. The coumarins contained in Peucedani Radix were quantitatively analyzed using HPLC-DAD to develop a simultaneous determination for the quality control of A. decursiva and P. praeruptorum. For quantitative analysis, four major coumarins contained in these medicinal plants were assessed. Nodakenin (1), nodakenetin (2), praeruptorin A (3), and praeruptorin B (4) were separated with a Phenomenex Luna C18 column (5 µm, 4.6 × 250 mm) under the gradient conditions using distilled water with 0.1% phosphoric acid and acetonitrile with 0.1% phosphoric acid as the mobile phase, at a flow rate of 1.0 ml/min and a detection wavelength of 330 nm. This method was fully validated for linearity, accuracy, precision, recovery, and limit of detection and quantification. As a result, A. decursiva and P. praeruptorum were clearly classified by the quantification of four major coumarins in extracts. Also, the pattern recognition analysis based on HPLC indicates that all of the samples were largely clustered into two groups. Therefore, it is possible to distinguish between A. decursiva and P. praeruptorum and contribute to quality control.

Multiple transcripts of anoctamin genes expressed in the mouse submandibular salivary gland

PURPOSE: Salivary fluid formation is primarily driven by Ca2+-activated, apical efflux of chloride into the lumen of the salivary acinus. The anoctamin1 protein is an anion channel with properties resembling the endogenous calcium-activated chloride channels. In order to better understand the role of anoctamin proteins in salivary exocrine secretion, the expression of the ten members of the anoctamin gene family in the mouse submandibular gland was studied. METHODS: Total RNA extracted from mouse submandibular salivary glands was reverse transcribed using primer pairs to amplify the full-length coding regions of each anoctamin gene and was subcloned into plasmid vectors for DNA sequencing. Alternative splice variants were also screened by polymerase chain reaction using primer pairs that amplified six overlapping regions of the complementary DNA of each anoctamin gene, spanning multiple exons. RESULTS: Multiple anoctamin transcripts were found in the mouse submandibular salivary gland, including full-length transcripts of anoctamin1, anoctamin3, anoctamin4, anoctamin5, anoctamin6, anoctamin9, and anoctamin10. Exon-skipping splicing in the N-terminal exons of the anoctamins1, anoctamin5, and anoctamin6 genes resulted in multiple alternative splice variants. No expression of anoctamin2, anoctamin7, or anoctamin8 was found. CONCLUSIONS: The predominant anoctamin transcript expressed in the mouse submandibular gland is anoctamin1ac. The chloride channel protein produced by anoctamin1ac is likely responsible for the Ca2+-activated chloride efflux, which is the rate-limiting step in salivary exocrine secretion.

Two Cases of Methimazole-Induced Insulin Autoimmune Syndrome in Graves' Disease

We report here the cases of two females with Graves' disease who developed insulin autoimmune syndrome after treatment with methimazole. The patients exhibited a sudden altered mental state after treatment with methimazole for approximately 4 weeks. Patients had hypoglycemia with serum glucose below 70 mg/dL, and laboratory findings showed both high levels of serum insulin and high titers of insulin autoantibodies. The two women had never been exposed to insulin or oral antidiabetic agents, and there was no evidence of insulinoma in imaging studies. After glucose loading, serum glucose, and total insulin levels increased abnormally. One of the patient was found to have HLA-DRB1*0406, which is known to be strongly associated with methimazole-induced insulin autoimmune syndrome. After discontinuation of methimazole, hypoglycemic events disappeared within 1 month. Insulin autoantibody titer and insulin levels decreased within 5 months and there was no further development of hypoglycemic events. We present these cases with a review of the relevant literature.

The Relationship Between Psychosocial Stress and Allergic Disease Among Children and Adolescents in Gwangyang Bay, Korea / 예방의학회지

OBJECTIVES: Stress is considered a causal factor in many diseases, allergic disease being one of them. The prevalence of allergic disease is increasing in Korea, but the relationship between allergic symptoms and stress is not empirically well known. We aimed to evaluate the relationship between allergy-related symptoms and stress in children and adolescents. METHODS: We investigated 698 children and adolescents living in Gwangyang Bay, Korea, using a multi-stage cluster sampling method. Using the International Study of Asthma and Allergies in Childhood and the Psychosocial Well-being Index, these subjects were surveyed on allergy-related symptoms and psychosocial stressors in their lives, respectively. We used a multivariate logistic analysis for odds ratios for the complaint rate of allergic symptoms, after adjusting for age, gender, household income, body mass index, and residence. RESULTS: After adjustments, lifetime rhinitis (odds ratio [OR], 1.024), rhinoconjunctivitis (OR, 1.090), diagnosis of itchy eczema (OR, 1.040), treatment of itchy eczema (OR, 1.049), 12-month allergic conjunctivitis (OR, 1.026), diagnosis of allergic conjunctivitis (OR, 1.031), and treatment of allergic conjunctivitis (OR, 1.034) were found to be significantly associated with stress. CONCLUSIONS: Our results support the notion that there is a relationship between stress and allergic symptoms in children and adolescents. Further research into any causal relationship between stress and allergies, as well as preventative public health plans for decreasing stress in children and adolescents are needed.

Factors Influencing Health Literacy in Community-Dwelling Adults

BACKGROUND: Health illiteracy is a problem often unrecognized by health care providers. It influences medical costs and the health status of adults. The purposes of this study were to determine the level of health literacy in community-dwelling adults and to identify the factors influencing it. METHODS: A cross-sectional survey was conducted in Seoul, Gyeonggi and Chungcheong province. A total of 420 adults aged 18 or older were interviewed by trained nursing students between November 1 to December 30, 2011. Health literacy was measured using the Short Test of Functional Health Literacy in Adults. Data were analyzed using the PASW 18.0 program. RESULTS: The mean score of health literacy was 50.64+/-19.18. In the multiple linear regression analysis, health literacy was significantly associated with education (beta=0.17, P=0.001), alcohol use (beta=-0.12, P=0.010), and perceived health status (beta=0.11, P=0.029). These factors accounted for about 7% of health literacy. CONCLUSIONS: Health literacy is a very important public health issue. Our findings showed that educational level, alcohol use and perceived health status should be considered when assessing this issue in patients. Furthermore, the development of a standardized Korean assessment tool for health literacy and specified interventions for enhancing health literacy are needed to improve health outcomes.

Application of a Loss of Resistance Syringe for Obtaining the Adequate Cuff Pressures of Endotracheal Intubated Patients in an Emergency Department

PURPOSE: The endotracheal tube cuff pressure must be kept within the optimal range. This study compared the usefulness of the conventional pilot balloon palpation technique using a 10 cc disposable syringe and passive release technique using a Loss of Resistance (LOR) syringe for obtaining adequate intracuff pressures of endotracheal intubated patients in an emergency department. METHODS: This was a prospective, observational study, conducted at the emergency department of a university teaching hospital. Patients who required endotracheal intubation in an emergency department were enrolled in this study. The patients were divided into two groups: one group who underwent the pilot balloon palpation technique using a conventional syringe (group A, n=40) and the other group who underwent the passive release technique using a LOR syringe technique (group B, n=40). The amount of air that infused into the cuff and the cuff pressure were measured. RESULTS: The mean cuff pressures of groups A and B were 41.0+/-23.7 cmH2O and 23.7+/-16.5 cmH2O, respectively. The mean cuff pressure of group A were significantly higher than group B (p=0.002). The mean air volume that infused into the cuff of groups A and B were 8.6+/-2.6 ml and 7.6+/-2.4 ml, respectively, showing no significant difference (p=0.688). The proportion of the optimal cuff pressure (Normal range: 22-32 cmH2O) of group A (9/40, 22.5%) was higher than that of group B (3/40, 7.5%). CONCLUSION: The range of air volumes and pressures for cuff inflation varied and it may not be possible to obtain the appropriate pressure using the LOR syringe technique. The endotracheal tube cuff pressure must be kept within the optimal range using a pressure monitor control inflator.

Association of Prescribed Drug intoxication and Neuropsychiatric history

PURPOSE: The purpose of this study was to investigate the relationship between patients with a neuropsychiatric history and features of their suicide attempt, in order to analyze the risk associated with psychiatrist prescribed drugs. METHODS: We retrospectively investigated cases of intentional overdose drug ingestion in patients greater than 14 years of age who visited OO emergency medical center between January 1, 2008 and July 31, 2010. We evaluated patient medical records to ascertain their age, sex, neuropsychiatric history, and components of ingested intoxicant. Information regarding any suicide reattempt was obtained after discharge through follow up telephone survey. SPSS version 13.0 was used for statistical analysis. Fisher's exact test was performed with p<0.05 considered statistically significant. RESULTS: Ninety-six of 209 patients (46%) had a past history of psychological problems. Among those 96 patients with a history of psychological problems, 46(48%) used an overdose of the medicine prescribed by their psychologist in order to attempt suicide. However, for patients without a history of psychological problems, intoxication by neuropsychiatric drugs was insignificant. Neuropsychiatric patients required greater follow up care after discharge and exhibited significantly more suicide reattempts. CONCLUSION: This study revealed that patients sometimes use the medicine prescribed by their psychologist to attempt suicide. Therefore, an exhaustive plan to control the medicines prescribed to psychiatric patients should be established.

Cardiac Tamponade Due to Suture Material and this Manifested as Convulsion

Cardiac tamponade is a potentially acute, life threatening emergency that can cause death if it is not promptly diagnosed and treated. Cardiac tamponade is a comparatively uncommon presentation to the emergency department and it is usually associated with penetrating trauma. We report here on a case of cardiac tamponade due to suture material that was used for colectomy ten years previously. A 17-year-old male was admitted to an emergency department with a complaint of loss of consciousness and convulsion. He also complained of chest pain, dyspnea and hypotension. After a while, he displayed cyanosis and his jugular veins were distended. The emergency echocardiogram showed a large amount of pericardial effusion with features of tamponade. Pericardiocentesis was immediately performed. Although 800 cc of fresh blood was drained from the pericardial cavity, his bleeding wouldn't stop. So, the patient was moved immediately to the operation room, and pericardiectomy and median sternotomy were performed. The surgeon found that the foreign suture material had penetrated the pericardium and he successfully removed it. The removed foreign body was a bundle of thread. The patient was discharged without any complications after 9 days.

A Case of the Symptomatic Bradycardia Treated with Norepinephrine at an ED

Symptomatic bradycardia might be regarded as a serious emergency disease and it requires prompt emergency treatments. The American Heart Association has recommended transcutaneous pacing as a gold standard of treatment and also atropine, epinephrine or dopamine as the first line drugs. We report here on a case of symptomatic bradycardia that was treated with norepinephrine and the patient was not treated with pacing, atropine and dopamine.

MR Imaging of Aggressive Angiomyxoma of the Female Pelvis: Case Report

Aggressive angiomyxoma is a rare tumor that predominantly occurs in the female genital tract. Because of its high tendency for local recurrence, preoperative diagnosis is important to ensure wide excision and reduce recurrence risk. But it is often misdiagnosed due to its rarity. We present two cases of aggressive angiomyxoma: a recurrent case that was initially misdiagnosed and a preoperatively diagnosed case.

Two Cases of Diabetic Gastropathy in Pregnant Diabetic Women / 대한산부인과학회잡지

Gastrointestinal complications of diabetes mellitus include diarrhea, constipation, small bowel bacterial overgrowth, and gastropathy. Diabetic gastropathy is well-described motility disorder probably caused by an automatic neuropathy of the upper gastrointestinal tract. Diabetic gastropathy is rarely discrbed in pregnant women. We have experienced two cases of diabetic gastropathy in pregnant diabetic women. We report these cases with a review of literatures.

Relationship between HPV Typing and Expressions of p16 and p53 in Cervical Neoplasia / 대한산부인과학회잡지

OBJECTIVE: To determine the relationships among the grade of cervical pathology, HPV infection status, p16 expression, and p53 expression in cervical neoplasia. METHODS: From July of 2002 to June of 2003, authors did HPV typing with HPV Oligonucleotide Microarray DNA Chip (Biomed, Seoul, Korea), p16 and p53 immunochemical staining in cervical tissues obtained from conization or hysterectomy RESULTS: 41 patients were enrolled in this study. There were 16 normal (WNL) patients, 9 CIN patients, 8 microinvasive cancer (MIC) patients, and 8 invasive cervical cancer (ICC) patients. The HPV infection rate in WNL, CIN, MIC and ICC were 0%, 100%, 50% and 50% retrospectively. The expression rate of p16 was 13% in WNL, 67% in CIN, 50% in MIC and 100% in ICC. That of p53 was 0% in WNL, 67% in CIN, 75% in MIC and 100% in ICC. With regard to p16 staining in HPV negative cases, negative staining was 67%, weakly positive was 13% and strongly positive was 21%. But in HPV-infected cases, negative staining of p16 was 29%, weakly positive was 53% and strongly positive was 18%. With regard to p53 staining in HPV negative cases, without HPV infection, negative staining was 62%, weakly positive was 33% and strongly positive was 4%. But in HPV infected cased, negative staining of p53 was 18%, weakly positive was 53% and strongly positive was 29%. Finally, in p16 negative cases, negative staining of p53 was 62%, weakly positive was 38% and strongly positive was 0%. But in p16 weakly positive cases, with weakly positive p16, negative staining of p53 was 33%, weakly positive was 42%, and strongly positive was 25%. In p16 strongly positive cases, negative staining of p53 was 13%, weakly positive was 50% and strongly positive was 38%. CONCLUSION: There were significant associations among grades of cervical pathology, HPV infection, p16 and p53 expression. Thus there might be some possibilities that expression of p16 and p53 induced by E7 and E6 proteins of HPV can impact on the tumorigenesis.