Cytochrome P450 endoplasmic reticulum-associated degradation (ERAD): therapeutic and pathophysiological implications

The hepatic endoplasmic reticulum (ER)-anchored cytochromes P450 (P450s) are mixed-function oxidases engaged in the biotransformation of physiologically relevant endobiotics as well as of myriad xenobiotics of therapeutic and environmental relevance. P450 ER-content and hence function is regulated by their coordinated hemoprotein syntheses and proteolytic turnover. Such P450 proteolytic turnover occurs through a process known as ER-associated degradation (ERAD) that involves ubiquitin-dependent proteasomal degradation (UPD) and/or autophagic-lysosomal degradation (ALD). Herein, on the basis of available literature reports and our own recent findings of as well as experimental studies, we discuss the therapeutic and pathophysiological implications of altered P450 ERAD and its plausible clinical relevance. We specifically (i) describe the P450 ERAD-machinery and how it may be repurposed for the generation of antigenic P450 peptides involved in P450 autoantibody pathogenesis in drug-induced acute hypersensitivity reactions and liver injury, or viral hepatitis; (ii) discuss the relevance of accelerated or disrupted P450-ERAD to the pharmacological and/or toxicological effects of clinically relevant P450 drug substrates; and (iii) detail the pathophysiological consequences of disrupted P450 ERAD, contributing to non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH) under certain synergistic cellular conditions.

High Prevalence of Rotavirus G4P6 Genotypes among Neonates in Two Korean Hospitals / 대한임상미생물학회지

BACKGROUND: The introduction of rotavirus vaccines has decreased the prevalence of rotavirus infections and might have changed the distribution of rotavirus genotypes. However, neonates are not eligible for vaccination and, therefore, are at risk for rotavirus infection while in the hospital nursery or neonatal intensive care unit. Our aim was to evaluate the shift of genotypes of group A rotavirus strains among neonates cared for in two geographically distant hospitals in Korea. METHODS: Analysis of rotavirus P and G genotypes was performed for 63 neonates (27 neonates in Seoul and 36 neonates in Busan) admitted to two hospitals between 2011 and 2013. RESULTS: Among the 63 tested neonates less than one month of age, 61 (96.8%) were infected with genotype G4P[6]. CONCLUSION: This study identified G4P[6] as the most frequently isolated genotypes among neonates in Korea; therefore, prevention of the G4P[6] genotype should be considered for neonates.

Early Prophylactic versus Late Selective Use of Surfactant for Respiratory Distress Syndrome in Very Preterm Infants: A Collaborative Study of 53 Multi-Center Trials in Korea

Pulmonary surfactant (PS) therapy was proven to be highly successful for the treatment of respiratory distress syndrome in premature infants. As a results, early prophylactic (EP) PS therapy has been introduced recently in Europe, the US and Korea. However, no multi-center study was compared EP and late selective (LS) PS therapies in Korea. We performed a retrospective multi-center study to compare the outcomes of EP and LS PS therapies in very preterm infants. We analyzed clinical morbidity and mortality for 1,291 infants in 2010 (LS group) and 1,249 infants in 2011 (EP group); the infants were born <30 weeks of gestation and had birth weight < or =1,250 g, and were chosen from 53 neonatal intensive care units in Korea. Compared to the LS group (22.5%), the overall mortality was better in the EP group (19.9%) and there was no increased need for retreatment.There were additional benefits in the EP group such as fewer associated complications. To the best of knowledge, our study is the first nationwide Korean study to compare the outcomes of EP and LS therapies, and it provides evidences that EP PS therapy is important in very preterm infants to improve for survival and reduce morbidities.

Early Prophylactic versus Late Selective Use of Surfactant for Respiratory Distress Syndrome in Very Preterm Infants: A Collaborative Study of 53 Multi-Center Trials in Korea

Pulmonary surfactant (PS) therapy was proven to be highly successful for the treatment of respiratory distress syndrome in premature infants. As a results, early prophylactic (EP) PS therapy has been introduced recently in Europe, the US and Korea. However, no multi-center study was compared EP and late selective (LS) PS therapies in Korea. We performed a retrospective multi-center study to compare the outcomes of EP and LS PS therapies in very preterm infants. We analyzed clinical morbidity and mortality for 1,291 infants in 2010 (LS group) and 1,249 infants in 2011 (EP group); the infants were born <30 weeks of gestation and had birth weight < or =1,250 g, and were chosen from 53 neonatal intensive care units in Korea. Compared to the LS group (22.5%), the overall mortality was better in the EP group (19.9%) and there was no increased need for retreatment.There were additional benefits in the EP group such as fewer associated complications. To the best of knowledge, our study is the first nationwide Korean study to compare the outcomes of EP and LS therapies, and it provides evidences that EP PS therapy is important in very preterm infants to improve for survival and reduce morbidities.

Overgrowth Syndrome with 9q22.3 Microdeletion Detected by Microarray Comparative Genomic Hybridization

Microdeletion of 9q22.3 is a rare chromosomal disorder characterized by body overgrowth, facial dysmorphic features and psychomotor delay. The presence of genomic microdeletion or microdu-plication can not be identified by the conventional chromosomal analysis. Microarray comparative genomic hybridization (CGH) is a newly developed molecular cytogenetic technique that enables the identification of minute copy number variation (CNV) in the human genome. Here, we report a case of microdeletion in the 9q22.31-q22.33 region, which included a patched homolog 1 (PTCH1) gene, as detected by CGH and confirmed by fluorescence in situ hybridization (FISH) analyses in a neonate with prenatal onset of macrosomia, dysmorphism, and muscle hypotonia. To the best of our knowledge, this is the first case report of 9q22.3 microdeletion detected by CGH in Korea.

Comparison of Stress Levels Arising in Clinical Settings, Stress Coping Mechanisms and Clinical Competency between Male and Female Nursing Students / 한국간호교육학회지

PURPOSE: This study compares differences between male and female nursing students in terms of the level of stress experienced during clinical practice settings, stress coping mechanisms and clinical competency. METHOD: The study population was nursing students who had taken a clinical practice course at one of five general hospitals in city D. Data were collected from September 15 to October 15, 2013 through a structured self-administered questionnaire survey given to 319 subjects (156 male students, 163 female students) enrolled in a statistical analysis course. RESULTS: Male nursing students experienced lower stress levels during clinical practice and higher clinical competency than female students. In terms of stress coping mechanisms, male students were significantly more active than female students in deploying coping strategies. Among all students, students with higher clinical practice stress and concomitant stress coping scores exhibited higher clinical competency. CONCLUSION: Noticeable differences between male and female nursing students in terms of stress levels arising in the course of clinical practice, stress coping strategies, and clinical competency were revealed. However, further identification of specific stressful situations for male and female nursing students and subsequent management of clinical training circumstances are required.

The effects of dexamethasone on the apoptosis and osteogenic differentiation of human periodontal ligament cells

PURPOSE: The purpose of the current study was to examine the effect of dexamethasone (Dex) at various concentrations on the apoptosis and mineralization of human periodontal ligament (hPDL) cells. METHODS: hPDL cells were obtained from the mid-third of premolars extracted for orthodontic reasons, and a primary culture of hPDL cells was prepared using an explant technique. Groups of cells were divided according to the concentration of Dex (0, 1, 10, 100, and 1,000 nM). A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed for evaluation of cellular viability, and alkaline phosphatase activity was examined for osteogenic differentiation of hPDL cells. Alizarin Red S staining was performed for observation of mineralization, and real-time polymerase chain reaction was performed for the evaluation of related genes. RESULTS: Increasing the Dex concentration was found to reduce cellular viability, with an increase in alkaline phosphatase activity and mineralization. Within the range of Dex concentrations tested in this study, 100 nM of Dex was found to promote the most vigorous differentiation and mineralization of hPDL cells. Dex-induced osteogenic differentiation and mineralization was accompanied by an increase in the level of osteogenic and apoptosis-related genes and a reduction in the level of antiapoptotic genes. The decrease in hPDL cellular viability by glucocorticoid may be explained in part by the increased prevalence of cell apoptosis, as demonstrated by BAX expression and decreased expression of the antiapoptotic gene, Bcl-2. CONCLUSIONS: An increase in hPDL cell differentiation rather than cellular viability at an early stage is likely to be a key factor in glucocorticoid induced mineralization. In addition, apoptosis might play an important role in Dex-induced tissue regeneration; however, further study is needed for investigation of the precise mechanism.

Occurrence and characterization of oseltamivir-resistant influenza virus in children between 2007-2008 and 2008-2009 seasons / 소아과

PURPOSE: There was a global increase in the prevalence of oseltamivir-resistant influenza viruses during the 2007-2008 influenza season. This study was conducted to investigate the occurrence and characteristics of oseltamivir-resistant influenza viruses during the 2007-2008 and 2008-2009 influenza seasons among patients who were treated with oseltamivir (group A) and those that did not receive oseltamivir (group B). METHODS: A prospective study was conducted on 321 pediatric patients who were hospitalized because of influenza during the 2007-2008 and 2008-2009 influenza seasons. Drug resistance tests were conducted on influenza viruses isolated from 91 patients. RESULTS: There was no significant difference between the clinical characteristics of groups A and B during both seasons. Influenza A/H1N1, isolated from both groups A and B during the 2007-2008 and 2008-2009 periods, was not resistant to zanamivir. However, phenotypic analysis of the virus revealed a high oseltamivir IC50 range and that H275Y substitution of the neuraminidase (NA) gene and partial variation of the hemagglutinin (HA) gene did not affect its antigenicity to the HA vaccine even though group A had a shorter hospitalization duration and fewer lower respiratory tract complications than group B. In addition, there was no significant difference in the clinical manifestations between oseltamivir-susceptible and oseltamivir-resistant strains of influenza A/H1N1. CONCLUSION: Establishment of guidelines to efficiently treat influenza with oseltamivir, a commonly used drug for treating influenza in Korean pediatric patients, and a treatment strategy with a new therapeutic agent is required.

A case of bilateral submandibular gland mucoceles in a 16-month-old child / 소아과

Mucoceles are common benign cystic lesions of the oral cavity that develop following extravasation or retention of mucous material from the major or minor salivary glands. Mucoceles are usually located in the lower lip (60 to 70% of cases), and the floor of the mouth is only involved in 6 to 15% of cases. Submandibular gland mucocele is extremely rare but should be considered in the differential diagnosis of swelling at the submandibular triangle in young children. We present the rare case of a 16-month-old child who was diagnosed with bilateral submandibular gland mucocele, presenting as serial swellings in both submandibular regions. We removed the cystic mass with the submandibular and sublingual glands to prevent recurrence.

A case of bilateral submandibular gland mucoceles in a 16-month-old child / 소아과

Mucoceles are common benign cystic lesions of the oral cavity that develop following extravasation or retention of mucous material from the major or minor salivary glands. Mucoceles are usually located in the lower lip (60 to 70% of cases), and the floor of the mouth is only involved in 6 to 15% of cases. Submandibular gland mucocele is extremely rare but should be considered in the differential diagnosis of swelling at the submandibular triangle in young children. We present the rare case of a 16-month-old child who was diagnosed with bilateral submandibular gland mucocele, presenting as serial swellings in both submandibular regions. We removed the cystic mass with the submandibular and sublingual glands to prevent recurrence.

Exogenous Surfactant Therapy in Neonates with Respiratory Failure Due to Pulmonary Hemorrhage / 대한주산의학회잡지

PURPOSE: The study assessed the effect of exogenous surfactant on oxygen and ventilator requirements and long-term prognosis of neonates with respiratory deterioration due to pulmonary hemorrhage. METHODS: From March 2007 to March 2011, neonates with respiratory deterioration due to pulmonary hemorrhage in Saint Mary's Hospital were identified from the database. Among 20 patients with pulmonary hemorrhage, 10 patients received surfactant therapy and the other 10 did not. We compared outcomes of these two groups. The primary outcome was changes in respiratory status following surfactant therapy, as reflected by oxygen index (OI), fractional inspired oxygen (FiO2) and mean airway pressure (MAP). Secondary outcomes were the duration of ventilation, nasal continuous positive airway pressure (NCPAP), oxygen inhalation days, progression to severe bronchopulmonary dysplasia or death, and retinopathy of prematurity, which required laser therapy. RESULTS: Surfactant treatment group had higher FiO2, MAP, and OI than the non-treatment group during pulmonary hemorrhage. But, there were no differences in FiO2, MAP, and OI after 2 hours of pulmonary hemorrhage between the two groups. There were no significant side effects during surfactant treatment in the surfactant treatment group. Surfactant treatment group had fewer ventilator days, NCPAP days (P<0.05) and a tendency towards shorter oxygen days than the non-treatment group (P=0.09). Also, the surfactant treatment group had less severe bronchopulmonary dysplasia (BPD) or death than the non-treatment group (10% vs. 70%, P<0.05). CONCLUSION: Exogenous surfactant appears to be a useful adjunctive therapy with neonates of severe respiratory deterioration due to pulmonary hemorrhage. Also, exogenous surfactant treatment dose not seen to be associated with any significant side effects during the therapy.

Surfactant Replacement Therapy for RDS: a Collaborative Study of 72 Multi-center Trials in Korea (2010) and a Review of Korean Experiences over 20 Years

In Korea, pulmonary surfactant (PS) replacement therapy in respiratory distress syndrome (RDS) was started in 1991 since when Surfacten(R) was imported from Japan. At the present time, Surfacten(R), Newfactan(R), Curosurf(R), and Infasurf(R) are available in Korea. The governmental health insurance covers the expense for multiple dose treatment since 2002 and the early prophylactic treatment (BW: <1,250 g or GP: <30 wks) since 2011. We undertook a multi-institutional collective study to evaluate the outcomes of PS over past 20 years in Korea (Period-I; 1990/91, P-II; 1996, P-III; 2002, and P-IV; 2007, P-V; 2010). There were 60 RDS neonates with PS treatment in P-I (16 hospitals), 1,179 in P-II (64), 1,595 in P-III (62), 1,921 in P-IV (57), and 3,160 in P-V (72). Decreased mortality rate, defined as the percentage of neonates who died within 28 days of birth, was seen between periods, P-V vs P-I, II, III, and IV (mortality rate: 10.1% vs. 40.0%, 30.0%, 18.7%, and 14.3%). We conclude that PS therapy contributed to improve remarkable outcome in RDS neonates over the last 20 years in Korea. However, more efforts should be made to optimize PS therapy for better outcome. Multiple PS doses for relapse and poor response, early prophylactic use, and better supportive care for pre-term infants are mandatory.

Oseltamivir efficacy, side effects, and safety in children with influenza / 소아과

PURPOSE: Although oseltamivir is widely used for treatment of influenza, few clinical studies of its efficacy and resistance have been performed in Korea. We evaluated the safety, side effects, and efficacy of oseltamivir treatment in Korean pediatric patients. METHODS: We analyzed 321 children diagnosed with influenza at Busan St. Mary's Medical Center, Korea, between January 2008 and June 2008 (first study period) and November 2008 and January 2009 (second study period). Patients were divided into two groups: those receiving oseltamivir treatment for 5 days and those receiving only symptomatic treatment. We investigated clinical symptoms, side effects, and resistance to oseltamivir. We also identified influenza strains and evaluated resistance to oseltamivir using an influenza virus culture. RESULTS: One hundred eighty-six patients were assigned to the treatment group, and 135 were assigned to the control group. The treatment group showed shorter admission duration (4.4 days) compared with controls (5.0 days) (P=0.000) and had fewer lower respiratory tract complications compared with controls (P<0.05). No significant statistical difference in the virus antigenic type was observed between the groups. In the first study period, virus culture showed influenza B (41.7% vs. 49.6%), A/H3N2 (7.9% vs. 8.4%), and A/H1N1 (9.4% vs. 6.5%). In the second study period, only A/H1N1 (55.3% vs. 50.0%) was isolated, except for one case of A (H3N2) in the treatment group. No differences in short- and long-term side effects, including neuropsychologic side effects, were noted between groups. There was no resistance to oseltamivir before or after treatment in the first study period. CONCLUSION: Based on our results, we suggest that osetalmivir therapy in pediatric patients is effective.

High-dose caspofungin salvage in a very-low-birth-weight infant with refractory candidemia / 소아과

Candidiasis is one of the most common causes of late-onset infection among very-low-birth-weight infants (VLBW) in most neonatal intensive care units and is associated with significant morbidity and mortality. Standard therapy consists of the administration of amphotericin B, amphotericin B complex, and fluconazole. In many cases, candidiasis is not easily eradicated, despite the administration of these drugs. We report our experience of the addition of high-dose caspofungin to the conventional antifungal drugs in a VLBW infant with refractory candidemia.

Effect of Neonatal Developmental Intervention Program (NDT) on Motor Development and Growth in Premature Infants

PURPOSE: The aim of this study was to identify the effects of neonatal developmental intervention program (NDT) in promoting motor development and growth and to determine the usefulness of Hammersmith Neonatal Neurological Examination (HNNE) and Neonatal Behavioral Assessment Scale (NBAS) in premature infants. METHODS: We performed NDT on selected premature infants (PI, n=42) and compared them with the full term control group (FC, n=20). NDT protocol and development assessment (HNNE, NBAS) were manipulated by the physical therapist in the neonatal intensive care unit. The data of this study were collected prospectively. RESULTS: The PI with GA or =34 weeks but less than 37 weeks (LPI) subgroup. The average scores improved significantly in the PI group between the 1st, 2nd, and 3rd assessment by repeated measure (P=0.000). Also, the PI group showed significantly higher total scores and average score at 40 weeks postmenstrual age, P=0.000, respectively than in the FC group. The LPI subgroup showed more weight gain and change in the head circumference between the 1st and 3rd assessment by repeated measure, respectively, P<0.05. The PI group showed no apnea, bradycardia and late sepsis associated with intervention and assessment. CONCLUSION: The NDT might be a safe and useful intervention to promote motor and growth outcomes in premature infants. Also, the HNNE and NBAS might be safe and useful tools for assessing neurodevelopment in premature infants.

Effect of Neonatal Developmental Intervention Program (NDT) on Motor Development and Growth in Premature Infants

PURPOSE: The aim of this study was to identify the effects of neonatal developmental intervention program (NDT) in promoting motor development and growth and to determine the usefulness of Hammersmith Neonatal Neurological Examination (HNNE) and Neonatal Behavioral Assessment Scale (NBAS) in premature infants. METHODS: We performed NDT on selected premature infants (PI, n=42) and compared them with the full term control group (FC, n=20). NDT protocol and development assessment (HNNE, NBAS) were manipulated by the physical therapist in the neonatal intensive care unit. The data of this study were collected prospectively. RESULTS: The PI with GA or =34 weeks but less than 37 weeks (LPI) subgroup. The average scores improved significantly in the PI group between the 1st, 2nd, and 3rd assessment by repeated measure (P=0.000). Also, the PI group showed significantly higher total scores and average score at 40 weeks postmenstrual age, P=0.000, respectively than in the FC group. The LPI subgroup showed more weight gain and change in the head circumference between the 1st and 3rd assessment by repeated measure, respectively, P<0.05. The PI group showed no apnea, bradycardia and late sepsis associated with intervention and assessment. CONCLUSION: The NDT might be a safe and useful intervention to promote motor and growth outcomes in premature infants. Also, the HNNE and NBAS might be safe and useful tools for assessing neurodevelopment in premature infants.

The Effects of Aromatherapy and Hand Massage on Anxiety, Sleep, and Depression in the Female Aged at Residential Facilities

PURPOSE: This study adopted the non-equivalent control group pre-posttest design in order to examine the effects of aromatherapy and hand massage on anxiety, sleep, and depression in the female aged at residential facilities. METHODS: This study was executed with the female aged at two residential facilities divided into an experimental group (n=35) and a control group (n=37). Data were collected by checking changes in the anxiety, sleep, and depression of the subjects who had received aromatherapy and hand massage ten times for two weeks. The data were analyzed using Chi-square test and ANCOVA (Analysis of Covariance). RESULTS: The condition of anxiety and depression of the experimental group who had received aromatherapy and hand massage was significantly lower than that of the control group. Also, the condition of sleep of experimental group was significantly higher than that of the control group. CONCLUSION: The aromatherapy and hand massage program had a positive effect on institutionalized elders' anxiety, sleep, and depression pattern.

The Importance and the Need of Early Pulmonary Surfactant Therapy in Premature Infant with Respiratory Distress Syndrome

Pulmonary surfactant (PS) therapy in premature infants has a remarkable impact on improving survival and outcomes in neonatal respiratory distress syndrome (RDS). Early PS therapy involves instillation of PS upon delivery of very premature infants or if there is evidence of RDS, such as an increased requirement of oxygen 2 hours after birth, especially in infants <30 weeks gestation. Early PS treatment in very premature infants results in a significant reduction in the severity of RDS, mortality, and incidence of pneumothorax, pulmonary interstitial emphysema, and bronchopulmonary dysplasia in comparison with late PS treatment. According to European and American consensus guidelines on the management of neonatal RDS, early PS instillation should be considered for infants <30 weeks gestation, infants with a birth weight <1,250 g, or if the mother has not received antepartum corticosteroids. We suggest that the Korean health insurance policy on RDS be modified so that PS can be used for better clinical outcomes of very premature infants.

Liver Abscess Associated with Maternal Perinatal Infection in a Premature Infant

Neonatal liver abscesses are rare, carry a high mortality rate, and are difficult to diagnose. The diagnosis of liver abscesses in the neonate cannot be established from the clinical presentation alone. Risk factors for liver abscesses in neonates are maternal infection, sepsis, umbilical venous catheterization, omphalitis, and necrotizing enterocolitis. In this report, we describe a preterm infant (32 weeks, 1,580 g) who presented with abdominal distension, respiratory difficulties, and a persistent inflammatory response in spite of broad spectrum antibiotic treatment; a large (6x5 cm) solitary pyogenic liver abscess was identified at 9 days of age. It appeared that the liver abscess had originated in the uterus and umbilical venous catheterization facilitated its spread. Percutaneous drainage under abdominal ultrasound guidance was performed and prolonged antibiotics were treated for 5 weeks, effecting a cure.

Bioluminescence Imaging of Chondrocytes in Rabbits by Intraarticular Injection of D-Luciferin / 핵의학분자영상

PURPOSE: Luciferase is one of the most commonly used reporter enzymes in the field of in vivo optical imaging. D-luciferin, the substrate for firefly luciferase has very high cost that allows this kind of experiment limited to small animals such as mice and rats. In this current study, we validated local injection of D-luciferin in the articular capsule for bioluminescence imaging in rabbits. MATERIALS AND METHODS: Chondrocytes were cultured and infected by replication-defective adenoviral vector encoding firefly luciferase (Fluc). Chondrocytes expressing Fluc were injected or implanted in the left knee joint. The rabbits underwent optical imaging studies after local injection of D-luciferin at 1, 5, 7, 9 days after cellular administration. We sought whether optimal imaging signals was could be by a cooled CCD camera after local injection of D-luciferin. RESULTS: Imaging signal was not observed from the left knee joint after intraperitoneal injection of D-luciferin (15 mg/kg), whereas it was observed after intraarticular injection. Photon intensity from the left knee joint of rabbits was compared between cell injected and implanted groups after intraarticular injection of D-luciferin. During the period of imaging studies, photon intensity of the cell implanted group was 5-10 times higher than that of the cell injected group. CONCLUSION: We successfully imaged chondrocytes expressing Fluc after intraarticular injection of D-luciferin. This technique may be further applied to develop new drugs for knee joint disease.