Oxidative Stress Modulates the Expression Pattern of Peroxiredoxin-6 in Peripheral Blood Mononuclear Cells of Asthmatic Patients and Bronchial Epithelial Cells

PURPOSE: Reduction-oxidation reaction homeostasis is vital for regulating inflammatory conditions and its dysregulation may affect the pathogenesis of chronic airway inflammatory diseases such as asthma. Peroxiredoxin-6, an important intracellular anti-oxidant molecule, is reported to be highly expressed in the airways and lungs. The aim of this study was to analyze the expression pattern of peroxiredoxin-6 in the peripheral blood mononuclear cells (PBMCs) of asthmatic patients and in bronchial epithelial cells (BECs).METHODS: The expression levels and modifications of peroxiredoxin-6 were evaluated in PBMCs from 22 asthmatic patients. Phosphorylated and acetylated peroxiredoxin-6 in hydrogen peroxide-treated human BECs was detected using immunoprecipitation analysis. The expression level of peroxiredoxin-6 was also investigated in BECs treated with hydrogen peroxide. Cycloheximide and proteasome inhibitors were used to determine whether peroxiredoxin-6 is degraded by proteasomes.RESULTS: Peroxiredoxin-6 expression was significantly reduced in the PBMCs of asthmatic patients compared to control subjects. Distinct modification patterns for peroxiredoxin-6 were observed in the PBMCs of asthmatic patients using 2-dimensional-electrophoresis. The levels of phosphorylated serine and acetylated lysine in peroxiredoxin-6 were significantly increased in the BECs following hydrogen peroxide treatment. The level of peroxiredoxin-6 expression was reduced in hydrogen peroxide-stimulated BECs, presumably due to proteasomes.CONCLUSIONS: The expression of peroxiredoxin-6, which is down-regulated in the immune cells of asthmatic patients and BECs, can be modified by oxidative stress. This phenomenon may have an effect on asthmatic airway inflammation.

Fluoxetine Induces Apoptotic and Oxidative Neuronal Death Associated with The Influx of Copper Ions in Cultured Neuronal Cells / 전남의대학술지

We examined the effect of fluoxetine, a selective serotonin reuptake inhibitor antidepressant, on neuronal viability in mouse cortical near-pure neuronal cultures. Addition of fluoxetine to the media for 24 hours induced neuronal death in a concentration-dependent manner. To delineate the mechanisms of fluoxetine-induced neuronal death, we investigated the effects of trolox, cycloheximide (CHX), BDNF, z-VAD-FMK, and various metal-chelators on fluoxetine-induced neuronal death. Neuronal death was assessed by MTT assay. The addition of 20 µM fluoxetine to the media for 24 hours induced 60–70% neuronal death, which was associated with the hallmarks of apoptosis, chromatin condensation and DNA laddering. Fluoxetine-induced death was significantly attenuated by CHX, BDNF, or z-VAD-FMK. Treatment with antioxidants, trolox and ascorbate, also markedly attenuated fluoxetine-induced death. Interestingly, some divalent cation chelators (EGTA, Ca-EDTA, and Zn-EDTA) also markedly attenuated the neurotoxicity. Fluoxetine-induced reactive oxygen species (ROS) generation was measured using the fluorescent dye 2′,7′-dichlorofluorescin diacetate. Trolox and bathocuproine disulfonic acid (BCPS), a cell membrane impermeable copper ion chelator, markedly attenuated the ROS production and neuronal death. However, deferoxamine, an iron chelator, did not affect ROS generation or neurotoxicity. We examined the changes in intracellular copper concentration using a copper-selective fluorescent dye, Phen Green FL, which is quenched by free copper ions. Fluoxetine quenched the fluorescence in neuronal cells, and the quenching effect of fluoxetine was reversed by co-treatment with BCPS, however, not by deferoxamine. These findings demonstrate that fluoxetine could induce apoptotic and oxidative neuronal death associated with an influx of copper ions.

Chicken RNA-binding protein T-cell internal antigen-1 contributes to stress granule formation in chicken cells and tissues

T-cell internal antigen-1 (TIA-1) has roles in regulating alternative pre-mRNA splicing, mRNA translation, and stress granule (SG) formation in human cells. As an evolutionarily conserved response to environmental stress, SGs have been reported in various species. However, SG formation in chicken cells and the role of chicken TIA-1 (cTIA-1) in SG assembly has not been elucidated. In the present study, we cloned cTIA-1 and showed that it facilitates the assembly of canonical SGs in both human and chicken cells. Overexpression of the chicken prion-related domain (cPRD) of cTIA-1 that bore an N-terminal green fluorescent protein (GFP) tag (pntGFP-cPRD) or Flag tag (pFlag-cPRD) induced the production of typical SGs. However, C-terminal GFP-tagged cPRD induced notably large cytoplasmic granules that were devoid of endogenous G3BP1 and remained stable when exposed to cycloheximide, indicating that these were not typical SGs, and that the pntGFP tag influences cPRD localization. Finally, endogenous cTIA-1 was recruited to SGs in chicken cells and tissues under environmental stress. Taken together, our study provide evidence that cTIA-1 has a role in canonical SG formation in chicken cells and tissues. Our results also indicate that cPRD is necessary for SG aggregation.

Changes in postharvest physio-biochemical characteristics and antioxidant enzymes activity of cut alsteroemeria aurantiaca flower as affected by cycloheximide, coconut water and 6-benzyladenine / Alterações nas características físico-bioquímicas pós-colheita e atividades de enzimas antioxidantes da flor de alstroemeria aurantiaca cortada como afetada pela cicloheximida, água de coco e 6-benziladenina

Early leaf yellowing in cut alstroemeria (Alstroemeria aurantiaca) flowers before flower development and petal abscission is an important limiting postharvest quality and vase life factors. Early leaf senescence reduces postharvest longevity of cut flowers and promotes petal's wilting. A study was made to evaluate the response of cut alstroemeria flowers at varying concentrations of cycloheximide (CHI) (50, 100 and 200 mg l-1), coconut water (5, 10 and 20%) and 6-benzyladenine (BA) (50, 100 and 200 mg l-1). CHI, coconut water and BA extended the vase life at all concentrations compared to the control, but coconut water at 5% concentration (with 17.39 days) was the most effective treatment. Control cut flowers showed the least vase life (10.76 days). Ethylene production in cut flowers promoted flower senescence. All concentrations of CHI, coconut water and BA delayed ethylene production compared to the control. Treatment of cut flowers with coconut water at concentration of 5% maintained the highest fresh weight of flowers and increased the content of water uptake. The chlorophyll degradation was significantly reduced by the application of CHI, coconut water and BA. The maximum content of membrane's lipid peroxidation and antioxidant enzymes activity (super oxide dismutase and peroxidase) was obtained in control cut flowers. Thus, 5% fresh coconut water has the potential to be applied as vase solution (preservative medium) due to prolongs of cut alstroemeria flowers.

O amarelecimento precoce das folhas em flores de alstroemeria (Alstroemeria aurantiaca) cortadas antes do desenvolvimento floral e da abscisão de pétalas é um importante limitante da qualidade pós-colheita e dos fatores de vida do vaso. A senescência precoce da folha reduz a longevidade pós-colheita das flores cortadas e promove o murchamento da pétala. Um estudo foi realizado para avaliar a resposta de flores de alstroemeria cortadas em diferentes concentrações de cicloheximida (CHI) (50, 100 e 200 mg l-1), água de coco (5, 10 e 20%) e 6-benziladenina (BA) 50, 100 e 200 mg l-1). CHI, água de coco e BA prolongou a vida do vaso em todas as concentrações em comparação com o controle, mas a água de coco a 5% de concentração (com 17,39 dias) foi o tratamento mais eficaz. As flores cortadas de controlo mostraram a menor vida útil do vaso (10,76 dias). A produção de etileno em flores cortadas promoveu a senescência da flor. Todas as concentrações de CHI, água de coco e BA atrasaram a produção de etileno em comparação com o controle. O tratamento de flores cortadas com água de coco a uma concentração de 5% manteve o maior peso fresco de flores e aumentou o conteúdo de absorção de água. A degradação da clorofila foi significativamente reduzida pela aplicação de CHI, água de coco e BA. O teor máximo de atividade de enzimas antioxidantes e de peroxidação lipídica da membrana (super óxido dismutase e peroxidase) foi obtido em flores cortadas de controle. Assim, 5% de água de coco fresca tem potencial para ser aplicada como solução de vaso (meio de conservação) devido a prolongamentos das flores de alstroemeria cortadas.

NADPH Oxidase Mediates β-Amyloid Peptide-Induced Neuronal Death in Mouse Cortical Cultures / 전남의대학술지

β-Amyloid peptide (Aβ) is the main component of senile plaques in patients with Alzheimer's disease, and is known to be a main pathogenic factor of the disease. Recent evidence indicates that activation of NADPH oxidase (NOX) in microglia or astrocytes may be a source of Aβ-induced reactive oxygen species (ROS). We investigated the role of neuronal NOX in Aβ-induced neuronal death in mouse mixed cortical cultures. Cell death was assessed by measuring lactate dehydrogenase efflux to bathing media 24 or 48 hr after exposure to Aβ₂₅₋₃₅, a fragment of Aβ with an equivalent neurotoxic effect. Aβ₂₅₋₃₅ induced neuronal death in concentration- and time- dependent manners with apoptotic features. Neuronal death was significantly attenuated, not only by anti-apoptotic drugs, such as z-VAD-fmk and cycloheximide, but also by antioxidants, such as trolox, ascorbic acid, and epigallocatethin gallate. We also demonstrated that treatment with 20 µM Aβ₂₅₋₃₅ increased fluorescent signals in mixed cortical cultures, but produced only weak signals in pure astrocyte cultures in the presence of 2',7'-dichlorofluorescin diacetate (DCF-DA), an indicator for intracellular ROS. Increased DCF-DA fluorescence was markedly inhibited, not only by trolox, but also by selective NOX inhibitors, such as apocynin and AEBSF. Western blot analyses revealed that Aβ₂₅₋₃₅ increased the expression of gp91phox, a main subunit of NOX in cells. The above antioxidants, apocynin, and AEBSF significantly attenuated neuronal death induced by Aβ₂₅₋₃₅. Furthermore, the gp91phox-specific siRNA-based knockdown of NOX significantly inhibited neuronal death. These results suggest that activation of neuronal NOX is involved in Aβ25-35-induced neuronal death.

NADPH Oxidase Mediates β-Amyloid Peptide-Induced Neuronal Death in Mouse Cortical Cultures / 전남의대학술지

β-Amyloid peptide (Aβ) is the main component of senile plaques in patients with Alzheimer's disease, and is known to be a main pathogenic factor of the disease. Recent evidence indicates that activation of NADPH oxidase (NOX) in microglia or astrocytes may be a source of Aβ-induced reactive oxygen species (ROS). We investigated the role of neuronal NOX in Aβ-induced neuronal death in mouse mixed cortical cultures. Cell death was assessed by measuring lactate dehydrogenase efflux to bathing media 24 or 48 hr after exposure to Aβ₂₅₋₃₅, a fragment of Aβ with an equivalent neurotoxic effect. Aβ₂₅₋₃₅ induced neuronal death in concentration- and time- dependent manners with apoptotic features. Neuronal death was significantly attenuated, not only by anti-apoptotic drugs, such as z-VAD-fmk and cycloheximide, but also by antioxidants, such as trolox, ascorbic acid, and epigallocatethin gallate. We also demonstrated that treatment with 20 µM Aβ₂₅₋₃₅ increased fluorescent signals in mixed cortical cultures, but produced only weak signals in pure astrocyte cultures in the presence of 2',7'-dichlorofluorescin diacetate (DCF-DA), an indicator for intracellular ROS. Increased DCF-DA fluorescence was markedly inhibited, not only by trolox, but also by selective NOX inhibitors, such as apocynin and AEBSF. Western blot analyses revealed that Aβ₂₅₋₃₅ increased the expression of gp91phox, a main subunit of NOX in cells. The above antioxidants, apocynin, and AEBSF significantly attenuated neuronal death induced by Aβ₂₅₋₃₅. Furthermore, the gp91phox-specific siRNA-based knockdown of NOX significantly inhibited neuronal death. These results suggest that activation of neuronal NOX is involved in Aβ25-35-induced neuronal death.

Cytotoxic Effects of Gallic Acid and its Derivatives Against HIV-I-infected Microglia

In the previous study, we found that flavonoids and ginsenosides exhibited high eliminate rates of human immunodeficiency virus type 1 (HIV-1) D3-transfected macrophages. Based on these findings, here we synthesized the derivatives of gallic acid, including methyl gallate, methyl 4-O-methyl gallate, methyl 3,4-O-dimethyl gallate, and methyl 3,4,5-O-trimethyl gallate and measured their cellular toxic effects against HIV-1-infected macrophages. Of these, treatment with methyl 4-O-methyl gallate in the presence of lipopolysaccharide (LPS) and cycloheximide (CHX) most effectively eliminated HIV-1-transfected cytoprotective human microglial CHME5 cells and HIV-1-D3-infected human primary macrophages. Furthermore, these strongly inhibited LPS/CHX-induced phosphorylation of phosphoinositide 3-kinase (PI3K), pyruvate dehydrogenase lipoamide kinase isozyme 1 (PDK1), Akt, and glycogen synthase kinase-3β (GSK-3β) in the Tat-transfected cells and HIV-1-D3-infected human primary macrophages. These findings suggest that methyl 4-O-methyl gallate may be a promising candidate for eliminating HIV-1 infected macrophages by blocking PI3K/Akt signaling pathway.

Brief low Mg2+o-induced Ca2+ spikes inhibit subsequent prolonged exposure-induced excitotoxicity in cultured rat hippocampal neurons

Reducing [Mg2+]o to 0.1 mM can evoke repetitive [Ca2+]i spikes and seizure activity, which induces neuronal cell death in a process called excitotoxicity. We examined the issue of whether cultured rat hippocampal neurons preconditioned by a brief exposure to 0.1 mM [Mg2+]o are rendered resistant to excitotoxicity induced by a subsequent prolonged exposure and whether Ca2+ spikes are involved in this process. Preconditioning by an exposure to 0.1 mM [Mg2+]o for 5 min inhibited significantly subsequent 24 h exposure-induced cell death 24 h later (tolerance). Such tolerance was prevented by both the NMDA receptor antagonist D-AP5 and the L-type Ca2+ channel antagonist nimodipine, which blocked 0.1 mM [Mg2+]o-induced [Ca2+]i spikes. The AMPA receptor antagonist NBQX significantly inhibited both the tolerance and the [Ca2+]i spikes. The intracellular Ca2+ chelator BAPTA-AM significantly prevented the tolerance. The nonspecific PKC inhibitor staurosporin inhibited the tolerance without affecting the [Ca2+]i spikes. While Go6976, a specific inhibitor of PKCalpha had no effect on the tolerance, both the PKCepsilon translocation inhibitor and the PKCzeta pseudosubstrate inhibitor significantly inhibited the tolerance without affecting the [Ca2+]i spikes. Furthermore, JAK-2 inhibitor AG490, MAPK kinase inhibitor PD98059, and CaMKII inhibitor KN-62 inhibited the tolerance, but PI-3 kinase inhibitor LY294,002 did not. The protein synthesis inhibitor cycloheximide significantly inhibited the tolerance. Collectively, these results suggest that low [Mg2+]o preconditioning induced excitotoxic tolerance was directly or indirectly mediated through the [Ca2+]i spike-induced activation of PKCepsilon and PKCxi, JAK-2, MAPK kinase, CaMKII and the de novo synthesis of proteins.

Clinical Study of Onychomycosis Observed in Children (2005~2014) / 대한피부과학회지

BACKGROUND: Pediatric onychomycosis has been previously investigated; however, the specific causative agents of onychomycosis in Korean children have not been reported. OBJECTIVE: This study aimed to determine the most common causative agents of onychomycosis in Korean children. METHODS: We reviewed the medical records of 149 pediatric patients (<18 years of age) referred for fungal cultures because of a clinical suspicion of onychomycosis between 2005 and 2014 at our clinic. Patient specimens were cultured on Sabouraud's dextrose agar with and without cycloheximide. RESULTS: Onychomycosis was clinically suspected in 149 children. Of the 44 patients with onychomycosis, confirmed by culture, 72.7% had toenail onychomycosis, 22.7% had fingernail onychomycosis, and 4.5% had toenail and fingernail onychomycosis. The male-to-female patient ratio was 1.93:1. Fourteen (31.8%) children had concomitant tinea pedis, and 12 (27.2%) had family members with tinea pedis or onychomycosis. Distal and lateral subungual onychomycosis were the most common (68%) clinical types. Trichophyton rubrum was the most frequently isolated pathogen (66.7%), followed by Candida albicans (14.8%), Microsporum canis (11.1%), Candida parapsilosis (3.7%), and Candida tropicalis (3.7%). Candida albicans was the most commonly isolated pathogen (50.0%) in fingernail onychomycosis. CONCLUSION: Pediatric onychomycosis is more common than most people think. Thus, we suggest the need for a careful mycological examination of children with suspected onychomycosis.

Neck circumference as a potential marker of metabolic syndrome among college students / Circunferência do pescoço como possível marcador para síndrome metabólica em universitários / La circunferencia del cuello como posible indicador del síndrome metabólico en universitarios

OBJECTIVE: to relate neck circumference with metabolic syndrome and its criteria among college students. METHOD: cross-sectional study conducted with 702 college students in Fortaleza, CE, Brazil from September 2010 to June 2011. Socio-demographic data, waist circumference and neck circumference were collected together with blood pressure, fasting blood sugar, triglyceride levels, and HDL-C. RESULTS: 1.7% of the studied sample presented metabolic syndrome. Of these, 58.3% presented altered neck circumference (p<0.006). As neck circumference decreases, pressure levels improve (p<0.001). Additionally, college students with high fasting blood sugar (p=0.003) and high triglyceride levels (p<0.001) presented higher values of neck circumference. CONCLUSION: neck circumference is a potential predictive marker in the detection of metabolic syndrome and its components among college students. .

OBJETIVO: relacionar a circunferência do pescoço com a síndrome metabólica e seus critérios em universitários. MÉTODO: estudo transversal, realizado com 702 universitários de Fortaleza, CE, Brasil, no período de setembro de 2010 a junho de 2011. Coletaram-se dados sociodemográficos, circunferência da cintura, circunferência do pescoço, níveis de pressão arterial e glicemia plasmática de jejum, triglicerídeos e lipoproteína de alta densidade. RESULTADOS: 1,7% da amostra investigada tinha a síndrome metabólica. Desses, 58,3% apresentaram circunferência do pescoço alterada (p<0,006). Na medida em que decresce a circunferência do pescoço, os valores pressóricos dos universitários melhoram (p<0,001). Também, observou-se que universitários com valores de glicemia de jejum plasmática (p=0,003) e triglicerídeos (p<0,001) elevados apresentaram maiores valores de circunferência do pescoço. CONCLUSÃO: a circunferência do pescoço mostrou-se um possível marcador preditivo para detecção da síndrome metabólica e seus componentes em universitários. .

OBJETIVO: relacionar la circunferencia del cuello con el síndrome metabólico y sus criterios en universitarios. MÉTODO: estudio transversal realizado con 702 universitarios de Fortaleza-CE, Brasil, en el período de septiembre de 2010 a junio de 2011. Se recolectaron datos sociodemográficos, circunferencia de la cintura, circunferencia del cuello, niveles de presión arterial y glucemia plasmática de ayuno, triglicéridos y HDL-C. RESULTADOS: 1,7% de la muestra investigada tenían el síndrome metabólico. De estos, 58,3% presentaron circunferencia del cuello alterada (p<0,006). A medida que decrece la circunferencia del cuello mejoran los valores de la presión de los universitarios (p<0,001). También, se observó que los universitarios con valores de glucemia de ayuno plasmática (p=0,003) y triglicéridos (p<0,001) elevados presentaron mayores valores de circunferencia del cuello. CONCLUSIÓN: la circunferencia del cuello se mostró un posible indicador de predicción para la detección del síndrome metabólico y sus componentes, en universitarios. .

Compounds used to produce cloned animals are genotoxic and mutagenic in mammalian assays in vitro and in vivo

The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.

Protective Effects of Inducible HO-1 on Oxygen Toxicity in Rat Brain Endothelial Microvessel Cells

BACKGROUND: Reperfusion in ischemia is believed to generate cytotoxic oxidative stress, which mediates reperfusion injury. These stress conditions can initiate lipid peroxidation and damage to proteins, as well as promote DNA strand breaks. As biliverdin and bilirubin produced by heme oxygenase isoform 1 (HO-1) have antioxidant properties, the production of both antioxidants by HO-1 may help increase the resistance of the ischemic brain to oxidative stress. In the present study, the survival effect of HO-1 was confirmed using hemin. METHODS: To confirm the roles of HO-1, carbon monoxide, and cyclic guanosine monophosphate further in the antioxidant effect of HO-1 and bilirubin, cells were treated with cycloheximide, desferoxamine, and zinc deuteroporphyrin IX 2,4 bis glycol, respectively. RESULTS: HO-1 itself acted as an antioxidant. Furthermore, iron, rather than carbon monoxide, was involved in the HO-1-mediated survival effect. HO-1 activity was also important in providing bilirubin as an antioxidant. CONCLUSION: Our results suggested that HO-1 helped to increase the resistance of the ischemic brain to oxidative stress.

A Clinical and Mycological Study of Onychomycosis in the Elderly Over 10 Years (2001-2010) / 대한의진균학회지

BACKGROUND: As the life expectancy has risen globally because of the advance of medicine, onychomycosis in the elderly has been increasing with higher concerns over nails. Onychomycosis has been studied quite extensively, however, few reports on onychomycosis in a geriatric Korean population have been available. OBJECTIVE: The purpose of this study was to investigate the clinical features of onychomycosis in the elderly compared with other age groups and to identify the etiological agents during 10-year period. METHODS: A total of 629 patients over 65 years of age had been diagnosed with onychomycosis during a 10-year period (2001-2010). The etiological agents were identified by cultures on Sabouraud's dextrose agar with and without cycloheximide. Nondermatophytic molds and yeasts were considered as pathogens, if the identical fungal elements were observed at the initial direct microscopy and repeatedly in specimen-yielding cultures at a follow-up visit. RESULTS: The 629 elderly patients represented 22.1% of all onychomycosis patients. Toenails were involved in 567 (90.1%) patients; fingernails in 39 (6.2%); both toenails and fingernails in 23 (3.7%). The ratio of male to female was 1.01:1. Associated systemic diseases were found in 327 (52.0%) cases. Distal and lateral subungual onychomycosis (80.2%) was the most common clinical type of onychomycosis, followed by TDO (10.7%), SWO (6.2%) and PSO (2.9%). TDO was increasing significantly in the elderly. Organisms causing onychomycosis were dermatophytes (76.5%), yeasts (14.3%) and nondermatophytic molds (9.2%). The most common cause of onychomycosis in the elderly was Trichophyton rubrum. Nondermatophytic molds were more frequently responsible for onychomycosis in the elderly. CONCLUSION: Onychomycosis has been increased in the elderly and there are many differences from other age groups in aspects of clinical features, associated diseases and etiologic agents. Therefore, we suggest the need of a careful mycological examination in the elderly patients with onychomycosis.

Application of chemical activation to in vitro fertilization by round spermatid injection in mice / 中华男科学杂志

<p><b>OBJECTIVE</b>To search for an optimal activation protocol by comparing the chemical activation effects of single-activator and combined activation protocols on mouse oocytes following injection of round spermatids (ROSI) from spermatogenic cells cultured in vitro.</p><p><b>METHODS</b>Using different concentrations of ethanol, ionomycin (Ion), calcium ionophore A23187 (CIA), strontium chloride (SrCl2), cycloheximide (CHX), and 6-dimethylaminopurine (6-DMAP) , we activated post-ROSI oocytes for different times, and activated them by combined protocols at optimal concentrations and action times according to different activation channels. We compared the activation effects of single-activator and combined activation protocols by comparing the rates of fertilization, cleavages, and morulas and blastocysts.</p><p><b>RESULTS</b>With a single activator, the optimal protocols of different activators were as follows: 7% ethanol for 6 min, 5 micromol/L CIA for 5 min, 5 micromol/L Ion for 5 min, 2 mmol/L 6-DMAP for 2 h, 10 mmol/L SrCl2 for 1.5 h, and 10 microg/ml CHX for 1.5 h, among which 10 mmol/L SrCl2 for 1.5 h achieved the highest rate of morulas and blastocysts, significantly better than CHX (P < 0.05) but with no remarkable difference from other activators. The ethanol + 6-DMAP group showed a significantly higher rate of morulas and blastocysts (29.63%) than all other combined activation groups and single-activator groups except SrCl2 (P < 0.05), and it also exhibited higher rates of normal fertilization, cleavages and morula than the SrCl2 group, but with no significant difference.</p><p><b>CONCLUSION</b>The single-activator 10 mmol/L SrCl2 for 1.5 h and the combined activation of 7% ethanol for 6 min + 2 mmol/L 6-DMAP for 2 h are the optimal protocols for chemical activation of mouse oocytes following ROSI, and the combined activation of ethanol + 6-DMAP is even superior to the single-activator protocol.</p>

Onychomycosis in Children: An Experience of 59 Cases

BACKGROUND: Although tinea unguium in children has been studied in the past, no specific etiological agents of onychomycosis in children has been reported in Korea. OBJECTIVE: The purpose of this study was to investigate onychomycosis in Korean children. METHODS: We reviewed fifty nine patients with onychomycosis in children (0~18 years of age) who presented during the ten-year period between 1999 and 2009. Etiological agents were identified by cultures on Sabouraud's dextrose agar with and without cycloheximide. An isolated colony of yeasts was considered as pathogens if the same fungal element was identified at initial direct microscopy and in specimen-yielding cultures at a follow-up visit. RESULTS: Onychomycosis in children represented 2.3% of all onychomycosis. Of the 59 pediatric patients with onychomycosis, 66.1% had toenail onychomycosis with the rest (33.9%) having fingernail onychomycosis. The male-to-female ratio was 1.95:1. Fourteen (23.7%) children had concomitant tinea pedis infection, and tinea pedis or onychomycosis was also found in eight of the parents (13.6%). Distal and lateral subungual onychomycosis was the most common (62.7%) clinical type. In toenails, Trichophyton rubrum was the most common etiological agent (51.3%), followed by Candida albicans (10.2%), C. parapsilosis (5.1%), C. tropicalis (2.6%), and C. guilliermondii (2.6%). In fingernails, C. albicans was the most common isolated pathogen (50.0%), followed by T. rubrum (10.0%), C. parapsilosis (10.0%), and C. glabrata (5.0%). CONCLUSION: Because of the increase in pediatric onychomycosis, we suggest the need for a careful mycological examination of children who are diagnosed with onychomycosis.

Onychomycosis in Children: An Experience of 59 Cases

BACKGROUND: Although tinea unguium in children has been studied in the past, no specific etiological agents of onychomycosis in children has been reported in Korea. OBJECTIVE: The purpose of this study was to investigate onychomycosis in Korean children. METHODS: We reviewed fifty nine patients with onychomycosis in children (0~18 years of age) who presented during the ten-year period between 1999 and 2009. Etiological agents were identified by cultures on Sabouraud's dextrose agar with and without cycloheximide. An isolated colony of yeasts was considered as pathogens if the same fungal element was identified at initial direct microscopy and in specimen-yielding cultures at a follow-up visit. RESULTS: Onychomycosis in children represented 2.3% of all onychomycosis. Of the 59 pediatric patients with onychomycosis, 66.1% had toenail onychomycosis with the rest (33.9%) having fingernail onychomycosis. The male-to-female ratio was 1.95:1. Fourteen (23.7%) children had concomitant tinea pedis infection, and tinea pedis or onychomycosis was also found in eight of the parents (13.6%). Distal and lateral subungual onychomycosis was the most common (62.7%) clinical type. In toenails, Trichophyton rubrum was the most common etiological agent (51.3%), followed by Candida albicans (10.2%), C. parapsilosis (5.1%), C. tropicalis (2.6%), and C. guilliermondii (2.6%). In fingernails, C. albicans was the most common isolated pathogen (50.0%), followed by T. rubrum (10.0%), C. parapsilosis (10.0%), and C. glabrata (5.0%). CONCLUSION: Because of the increase in pediatric onychomycosis, we suggest the need for a careful mycological examination of children who are diagnosed with onychomycosis.

Quantitative in Vivo Detection of Brain Cell Death after Hypoxia Ischemia Using the Lipid Peak at 1.3 ppm of Proton Magnetic Resonance Spectroscopy in Neonatal Rats

This study was performed to determine the accuracy of proton magnetic spectroscopy (1H-MRS) lipid peak as a noninvasive tool for quantitative in vivo detection of brain cell death. Seven day-old Sprague Dawley rats were subjected to 8% oxygen following a unilateral carotid artery ligation. For treatment, cycloheximide was given immediately after hypoxic ischemia (HI). Lipid peak was measured using 1H-MRS at 24 hr after HI, and then brains were harvested for fluorocytometric analyses with annexin V/propidium iodide (PI) and fluorescent probe JC-1, and for adenosine-5'-triphosphate (ATP) and lactate. Increased lipid peak at 1.3 ppm measured with 1H-MRS, apoptotic and necrotic cells, and loss of mitochondrial membrane potential (DeltaPsi) at 24 hr after HI were significantly improved with cycloheximide treatment. Significantly reduced brain ATP and increased lactate levels observed at 24 hr after HI showed a tendency to improve without statistical significance with cycloheximide treatment. Lipid peak at 1.3 ppm showed significant positive correlation with both apoptotic and necrotic cells and loss of DeltaPsi, and negative correlation with normal live cells. Lipid peak at 1.3 ppm measured by 1H-MRS might be a sensitive and reliable diagnostic tool for quantitative in vivo detection of brain cell death after HI.

A Clinico-mycological Study of Onychomycosis with Dermatophytoma / 대한의진균학회지

BACKGROUND: Although there have been many studies about onychomycosis, the study about onychomycosis with dermatophytoma has not been reported yet in Korea. OBJECTIVE: The purpose of this study was to investigate the clinical characterictics and treatment strategies of the onychomycosis with dermatophytoma compare with the other onychomycosis. METHODS: In the 5-year period 2007-2011, we reviewed forty five patients with toenail onychomycosis with dermatophytoma, proven by direct potassium hydroxide examination. The etiological agents were identified by cultures on Sabouraud's dextrose agar with and without cycloheximide. To confirm dermatophytoma, we performed histopathologic evaluation of the nail plate by nail clipping. RESULTS: Toenail onychomycosis with dermatophytoma were 2.9% of all onychomycosis. Among the age groups, the incidence rate was highest in the sixties (24.4%). The ratio of male to female patients was 1:1.1. The frequency of associated disease was highest in diabetes mellitus (17.7%). The right great toenail was most common affected nails. Distal and lateral subungual onychomycosis (88.9%) was the most common clinical type. The round lesion was most common clinical features of affected area (66.7%), followed by linear lesion (33.3%). Trichophyton rubrum was most common etiological agent (57.8%). The partial removal of the tonail combined with oral and topical antifungal agent was most common in treatment of onychomycosis with dermatophytoma. CONCLUSION: Because of the increase in onychomycosis with dermatophytoma, and its relative resistance to the conventional treatment of onychomycosis, we suggest the need of a careful mycological examination to diagnose the dermatohpytoma in patients with onychomycosis, and we also propose more aggressive treatment strategy is required to treat dermatophytoma.

Onychomycosis Caused by Chaetomium globosum

Onychomycosis is usually caused by dermatophytes, but some nondermatophytic molds and yeasts are also associated with invasion of nails. The genus Chaetomium is a dematiaceous nondermatophytic mold found in soil and plant debris as a saprophytic fungus. We report the first Korean case of onychomycosis caused by Chaetomium globosum in a 35-year-old male. The patient showed brownish-yellow discoloration and subungual hyperkeratosis on the right toenails (1st and 5th) and left toenails (1st and 4th). Direct microscopic examination of scraping on the potassium hydroxide preparation revealed septate hyphae and repeated cultures on Sabouraud's dextrose agar (SDA) without cycloheximide slants showed the same fast-growing colonies, which were initially velvety white then turned to dark gray to brown. However, there was no growth of colony on SDA with cycloheximide slants. Brown-colored septated hyphae, perithecia and ascospores were shown in the slide culture. The DNA sequence of internal transcribed spacer region of the clinical sample was a 100% match to that of C. globosum strain ATCC 6205 (GenBank accession number EF524036.1). We confirmed C. globosum by KOH mount, colony, and light microscopic morphology and DNA sequence analysis. The patient was treated with 250 mg oral terbinafine daily and topical amorolfine 5% nail lacquer for 3 months.

Contamination of Slippers in Operating Room by Dermatophytes and Effect of Antifungal Agent (Terbinafine) Disinfection / 대한의진균학회지

BACKGROUND: It is known that floor, clothes, shoes and slippers of common uses are the sources of infection by dermatophytes. However there hasn't been any report about the culture of dermatopytes from slippers in operating room. OBJECTIVE: The purpose of this study was to evaluate the contamination status of the slippers in operating room by dermatophytes and the effect of antifungal agent disinfection. METHODS: The samples were collected from 240 pairs of slippers that were used in common at operating room of Daegu Catholic University Medical Center with scrapping method. The collected samples were cultured on the media with chloramphenicol (500 mg/L) and cycloheximide (500 mg/L) to control the growth of nondermatophytic fungi. The same collection and culture was done again after the antifungal agent (terbinafine) disinfection. RESULTS: Dermatophytes were isolated from 22 (9.2%) pairs of slippers from a total of 240 pairs before the treatment, and 9 (3.8%) pairs after the treatment. There was significant difference in isolation rate between the slippers before the treatment and after the treatment (p=0.016). Trichophyton (T.) mentagrophytes and T. rubrum were isolated from the slippers and T. mentagrophytes (54.8%) was the most common isolated fungus. CONCLUSION: About ten percent of slippers in operation room were contaminated by dermatophytes. Regular antifungal agent disinfection on slippers in operating room will help to decrease in the prevalence of dermatophytes growth and prevent the nosocomial infection.