OBJECTIVE: To determine the effects of extracellular ATP on mitochondrial function and apoptosis during human luteinized granulosa cell cultures. METHODS: The addition of various concentrations of ATP (0, 0.1, 0.25, 0.5, 0.75 mM) to luteinized granulosa cells obtained during In vitro fertilization ovum pickup procedures. After culture for 24 hours, purinoceptor activity and functional changes in mitochondria were measured by patch clamp, flow cytometry, and confocal microscopy. RESULTS: Measurement by patch clamp of the granulosa cell membrane potential after ATP addition to cultured granulosa cells showed that both the inward and outward currents were expressed. After treatment of the granulosa cells with JC-1, measurement of the mitochondrial activity by confocal microscopy showed that the with increasing concentrations of ATP the relative ratio of undamaged mitochondria (red/green ratio) tended to decrease (P=0.027). After double staining of the cultured granulosa cells with Annexin V and Propidium Iodide, quantitative flow cytometry analysis showed that apoptosis increased with increasing concentrations of ATP (7.88%, 8.44%, 11.40%, 13.52%, 18.57%). CONCLUSION: The results of this study shows that apoptosis of granulosa cells increases with increasing extracellular ATP concentrations in cultured human luteinized granulosa cells. This is observed to be a consequence of cell membrane purinoceptor activity and functional changes in the mitochondria. It is therefore thought that remodelling processes of the ovarian tissue is regulated by neuroendocrine factors of the extracellular ATP.
Adenosine Triphosphate ; Annexin A5 ; Apoptosis* ; Cell Membrane ; Female ; Fertilization in Vitro ; Flow Cytometry ; Granulosa Cells* ; Humans* ; Lutein* ; Membrane Potentials ; Microscopy, Confocal ; Mitochondria* ; Ovum ; Propidium ; Receptors, Purinergic

BACKGROUND: The intravenous administration of indigo carmine has been reported to produce transiently increased blood pressure in patients. The goal of this in vitro study was to examine the effect of indigo carmine on phenylephrine-induced contractions in an isolated rat aorta and to determine the associated cellular mechanism with particular focus on the endothelium-derived vasodilators. METHODS: The concentration-response curves for phenylephrine were generated in the presence or absence of indigo carmine. Phenylephrine concentration-response curves were generated for the endothelium-intact rings pretreated independently with a nitric oxide synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME), a cyclooxygenase inhibitor, indomethacin, and a low-molecular-weight superoxide anion scavenger, tiron, in the presence or absence of indigo carmine. The fluorescence of oxidized dichlorofluorescein was measured in rat aortic vascular smooth muscle cells cultured in the control, indigo carmine alone and tiron plus indigo carmine. RESULTS: Indigo carmine (10(-5) M) increased the phenylephrine-induced maximum contraction in the endothelium-intact rings with or without indomethacin, whereas indigo carmine produced a slight leftward shift in the phenylephrine concentration-response curves in the endothelium-denuded rings and L-NAME-pretreated endothelium-intact rings. In the endothelium-intact rings pretreated with tiron (10(-2) M), indigo carmine did not alter phenylephrine concentration-response curves significantly. Indigo carmine (10(-5) M) increased the fluorescence of oxidized dichlorofluorescein in the vascular smooth muscle cells, whereas tiron abolished the indigo carmine-induced increase in oxidized dichlorofluorescein fluorescence. CONCLUSIONS: Indigo carmine increases the phenylephrine-induced contraction mainly through an endothelium-dependent mechanism involving the inactivation of nitric oxide caused by the increased production of reactive oxygen species.
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt ; Administration, Intravenous ; Animals ; Aorta ; Blood Pressure ; Contracts ; Fluorescence ; Humans ; Indigo Carmine ; Indoles ; Indomethacin ; Muscle, Smooth, Vascular ; Nitric Oxide ; Nitric Oxide Synthase ; Phenylephrine ; Prostaglandin-Endoperoxide Synthases ; Rats ; Reactive Oxygen Species ; Superoxides

OBJECTIVE: Attention deficit hyperactivity disorder (ADHD) has a strong genetic basis, and the relationship between the allele frequency distribution and ADHD has been researched actively. We investigated the association between the DRD4 genotype and ADHD patients in Korea, in comparison with case control and family based control groups. METHODS: The study enrolled 118 patients diagnosed with ADHD according to the DSM-IV diagnostic criteria and their parents (n = 70), and 84 normal children were recruited as controls. The clinical features of the patients were confirmed using the Korean versions of the Child Behavior Checklist (CBCL), Conners' parent rating scale, Attention Deficit Diagnostic System, and Spielberger state/trait anxiety scale. Blood samples were taken from the 272 subjects. DNA was extracted from blood lymphocytes. PCR was performed to examine DRD4 polymorphisms. Allele and genotype frequencies were compared using the Chi-square test for the casecontrol analysis and the transmission disequilibrium test (TDT) for the family-based analyses. RESULTS: In comparing the ADHD transmitted group with the not transmitted group, no significant differences were seen in the DRD4 genotype, allele distribution, or ADHD. CONCLUSION: However, there was a trend to an association between the DRD4 genotype, allele distribution, and ADHD for the case-control analysis. Follow-up studies with more patients or pure subgroups are needed.
Alleles ; Anxiety ; Attention Deficit Disorder with Hyperactivity ; Case-Control Studies ; Checklist ; Child ; Child Behavior ; Diagnostic and Statistical Manual of Mental Disorders ; DNA ; Gene Frequency ; Genotype ; Humans ; Korea ; Lymphocytes ; Parents ; Polymerase Chain Reaction

OBJECTIVES: The aim of the current study is to test for the association of DRD4 exon III VNTR polymorphism with attention-deficit hyperactivity disorder(ADHD) in Korean children population. METHODS: 113 Korean children with ADHD, 102 parents, and 133 control subjects participated with the current study. The distribution of genotypes and alleles of DRD4 exon III VNTR in children with ADHD was compared with that in control subjects. In addition, 69 children with ADHD and their parents were analyzed using the transmission disequilibrium test(TDT). RESULTS: We could not find any significant differences in the distribution of genotypes and alleles at DRD4 exon III VNTR polymorphism between children with ADHD and control subjects. In addition, there was no preferential transmission of long allele of DRD4 exon III VNTR polymorphism. CONCLUSION: These results suggest that DRD4 is not associated with ADHD in Korean children population.
Alleles ; Child* ; Exons* ; Genetic Association Studies ; Genotype ; Humans ; Parents

OBJECTIVE: The purpose of this study was to investigate the association between the T102C polymorphism in the serotonin 2A receptor gene and attention-deficit/hyperactivity disorder (ADHD) in Korean patients. METHODS: A total of 189 Korean children with ADHD as well as both parents of the ADHD children and 150 normal children participated in this study. DNA was extracted from blood samples from all of the subjects, and genotyping was conducted. Based on the allele and genotype information obtained, case-control analyses were performed to compare the ADHD and normal children, and Transmission disequilibrium tests (TDTs) were used for family-based association testing (number of trios=113). Finally, according to the significant finding which was showed in the case-control analyses, the results of behavioral characterastics and neuropsychological test were compared between ADHD children with and without the C allele. RESULTS: In the case-control analyses, statistically significant differences were detected in the frequencies of genotypes containing the C allele (chi2=4.73, p=0.030). In the family-based association study, TDTs failed to detect linkage disequilibrium of the T102C polymorphism associated with ADHD children. In the ADHD children, both the mean reaction time and the standard deviation of the reaction time in the auditory continuous performance test were longer in the group with the C allele compared to the group without the C allele. CONCLUSION: The results of this study suggest that there is a significant genetic association between the T102C polymorphism in the serotonin 2A receptor gene and ADHD in Korean children.
Alleles ; Case-Control Studies ; Child ; DNA ; Genotype ; Humans ; Linkage Disequilibrium ; Neuropsychological Tests ; Parents ; Reaction Time ; Receptor, Serotonin, 5-HT2A ; Serotonin

OBJECTIVES : We investigated the relationship between periventricular and deep white matter hyperintensity and cognitive function in Alzheimer's disease (AD) and mild cognitive impairment (MCI). METHODS : T2-weighted MRI scans were performed in 41 subjects with AD 38 subjects with mild cognitive impairment and 38 control subjects. Periventricular and deep white matter hyperintensities were rated on a Fazekas 0-3 scale by a medical specialist of the department of radiology blind to clinical diagnosis. Cognitive function was assessed by using Cognitive Assessment and Reference Diagnoses System. RESULTS : No significant differences between demographic characteristics and vascular risk factors were revealed comparing AD, MCI and controls. The frequencies of AD were significantly higher than those of MCI and normal control in Grade 2 and 3 of periventricular hyperintensity and Grade 3 of deep white matter hyperintensity. The scores of amnesia, executive function and attention were significantly lower in Grade 2 and 3 of periventricular hyperintensity than in Grade 0 and 1. The scores of attention were significantly lower in Grade 3 of deep white matter hyperintensity than in Grade 0, 1 and 2. CONCLUSION : Periventricualr hyperintensities are associated with cognitive decline in amnesia, executive function and attention, while deep white matter hyperintensities are associated with cognitive decline in attention.
Alzheimer Disease ; Amnesia ; Executive Function ; Humans ; Magnetic Resonance Imaging ; Mild Cognitive Impairment ; Risk Factors ; Specialization

OBJECTIVES : We investigated the relationship between periventricular and deep white matter hyperintensity and cognitive function in Alzheimer's disease (AD) and mild cognitive impairment (MCI). METHODS : T2-weighted MRI scans were performed in 41 subjects with AD 38 subjects with mild cognitive impairment and 38 control subjects. Periventricular and deep white matter hyperintensities were rated on a Fazekas 0-3 scale by a medical specialist of the department of radiology blind to clinical diagnosis. Cognitive function was assessed by using Cognitive Assessment and Reference Diagnoses System. RESULTS : No significant differences between demographic characteristics and vascular risk factors were revealed comparing AD, MCI and controls. The frequencies of AD were significantly higher than those of MCI and normal control in Grade 2 and 3 of periventricular hyperintensity and Grade 3 of deep white matter hyperintensity. The scores of amnesia, executive function and attention were significantly lower in Grade 2 and 3 of periventricular hyperintensity than in Grade 0 and 1. The scores of attention were significantly lower in Grade 3 of deep white matter hyperintensity than in Grade 0, 1 and 2. CONCLUSION : Periventricualr hyperintensities are associated with cognitive decline in amnesia, executive function and attention, while deep white matter hyperintensities are associated with cognitive decline in attention.
Alzheimer Disease ; Amnesia ; Executive Function ; Humans ; Magnetic Resonance Imaging ; Mild Cognitive Impairment ; Risk Factors ; Specialization

PURPOSE: To evaluate the usefulness of CT scanning in determining the etiology of spontaneous rupture of the kidney. MATERIALS AND METHODS: We retrospectively analyzed the CT findings of spontaneous rupture of the kidney in eleven patients, Four were male and seven were female, and they were aged between 20 and 71 (mean, 46.6) years. Both pre- and post-contrast enhanced CT scanning was performed in all patients. RESULTS: Spontaneous renal rupture was induced in seven cases by neoplasms (three angiomyolipomas, three renal cell carcinomas, and one metastatic choriocarcinoma), in three cases by infection or inflammation (acute and chronic pyelonephritis, and renal abscess), and in one, by renal cyst. Common CT findings of rupture of the kidney were the accumulation of high density fluid in the perirenal and anterior pararenal space, and in homogeneous irregular low density of renal parenchyma and the rupture site. Angoimyolipoma showed fat and an angiomatous component in the lesion, while acute and chronic pyelonephrities revealed thinning of the renal parenchyma and an irregular renal outline. Renal cell carcinoma showed a dense soft tissue mass in the parenchyma. Well-defined, round low-density lesions were noted in the case of renal cyst and renal abscess. CONCLUSION: CT is very useful in diagnosing and determining the etiology of non-traumatic spontaneous rupture of the kidney and plays an important role in the evaluation of emergency cases.
Abscess ; Angiomyolipoma ; Carcinoma, Renal Cell ; Emergencies ; Female ; Humans ; Inflammation ; Kidney* ; Male ; Pyelonephritis ; Retrospective Studies ; Rupture ; Rupture, Spontaneous* ; Tomography, X-Ray Computed

OBJECTIVE: Arsenic trioxide (As2O3) is known to have potent anti-vascular activity and significantly suppress solid tumor growth. The present study was conducted to investigate the vascular shutdown effects of a novel arsenic compound, tetraasrsenic oxide (As4O6), in comparison with As2O3 using cervical cancer animal model. METHODS: Mice tumor challenge model was used C57BL/6 mice transplanted with TC-1 cells. After the growth of tumors was reached up 200~250 mm3, mice were divided into 3 groups randomly for control and treatment of either As2O3 or As4O6. As2O3 and As4O6 was treated by i.p. injection. The tumor size was caliperated in twice for weeks and anti-vascular effect were assessed by Evans blue extraction assay and Hoechst 33342 staining. In tumor tissue, histopathological feaure was obserevd by hematoxylin and eosin (H&E) staining. RESULTS: In mice treated with either As2O3 and As4O6 (i.p.), both of As2O3 and As4O6 was significantly suppressed the tumor growth compared with control group. Moreover, effect of As4O6 is more pronounced. These tumor growth inhibition is led to the massive necrosis and vacular shutdown in tumor tissue. CONCLUSION: This study suggests that As4O6 may have potential anticancer activity via vascular shutdown in C57BL/6 mice transplanted with TC-1 cells.
Animals ; Arsenic ; Arsenicals ; Benzimidazoles ; Eosine Yellowish-(YS) ; Evans Blue ; Hematoxylin ; Mice ; Models, Animal ; Necrosis ; Oxides ; Transplants ; Uterine Cervical Neoplasms

OBJECTIVE: Autophagy plays a vital role in homeostasis by combining organelles and cellular proteins with lysosome under starvation conditions. In addition, autophagy provides tumor cells with a source of energy. Continued autophagy will induce cells death. Here we aim to see if autophagic induction has an effect on conventional chemotherapeutic agents. METHODS: Rapamycin, or mammalian target of rapamycin and paclitaxel, apoptosis-inducing agents were used autophagy in HeLa cervical cancer cells. RESULTS: Growth inhibition of cells was not observed after the application of 0, 10, 20 nM of paclitaxel with or without rapamycin. Using a 5 nM concentration of paclitaxel, rapamycin administration inhibited cell growth significantly compared to no treatment. This implies the synergic antitumor effect of paclitaxel and rapamycin. Paclitaxel itself did not show any autophagic effect on cells but did show cell apoptosis by flow cytometry. Light chain 3, a microtubule-associated protein, which reflect autophagy, was increased with 5 nM of paclitaxel after pretreatment with 10 nM of rapamycin. CONCLUSION: These findings suggest that the autophagic inducer, rapamycin, can potentiate autophagic cell death when added as an apoptosis-inducing chemotherapeutic agent. In conclusion, the control of autophagy may be a future target for chemotherapy.
Apoptosis ; Autophagy ; Cell Death ; Flow Cytometry ; Homeostasis ; Light ; Lysosomes ; Organelles ; Paclitaxel ; Proteins ; Sirolimus ; Starvation ; TOR Serine-Threonine Kinases ; Uterine Cervical Neoplasms