BACKGROUND: Atherosclerosis is the most important disease that may cause ischemic syndrome in many organs including heart. It is supposed that apoptosis of vascular smooth muscle cells(VSMCs) is closely related to the progression and rupture of atheromatous plaque. Recent studies have documented evidence for elevated level of nitric oxide(NO) within advanced human atheroma and evidence of regression of atheroma by NO. So this study is designed to evaluate whether exogenous NO from NO donors can induce apoptosis of cultured rat VSMCs and which proapoptotic gene(s) is involved in this type of apoptosis. METHODS: Rat VSMCs were cultured and used for experiment at passage 5 through 7. For NO donor, sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine(SNAP) of 0.5, 1, 2, 4mM were exposed to subconfluent VSMCs. The cells were harvested at 6, 12, 24, 48, 72hours after exposure of NO donors. Apoptosis was to be identified by 4, 6-diamidino-2-phenylindole dihydrochloride(DAPI) staining of nuclei and in-situ nick end labeling(TUNEL). The amount of fragmented DNA was analyzed semiquantitatively by diphenylamine(DPA) assay. Immunocytochemical(ICC) staining and western bolt analyses were designed to detect apoptosisrelated gene products, such as Bax-a, Fas and Bcl-2. RESULTS: 1) Decreased mitotic activity was shown after 12 hours exposure of exogenous NO donors, and condensation and margination of chromatin was identified agter 24 hours exposure, by DAPI staining. 2) Percent DNA fragmentation assessed by DPA method was 0,2,9,48,45% at 0,6,12,24,48 hours after exposure of 2mM of NO donors respectively. 3) The expression of Bax-a and Bcl-2 proteins was demonstrated in apoptotic cells by ICC staining. 4) The expression of Bax-a protein in cells under 24 hours exposure of NO donors was elevated by more than 18% of control level on densitometric analysis of western blot. The level of Bcl-2 was suppressed by 26% of control. So, Bax-a/Bcl-2 ratio in cells under exposure of NO donors was elevated to 2.0 from 1.2 of control level. CONCLUSIONS: Exogenous NO from NO donors can induce apoptosis of cultured rat VSMCs, and it is considered that bax-a and bcl-2 genes are involved in this type of apoptosis.
Animals ; Apoptosis* ; Atherosclerosis ; Blotting, Western ; Chromatin ; DNA ; DNA Fragmentation ; Genes, bcl-2 ; Heart ; Humans ; Muscle, Smooth, Vascular* ; Nitric Oxide ; Nitroprusside ; Plaque, Atherosclerotic ; Rats* ; Rupture ; Tissue Donors

We previously reported that the p53 tumor suppressor protein plays an essential role in the induction of tetraploid G1 arrest in response to perturbation of the actin cytoskeleton, termed actin damage. In this study, we investigated the role of p53, ataxia telangiectasia mutated protein (ATM), and catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) in tetraploid G1 arrest induced by actin damage. Treatment with actin-damaging agents including pectenotoxin-2 (PTX-2) increases phosphorylation of Ser-15 and Ser-37 residues of p53, but not Ser-20 residue. Knockdown of ATM and DNA-PKcs do not affect p53 phosphorylation induced by actin damage. However, while ATM knockdown does not affect tetraploid G1 arrest, knockdown of DNA-PKcs not only perturbs tetraploid G1 arrest, but also results in formation of polyploidy and induction of apoptosis. These results indicate that DNA-PKcs is essential for the maintenance of actin damage induced-tetraploid G1 arrest in a p53-independent manner. Furthermore, actin damage-induced p53 expression is not observed in cells synchronized at G1/S of the cell cycle, implying that p53 induction is due to actin damage-induced tetraploidy rather than perturbation of actin cytoskeleton. Therefore, these results suggest that p53 and DNA-PKcs independently function for tetraploid G1 arrest and preventing polyploidy formation.
Actins/*metabolism ; Apoptosis ; Catalytic Domain ; Cell Cycle Proteins/genetics/*metabolism ; Cell Line ; Cell Line, Tumor ; DNA-Activated Protein Kinase/chemistry/genetics/*metabolism ; DNA-Binding Proteins/genetics/*metabolism ; Furans/pharmacology ; *G1 Phase ; Gene Knockdown Techniques ; Humans ; Phosphorylation/drug effects ; Protein-Serine-Threonine Kinases/genetics/*metabolism ; Pyrans/pharmacology ; Tumor Suppressor Protein p53/*metabolism ; Tumor Suppressor Proteins/genetics/*metabolism

We report a case of Strongyloides stercoralis infection in a 85-year-old male patient who had complained of poor oral intake, diarrhea, and upper abdominal pain for 6 months. At admission, he showed severe eosinophilia in peripheral blood. Rhabditiform larvae were detected in the stool examination on the 15th admission day and developed into filariform larvae with a notched tail after stool culture by the Harada-Mori method. The patient received albendazole therapy for 7 days but no improvement were observed and he fell into pulmonary edema and coma.
Abdominal Pain ; Aged, 80 and over ; Albendazole ; Coma ; Diarrhea ; Eosinophilia* ; Humans ; Larva ; Male ; Pulmonary Edema ; Strongyloides stercoralis* ; Strongyloides*

NF-Y transcription factor binds to CCAAT boxes on promoters of cell cycle regulatory genes such as cdc2, cyclin B, cdc25C, and cyclin A. We previously reported that the DNA binding activity of NF-Y is regulated by p53-p21-cdk2 pathway. CBF/HSP70 was originally identified as a transcription factor binding to the CCAAT box on the hsp70 promoter and mediates transcription repression of hsp70 pro- moter by p53. Recently it was demonstrated that CBF/HSP70 interacts and cooperates with NF-Y. In this study, we found that p53 represses the transcription of CBF/HSP70. Since transactivation ability of NF-Y is regulated in a cell cycle-dependent manner, we examined the transcription of CBF/HSP70 during the cell cycle. After synchronization of a human bladder carcinoma cell lacking functional p53 at early S phase, we infect the cells with adenovirus encoding p53. Cells infected with control virus progressed to S and G2 after release from the arrest. In contrast, cells expressing p53 enter S and G2 phases, but arrest at G2/M. The expression of CBF/HSP70 was induced at S/G2 phase in cells infected with a control virus, but kept to be repressed in cells expressing p53. Thus, these results suggest that p53 suppresses the expression of cell cycle regulatory genes though inhibiting both CCAAT binding factors, CBF/HSP70 and NF-Y.
CCAAT-Binding Factor/*metabolism ; Cell Cycle ; Cell Line, Tumor ; *Down-Regulation ; HSP70 Heat-Shock Proteins/*metabolism ; Humans ; Protein Binding ; Research Support, Non-U.S. Gov't ; *Transcription, Genetic ; Tumor Suppressor Protein p53/genetics/*metabolism

Oocyte in vitro maturation (IVM) is an assisted reproductive technology in which oocytes are retrieved from the antral follicles of unstimulated or minimally stimulated ovaries. IVM of human oocytes has emerged as a promising procedure. This new technology has advantages over controlled ovarian stimulation such as reduction of costs, simplicity, and elimination of ovarian hyperstimulation syndrome. By elimination or reduction of gonadotropin stimulation, IVM offers eligible infertile couples a safe and convenient form of treatment, and IVM outcomes are currently comparable in safety and efficacy to those of conventional in vitro fertilization. IVM has been applied mainly in patients with polycystic ovary syndrome or ultrasound-only polycystic ovaries, but with time, the indications for IVM have expanded to other uncommon situations such as fertility preservation, as well as to normal responders. In this review, the current clinical experiences with IVM will be described.
Family Characteristics ; Female ; Fertility Preservation ; Fertilization in Vitro ; Gonadotropins ; Humans ; In Vitro Oocyte Maturation Techniques ; Infertility ; Oocytes ; Ovarian Hyperstimulation Syndrome ; Ovary ; Ovulation Induction ; Polycystic Ovary Syndrome ; Reproductive Techniques, Assisted

BACKGROUND: Familial hypercholesterolemia(FH) is an autosomal dominant inharited disorder. Total cholesterl level of FH heterozygotes is two to fourfold higher than that of normal population. Substained hypercholesterolemia results in cholesterol deposition on various organs or tissues and Achilles tendon xanthoma due to cholesterol deposition is one of the specific clinical findings of FH. One of the lipid lowering drugs, 3-hydroxy-3-methylglutaryl coenzyme A(HMG Co-A) reductase inhibitor effectively lowers the blood cholesterol level in patients with FH, but whether the cholesterol deposition can be regressed by the lipid lowering drugs is rarely reported. This study attemted to determine whether the tendon xanthoma can be regressed by lipid lowering drugs commonly used in patients with FH. METHODS: We analyzed procepectively the serum lipid levels of patients with heterozygous FH before and after lipid lowering therapy with HMG Co-A reductase inhibitor alone(Lovastatin or Pravastatin) or in combination with bile acid sequestrating resin(Cholestyramine). The Achilles tendon thickness was measured radiographically by using soft tissue technique. RESULT: Total 18 patients with heterozygotes FH(M : F=8 :10, mean age; 51.7+/-9.0 years) were treated with the HMG Co-A reductase inhibitor alone or combined with bile acid sequestrating resin and followed for mean 31.9+/-11.9 months. During that period, serum total cholesterol and low density lipoprotein cholesterol significantly fell from 329+/-42 mg/dl to 230+/-29 mg/dl and from 246+/-56 mg/dl to 151+/-28 mg/dl, respectively(p<0.001). Serum high density lipoprotein level increased and maintained 15.3% higher than basal level(p<0.01). Achilles tendon thickness decreased significantly from 13.3+/-3.1 mm to 11.9+/-3.2 mm(p<0.001) with percent reduction of 9.8+/-10.5%(range; 3.1-36.4%). The amount of change of tendon thickness was significantly correlated only with percent reductionof LDL(p=0.029) and female sex(p0.020) on univariate analysis, but it was found to be significantly correlated only with percent reduction of LDL on multivariate analysis(r=0.514,p=0.029). CONCLUSION: Achilles tendon xanthoma can be regressed by effective lipid lowering therapy with HMG Co-A reductase inhibitor alone or with bile acid sequestrating resin in patients with heterozygous FH. the regression of tendon xanthoma is likely to be related to reduction of serum LDL.
Achilles Tendon* ; Bile* ; Cholesterol ; Cholesterol, LDL ; Female ; Heterozygote ; Humans ; Hypercholesterolemia ; Hyperlipoproteinemia Type II* ; Lipoproteins ; Oxidoreductases* ; Tendons ; Xanthomatosis*

PURPOSE: Iodine has been used for the prevention or management of wound infection as a topical agent. Although iodine was widely used mainly by Betadine(R) and cadexomer iodine, there was no comparative study on the efficacies of dressing methods of iodine. And also it's wound healing effect was not yet clear. The purpose of this study is to compare antibacterial effects and wound healing effects associated with various dressing methods of iodine on infected full thickness skin defect in the mouse. METHODS: One full thickness skin defects in the mice (n=60) were developed on the back and left open for twenty-four hours. Sixty mice were divided into four groups : group S(dressing with Betadine(R) soaking, n=15), group T (dressing with Betadine(R) topping, n=15), group I(dressing with Iodosorb(R), n=15), group G(control group, dressing with dry gauze, n=15). The size of the wound defects and the grades of wound healing were evaluated in 4, 7, 10 days, and antibacterial effect was evaluated with restricted zone in Mueller Hinton agar by disk diffusion method. RESULTS: After the wound was left open for twenty-four hours, many Staphylococcus aureus were cultured. The wound defect size was decreased in order of Betadine(R) soaking, Iodosorb(R), Betadine(R) topping and gauze dressing group in all days, but difference among experimental groups was not statistically significant. The grade score of wound healing was increased in order of Betadine(R) soaking, Iodosorb(R), Betadine(R) topping and gauze dressing group, and the difference was statistically significant. Antibacterial effect for S. aureus was increased in order of Iodosorb(R), Betadine(R) soaking, Betadine(R) topping and gauze dressing group, and the difference was statistically significant. CONCLUSION: Selection of the effective dressing method of iodine for infected wounds remains a controversial decision. According to this study, Iodosorb(R) may be most effective method for antibacterial effect and Betadine(R) soaking may be most effective method for infected wound healing. However, further study is necessary to evaluate the clinical efficacy of dressing methods of iodine and to search for the mechanisms that explain their effects.
Agar ; Animals ; Bandages ; Diffusion ; Iodine ; Iodophors ; Mice ; Skin ; Staphylococcus aureus ; Wound Healing ; Wound Infection

PURPOSE: Many topical agents had been used for contaminated wound treatment. Although antimicrobial ointments were widely used as topical agents, their comparative antibacterial and wound healing effects were largely unreported. The purpose of this study was to compare antibacterial effects and wound healing effects of common topical ointments on infected full thickness skin defect in the mouse. METHODS: One full thickness skin defects in the mice(n=60) were developed on the back and left open for twenty-four hours. Sixty mice were divided into four groups: group B(dressing with Bactroban(R), n=15), group I (dressing with Iodosorb(R), n=15), group T(dressing with Terramycin(R), n=15), group G(control group, dressing with dry gauze, n=15). The size of wound defects and the grades of wound healing were evaluated at 4, 7, 10 days, and antibacterial effect was evaluated with restricted zone in Mueller Hinton agar by disk diffusion method. RESULTS: After the wound was left open for twenty-four hours, many Staphylococcus aureus was cultured. The wound defect size was decreased in order of Bactroban(R)(B), Iodosorb(R)(I), Terramycin(R) and gauze dressing group in all days, but difference among experimental groups was not statistically significant. The grade score of wound healing was increased in order of Bactroban(R), Iodosorb(R), Terramycin(R) and gauze dressing group, and the difference was statistically significant. Antibacterial effect for was increased in order of Bactroban(R), Iodosorb(R), Terramycin(R) and gauze dressing group, and the difference was statistically significant. CONCLUSION: Topical antimicrobial ointments application was effective against wound infections by S. aureus. Bactroban(R) may be an optimal topical treatment for infected wounds according to this study. However, further study is necessary to evaluate the clinical efficacy of antimicrobial ointments and to search for the mechanisms that explain their effects.
Agar ; Animals ; Bandages ; Diffusion ; Mice ; Ointments ; Skin ; Staphylococcus aureus ; Wound Healing ; Wound Infection

PURPOSE: With increasing frequency, Yersinia enterocolitica is being recognized as an important bacterial cause of acute gastrointestinal infection with abdominal pain. In addition, the association of Y. enterocolitica infections with acute appendicitis has been suggested. This study was undertaken to ascertain whether Y. enterocolitica is a possible infectious agent in acute appendicitis. METHODS: Between December 2007 and April 2008, 162 patients who underwent appendectomy for presumed appendicitis, enrolled in this prospective study. After surgical excision of appendix, a portion of each specimen was cultured for Y. enterocolitica with highly selective media (Cefsulodin-Irgasan-Novobiocin agar). RESULTS: Pathologically, 150 of the patients had appendicitis and 12 patients had normal appendices. Only one of the 150 patients (0.7%) with appendicitis was found to be culture positive for Y. enterocolitica, while it was not detected from normal appendices. CONCLUSION: The authors were unable to implicate Y. enterocolitica as a major pathogen in acute appendicitis within the Seoul area. However, we thought there to be more need for investigation for association of Y. enterocolitica with acute appendicitis over a broader area and season.
Abdominal Pain ; Appendectomy ; Appendicitis ; Appendix ; Humans ; Prospective Studies ; Seasons ; Yersinia ; Yersinia enterocolitica

The effect of artificial oocyte activation (AOA) with a calcium ionophore on intracytoplasmic morphologically selected sperm injection (IMSI) was examined in patients with histories of repeated failed implantation attempts. Four singleton pregnancies and one twin pregnancy were obtained after embryos transfer (5/14, 35.7%). Therefore, AOA combined with IMSI can be considered an option for cycles with a fertilization defect and recurrent implantation failures.
Calcium ; Embryonic Structures ; Fertilization* ; Humans ; Oocytes ; Pregnancy ; Pregnancy, Twin ; Spermatozoa