No abstract available.
Emergencies* ; Emergency Service, Hospital* ; Hospitals, General* ; Humans

Nitric oxide (NO) has been emerged as an important intracellular and intercellular regulatory molecule having functions as diverse vasodilatation, neural communication, and host defense. In the immune system, NO produced by activated macrophage or neutrophil is known to kill tumor cells as a defense molecule. In addition, recent reports demonstrated that NO could interact with superoxide to generate peroxynitrite (ONOO-), an anion and a potent oxidant, in macrophages or other cellular systems. The production of peroxynitrite has been recognized to be associated with the activation and expression of inducible NO synthase (iNOS). In this study, to evaluate the role of NO and peroxynitrite in murine bladder tumor cells, the author investigate the effect of NO and peroxynitrite on the viability, cytotoxicity, and DNA fragmentation of MBT-2 cells. The results are as followings: 1. Activated macrophages treated with INF-r, LPS, or INF-r+ LPS showed increment of nitrite (NO2) production and cytotoxicity against MBT-2 cells in a dose dependent manner. However, treatment with NGMMA, a NOS inhibitor, decreased NO2- production and cytotoxicity. 2. Treatment with SNP, a nitric oxide donor, increased NO2 production and DNA fragmentation (%), but decreased viability (%) of MBT-2 cells in a concentration dependent manner. 3. Treatment with peroxynitrite increased cytotoxicity and DNA fragmentation, but decreased viability of MBT-2 cells in a concentration dependent manner. 4. NO- and peroxynitrite-mediated increment of cytotoxicity in MBT-2 cells was corresponded to the programmed cell death, apoptosis. Taken together, these data indicate that NO and peroxynitrite elaborated from macrophages or other cellular systems may increase the cytotoxicity of MBT-2 cells via the mechanism of apoptosis.
Apoptosis* ; Cell Death ; Cell Line* ; DNA Fragmentation ; Humans ; Immune System ; Macrophages ; Neutrophils ; Nitric Oxide Synthase ; Nitric Oxide* ; Peroxynitrous Acid* ; Superoxides ; Tissue Donors ; Urinary Bladder Neoplasms ; Urinary Bladder* ; Vasodilation

Intravesical bacillus calmette-guerin( BCG) therapy for superficial bladder carcinoma is believed to exert its antitumor effects through immune mechanisms which have yet to be more clearly defined. Recently, BCG infection is known to induce nitric oxide(NO) production by macrophages through a T cell mediated process. NO is known to be microbicidal and tumoricidal. Therefore, we studied the effects of intravesical BCG instillation on the induction of inducible NO synthase(iNOS) which is responsible for the production of NO in the vesical tissue of rat Forty Sprauge-Dawley female rats were equally divided into 5 groups. In group 1, normal saline( 0.85 ml/kg) was intravesically instilled one time. In group 2 and group 3, BCG of Pasteur strain(2 mg/kg, normal saline 0.85 ml/kg) was instilled one time and 3 times weekly respectively. In group 4, 10-fold dose of the strain( 20 mg/kg, 0.86 ml/kg) and in group 5, 1/10-fold dose of the strain (0.2 mg/kg, 0.85 ml/kg) were instilled one time respectively. We sacrificed two rats to excise the bladders in each group 1, 3, 7, and 14 days after the instillation( after the last instillation in group 3). iNOS mRNA was not detected in the vesical tissues from the rats of group 1, whereas it was strongly detected in all the vesical tissues from the rats in group 2, 3. or 4. More iNOS mRNA was detected 14 days after the instillation in group 3 than group 2 or 4. In group 5, iNOS mRNA was weakly detected 1, 3, and 7 days and not detected 14 days after the instillation. Our results indicate that intravesical BCG instillation of rat induces the expression of iNOS mRNA in the vesical tissue and suggest that the duration and degree of iNOS mRNA expression is dependent on the dose of BCG and the frequency of the instillations. On the basis of these observations, we conclude that adequate dose and frequency are required for effective treatment of superficial bladder carcinoma in the intravesical BCG therapy.
Animals ; Bacillus ; Female ; Humans ; Macrophages ; Mycobacterium bovis* ; Nitric Oxide Synthase Type II* ; Rats* ; RNA, Messenger ; Urinary Bladder

The most common etiology for female pseudohermaphroditism is congenital adrenal hyperplasia, which accounts for more than 60 percent of children with ambiguous genitalia, and is treated with cortisol replacement and surgical correction of ambiguous genitalia. Flap vaginoplasty, the inverted U-Shaped type has been applied worldwide to the patient with low vaginal entry. The most frequent complication of the operation is contraction of the new vaginal introitus as a result of ischemic and fibrotic changes in the overlapping suture line between the flap and posterior vaginal wall. Maintenance of a good blood supply for the flap and tension free anastomosis should always be kept in mind to avoid this complication. We experienced a vaginoplasty with labioscrotal flap instead of the inverted U-shaped flap and achieved a good result in a 14-year-old girl with low vaginal entry due to congenital adrenal hyperplasia. The labioscrotal flap seems to be more suitable than inverted U-shaped flap for vaginoplasty because the labioscrotal skin is more elastic and more easily elongated than the perineal skin.
46, XX Disorders of Sex Development* ; Adolescent ; Adrenal Hyperplasia, Congenital* ; Child ; Disorders of Sex Development ; Female* ; Humans ; Hydrocortisone ; Skin ; Sutures

Recently, BCG infection is known to induce nitric oxide(NO) production by macrophages through T cell mediated process and NO is known to be microbicidal and tumoricidal. There are several strains of BCG which are commercially available and vary in the number, pathogenicity, viability, and immunogenicity of organisms. Therefore, we wanted to know if there are any differences between three different strains of BCG(Pasteur, Connaught or Tice strain) on the induction of inducible NO synthase(iNOS) and the histological changes in vesical tissue after intravesical instillation of BCG in rats. Thirty two Sprauge-Dowley female rats were equally divided into 4 groups. In group 1, normal saline(0.85 ml/kg) was intravesically instilled one time. In group 2, 3, and 4, BCG of Pasteur strain(2mg/kg, normal saline 0.85ml/kg), Connaught strain(1.35mg/kg, normal saline 0.85ml/kg), Tice strain(0.21mg/kg, normal saline 0.85ml/kg) was instilled one time, respectively. The bladders were excised from each group on day 1, 3, 7, and 14 after BCG instillation. iNOS mRNA was not detected in the vesical tissues of control group, whereas it was strongly detected in group 2, 3, or 4. Also, iNOS mRNA was more strongly detected on day 1, 3, and 7 after intravesical BCG instillation than day 14 in the vesical tissues of group 2, 3, and 4. Histologic findings were well related with expression of iNOS mRNA. Our results indicate that intravesical BCG instillation of rat induces expression of iNOS mRNA in the vesical tissue accompanying the infiltration of inflammatory cells and suggest that all of the 3 strains of BCG including Pasteur, Connaught, and Tice are good at inducing expression of iNOS mRNA without significant differences between the strains.
Administration, Intravesical* ; Animals ; Female ; Gene Expression* ; Humans ; Macrophages ; Mycobacterium bovis* ; Nitric Oxide Synthase Type II* ; Rats* ; RNA, Messenger ; Urinary Bladder ; Virulence

Transforming growth factor-B1(TGF-B1) has many fundamental biological processes including cell growth, extracellular matrix deposition and degradation, and inflammatory responses. TGF-B1 is released by platelet and inflammatory cells, and it affects all phases of wound healing after injury. It contributes to the regulation of fibroblast chemotaxis and proliferation, and also controls the synthesis and degradation of extracellular matrix necessary for tissue repair. Clinically, scar tissue formation and subsequent stricture after urethral injury frequently results in troublesome problems to urologists. In the phase I study of this report, we intended to how the histological changes and the involvement of TGF-B1 in the formation of stricture in injured urethrae of rats. We injured urethrae of 24 adult male Sprauge-Dawley rats(200-250 g.) by urethrotome and Dormia basket and then observed histological changes and analysed TGF-f, mRNA levels of the injured urethrae by Northern blot. Northern b1ot analysis showed that TGF-t, mRNA was much expressed on day 1,3,5 after injury. Fibroblasts and deposition of extracellular matrix were markedly increased on day 5. Reepithelialization was completed and urethral lumen was narrowed on day 10. In the phase II study, we tried to know that antisense TGF-B1 oligodeoxynucleotides(ODNs) could inhibit TGF-B1 expression and the formation of stricture in injured urethrae of rats. We injured urethrae of rats and treated the urethral injury with the application of antisense TGF-B1 ODNs. Northern blot analysis showed that TGF-B1 mRNA was little expressed in the urethrae treated with the antisense on day 1,3 after injury. Comparing to the antisense-nontreated urethrae, the antisense-treated urethrae showed decrease of submucosal thickening and maintained normal sized urethral lumens on day 14, 21 after injury. In conclusion, increase of TGF-B1 mRNA in injured urethrae of rats suggests that TGF-B1 could play an important role in repair mechanism. With application of antisense TGF-B1 ODNs in injured urethrae of rats, the expression of TGF-B1 can be inhibited and also the formation of stricture prevented.
Adult ; Animals ; Biological Processes ; Blood Platelets ; Blotting, Northern ; Chemotaxis ; Cicatrix ; Constriction, Pathologic* ; Extracellular Matrix ; Fibroblasts ; Humans ; Male ; Oligodeoxyribonucleotides* ; Rats* ; RNA, Messenger ; Transforming Growth Factor beta1* ; Urethra ; Urethral Stricture ; Wound Healing

Fibronectin(Fn) is a large, multidomain glycoprotein, which exists in a soluble form in plasma and an insoluble fibrillar form in extracellular matrices. Fn affects many aspects of cellular responses. However, it is not known whether Fn could activate macrophages for the tumor cell killing. We report that Fn induces the tumoricidal activity of macrophages for murine bladder tumor(MBT-2) cell. Tumoricidal activity was determined by 3[H]-thymidine uptake of MBT-2 cell. Fn alone had no effect, whereas recombinant interferon- r(IFN- r) weakly induced the tumoricidal activity of macrophages for MBT-2 cells. However combination of Fn with recombinant IFN-r synergized to activate macrophages to kill MBT-2 cells in a dose dependent manner. At this point nitric oxide(NO) was secreted by activated macrophages, and the secretion of NO and tumoricidal activity of macrophage were inhibited in the presence of NG-monomethyl- L-arginine(NGMMA), a competitive inhibitor of NO synthase(NOS). Fn has various cell binding sites. The Arg-Gly-Asp(RGD) sequence present in the central cell binding domain of Fn is the prototype of these sites. Engineered fibronectin(eFn) is formed by RGD-rich sequence. Combination of eFn, instead of Fn, with recombinant IFN- T resulted in more powerful activation of macrophage in tumor cell killing than Fn. In conclusion, our results demonstrate that Fn acts as a modulator of macrophage activation for tumor cell killing induced by IFN-r via a process involving L-arginine dependent NO production. Especially, RGD sequence of Fn has important role for tumoricidal activity of macrophage. Although the precise mechanism of Fn to promote NO synthesis induced by IFN-r remains to be further elucidated, Fn-mediated macrophage adhesion by specialized cell surface receptors and activation of intracellular signals might be important in the development of macrophage activation.
Arginine ; Binding Sites ; Extracellular Matrix ; Fibronectins* ; Glycoproteins ; Homicide ; Macrophage Activation ; Macrophages* ; Nitric Oxide ; Plasma ; Receptors, Cell Surface ; Urinary Bladder

Inducible nitric oxide(NO) is understood as a tumoricidal molecule. In animal study, urinary NO excretion after intravesical BCG instillation is recently known. In this study, the authors investigated the pattern of urinary nitrite (stable oxidizide form of nitric oxide) excretion according to the dosage of instilled BCG or number of BCG instillation. According to the dosage of instilled BCG(Pasteur strain :8.l5 x 1000000CFU/mg), 32 Sprauge-Dawley female rats were di-vided into 4 Groups(Group I: control, Group II: BCG1/l0X, Group III: BCGlX. Group IV :BCG10X). Also, each Group was subdivided into a one-time instillation subgroup(one time instillation at the first day) and a six-time instillation subgroup( weekly instillation for six weeks). The urinary nitrite in 24 hours' urine was measured quantitatively in each group by Titertek Multiscan MCC/340 ELISA Reader on an every other day basis after BCG instillation until decreased to control level. Intravesical BCG instillation of the rats led to significant urinary nitrite excretion in Group II (BCG 1/10X:one-time ;30+/-12, six-time ;52+/-6), Group III(BCG 1X,57+/-8, 62+/-2uM/L), Group IV (BCG 10X;65+/-2, 80+/-5uM/L) compared to Group I (control;22+/-4, 21+/-3uM/L) respectively( p<0.05). There was no statistical difference in the increment of urinary nitrite excretion between Group II, Group III and Group IV. The urinary nitrite excretion increased significantly in Group II and Group IV of six-time instillation subgroup compared to one-time instillation subgroup(p <0.05). BCG instillation resulted in significant excretion of urinary nitrite at the first day, which reached peak urinary level at the period of the 3rd to 9th day and the 11th day after instillation in one-time and six-time instillation subgroup respectively. The excretion also remained increased for 33 days and 44 days in two subgroups respectively. The preservation of the urine at -40'C for 4-weeks had no influence on the urinary nitrite concentration. The study demonstrates the pattern of urinary excretion according to the dosage of instilled BCG or number of instillations in addition to disclosing good effects of BCG on the excretion of urinary nitrite in rats. The following can be deduced from this study. Intravesical BCG instillation using only one tenth of the usual dose of which the BCG is used for treatment of superficial bladder tumor, and six-times resulted in effective excretion of urinary nitrite in rats.
Animals ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Mycobacterium bovis* ; Nitric Oxide* ; Rats* ; Urinary Bladder Neoplasms

BACKGROUND: The purpose of this stady is to define the clinical characteristics of Tsutsugamushi disease. METHODS: We analyzed 12 cases of tsutsugamushi disease admitted to BAG AE general hospital from 1999. Oct. 1st to Dec. 10. Tsutsugamushi disease was diagnosed by indirect immunofluorescent antibody test. RESULTS: 1) Among them, 8 cases were female and 4 cases were male. The prevalence was predominantly high in female. 2) The age distribution ranged from 40 to 80 years old. The high incidence of age was 70's and 80's. 3) The peak incidene was showed between October and November.(91.6%). 4) The most frequent symptoms were fever and chills(91.6%). 5) Rash and eschar were observed in 83.3%(10 of 12) both. 6) Hepatosplenomegaly and lymphadenopathy were also observed in 25%(3 of 12), 16.6%(2 of 12) respectively. 7) The Laboratory finding showed the elevation of AST 66.6% (8 of 12), ALT 58.3%(7 of 12), alkaline phosphatase 25%(3 of 12) and LDH 25%(3 of 12). Albuminuria and hypoalbuminuria were also observed in 25%(3 of 12), 58.3%(7 of 12) respectively. 8) The therapeutic efficacy of doxycycline was excellent. CONCLUSION: Tsutsugamushi disease which appeared in Pyoungtack area showed typical clinical pattern and was effective to doxycycline treatment.
Age Distribution ; Aged, 80 and over ; Albuminuria ; Alkaline Phosphatase ; Doxycycline ; Exanthema ; Female ; Fever ; Gyeonggi-do* ; Hospitals, General ; Humans ; Incidence ; Lymphatic Diseases ; Male ; Prevalence ; Scrub Typhus*

PURPOSE: Recently, in assisted reproductive technologies(ART) programs theme Is an increasing Interest in the use of agents for the enhancement of sperm motility for assisted fertilization. In an attempt to improve the motility at the cryopreserved human semen and hence the fertilizing capacity of asthenospermic semen samples, different semen preparation techniques have been attempted and the effects of chemical stimulants as nitric oxide(NO) have been studied extensively. Superoxide anions cause lipid peroxidative damage to cell membrane phospholipids, and sperm are known to be particularly susceptible to lipid peroxidation. Such sperm with damaged membranes are impaired functionally. Recently, peroxynitrite, an anion and a potent oxidant, generated by the interaction of nitric oxide and superoxide anions has been demonstrated In macrophages and other cellular systems. Also this anion cause lipid peroxidative damage to cell membrane phospholiplds. We therefore Investigated whether NO and peroxynitrite have the roles to modulate sperm motility and to affect Its viability. MATERIALS AND METHODS: Normal human semen samples(as per World health Organization (WHO) criteria) were obtained after 3day period of abstinence by donors. The samples(n=5) were incubated with either sodium nitroprusside(SNP; 0.1, 05, 1 or 2mM) or peroxynitrite (10, 50 or 100 micrometer ) and the percent viability and motility were assessed at various time inteval up to 4hr. The human semen samples were treated with N-acetyl-L-cystein(NAC;10mM), SNP (0.5mM), phorbol myristate acetate(PMA; 100nM), of SNF plus PMA. Both superoxide and peroxynitrite release were measured directly by chemiluminometer. Percent viability and motility were assessed at 4hr of Incubation. A sample of each aliquot was placed in a Mauler chamber for videomicrography Percent motility were analyzed by using the sperm analysis imaging system. The sperm vlability was assessed by flow cytometer using LIVE/DEAD sperm viability kIt. The production of superoxide and peroxynitrite were measured by the method of chemiluminescence assay. Result : All results represent a mean +/-SEM, n=5. Treatment of human semen samples for 4hr with SNP, a NO generating agent, significantly decreased sperm motility and viability in high concentration [relative motility(% of control); 38 +/-4 and 30 +/-5, relative viability; 42 +/-4 and 30 +/-3 by 1 and 2mM of SNP]. In the presence of low concentration SNP(0.5mM), the sperm viability was not significantly affected(82 +/-3), whereas the sperm motility was affected(64 2). SNP(0.5mM) also decreased sperm motility(80 +/-2 at 2hr 64 +/-3 at 4hr, 44 +/-3 at 6hr, and 38 +/-4 at 8hr) in a time dependent manner. Since it was demonstrated that superoxide anions are one of the common source of lipid peroxidation, we investigated whether superoxide anions produced by human semens could Interact wlth NO to generate peroxynitrite. Adding N-acetyl-L-cystein(NAC) to the human semen samples partially blocked spontaneous release of superoxide, whereas PMA augmented the release of superoxide from human semen samples (control:0.9 106 0.3, NAC: 0.5 106 +/-0.4, and PMA: 2.5 106 +/-0.4photons/60min). The production of superoxide was corresponded with the production of peroxynltrite(control: 1.0 104CPM, SNP: 3.8 106CPM, SNP plus PMA. 12chi106CPM). In addition, SNP in combination with PMA(65 +/-3) markedly decreased sperm motility than that of SNP alone(77 +/-2.5) at 4hr, implying that nitric oxide might inhibit sperm motility via the formation of peroxynitrite In human semen samples. Exogenous peroxynitrite also decreased sperm motility in a dose dependent manner(10 micrometer : 64 +/-2, 50rM: 53 +/-3, and 1 0 micrometer of peroxynitrite: 23 4). CONCLUSIONS: These results suggest that NO inhibits sperm motility via the formation of peroxynltrite and further demonstrate that NO-induced inhibition of sperm motility is depended on the production of superoxide from human semens because peroxynitrite is generated by the interaction of NO and superoxide.
Cell Membrane ; Fertilization ; Humans ; Lipid Peroxidation ; Luminescence ; Macrophages ; Membranes ; Microscopy, Video ; Myristic Acid ; Nitric Oxide* ; Peroxynitrous Acid* ; Phospholipids ; Semen ; Sodium ; Sperm Motility* ; Spermatozoa* ; Superoxides ; Tissue Donors ; World Health Organization