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 majority of nasal fractures have been managed by using closed reduction, intranasal packing and external splinting. However, in comminuted nasal bone fractures, the conventional closed reduction may be inadequate to reduce the fracture segments accurately, and insufficient to prevent secondary nasal deformity. In these cases, open reduction and interfragment wire fixation was recommended for accurate reduction, and has been mainstay of treatment modality. Furthermore, in nasoethmoid orbital fractures, anatomic reduction of fractured nasal bone, medial and inferior orbital rim segments to reconstruct nasofrontal buttress and transnasal wiring to prevent telecanthus were essential. But, the interfragment wire fixation is difficult and time-consuming procedure. The care must be taken to fix small fracture segments. It is also difficult to obtain bony support due to extensive dissection of periosteum, and to achieve rigid fixation on three- dimensional space, causing depression of bony contour. From April 1998 to August 1999, we used malleable mesh microplates for treatment of 3 comminuted nasal bone fractures and 3 nasoethmoid orbital fractures. During the follow up period of 8 months to 24 months, all of six patients had successful cosmetic result without complications. There was no recurrent depression of bony contour, no secondary nasal deformity, no displacement of microplates and no palpable, externally shown hardwares. The use of mesh microplates is reliable and useful method for the treatment of comminuted nasal fractures because it is relatively simple procedure and achieves rigid fixation on three-dimensional space without postoperative temporary nasal packing which may cause patient's discomfort. Furthermore, in nasoethmoid orbital fractures, correction of telecanthus can be done without application of transnasal wiring.
Congenital Abnormalities ; Depression ; Follow-Up Studies ; Humans ; Nasal Bone* ; Orbit ; Orbital Fractures ; Periosteum ; Splints

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

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

PURPOSE: To evaluate the eligibility of laparoscopic surgery for the treatment of renal cysts, we analysed our clinical results of laparoscopic renal cyst marsupialization in 9 patients. MATERIALS AND METHODS: Between January 1994 and February 1997, a total of 9 patients with renal cyst underwent laparoscopic surgery. They were 2 men and 7 women 42 to 67 years old. Mean cyst size of the patient was 7.3cm(5-10cm). Presenting symptoms were flank pain in 8 and palpable mass in 1. One patient had previously undergone ultrasonography guided percutaueous aspiration and ablation. One patient had bilateral renal cyst. Initially the procedures had been performed via the transperitoneal approach in 4 patients after that we attempted retroperitoneal access for laparoscopy in 5 patients(6 renal units). During the operation cyst fluid was obtained for cytologic examination and cyst walls were excised and sent for pathological examination. RESULTS: Mean operation time was 2 hours and 14 minutes. Mean hospital stay was 5.9 days. Perioperative complications were pain(20%), bleeding(10%), subcutaneous emphysema(20%) and pneumothorax(10%). Bleeding was minimal but 2 units of packed red cell were needed in 1 patient. Conversion to open surgery from laparoscopic procedure was needed in 1 patient. Biopsy results were negative for carcinoma. All patients were asymptomatic at a mean follow up of 12.5 months. CONCLUSIONS: Laparoscopic renal cyst marsupialization for symptomatic renal cysts seems to be a safe and effective alternative to open surgery.
Aged ; Biopsy ; Conversion to Open Surgery ; Cyst Fluid ; Female ; Flank Pain ; Follow-Up Studies ; Hemorrhage ; Humans ; Laparoscopy ; Length of Stay ; Male ; Ultrasonography

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

PURPOSE: Taxol, an anticancer drug, blocks cell division by stabilizing microtubules. However, taxol has distinct cell-cycle-independent effects. For example, taxol and interferon gamma(IFN-gamma) induce tumoricidal activity of murine peritoneal macrophages. This study was designed to know whether taxol has indirect tumoricidal effect on murine bladder tumor-2(MBT-2) cells besides its direct cytotoxicity. MATERIALS AND METHODS: The original stock of C57BL/6 mice were used at 8 to 12 weeks of age. Macrophages were obtained by peritoneal lavage from the mice which had been treated with thioglycollate. The tumor target cells were MBT-2 cell line. MBT-2 cells were cultivated in different concentration of taxol for various times and the growth of MBT-2 cells were tested. Tumoricidal activitiy was measured by indirect methylthiazol-2-yl-2,5-diphenyl tetrazolium bromide(MTT) assay after co-cultures of stimulated macrophage and MBT-2 cells with taxol, INF-gamma, lipopolysaccharide(LPS) or with combination of taxol and INF-gamma or LPS and INF-gamma. Nitric oxide(NO) formation was measured by Griess method under the same conditions. Effect of NG -monomethyl-L-arginine(NGMMA) on nitrite formation and cytotoxicity toward MBT-2 cells were also evaluated. RESULTS: Significant retardation of cell growth was observed after treatment of tumor cells with taxol in a dose dependent manner but does not affect cell viability. Taxol(19+/-2%) or LPS(19+/-4%) alone weakly activated macrophages to kill MBT-2 cell lines, whereas combinations of taxol(77+/-3%) or LPS(75+/-4%) with IFN-gamma(control: 2%, IFN-gamma:18+/-3%) synergized to activate macrophages to kill tumor cells in a dose dependent manner. Taxol(20+/-5microM), LPS(15+/-5microM) or IFN-gamma(25+/-3microM) alone induced small amounts of NO secretion but the combinations of either taxol and INF-gamma(73+/-5microM) or LPS and IFN-gamma(77+/-5microM) synergistically induced large amounts of NO secretion. The production of NO(control:<5, IFN-gamma+ taxol: 73+/-5microM, IFN-gamma+ taxol + NGMMA: 15+/-3microM) and tumor cell killing(control: 2%, IFN-gamma+ taxol: 77+/-3%, IFN-gamma+ taxol + NGMMA: 20+/-3%) were blocked in the presence of NGMMA, a competitive inhibitor of NO synthase. CONCLUSIONS: Collectively, the data indicate that taxol is directly non-cytotoxic for MBT-2 cells via its effect on micrtubules but indirectly activates macrophages to kill MBT-2 cells probably via NO secertion.
Animals ; Cell Division ; Cell Line ; Cell Survival ; Coculture Techniques ; Interferons ; Macrophages ; Macrophages, Peritoneal ; Mice ; Microtubules ; Nitric Oxide Synthase ; Nitric Oxide* ; Paclitaxel* ; Peritoneal Lavage ; Urinary Bladder*

PURPOSE: Taxol, an anticancer drug, blocks cell division by stabilizing microtubules. However, taxol has distinct cell-cycle-independent effects. For example, taxol and interferon gamma(IFN-gamma) induce tumoricidal activity of murine peritoneal macrophages. This study was designed to know whether taxol has indirect tumoricidal effect on murine bladder tumor-2(MBT-2) cells besides its direct cytotoxicity. MATERIALS AND METHODS: The original stock of C57BL/6 mice were used at 8 to 12 weeks of age. Macrophages were obtained by peritoneal lavage from the mice which had been treated with thioglycollate. The tumor target cells were MBT-2 cell line. MBT-2 cells were cultivated in different concentration of taxol for various times and the growth of MBT-2 cells were tested. Tumoricidal activitiy was measured by indirect methylthiazol-2-yl-2,5-diphenyl tetrazolium bromide(MTT) assay after co-cultures of stimulated macrophage and MBT-2 cells with taxol, INF-gamma, lipopolysaccharide(LPS) or with combination of taxol and INF-gamma or LPS and INF-gamma. Nitric oxide(NO) formation was measured by Griess method under the same conditions. Effect of NG -monomethyl-L-arginine(NGMMA) on nitrite formation and cytotoxicity toward MBT-2 cells were also evaluated. RESULTS: Significant retardation of cell growth was observed after treatment of tumor cells with taxol in a dose dependent manner but does not affect cell viability. Taxol(19+/-2%) or LPS(19+/-4%) alone weakly activated macrophages to kill MBT-2 cell lines, whereas combinations of taxol(77+/-3%) or LPS(75+/-4%) with IFN-gamma(control: 2%, IFN-gamma:18+/-3%) synergized to activate macrophages to kill tumor cells in a dose dependent manner. Taxol(20+/-5microM), LPS(15+/-5microM) or IFN-gamma(25+/-3microM) alone induced small amounts of NO secretion but the combinations of either taxol and INF-gamma(73+/-5microM) or LPS and IFN-gamma(77+/-5microM) synergistically induced large amounts of NO secretion. The production of NO(control:<5, IFN-gamma+ taxol: 73+/-5microM, IFN-gamma+ taxol + NGMMA: 15+/-3microM) and tumor cell killing(control: 2%, IFN-gamma+ taxol: 77+/-3%, IFN-gamma+ taxol + NGMMA: 20+/-3%) were blocked in the presence of NGMMA, a competitive inhibitor of NO synthase. CONCLUSIONS: Collectively, the data indicate that taxol is directly non-cytotoxic for MBT-2 cells via its effect on micrtubules but indirectly activates macrophages to kill MBT-2 cells probably via NO secertion.
Animals ; Cell Division ; Cell Line ; Cell Survival ; Coculture Techniques ; Interferons ; Macrophages ; Macrophages, Peritoneal ; Mice ; Microtubules ; Nitric Oxide Synthase ; Nitric Oxide* ; Paclitaxel* ; Peritoneal Lavage ; Urinary Bladder*