Effects of L-ascorbic acid on the production of pro-inflammatory and anti-inflammatory cytokines in C57BL/6 mouse splenocytes / 고신대학교의과대학학술지

OBJECTIVE: The imbalance between pro-inflammatory and anti-inflammatory cytokines may underlie different pain states. Although ascorbic acid is the most important physiological antioxidant that affects host defense mechanisms and immune homeostasis, there is limited information on the effects of ascorbic acid on the production of cytokines. METHODS: In this study, we investigated the in vitro effect of L-ascorbic acid (AA) on the production of pro-inflammatory and anti-inflammatory cytokines by stimulating C57BL/6 mouse splenocytes with the polyclonal activators lipopolysaccharide or concanavalin A. RESULTS: AA significantly downregulated the expression of IL-6, IL-12, and TNF-alpha at 48 h and 72 h in mouse splenocytes treated with a combination of polyclonal activators and AA. AA treatment also resulted in upregulation of IL-4 and IL-10 at 72 h. These findings demonstrated that AA significantly potentiated production of anti-inflammatory cytokines whereas there was an inverse association between AA and expression of pro-inflammatory cytokines in mouse splenocytes. CONCLUSION: AA may have potential applications in the reduction of inflammatory pain because of its function in modulating the production of cytokines. However, further in vivo investigations are necessary to elucidate the mechanisms involved.

Analysis of Clarithromycin Resistance of Helicobacter pylori Isolated in Korea

Forty-four strains of Helicobacter pylori were isolated from Kosin Medical Center were tested of resistance to antimicrobial agents, and the mechanism of resistance to clarithromycin was investigated. We determined the MICs of amoxicillin, amoxicillin/clavulanic acid, clarithromycin, and metronidazole by agar and broth dilution method. To detect the mutations of 23S rRNA which is associated with clarithromycin resistance, a 3'-mismatched polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis with restriction enzymes BbsI and BsaI were performed. The nucleotide sequence of 23S rRNA was determined. All H. pylori strains appeared to be susceptible to amoxicillin/clavulinic acid, but 2.3% of strains (1 strain) are resistant to amoxicillin, 13.6% (6 strains) to clarithromycin, and 15.9% (7 strains) to metronidazole. No PCR products was observed by the 3'-mismatched PCR. A 291 bp of PCR product was not digested by BbsI, but was digested by BsaI, which was a characteristic of the A2143G point mutation in the 23S rRNA gene. The nucleotide sequencing analysis revealed that all resistant strains had A2143G, T2182C, and T2244C mutations in 23S rRNA gene.

Rapid Detection of Mycoplasma pneumoniae and Antimicrobial Susceptibilities of the M. pneumoniae Isolates

The throat swabs obtained from 1,098 adults and 432 children patients with respiratory diseases were examined for Mycoplasma pneumoniae infection detected by culture and polymerase chain reaction (PCR). Antimicrobial susceptibilities of the resulting 60 M. pneumoniae isolates were evaluated by testing minimum inhibitory concentrations (MICs) of erythromycin, minocycline, tetracycline, josamycin, sparfloxacin, ofloxacin, and ciprofloxacin by a broth micro-dilution method. In a preliminary screening, the detection rate of M. pneumoniae by PCR was 29.2% (277/948) for the adults and 28.3% (90/318) for the children. In the second survey, the isolation rate of M. pneumoniae by culture was 29.3% (44/150) for the adults, and 14.0% (16/114) for the children. The PCR detection rate was 36.7% (55/150) for the adults and 23.7% (27/114) for the children. The MIC90s of the M. pneumoniae isolates were 0.015 mg/ml for erythromycin, lower than 0.03 mg/ml for josamycin, 0.06 mg/ml for sparfloxacin and minocycline, 0.12 mg/ml for tetracycline, 0.5 mg/ml for ofloxacin and CFC-222, and 1.0 mg/ml for ciprofloxacin. The isolates were susceptible to erythromycin, josamycin, sparfloxacin, minocycline, tetracycline, and ofloxacin, but the 63.3% of them was resistant to ciprofloxacin. These results indicate that the PCR method has a significant potential as a rapid and sensitive method for early detection of M. pneumoniae infection in clinical specimens as compared with the culture method, but the PCR method could not provide any information concerning the biological chracteristics of M. pneumoniae strains. Erythromycin, josamycin, sparfloxacin, minocycline, and tetracycline could be recommended as the antimicrobial agents of choice in Korea.

Production Of Gm-Csf And Tgf-beta 1 In Irradiated Human Gingival Fibroblasts Cultured With Lipopolysaccharide

PURPOSE: Irradiation in the oral cancer patients causes early and late complications such as intraoral mucositis and fibrosis, with a various expression of GM-CSF and TGF-beta. The purpose of this study was to investigate the production of GM-CSF and TGF-beta1 by the irradiated human gingival fibroblasts cultivated with lipopolysaccharide. MATERIALS AND METHODS: Irradiated (total dose, 60 Gy) human gingival fibroblasts were incubated with LPS. Culture supernatants that were collected at 24, 48, and 72 hours were assessed for GM-CSF and TGF-beta1 by enzyme-linked immunosorbent assay. RESULTS: 1. GM-CSF production in nomal gingival fibroblasts was increased with incubation time, but decreased with incubation time in irradiated gingival fibroblasts. GM-CSF production in both normal and irradiated gingival fibroblasts induced with LPS was higher than the control. 2. TGF-beta1 production in normal gingival fibroblasts was decreased after 24 hours, but, it was increased until 48 hours in irradiated gingival fibroblasts. TGF-beta1 production in normal gingival fibroblasts exposed with LPS was higher than the control. Conversely, It was lower than the control in irradiated gingival fibroblasts exposed with LPS. CONCLUSION: This indicates that irradiation in gingival fibroblasts may play an important role in radiation-induced intraoral mucositis and fibrosis. However, LPS decreases the production of TGF-beta1 in the irradiated gingival fibroblasts.

Detection and Isolation of Genital Mycoplasmas from Public Toilet Bowls

Genital mycoplasmas are sexually transmitted. There are considerable public concern that causative agents of sexually transmitted diseases might be transmitted nonsexually through public restrooms. In the present study, Mycoplasma hominis, Ureaplasma urealyticum and M. penetrans among genital mycoplasmas were identified in 100 public restroom toilet bowls (50 men's and 50 women's public restrooms, each). Mycoplasmas were genotypically identified by two methods; (1) PCR of primary selective culture and (2) direct PCR of original specimens before primary selective culture. From 50 men's public restrooms, M. hominis, U. urealyticum and M. penetrans were identified from PCR of primary selective cultures in 6%, 4% and 0% of the specimens, respectively and M. hominis and U. urealyticum was codetected in 2% of those. And M. hominis, U. urealyticum and M. penetrans were identified by direct PCR in 20%, 16% and 0% of the original specimens, respectively and co-detection rate of M. hominis and U. urealyticum was 4% in those. From 50 women's public restrooms, 38% was positive for M. hominis, 14% for U. urealyticum, 0% for M. penetrans and 10% for both U. urealyticum and M. penetrans by PCR of primary selective culture. And 50% was positive for M. hominis, 46% for U. urealyticum and 0% for M. penetrans and 34% for both M. hominis and U. urealyticum by direct PCR of the original specimens. These results indicate that the genital mycoplasmas can survive for considerable duration in toilet bowels, and might be transmitted by through public restrooms.

Relationship of Serum Interleukin-6 and Liver Regeneration after Hepatectomy in N-nitrosodi ethylamine-induced Cirrhotic Rats

PURPOSE: The evolution of cirrhosis from chronic inflammatory liver disease represents a dysmorphogenic "response to injury". It is important to understand how inflammatory cytokines, known to be associated with such responses, influence the growth of different cell populations within the liver. The purpose of this work is to establish a role of serum interleukin-6 (IL-6) in liver regeneration following partial hepatectomy in N-nitrosodieth ylamine (DEN)-induced cirrhotic rats. METHODS: Male Sprague-Dawley rats were used for this study. Liver cirrhosis was induced using DEN (100 mg/kg) given once a week for 6 weeks. In Group I (n=18), 70% partial hepatectomy was accomplished and then the resected liver weight, regenerated liver weight, serum IL-6, and serum GOT/GPT was determined on postoperative days 1, 2, and 4 and at intervals. In Group II (n=19), partial hepatectomy was carried out and Laennec, a hepatocyte growth promoter, was injected on preoperative 1 day and postoperative days 1 and 2. RESULTS: The value of serum GOT in Group I was 415 IU/ml on the first postoperative day and peaked at 1870 IU/ml on the third day. In Group II, the level of serum GOT was 404 IU/ml on the first postoperative day and peaked at 593 IU/ml on the third day, then decreased gradually thereafter. The value of serum IL-6 was 106.54 pg/ml on the first postoperative day, 130.59 pg/ml on the 14th postoperative day in Group I, however in Group II, it was 40 pg/ml on thefirst postoperative day and then decreased to 29.18 pg/ml on the 14th postoperative day. The percentages of regenerated weights of liver at intervals following the 70% partial hepatectomy was 55.1% on the first postoperative day, and 102.3% on the 4th week in Group I and 60.4% on the first postoperative day, 95.8% on the first week postoperatively, and 116.1% on the 2nd week in Group II. CONCLUSION: As the value of serum IL-6 was sustained below 40 pg/ml, which was the value on the first postopeative day following partial hepatectomy with Laennec treatment, the resected liver was rapidly regenerated and restored to normal liver function. In cirrhotic liver, regenerative activity was related to serum IL-6 level, so downregulation of serum IL-6 might be helpful to the regeneration of resected liver.

Production of Transforming Growth Factor-beta1 in Human Fibroblasts Induced with Bacterial Toxins

TGF-beta1 is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to wound healing. The objective of this study is to investigate production of TGF-beta1 in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of TGF-beta1 which may be responsible for wound healing The fibroblasts were originated from facial dermis and hypertrophic scar in 26 year-old male patient. In the presence of LPS(0.0l microgram, 0.1 microgram, 1.0 microgram), SEB(0.0l microgram, 0.1 microgram, 1.0 microgram) respectively, cells(5x103ml) were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, cells(2.5x105ml) were cultivated in EMEM with LPS(0.01, 0.1 and 1.0 microgram), SEB(0.01, 0.1 and 1.0 microgram) respectively and LPS(0.1 microgram) and SEB(0.1 microgram) in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and TGF-beta1 was assayed in duplicate. The results were as follows. 1. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of TGF-beta1 was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of TGF-beta1 was increased very significantly at 1 day after incubation, compared with the control. 2. In hypertrophic scar fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation did not occur at 1 day after incubation, compared with the control. In SEB and LPS exposure in combination, the production of TGF-beta1 was increased very significantly at 1 day after incubation, compared with the control. However, the production of TGF-beta1 did not occur in SEB and LPS exposure respectively. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of TGF-beta1 very significantly and both fibroblasts have different phenotype each other in this regard. This data suggest that the significant production of TGF-beta1 may develope abnormal wound healing associated with tissue fibroproliferative disorder, such as hypertrophic scar and keloid formation.

Prevalence of Ureaplasma Urealyticum in Ovarian or Cervical Cancer

No Abstract Available.

Effects of Mycoplasmal Antigens on Production of Tuansforming Growth Factor-beta1 in Human Tumor Cells

No Abstract Available.

Quantitative Analysis of Transforming Growth Factor-beta1 in Human Fibroblasts Induced with Staphylococcus enterotoxin B and Lipopolysaccharide

Production of IL - 6 and IL - 8 in Human Fibroblasts Stimulated with Mycoplasma Lysates and Bacterial Toxins

Cytokines are hormone-like proteins which mediate and regulate inflammatory and immune responses. The purpose of this study was to investigate the effect of lipopolysaccharide (LPS), Staphylococcus enterotoxin B (SEB), and Mycoplasma lysates on regulation of IL-6 and IL-8 production by human nasal fibroblasts. Primary cultured cells were incubated with LPS (1.0 ug/ml) from E.coli, SEB (1.0 ug/ml) from S.aureus, or Mycoplasma lysates (M.pneumoniae, Mp; M. fermentans, Mf; M. hominis, Mh, each 1.0 ug/ml). The culture supernatants were collected at 2, 6, and 24 hr and assessed for IL-6 and IL-8 production by enzyme-linked immunosorbent assay. The production of IL-6 in the culture supematant was downregulated by LPS, SEB, or Mycoplasma lysates. But IL-6 was upregulated by mixed exposure with Mp+LPS (2 hr), Mp+LPS+SEB (24 hr), Mf+LPS (24 hr), Mf+LPS+SEB (2 hr), Mh+LPS (24 hr), Mh+SEB (24 hr), or Mh+LPS+ SEB (24 hr). The production of IL-8 in the culture supematant was similar to that of IL-6 by same stimulants. But IL-8 was upregulated by mixed exposure with Mf+LPS+SEB (2 hr), Mh+LPS (24 hr), Mh+ SEB (24 hr), or Mh+LPS+SEB (24 hr). These studies show that costimulation of LPS or SEB with Mycoplasma whole cell lysates upregulates the production of IL-6 and IL-8.

Production of IL-6 and IL-8 in human fibroblasts stimulated with bacterial toxins

Studies on the Production of Nitric Oxide, Interleukin-1 and Interleukin-6 in Mice Exposed with Sarcoma 180 Cells / 대한해부학회지

This study was conducted to elucidate the biological role of the overproduced nitric oxide (NO), interleukin-l (IL-l), and interleukin-6 (IL-6) which are known to elicit inflammation, rheumatic arthritis, fever, septic shock or other fatal reactions. I investigated whether the Scarcoma 180 cells elicit NO, IL-l and IL-6 production in vivo and in vitro by measuring the NO, IL-1 and IL-6 level in the splenocyte adherent cell (AD), non-adherent cell (NAD) and whole cell (W) exposed to sarcoma 180 cells. I also measured the NO, IL-1 and IL-6 level in the plasma and peritoneal fluid of the sarcoma 180 cell-transplanted mice after 2 hours, 4 hours, 6 hours and 24 hours of incubation. 1. In the splenocyte exposed to sarcoma 180 cells, the NO production of AD and NAD increased after 2, 4, and 6 hours but decreased after 24 hours of incubation. In the whole cell, the NO production was variable; it showed increased level of synthesis at 2 hours, decreased level at 4 hours, increased level again at 6 hours, and decreased level at 24 hours of incubation. In the plasma of the sarcoma 180 cell-transplanted mice, the NO synthesis significantly increased from the 6 hours of incubation. In the peritoneal fluid, the NO production significantly increased until the 4 hours of incubation then decreased gradually. However, it showed higher level of NO production compared to the control group. 2. For the splenocyte AD cell exposed to saracoma 180 cells, the IL-1 level decreased after 6 hours of incubation. For the W cells, the IL-1 level decreased at 4 hours then increased until 24 hours of incubation. The NAD cell showed increased level of IL-I production from 2 to 24 hours. All these cells showed significantly increased level of IL-1 production compared to the control. In the plasma of the sarcoma 180 cell-transplanted mice, the IL-1 production increased more than twice the level of control from the beginning. In the peritoneal fluid, no IL-1 production was detected as in the control. 3. The IL-6 synthesis of the sarcoma 180 cell-exposed splenocyte singnificantly increased compared to the control: the AD cell showed increased level of IL-6 production after 4 hours of incubation. For the NAD cell, increased level of IL-6 was detected at 2 hours after the incubation. In the plasma of the sarcoma 180 cells transplanted mice, the IL-6 level at 2 hours after incubation was 64.22+/- 5.85 pg/ml. After 4 hours of incubation, the level decreased to 43.55+/-1.56 pg/ml. At six and 24 hours after the incubation, no IL-6 was detected. In the control, IL-6 production was not detected. In the peritoneal fluid, the IL-6 production level was 712.41+/-4.27 pg/ml after 2 hours and 225.71+/-9.74 pg/ml after 4 hours of incubation, producing singnificantly higher level of IL-6 compared to the control. After 6 hours of incubation, IL-6 level was 8.27+/-0.78 pg/ml. After 24 hours of incubation, it decreased to 1.38+/-0.39 pg/ml as time proceeds. After 24 hours of incubation, the IL-6 level was the same as the control. These results suggest that the mice which were exposed to sarcoma 180 cells, nitric oxide, interleukin-1 and interleukin-6 which may lead to inflammation, fever, sepsis and septic shock. This study also helps us better understanding of the role of cytokines in inflammatory reaction.

Antibiotic Susceptibility to Helicobacter pylori Isolated from Patients with Gastric Cancer

This investigation was evaluated for in vitro antibacterial activity against 169 Helicobacter pylori strains isolated from patients with gastric cancer in Pusan, Korea. The minimum inhibitory concentration (MIC) of 6 antibiotics was determined by broth microdilution method. The isolation rate of H. pylori was 39.3% in the patients with gastric cancer, and which was not observed any differences between male and female or age group. The MIC50amoxacillin, clarithromycin, and amoxacillin plus clarithromycin against H. pylori isolates was 4.0, 2.0, 0.5, 0.5, 0.5, 1.0, and 1.0 microgram/ml, respectively. The MIC50 of the metronidazole, erythromycin, ciprofloxacin, tetracycline, amoxacillin, clarithromycin, and amoxacillin plus clarithromycin against H. pylori isolates was 32.0, 16.0, 1.0, 1.0, 4.0, 16.0, and 8.0 microgram/ml, respectively. The prevalence of one kind of antibiotic resistant strains of H. pylori was 31.9% for metronidazole, 31.9% for erythromycin, 23.1% for clarithromycin, 11.2% for amoxacillin, 6.5% for ciprofloxacin, and 9.5% for amoxacillin and for clarithromycin. The prevalence of two kinds of antibiotic resistant strains of H. pylori was 8.3% for amoxacillin and clarithromycin, 4.1% for metronidazole and erythromycin, 1.3% for metronidazole and ciprofloxacin, 1.3% for erythromycin and ciprofloxacin. The prevalence of three kinds of antibiotic resistant strains of H. pylori was 5.9% for metronidazole, amoxaciltin, and ciprofloxacin, 2.4% for metronidazole, erythromycin, and ciprofloxacin. The prevalence of four kinds of antibiotic resistant strains of H. pylori was 1.3% for metronidazole, erythromycin, tetracycline, and ciprofloxacin.

Effect of cisplatin on sodium-dependent hexose transport in LLC-PK-1 renal epithelial cells

Cis-dichlorodiammine platinum II (Cisplatin), an effective chemotherapeutic agent, induces acute renal failure by unknown mechanisms. To investigate direct toxic effects of cisplatin on the renal proximal tubular transport system, LLC-PK-1 cell line was selected as a cell model and the sugar transport activity was evaluated during a course of cisplatin treatment. Cells grown to confluence were treated with cisplatin for 60 min, washed, and then incubated for up to 5 days. At appropriate intervals, cells were tested for sugar transport activity using alpha-methyl-D-(14C)glucopyranoside (AMG) as a model substrate. In cells treated with 100 micrometer cisplatin, the AMG uptake was progressively impaired after 3 days. The viability of cells was not substantially changed with cisplatin of less than 100 micrometer, but it decreased markedly with 150 and 200 micrometer. In cisplatin-treated cells, the Na+/-dependent AMG uptake was drastically inhibited with no change in the Na+/-independent uptake. Kinetic analysis indicated that Vmax was suppressed, but Km was not altered. The Na+/-dependent phlorizin binding was also decreased in cisplatin-treated cells. However, the AMG efflux from preloaded cells was not apparently retarded by cisplatin treatment. These data indicate that the cisplatin treatment impairs Na+/-hexose cotransporters in LLC-PK-1 cells and suggest strongly that defects in transporter function at the luminal plasma membrane of the proximal tubular cells constitute an important pathogenic mechanism of cisplatin nephrotoxicity.

Efficacy of Ciprofloxacin-soaked Collagen Shields for Treatment of Experimental Staphylococcus aureus Keratitis in Rabbits

In order to study the efficacy of a 0.2% ciprofloxacin-soaked collagen shield for the treatment of Staphylococcus aureus keratitis, an experimental study was performed on 40 eyes of 20 rabbits. Twenty hours after intrastromal injection with S. aureus, the rabbits were divided into four groups: Group 1(10 eyes) was treated with a 0.2% ciprofloxacin-soaked collagen shield with an additional instillation of 0.2% ciprofloxacin drops every 30 minutes for 4 hours; Group 2 (10 eyes) was treated with a 0.2% ciprofloxacin-soaked collagen shield for 4 hours; Group 3 (10 eyes) was treated with 0.2% ciprofloxacin drops every 30 minutes for 4 hours; Group 4 (10 eyes) received BSS every 30 minutes for 4 hours as a control. Bacterial killing was quantitated by culturing corneal homogenates and calculating the number of viable bacteria (colony-forming units) per cornea. Groups 1, 2, and 3 showed significantly reduced numbers of bacteria compared with the control (p0.05). These results suggested that a 0.2% ciprofloxacin-soaked collagen shield may be an effective and convenient mode of therapy for S. aureus keratitis.

Experimental Inhibitory, and Pharmacokinetic Stydy on The Development of Proliferative Vitreoretinopathy by Sustained Delivery of Intravitreal Fluoropyrimidenes

The efficacy of the sustained release-delivery system was tested in suppressing the development of proliferative vitreoretinopathy(PVR) with intravitreal Fluoropyrimidines(5-Fu: FU, Fluorouridine: FUD, 5-Fluoro-5'-monophosphate: FUMP). PVR was induced in one-hundred-eightyeight eyes of one-hundred-twenty rahbits by intravireal injection of homologous dermal fibroblasts(250.000 cells/0.1 ml). Each drugs were encapsulated with liposome(LFU. LFUD. LFUMP) or polymer(PFU. PFUD) and non-coated drugs wer e used as controls. 3 weeks after injections of fibroblasts, retina detached in 50.0% of control eyes. Among treated eyes, the detachment rates in percentage are as follows; FU 37.5, FUD 50.3, LFU 37.5, LFUD 37.5, PFU 16.7, PFUD 22.2. For the pharmacokinetic study. radiolaveled(-14C) drugs were used in liposome group and frozen vitreouses were measured by scintillatng counter; polymer group was measured by HPLC. The intravitreal half life(hour) of injected drugs were FU 3.2, FUMP 2.9, LFU 7.1, LFUMP 7.6, and PFU was exceptionally long(11.1 days).

Detection of contaminated mycoplasmas on the cultured cell lines

No abstract available.

The influence of linoleic acid and ursolic acid on mouse peritoneal macrophage activity / 대한면역학회지

No abstract available.

Effects of small water dropwort extract on celluar immune response of mice

No abstract available.