The development of new antimicrobial agents is essential for the effective treatment of diseases such as sepsis. We previously developed a new short peptide, Pap12-6, using the 12 N-terminal residues of papiliocin, which showed potent and effective antimicrobial activity against multidrug-resistant Gram-negative bacteria. Here, we investigated the antimicrobial mechanism of Pap12-6 and a newly designed peptide, Pap12-7, in which the 12th Trp residue of Pap12-6 was replaced with Val to develop a potent peptide with high bacterial selectivity and a different antibacterial mechanism. Both peptides showed high antimicrobial activity against Gram-negative bacteria, including multidrug-resistant Gram-negative bacteria. In addition, the two peptides showed similar anti-inflammatory activity against lipopolysaccharide-stimulated RAW 264.7 cells, but Pap12-7 showed very low toxicities against sheep red blood cells and mammalian cells compared to that showed by Pap12-6. A calcein dye leakage assay, membrane depolarization, and confocal microscopy observations revealed that the two peptides with one single amino acid change have different mechanisms of antibacterial action: Pap12-6 directly targets the bacterial cell membrane, whereas Pap12-7 appears to penetrate the bacterial cell membrane and exert its activities in the cell. The therapeutic efficacy of Pap12-7 was further examined in a mouse model of sepsis, which increased the survival rate of septic mice. For the first time, we showed that both peptides showed anti-septic activity by reducing the infiltration of neutrophils and the production of inflammatory factors. Overall, these results indicate Pap12-7 as a novel non-toxic peptide with potent antibacterial and anti-septic activities via penetrating the cell membrane.
Hepcidin, a peptide hormone synthesized mainly by the liver, has been implicated as a key regulator of iron homeostasis. Results from studies with experimental animal models suggested that hepcidin levels are related with body iron status, but little data is available in human subjects. This study was conducted to determine the relationship between serum pro-hepcidin levels, blood indexes of anemia, and dietary iron intake in female college students. Serum pro-hepcidin concentrations were measured by enzyme-linked immunosorbent assay in eighty-two women with $22.1\pm0.2$ years old. Dietary intake data were collected by using the 24-hour recall method for 3 days. Mean concentrations of serum pro-hepcidin were 85.1 ng/ml$\pm$6.1(s.d.) with the range of 13.6-295.7 ng/ml. The median value of serum pro-hepcidin in the study subjects was 70.3 ng/ml. Serum pro-hepcidin concentrations were positively correlated with hemoglobin concentrations (r=0.273, p=0.013), and also with hematocrit (r=0.291, p=0.008). To examine whether the level of dietary iron intake affects serum pro-hepcidin levels, study subjects were divided into two groups according to the amounts of daily iron intake. Serum pro-hepcidin concentrations were $22\%$ lower in groups with low iron intake (${\leq}10.1$ mg/day), compared to high-iron intake group (>10.1 mg/day) . In conclusion, these data, as in agreement with findings in mice, suggest that hepcidin plays an important role in regulating iron metabolism in the human body.
Objective : Diagnosing acute cerebral infarction is crucial in determining prognosis of stroke patients. Although many serologic tests for prompt diagnosis are available, the clinical application of serologic tests is currently limited. We investigated whether $S100{\beta}$, matrix metalloproteinase-9 (MMP-9), D-dimer, and heat shock protein 70 (HSP70) can be used as biomarkers for acute cerebral infarction. Methods : Focal cerebral ischemia was induced using the modified intraluminal filament technique. Mice were randomly assigned to 30-minute occlusion (n=10), 60-minute occlusion (n=10), or sham (n=5) groups. Four hours later, neurological deficits were evaluated and blood samples were obtained. Infarction volumes were calculated and plasma $S100{\beta}$, MMP-9, D-dimer, and HSP70 levels were measured using enzyme-linked immunosorbent assay. Results : The average infarction volume was $12.32{\pm}2.31mm^3$ and $46.9{\pm}7.43mm^3$ in the 30- and 60-minute groups, respectively. The mean neurological score in the two ischemic groups was $1.6{\pm}0.55$ and $3.2{\pm}0.70$, respectively. $S100{\beta}$, MMP-9, and HSP70 expressions significantly increased after 4 hours of ischemia (p=0.001). Furthermore, $S100{\beta}$ and MMP-9 expressions correlated with infarction volumes (p<0.001) and neurological deficits (p<0.001). There was no significant difference in D-dimer expression between groups (p=0.843). The area under the receiver operating characteristic curve (AUC) showed high sensitivity and specificity for MMP-9, HSP70 (AUC=1), and $S100{\beta}$ (AUC=0.98). Conclusion : $S100{\beta}$, MMP-9, and HSP70 can complement current diagnostic tools to assess cerebral infarction, suggesting their use as potential biomarkers for acute cerebral infarction.
Titania nanomaterials are widely used as cosmetics and dyes, however the impacts on human health are uncertain, We investigated the biodistribution of inhaled titania nanoparticles in rats, Methods Eight weeks-old SD rats were intubated and inhaled with 3 mg titania nanoparticles, twice a week, for 2 weeks, After inhalation, the rats were sacrificed and tissues or heart, lung. intestine, brain, and liver were obtained, We investigated the tissues with optical microscope (OM), transmission electron microscope (EM), scanning EM, And to analyze titania concentration of each tissue, we lysed the tissues with radioimmunoprecipitation assay (RlPA) lysis buffer or acid. Results Granulation tissues in lung were confirmed on the optical microscope, however the other organs had no abnormalities in OM images, In EM images, the rats which inhaled titania nanoparticles showed calcium deposition at heart, brain, and intestine, Titania concentration in lung was increased on the inhaled rat sacrificed I month after last exposure. Conclusion Inhaled titania nanoparticles is thought to be deposited and make inflammatory reaction in lung, and the deposition was not efficiently cleared over a month. However inhaled titania nanoparticles may rarely pass through the alveolus-blood barrier and distribute to other organs of the bod.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
/
v.19
no.2
/
pp.59-70
/
2006
Objective : This study was performed to investigate the Anti-Inflammatory Effects of WooBangHaeGiTang on the Allergic Contact dermatitis. Method : Contact hypersensitivity assay, video microscope, WBC count, RBU count, neutrophil ratio, lymphpcyte ratio and GOT & GPT were measurd. Result : 1. At Contact hypersensitivity Aasay, at 24 hours after Control group & Sample group showed appreciably more ear swelling than Normal group, at 48 hours after Control group showed more ear swelling than Normal group & Sample group, at 72 hours after Control group & Sample group showed more ear swelling than Normal group 2. Regarding the number of WBC, at 24 or 48 hours after There are no significant difference among Normal group, Control group. Sample group, at 72 hours after Sample group showed more WBC than Control group. 3. Regarding the number of RBC, at 24 hours after Control group showed more RBC than Normal group, at 48 hours after Control group & Sample group showed more RBC than Normal group. at 72 hours after Control group showed more RBC than Normal group. 4. Regarding the percentage of neutrophil in WBC, at 24 & 48 hours after Control group & Sample group showed higher percentage than Normal group. 5. Regarding the percentage of Lymphocyte in WBC, at 24 hours after Sample group showed lower percentage than Normal group, at 48 hours after Sample group & Control group showed lower percentage than Normal group. 6. Regarding the GOT & GPT in blood, the GOT & GPT are in Normal scale, so there are no Hepatotoxicity in Normal group, Control group, Sample group. Conclusions : According to above experiments, WooBangHaeGiTang have some anti allergy effects on allergic contact dermatitis.
Corticosteroids have long been used for anti-inflammatory, anti-rheumatoid and other purposes in hospital. These effects may be due to inhibit immune reaction. So the animal given corticosteroids was more susceptible to infection because of immunosuppressive effect of corticosteroids. The purpose of this study was to investigate the effects of prednisolone on the lymphocyte subset in the spleen, immunoglobulin in serum, spleen weight, thymus weight and total WBC in peripheral blood. Mice were randomized into 3 groups. Each group has 24 mice. The small dosage group were given by 4 mg/kg/day of prednisolone for 4 days and the large dosage group were given by 8 mg/kg/day respectively. Prednisolone was suspended in saline and orally administered. Mice in control group were given saline alone. Eight mice in each group were sacrificed every week after administration of predisolone. The weight of thymus and spleen were mesured immediately. Lymphocytes were taken from spleen and these cells were analysed by flow cytometry. Also the concentration of total immunoglobulins in serum were assayed by enzyme-linked immunosorbant assay (ELISA). T cell, T-helper cell and T-cytotoxic cell were all significantly (P<0.05) decreased at 1 week after administration of predisolone and at 2 weeks they recovered similarly to that of control. Population of B cell showed various distribution. The concentration of total immunoglobulins in serum was not changed significantly. The weight ratio of spleen to body decreased significantly (P<0.05) during predisolone administration but increased at 1 week later, Eventually the weight ratio was recovered to that of control at 2 weeks. The weight ratio of thymus to body decreased significantly (P<0.05) by prednisolone and recovered gradually up to normal ratio 2 weeks later.
Kim, Huyn-Sik;Huh, In-Hoi;Lee, Sang-Joon;Ann, Hyung-Soo
YAKHAK HOEJI
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v.38
no.6
/
pp.806-813
/
1994
These experiments were conducted to investigate the effects of Cervi cornu extract on lymphocyte blastogenesis in spleen, thymus, lymph node, born marrow cells of Balb/c mouse, haemagglutination reaction against sheep red blood cell (SRBC), plaque forming cell (PFC) assay against SRBC and IL-2 production. Lymphocyte blastogenesis was determined by $[^3H]-thymidine$ incorporation. According to the lymphcoyte blastogenesis test on the immune cell. Ceriv cornu extrat was showed a potent mitogenic activity on the spleen and lymph node cells, but had mild mitogenic activity on the thymus and born marrow cells. Mitogenic active component of Crevi cornu extract was identified to be materials where molecular weights are higher than 5,000 by membrane filteration method. Cervi cornu extrat was shown to increase mitogenic effect on the lipopolysaccharide (LPS)-stimulated spleen cells significantly, but decrease mitogenic effect on the Con A stimulated spleen cell at the concentration 0.3%, 1% and 3%. Ceriv cornu extract didn't show to be haemagglutination reaction and showed to inhibit the Con A-induced haemagglutination reaction against SREC. Result of SRBC-PEC test. Ceriv cornu extract significantly increase the number of PEC at the concentration of 0.1% and 1%. When IL-2 or IL-4 production was determined by proliferation of CTLL-2 cells. Ceriv cornu extract was not shown to stimulate the production of IL-2. From the above results, it is shown that Ceriv cornu extract increased antibody production by B cells, but nor IL-2 production by helper T cells.
Background: Due to the increase in morbidity and mortality rate, cancer has become an alarming threat to the human population worldwide. Since cancer is a progressive disorder, timely diagnosis would be helpful to prevent/stop cancer from progressing to severe stage. In Khyber Pakhtunkhwa, Pakistan, most of the time, tumors are diagnosed with endoscopy and biopsy; therefore rare studies exist regarding the diagnosis of gastrointestinal (GIT) carcinomas based on tumor markers, especially CEA. Objectives: This study made a comparative analysis of CEA in admitted hospitalized stomach and colon cancer patients diagnosed as GIT with biopsy. Materials and Methods: In this study, a total of 66 cases were included. The level of CEA was determined in the blood of these patients using ELISA technique. Results: Out of 66 patients, the level of CEA was high in 59.1% of the total, 60.7% in colon cancer patients and 57.9 % in stomach cancer patients. Moreover, the incidence of colorectal and stomach cancer was greater in males as compared to females. Patients were more of the age group of 40-60 and the level of CEA was comparatively higher in patients (51.5%) with histology which was moderately differentiated, than patients with well differentiated and poorly differentiated tumor histology. Conclusions: CEA level was high in more than 50% of the total patients. Moreover, CEA exhibited higher sensitivity for colon than stomach cancer.
Objective: To investigate the effect of epirubicin on soluble CD25 (sCD25) secretion by CD4+CD25+ regulatory T (Treg) cells isolated from diffuse large B-cell lymphoma (DLBCL) patients. Methods: Treg cells were isolated from the peripheral blood mononuclear cells isolated from the newly diagnosed DBLCL patients. The concentration of sCD25 in the supernatant was determined with a commercial sCD25 (IL-2R) enzyme-linked immunosorbent assay (ELISA) kit. The fluorescence intensity of CD25 was detected by flow cytometry. Results: Cell survival rate was significantly decreased along with the increase of epirubicin concentration after treatment for 24 h. There was also a significant difference in the concentration of sCD25 between the epirubicin group and the control group (P<0.01). A positive correlation between the Treg cells survival rate and the concentration of sCD25 was detected (r=0.993, P<0.01). When equal numbers of CD4+CD25+ Treg cells of the epirubicin group and the control group were cultured for another 24 h without epirubicin the CD25 fluorescence intensity on the surface of Treg cells was obviously higher in the epirubicin group than that in the control group (P<0.01), while the sCD25 concentration in the supernatant in the epirubicin group was significantly lower than that in the control group (P<0.05). Conclusion: Epirubicin may improve the body's immune functions by inhibiting the sCD25 secretion by Treg cells in DLBCL patients.
Background: Carcinoembryonic antigen (CEA) is well-known soluble tumor marker frequently detectable in peripheral blood of carcinoma patients and considered as good target for antigen-specific immunotherapy. In this study, we used a replication-deficient adenovirus containing CEA to study CTL induction in vitro after adenovirus-mediated gene transfer into DC. Methods: DC were obtained from mouse bone marrow and cultured with IL-4 and GM-CSF. For measuring CTL activity, splenocytes were harvested from the mice, which were immunized with DC that had been infected AdV-CEA or pulsed with CEA peptide. Untreated DC was used as a control. Splenocytes were re-stimulated in vitro with DC pulsed with CEA peptide for 7 days and CTL activity with CEA peptide-pulsed EL-4 cells were assessed in a standard $^{51}Cr$-release assay. The frequencies of antigen-specific cytokine-secreting T cell were determined with $mIFN-{\gamma}$ELISPOT. Results: DC infected with recombinant adenovirus expressing CEA induced CEA-specific CTL responses in vivo. Splenocyte induced from mice immunized with AdV-CEA-infected DC increase in the number of $IFN-{\gamma}$ secreting T cells compared with those from mice immunized with CEA peptide-pulsed DC. Conclusion: These results suggested that DC infected with recombinant adenovirus has advantages over other forms of vaccination and could provide an alternative approach vaccination therapies.
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