Research communications in molecular pathology and pharmacology最新文献

Pharmacokinetics of itraconazole was measured after 1-min intravenous infusion at a dose of 10 mg/kg to male New Zealand white rabbits. The terminal half-life of itraconazole was 524 min. Itraconazole was eliminated slowly in rabbits with total body clearance of 3.26 ml/min/kg. Blood partition of itraconazole between plasma and blood cells of rabbit blood was measured. Itraconazole reached equilibrium rapidly between plasma and blood cells of rabbit blood. The equilibrium plasma/blood cells concentration ratios were independent of initial rabbit blood concentrations of itraconazole, 1, 5, and 10 microg/ml; the mean value was 3.25. Tissue distribution of itraconazole was also measured after 1-min intravenous administration at a dose of 10 mg/kg to rats. The tissue-to-plasma (T/P) ratios of itraconazole were greater-than-unity in all rat tissues studied at both 1 and 24 h except in fat and stomach at 1 h. This indicated that rat tissues studied had a high affinity to itraconazole.

Once prostate cancer has metastasized, current treatment methods are generally ineffective. Due to the reported anti-tumor properties of specific nutrients, we investigated the effect of a unique formulation (NS) of lysine, proline, arginine, ascorbic acid, and epigallocatechin gallate on human prostate cancer cell lines: PC-3, DU145 (androgen insensitive) and LNCaP (androgen sensitive), by measuring cell proliferation, MMP expression, and invasion potential. Cell lines DU145, PC-3, and LNCaP were treated at near confluence with NS at various concentrations. Cell proliferation was measured by MTT assay after 24 hours, MMP expression was measured by gelatinase zymography in condition media, and invasion activity was measured by Matrigel. The nutrient mixture did not significantly inhibit PC-3 cell proliferation at 50 microg/ml, but showed significant antiproliferative effect at 500 ug/ml. When treated with NS, proliferation of LNCaP cells was inhibited by 80% of control at 100 microg/ml. NS showed dose-dependent inhibition of DU145 cell proliferation with 47% reduction at 1000 microg/ml. NS showed a dose-dependent inhibition of both MMP-2 and MMP-9 expression by PC-3 cells and MMP-9 expression by PMA-treated (200 ng/ml) DU145 cells. Neither MMP-2 nor MMP-9 gelatinolytic activity was detected in LNCaP cell culture. Invasion of DU145 and LNCaP cells through Matrigel was completely inhibited at 500 microg/ml and PC-3 at 1000 microg/ml. Inhibition of MMP expression and invasion suggests the mixture of nutrients studied is a potent, natural anticancer agent for the treatment of prostate cancer.

The antirheumatic effect of pirfenidone was compared with a positive control drug, oxyphenbutazone which is used in patients suffering from rheumatoid arthritis, in a double blind clinical trial in humans. The data collected in this pilot project revealed that pirfenidone was more effective (p < 0.025) than oxyphenbutazone in providing relief from arthritic pain. In addition, a greater number (p < 0.025) of patients reported favorable response to oral pirfenidone than oral oxyphenbutazone. However, there were no significant differences in the number of patients who dropped out from the trial and the number of patients who tolerated the drugs for 21 days of the trial between the pirfenidone and oxyphenbutazone groups. It was concluded from this pilot study that pirfenidone potentially offers a novel therapeutic modality for the management of rheumatoid arthritis with little or no adverse effects unlike steroidal and non-steroidal anti-inflammatory drugs which are frequently used for this chronic debilitating disease.

To investigate the effects of physical exercise on bone mass during the climacteric and menopausal period. The study group were 1123 postmenopausal Japanese women (mean: 55.4 +/- 3.7 years). Their current bone mineral density of lumbar vertebrae (L2-4) was analyzed taking the presence or absence of regular physical exercise, the type of exercise and its duration into consideration. Of the 1123 postmenopausal women, 643 (57.3%) were currently involved is some form of physical exercise on a regular basis. Bone mineral density did not differ significantly between the women exercising at present (1.035 +/- 0.08 g/cm2) and the women who had never been involved in regular physical exercise at any time in their life (1.089 +/- 0.08 g/cm2). The bone mineral density did not differ significantly in relation to the duration of physical activity (less than 6 months: 1.054 +/- 0.169 g/cm2; 6 months to 3 years: 1.049 +/- 0.128 g/cm2; over 5 years: 1.024 +/- 0.168 g/cm2). Although a majority of the postmenopausal women surveyed were involved in some form of physical activity, the practice of mild exercise at and around the time of perimenopause did not significantly increase bone mineral density.

Low-power laser irradiation (LPLI) has come into a wide range of use in medical field. Considering basic research, LPLI can enhance DNA synthesis and increases proliferation rate of human cells. But only a few data about the effects of LPLI on human liver or hepatoma cells are available. The cytoskeleton plays important roles in cell function and therefore is implicated in the pathogenesis of many human liver diseases, including malignant tumors. In our previous study, we found the stability of cytokeratin molecules in human hepatocytes was related to the intact microtubule network that was influenced by colchicine. In this study, we are going to search the effect of LPLI on proliferation of human hepatoma cell line HepG2 and J-5 cells. In addition, the stability of cytokeratin and synemin (one of the intermediate filament-associated proteins) were analyzed under the action of LPLI to evaluate the possible mechanism of LPLI effects on proliferation of human hepatoma cells. In experiment, HepG2 and J-5 cells were cultured in 24-well plate for 24 hours. After irradiation by 130 mW diode 808 nm GaAlAs continue wave laser in different time intervals, the cell numbers were counted. Western blot and immunofluorescent staining examined the expression and distribution of PCNA, cytokeratin and synemin. The cell number counting and PCNA expression were evaluated to determine the proliferation. The organization and expression of cytokeratin and synemin were studied to identify the stability of cytoskeleton affected by LPLI. The results revealed that proliferation of HepG2 and J-5 cells was inhibited by LPLI since the cell number and PCNA expression was reduced. Maximal effect was achieved with 90 and 120 seconds of exposure time (of energy density 5.85 J/cm2 and 7.8 J/cm2, respectively) for HepG2 and J-5, respectively. The decreased ratio of cell number by this dose of irradiation was 72% and 66% in HepG2 and J-5 cells, respectively. Besides that, the architecture of intermediate filaments in these cells was disorganized by laser irradiation. The expression of intermediate filament-associated protein, synemin, was also reduced. Two significant findings are raised in this study: (1) Diode 808 nm GaAlAs continuous wave laser has an inhibitory effect on the proliferation of human hepatoma cells line HepG2 and J-5. (2) The mechanism of inhibition might be due to down-regulation of synemin expression and alteration of cytokeratin organization that was caused by laser irradiation.

The p53 tumor suppressor protein is a biologically very important molecule. In addition to its central relevance in cancer, its function as an inducer of cell cycle checkpoint and apoptosis may be important in a number of cellular stress responses. However, studies of p53 interactions with other proteins have been hampered in large part by the low abundance of normal p53 in cells. Moreover, the detection of p53 in immune complexes is complicated by the presence of comigrating immunoglobulin chains in SDS-polyacrylamide gels. The method described herein, which utilizes protein A-horseradish peroxidase conjugates, in combination with chemiluminescent detection methods, allows ready sensitive detection of p53 protein in immune complexes with little interference by comigrating immunoglobulin chains. Using this method, pseudo-mutant form as one of confusing conformations of p53 mutant protein was able to be identified with the criteria as high basal level of p53 expression and immunodetection with PAb1620 in human cells.

P53-mediated cellular response has been known to play a crucial role in maintaining genomic stability of mammalian cells against genotoxic stresses. In our previous study, we showed that mild hyperthermia was sufficient to induce apoptosis on p53-dependent pathway in human lymphoid system (Seo et al., 1999), suggesting that mild hyperthermia might be useful for the reducing of genomic instability. However, there have been few reports to show the direct evidence on preventive role of p53 under mild hyperthermia against carcinogenic DNA damage. Here we first show the elimination of MMS-induced micronuclei (MN) as one of biomarkers of carcinogenic risk by p53 activation in human lymphoid cells in response to mild hyperthermia, strongly suggesting a possible protective role of mild hyperthermia in chromosomal stability against genotoxic stresses. Our data might support investigation of the clinical application of mild hyperthermia for the prevention of carcinogenesise in the human lymphoid system.

Osteoarthritis is thought to be induced by the aging-related loss of homeostatic balance between degeneration and repair mechanism around cartilage tissue in which inflammatory mediators such as reactive oxygen species, cytokines and prostaglandins are prone to overproduction under undesirable physiological conditions. Phlorotannins are unique polyphenolic compounds bearing dibenzo-1,4-dioxin skeleton which are not found in terrestrial plants but found only in some brown algal species such as Ecklonia and Eisenia families. Phlorotannin-rich extracts of Ecklonia cava including ventol showed significant antioxidant activities such as DPPH radical scavenging, ferric ion reducing power, peroxynitrite scavenging and inhibition of LDL oxidation, indicating their possible antioxidative interference both in onset and downstream consequences of osteoarthritis. Ventol also showed significant down regulation of PGE2 generation in LPS-treated RAW 246.7 cells, and significant inhibition of human recombinant interleukin-1alpha-induced proteoglycan degradation, indicating its beneficial involvement in pathophysiological consequences of osteoarthritis, the mechanism of which needs further investigation. Since ventol showed strong therapeutic potentials in arthritic treatment through several in vitro experiments, it is highly encouraged to perform further mechanistic and efficacy studies.

Interleukin (IL)-15, an inhibitor of tumor necrosis factor (TNF)-alpha, causes liver injury in mice. We determined levels of IL-15, IL-6, and IL-18 by enzyme-linked immunosorbent assays in 20 patients with acute hepatic failure and examined relationship between these proinflammatory cytokines and IL-15. A significant correlation was observed between the levels of IL-18 and IL-15 (p = 0.0118). IL-15 levels in the nonsurvivors were significantly higher than those in the survivors (p = 0.0357). Our results suggest that IL-15 overexpression may cause liver injury in human.

Cholestasis is one of the major liver diseases and results in progressive liver fibrosis and cirrhosis. In this study, the transcriptional response of the liver to common bile duct ligation in rats was examined by cDNA microarray analysis, and 134 genes for which expression was altered during long-term cholestasis were identified. Clustering analysis of these genes for multiple time-point data yielded 7 different patterns in which a large portion of the genes was classified into 3 clusters. Two clusters consisted of up-regulated genes, including genes that may be related to disruption of lipid metabolism and liver fibrosis observed in the early stage of cholestasis, and the other cluster consisted of down-regulated genes, including a gene that has been thought to be involved in the mechanism of cell protection against accumulation of bile acids. Since the expression patterns of these genes appear to reflect molecular features of cholestasis. Characterization of the genes identified in this study may shed further light on the physiological and pathological characteristics of long-term cholestasis.