BACKGROUND: Nitric Oxide (NO) is a multi-faceted molecule with dichotomous regulatory roles in many areas of biology. NO can promote apoptosis in some cells, whereas it inhibits apoptosis in other cell types. This study was performed to characterize NO-induced cell death in lung epithelial cells and to investigate the roles of cell death regulators including iron, bcl-2 and p53. METHODS: A549 cells were used for lung epithelial cells. SNP (sodium nitroprusside) and SNAP (S-nitroso-N-acetyl-penicillamine) were used for NO donor. Cytoxicity assay was done by MTT assay and crystal violet assay. Apoptotic assay was done by fluorescent microscopy after double staining with propidium iodide and hoecst 33342. Iron inhibition study was done with RBCs and FeSO4. For bcl-2 study, bcl-2 overexpressing cells (A549-bcl-2) were used and for p53 study, Western blot analysis and p53 functionally knock-out cells (A549-E6) were used. RESULTS: SNP and SNAP induced dose-dependent cell death in A549 cells and fluorescent microscopy revealed that SNAP induced apoptosis in low doses but necrosis in high doses while SNP induced exclusively necrotic cell death. Iron inhibition study using RBCs and FeSO4 significantly blocked SNAP-induced cell death. And also SNAP-induced cell death was blocked by bcl-2 overexpression. Finally, we found that SNAP activate p53 by Western blot analysis and that SNAP-induced cell death was decreased in the abscence of p53. CONCLUSION: In lung epithelial cells, NO can induce cell death, more precisely apoptosis in low doses and necrosis in high doses. And iron, bcl-2, and p53 play important roles in NO-induced cell death.
Apoptosis ; Biology ; Blotting, Western ; Cell Death* ; Epithelial Cells* ; Gentian Violet ; Humans ; Iron ; Lung* ; Microscopy ; Necrosis ; Nitric Oxide* ; Propidium ; Tissue Donors

The present study evaluated the effect of various dosages of soybean isoflavone extract on body weight changes, glucose tolerance and liver function in streptozotocin-induced diabetic rats. One group of normal rats (normal control) was fed an AIN-76-based experimental diet and four groups of diabetic rats were fed the same diet supplemented with four different levels of soybean isoflavone extract for seven weeks. The daily dosages of pure isoflavone for four diabetic groups were set to be 0 mg (diabetic control), 0.5 mg (ISO-I), 3.0 mg (ISO-II) and 30.0 mg (ISO-III) per kilogram of body weight, respectively. The daily consumption of isoflavone at the level of 3.0mg per kilogram of body weight resulted in the suppression of body weight loss and increased the survival rate of diabetic animals one and half times compared to that of the diabetic control group. Blood glucose levels in a fasting state and after the oral administration of glucose were significantly lower in the ISO-II group during the oral glucose tolerance test. The ISO-II group showed a tendency to elongate the gastrointestinal transit time. The activity of serum aminotransferases, indicator of liver function, was not negatively affected by any intake level of isoflavone. The present study demonstrated that the soybean isoflavone extract may be beneficial to diabetic animals by improving their glucose tolerance and suppressing weight loss without incurring hepatotoxicity at the daily dosage of 3.0 mg per kg of body weight.
Administration, Oral ; Animals ; Blood Glucose ; Body Weight ; Body Weight Changes ; Diet ; Fasting ; Gastrointestinal Transit ; Glucose Tolerance Test ; Glucose* ; Liver ; Rats* ; Soybeans* ; Streptozocin ; Survival Rate* ; Transaminases ; Weight Loss

The present study evaluated the effects of various dosages of soybean isoflavone extract on lipid profiles, lipid peroxidation and antioxidant activities in streptozotocin-induced diabetic rats. The one normal control group was fed an AIN-76-based experimental diet and four diabetic groups were fed the same diet, supplemented with four different levels of soybean isoflavone extract for seven weeks. The daily dosages of pure isoflavone for four diabetic groups were set to be 0 mg (diabetic control), 0.5 mg (ISO-I), 3.0 mg (ISO-II) and 30.0 mg (ISO-III) per kilogram of body weight, respectively. The plasma total cholesterol levels and the TBA-reactive substances contents in the liver and kidney were significantly lowered in ISO-II and ISO-III groups compared to those in the diabetic control group. The levels of plasma HDL-cholesterol, plasma vitamin A and hepatic superoxide dismutase were significantly increased in those two groups compared with the diabetic control group. The present study demonstrated the possibility that the diets supplemented with 3.0 mg and 30.0 mg of soybean isoflavone extract may have beneficial effects on the plasma lipids, tissue lipid peroxidation and partly on antioxidant system in diabetic animals and there were no significant differences between the ISO-II and ISO-III groups. The results suggest that the effective daily dosage level of isoflavone for improving lipid metabolism in diabetic rats may be above 3.0 mg per kilogram body weight.
Animals ; Body Weight ; Cholesterol ; Diet ; Kidney ; Lipid Metabolism ; Lipid Peroxidation ; Liver ; Plasma ; Rats ; Soybeans ; Superoxide Dismutase ; Vitamin A

OBJECTIVES: Many recent studies have suggested that several systemic conditions, such as obesity, hypertension, hyperlipidemia, and diabetes, are related to periodontitis. The aim of this study was to investigate whether any particular metabolic syndrome component correlates with the periodontal status. METHODS: This study analyzed data from 501 adults (age range, 30 to 64 years) who completed a general physical examination and an oral-health-related questionnaire between January 2014 and June 2014. Periodontitis was already used by the validity and reliability has been proven questionnaire. Multiple logistic regression analysis was used to assess the association between metabolic syndrome and the presence of chronic periodontitis. RESULTS: Among the 501 participants, 81 (16.2%) had metabolic syndrome and 91 (18.2%) had chronic periodontitis. The prevalence of chronic periodontitis was 28.40% (23/81) and 16.19% (68/420) in the participants with and without metabolic syndrome, respectively. The association remained significant after adjusting for sex, age, education, income, occupation, and smoking status; the adjusted odds ratio was 2.03 (95% confidence interval, 1.11 to 3.71). CONCLUSIONS: A significant association exists between metabolic syndrome components and the periodontal status.
Adult ; Chronic Periodontitis ; Education ; Humans ; Hyperlipidemias ; Hypertension ; Logistic Models ; Obesity ; Occupations ; Odds Ratio ; Periodontitis ; Physical Examination ; Prevalence ; Reproducibility of Results ; Smoke ; Smoking

Postmenopausal atrophic vaginitis, along with vasomotor symptoms and sleep disorders, is one of the most troublesome symptoms of menopause. However, many women do not manage this symptom properly due to insufficient knowledge of the symptoms or sexual embarrassment. With appropriate treatment, many postmenopausal women can experience relief from discomforts, including burning sensation or dryness of the vagina and dyspareunia. Topical lubricants and moisturizers, systemic and local estrogens, testosterones, intravaginal dehydroepiandrosterones (DHEAs), selective estrogen receptor modulators, and energy-based therapies are possible treatment modalities. Systemic and local estrogen therapies effectively treat genitourinary syndrome of menopause (GSM), but they are contraindicated in patients with breast cancer, for whom lubricants and moisturizers must be considered as the primary treatment. Intravaginal DHEA and ospemifene can be recommended for moderate to severe GSM; however, there is insufficient data on the use of intravaginal DHEA or ospemifene in patients with breast cancer, and further studies are needed. Energy-based devices such as vaginal laser therapy reportedly alleviate GSM symptoms; however, the U.S. Food and Drug Administration warning has recently been issued because of complications such as chronic pain and burning sensations of the vagina. To summarize, clinicians should provide appropriate individualized treatment options depending on women’s past history, symptom severity, and chief complaints.

Postmenopausal atrophic vaginitis, along with vasomotor symptoms and sleep disorders, is one of the most troublesome symptoms of menopause. However, many women do not manage this symptom properly due to insufficient knowledge of the symptoms or sexual embarrassment. With appropriate treatment, many postmenopausal women can experience relief from discomforts, including burning sensation or dryness of the vagina and dyspareunia. Topical lubricants and moisturizers, systemic and local estrogens, testosterones, intravaginal dehydroepiandrosterones (DHEAs), selective estrogen receptor modulators, and energy-based therapies are possible treatment modalities. Systemic and local estrogen therapies effectively treat genitourinary syndrome of menopause (GSM), but they are contraindicated in patients with breast cancer, for whom lubricants and moisturizers must be considered as the primary treatment. Intravaginal DHEA and ospemifene can be recommended for moderate to severe GSM; however, there is insufficient data on the use of intravaginal DHEA or ospemifene in patients with breast cancer, and further studies are needed. Energy-based devices such as vaginal laser therapy reportedly alleviate GSM symptoms; however, the U.S. Food and Drug Administration warning has recently been issued because of complications such as chronic pain and burning sensations of the vagina. To summarize, clinicians should provide appropriate individualized treatment options depending on women’s past history, symptom severity, and chief complaints.

OBJECTIVE: Diabetic nephropathy and ablation nephropathy are characterized by sclerotic processes in the glomeruli. To elucidate the site, degree and time-honored changes of glomerular sclerosis, morphometric analysis was performed using the experimental animals models. METHODS: The animals used were male Sprague Dowley rats and separated into 4 groups as young normal control, old control, streptozotocin-injected group, and right nephrectomized group. Chronologically kidney specimens were obtained after each treatment and processed to evaluated histologic changes. To evaluated the glomerular area, interstitial fibrosis and glomerular tuft fibrosis, the kidney specimens were fixed in Buin's solution, paraffin-embedded and 2 micrometer sections were Sirius red stained. To study the mesangial area, mesangial matrix area, glomerular basement membrane, and tubu lar basement membrane, the specimens were fixed in 2.5% glutaraldehyde, epon-embedded, double-stained and examined under the transmission electron microscope. All the specimens were analyzed morphometrically using the Image Pro Plus software. The obtained morphometric data were statistically analyzed to evaluate the differences of fibrotic processes and degree between experimental groups. RESULTS: Diabetic group revealed statistically significant increase of glomerular area from 8th week after streptozotocin injection to 24th week of experimental date. The parenchymal fibrosis and glomerular tuft fibrosis was prominent from the 2nd week of injection and steadily increased until the end of experimental date. The thickness of glomerular basement membrane was significantly increased even at the first week of injection and the tubular basement membrane also increased in thickness at the 3rd week of experiment. Ablation nephropathy model made by right nephrectomy showed increased glo merular area at the 7th week of ablation and the degree were intensified after 16th week of experimental date. The amount of stainable collagen in the renal parenchyme and glomerular tuft increased in the second week kidney sample and steadily increased thereafter until the end of experimental date. The increase of thickness of GBM and TBM also started to appear at the second week of operation. The old control also revealed fibrosis but the degree was less than the diabetic and ablation groups. Both diabetic and ablation nephropathy groups exhibited extensive increase of glomerular area, stainable colla gen, thickness of GBM and TBM at the end of experimental date and the ablation group revealed more extensive evidences of fibrosis without statistical significance. Comparison between the experimental groups were meaningless because the duration of the experimental manipulation was not the same. CONCLUSION: Glomerular and renal interstitial sclerosis and thickening of GBM and TBM are not the specific lesions of the diabetic glomerulopathy and are the common histologic changes occur in the kidney of partial parenchymal loss of any etiology. And it is suggested by this study that the common hemodynamic change involving the diabetic nephropathy, ablation nephropathy and physiologic aging is one of the important pathogenetic factors of glomerular sclerosis.
Aging ; Animals ; Basement Membrane ; Collagen ; Diabetic Nephropathies ; Fibrosis ; Glomerular Basement Membrane ; Glutaral ; Hemodynamics ; Humans ; Hyperglycemia* ; Kidney ; Male ; Models, Animal* ; Nephrectomy ; Rats* ; Sclerosis ; Streptozocin

OBJECTIVE: Diabetic nephropathy and ablation nephropathy are characterized by sclerotic processes in the glomeruli. To elucidate the site, degree and time-honored changes of glomerular sclerosis, morphometric analysis was performed using the experimental animals models. METHODS: The animals used were male Sprague Dowley rats and separated into 4 groups as young normal control, old control, streptozotocin-injected group, and right nephrectomized group. Chronologically kidney specimens were obtained after each treatment and processed to evaluated histologic changes. To evaluated the glomerular area, interstitial fibrosis and glomerular tuft fibrosis, the kidney specimens were fixed in Buin's solution, paraffin-embedded and 2 micrometer sections were Sirius red stained. To study the mesangial area, mesangial matrix area, glomerular basement membrane, and tubu lar basement membrane, the specimens were fixed in 2.5% glutaraldehyde, epon-embedded, double-stained and examined under the transmission electron microscope. All the specimens were analyzed morphometrically using the Image Pro Plus software. The obtained morphometric data were statistically analyzed to evaluate the differences of fibrotic processes and degree between experimental groups. RESULTS: Diabetic group revealed statistically significant increase of glomerular area from 8th week after streptozotocin injection to 24th week of experimental date. The parenchymal fibrosis and glomerular tuft fibrosis was prominent from the 2nd week of injection and steadily increased until the end of experimental date. The thickness of glomerular basement membrane was significantly increased even at the first week of injection and the tubular basement membrane also increased in thickness at the 3rd week of experiment. Ablation nephropathy model made by right nephrectomy showed increased glo merular area at the 7th week of ablation and the degree were intensified after 16th week of experimental date. The amount of stainable collagen in the renal parenchyme and glomerular tuft increased in the second week kidney sample and steadily increased thereafter until the end of experimental date. The increase of thickness of GBM and TBM also started to appear at the second week of operation. The old control also revealed fibrosis but the degree was less than the diabetic and ablation groups. Both diabetic and ablation nephropathy groups exhibited extensive increase of glomerular area, stainable colla gen, thickness of GBM and TBM at the end of experimental date and the ablation group revealed more extensive evidences of fibrosis without statistical significance. Comparison between the experimental groups were meaningless because the duration of the experimental manipulation was not the same. CONCLUSION: Glomerular and renal interstitial sclerosis and thickening of GBM and TBM are not the specific lesions of the diabetic glomerulopathy and are the common histologic changes occur in the kidney of partial parenchymal loss of any etiology. And it is suggested by this study that the common hemodynamic change involving the diabetic nephropathy, ablation nephropathy and physiologic aging is one of the important pathogenetic factors of glomerular sclerosis.
Aging ; Animals ; Basement Membrane ; Collagen ; Diabetic Nephropathies ; Fibrosis ; Glomerular Basement Membrane ; Glutaral ; Hemodynamics ; Humans ; Hyperglycemia* ; Kidney ; Male ; Models, Animal* ; Nephrectomy ; Rats* ; Sclerosis ; Streptozocin

BACKGROUND: Gemcitabine is a new anti-cancer agent for treating non-small cell lung cancer. Functioning as an antimetabolite, it induces anti-cancer effects by suppressing DNA synthesis after being incorporated into the DNA as a cytosine arabinoside analogue. When Gemcitabine is incorporated into the DNA, the p53 gene may be activated by induction of the DNA defect. However, there are a few studies on the molecular mechanisms of Gemcitabine-induced cell death. This study examined the role of p53 in Gemcitabine-induced cell death. METHODS: A549 and NCl-H358 lung cancer cells were used in this study. The cell viability test was done using a MTT assay at Gemcitabine concentrations of 10nM, 100nM, 1uM, 10uM and 100uM. A FACScan analysis with propium iodide staining was used for the cell cycle analysis. Western blot analysis was done to investigate the extent of p53 activation. For the functional knock-out of p53, stable A549-E6 cells and H358-E6 cells were transfected pLXSN-16E6SD which is over expresses the human papilloma virus E6 protein that constantly degrades p53 protein. The functional knock out of p53 was confirmed by Western blot analysis after treatment with a DNA damaging agent, doxorubicine. RESULTS: Gemcitabine exhibited cell toxicity in dose-dependent fashion. The cell cycle analysis resulted in an S phase arrest. Western blot analysis significant p53 activation in time-dependent manner. Gemcitabine-induced cytotoxicity was reduced by 20-30% in the A549-E6 cells and the 30-40% in H358-E6 cells when compared with the A549-neo and H358-neo control cells. CONCLUSION: Gemcitabine induces an S phase arrest, as expected for the anti-metabolite, and activates the p53 gene. Furthermore, p53 might play an important role in Gemcitabine-induced cell death. Further investigation into the molecular mechanisms on how Gemcitabine activates the p53 gene and its signaling pathway are recommended.
Humans ; Lung Neoplasms ; Genes, p53 ; Antimetabolites